肿瘤领域文献阅读汇编

People who have experienced cancer are integral partners in the research process and improving standard-of-care treatments. Advocates contribute insights from lived experience that help researchers identify impactful scientific priorities, improve study design, and enhance clinical trial participation. Although programs have been established to connect scientists and advocates to enhance discovery, translational, and clinical research, challenges remain that hinder engagement and sustained partnerships. In this article, we describe prevalent barriers and provide suggestions to improve bidirectional communication and collaboration, as strengthening researcher-advocate partnerships promotes impactful, inclusive, and patient-centered cancer research.

Multiple myeloma (MM) is a plasma-cell malignancy with extensive genomic and transcriptional heterogeneity, limiting disease classification and precision therapy. Here we generated a clinically annotated, population-scale, single-cell atlas of MM from 341 individuals spanning the disease and treatment continuum. We identified five recurrent malignant transcriptional archetypes and an orthogonal proliferative program associated with genomic features, therapeutic resistance and clinical outcomes. Validation in the independent CoMMpass cohort demonstrated robustness, prognostic relevance and portability across platforms. We developed a single-cell, target-discovery pipeline prioritizing malignant enrichment, cell-type specificity and tissue restriction, identifying FCRL2 as a plasma-restricted or B cell-lineage-restricted surface target expressed by malignant plasma cells. FCRL2-targeted chimeric antigen receptor T cells demonstrated antigen-specific activity in vitro and survival benefit in vivo. Together, these data provide a clinically actionable blueprint for patient stratification and precision target nomination in plasma-cell malignancies.

Ovarian cancer remains the most lethal gynaecological malignancy, largely due to late-stage diagnosis and the limited sensitivity of CA-125 for borderline and low-grade tumours. Dysregulated lipid metabolism may provide complementary diagnostic information. This study aimed to develop lipid-based biomarkers to improve the differential diagnosis of adnexal masses. We profiled total plasma and small extracellular vesicle (sEV) lipids in women with benign, borderline and malignant ovarian tumours, using healthy women as controls. Solid-phase extraction demonstrated superior reproducibility, lipid-class specificity and sEV lipid enrichment compared with liquid-liquid extraction. Combined with CA-125, both plasma and sEV lipid biomarkers outperformed CA-125 alone for distinguishing borderline and low-grade serous ovarian cancer from benign tumours (accuracy up to 0.89 vs. 0.67). In patients with normal CA-125 levels, sEV lipids provided the greatest diagnostic advantage, distinguishing invasive and borderline tumours from benign adnexal masses (AUROC up to 0.82). Generative artificial intelligence was used to simulate biomarker performance in synthetic cohorts for differential diagnosis (n = 10,000) and screening (n = 100,000). For differential diagnosis, sEV lipids combined with CA-125 outperformed plasma lipids for identifying endometrioid and mucinous ovarian cancer from benign adnexal masses. For screening, sEV lipids alone achieved accuracies of 0.91 and 0.94 for benign conditions and high-grade serous ovarian cancer, respectively. Simulations restricted to patients with normal CA-125 levels confirmed that sEV lipids provided the highest discriminative performance for low-grade serous ovarian cancer (AUROC up to 0.74), demonstrating their potential where CA-125 alone fails. To the best of our knowledge, this is the first study to directly compare matched plasma and sEV lipid profiles across ovarian cancer histotypes, demonstrating a diagnostic advantage for sEV lipids-particularly for borderline, low-grade serous and mucinous tumours, supporting their integration alongside CA-125 in future differential diagnosis and screening strategies.

Multiple myeloma remains incurable despite therapeutic advances. Longitudinal profiling of bone marrow and peripheral blood by Chander et al. reveals selective persistence of malignant plasma cell states, compartment-specific immune remodeling, incomplete humoral recovery after autologous stem cell transplantation, and divergent vaccine responses.

The pathobiology of multiple myeloma is influenced by cells of the bone marrow, but whether tumor support is organized in a spatially-defined tumor microenvironment (TME) remains unclear. Using spatial transcriptomics and imaging mass cytometry, we show that myeloma cells initially exist as scattered cells throughout the marrow, yet with disease evolution condense into a spatially-defined TME. Dense tumor nodules contain a cellular ecosystem distinct from the surrounding marrow, enriched for macrophages, conventional dendritic cells, and CD8⁺ T cells, as well as endothelial cells and THY1⁺ mesenchymal stromal cells. Conversely, neutrophil lineage cells are absent from this TME but interact with scattered myeloma cells outside of dense nodules. Presence of the dense tumor architecture at diagnosis confers a worse prognosis, emphasizing the relevance of spatial organization of the bone marrow. Together, these findings define a discrete spatial and cellular myeloma TME, that provides spatially-guided insights into patient stratification and disease pathobiology.

Prediction of the outcome of large B-cell lymphomas/high-grade B-cell lymphomas (DLBCLs/HGBCLs) is based on clinical parameters and molecular testing, for example, for rearrangements of MYC (MYC-R) and MYC-R in combination with BCL2 and BCL6 translocations (double/triple hit). However, the group of DLBCL/HGBCL with poor outcome is not confined to MYC-R lymphomas, and fluorescence in situ hybridization (FISH) testing for MYC-R misses several high-risk lymphomas. We aimed to understand if artificial intelligence (AI) trained to identify MYC-R will delineate a poor prognostic subgroup. We generated a collection of digital hematoxylin and eosin (H&E)-stained slides of DLBCL/HGBCL (N = 2018) annotated for MYC, BCL2, and BCL6 translocations. A multiple-instance deep learning AI model for the identification of MYC-R alone or as double/triple hit was established using 1035 H&E-stained slides and evaluated on an external test cohort (N = 499). A pretrained tumor tissue classifier improved reliability and interpretability by focusing the model on tumor areas. Our model score reflects a morphological "MYCness" in DLBCL/HGBCL and demonstrates a strong association with overall survival (OS) and progression-free survival (PFS) in the external test cohort. This was confirmed with an additional clinical test cohort (N = 484) without FISH labels. The AI model scores correlate with various molecular features of DLBCL/HGBCL, including BCL2 and MYC gene expression, and the high-grade gene expression signature, but also features of the tumor microenvironment. Multivariate analysis, adjusted for International Prognostic Index (IPI) factors, demonstrated the prognostic significance of our model in identifying high-risk cases for both PFS and OS.

We provide an overview of a recent study in Cancer Discovery by Leongamornlert and colleagues involving genomic profiling of 30 patients with myeloproliferative neoplasms using available blood or bone marrow samples to assess what factors underlie disease evolution or predict stability. See related article by Leongamornlert et al., p. 1843.

Messenger RNA (mRNA) cancer vaccines have progressed rapidly, with personalized platforms such as mRNA-4157 and BNT122 demonstrating feasibility, safety, and durable immune activity but limited scalability. Off-the-shelf constructs, including BNT111 and BNT113, enable faster, broader deployment, yet they risk reduced precision or immune tolerance. These complementary approaches reveal key translational dualities: personalization versus shared antigen (off-the-shelf), potency versus safety, and speed versus durability. This mini-review synthesizes emerging clinical evidence and outlines strategies such as modular vaccine design, prime-boost vaccination regimens, and adaptive trial frameworks to reconcile these trade-offs and advance scalable, durable mRNA vaccines for broad oncologic impact. By dissecting the competing design pressures that shape mRNA vaccine performance, this mini-review proposes integrative strategies, spanning modular architectures, prime-boost regimens, and adaptive trials, to reconcile immunologic potency with manufacturability and safety, charting a roadmap toward next-generation cancer vaccines.

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative treatment option for a significant proportion of patients with acute myeloid leukemia (AML), and it is generally recommended when the relapse risk without allo-HSCT outweighs the estimated non-relapse mortality significantly. While current recommendations for allo-HSCT are based on risk groups, there is considerable heterogeneity within these individual categories. We analyzed 2550 intensively treated AML patients aged 18-70 with cytogenetic and next-generation sequencing data from the HARMONY Alliance database, who did not receive allo-HSCT in first complete remission (CR1). A non-parametric machine learning (ML) model based on Bayesian Additive Regression Trees (BART) integrated clinical variables and genomic aberrations to provide individualized outcome estimations. External validation was performed in a cohort of 714 patients enrolled in UK-NCRI trials. The predictive performance of the HARMONY ML model, measured by the area under the time-dependent receiver operating curve (AUC(t)), was superior to European LeukemiaNet (ELN)2022 risk classification in estimating 5-year overall survival (0.741 vs. 0.700), 5-year relapse-free survival (0.752 vs. 0.705), and 5-year cumulative incidence of relapse (0.742 vs. 0.708), which was confirmed in the external validation cohort. Notably, the model revealed substantial heterogeneity within ELN2022 risk groups, identifying a significant proportion of favorable-risk patients with a predicted 5-year CIR > 40%, who could potentially benefit from allo-HSCT in CR1. The HARMONY ML model provides individualized risk prediction in intensively treated adult AML patients and supports more tailored therapeutic decisions regarding allo-HSCT in CR1, which should be further advanced by integrating measurable residual disease in the future.

Optimal consolidation therapy for patients with intermediate-risk acute myeloid leukemia (AML) in first complete remission (CR1) is controversial. Retrospective studies have suggested that the clearance of leukemia-associated mutations (LAMs) in CR1 may predict lower relapse risk and better outcomes with high-dose cytarabine (HiDAC) consolidation. We tested this hypothesis prospectively. We performed a phase II, multicenter study of intermediate-risk, transplant-eligible, de novo AML in patients 18-60 years of age who achieved a complete remission (CR) or CR with incomplete count recovery (CRi) after induction therapy. Tumor and normal whole-exome sequencing was performed at presentation to identify somatic LAMs (median ∼30 LAMs/patient). In remission marrow samples, LAM variant allele frequencies (VAFs) were then remeasured using a VAF cutoff of less than 2.5% to define clearance. Patients who met this LAM clearance threshold received HiDAC consolidation, whereas those with persistent LAMs (VAF ≥2.5%) were recommended to undergo allogeneic hematopoietic cell transplantation. The primary endpoint compared relapse-free survival (RFS) of intermediate-risk patients with complete LAM clearance to historical cohorts with intermediate-risk AML who received HiDAC-based regimens in CR1. To account for an unplanned interim assessment, the significance threshold for the primary analysis was 0.01. Among 100 patients who were evaluated, intermediate-risk patients who cleared all LAMs in CR1 (n=33) had a median RFS of 33.1 months (95% confidence interval, 11.7-NA) compared to a median RFS of 11.7 months in the historical cohort (n=239; 95% confidence interval, 9.9-15.6, P=0.015). Among patients with intermediate-risk AML, clearance of LAMs after induction, followed by HiDAC consolidation in CR1, was associated with longer RFS compared with similarly treated historical controls. Although this result did not meet the prespecified threshold for statistical significance, the reported association sets the stage for a randomized trial to further evaluate this strategy. (ClinicalTrials.gov number, NCT02756962.).

Altered adipose tissue biology plays a key role in the development of cardiovascular disease (CVD) and cancer and contributes to mortality. We assessed the association between obesity in people with HIV (PWH) at antiretroviral therapy (ART) initiation and the risk of clinical events. In this prospective cohort study, we included ART-naïve adults PWH enrolled in the Italian Cohort Naive Antiretrovirals (Icona) Foundation study cohort in Italy across 96 sites between January 1997 and July 2025. Participants were included if they initiated ART, had body mass index (BMI) > 18.5 kg/m2, and were free from AIDS, CVD, and cancer at ART initiation. Participants were classified as having obesity, defined as BMI ≥ 30 kg/m2, or not having obesity, defined as BMI 18.5-29.9 kg/m2, at ART initiation. The primary outcome was the first occurrence of CVD, cancer, or death. We performed a standard survival analysis with time-fixed covariates at baseline with a composite outcome of CVD/cancer/death. We hypothesized that age may be an effect measure modifier: results are presented after stratification by age (young [18-30], middle aged [31-60] and older [>60 years]). A total of 11,652 PWH (20.7% females, median age 38 years [Inter Quartile Range (IQR): 31, 46]) were included: 11,005 (94.4%) were people without obesity and 647 (5.6%) people with obesity. A total of 837 events were recorded: 122 CVD, 374 cancers and 341 deaths. By 15 years from ART initiation, the risk of CVD/cancer/death was 19.1% in people with obesity (95% Confidence Interval [CI]: 14.1-24.9%) vs. 12.4% in people without obesity (95% CI:11.3-13.4%, log-rank p = 0.0029). After controlling for confounding, the difference was attenuated (adjusted hazard ratio [aHR] 1.28 [95% CI:0.98-1.67], p = 0.073). Although we did not find evidence for interaction (p-value = 0.758) there was a trend for a larger effect among younger PWH: young (aHR 1.83 [95% CI 0.74, 4.56]), middle aged (aHR 1.49 [95% CI 1.11, 2.00]) and older (aHR 1.20 [95% CI 0.60, 2.41]). Obesity at ART initiation may be associated with a clinically meaningful increase in the risk of CVD, cancer, and death. We found weak evidence of a possible age-related gradient, with larger effect size in PWH under 60 years of age, which warrants further investigation. The Italian Cohort Naive Antiretrovirals (Icona) Foundation Study is supported by unrestricted grants from Gilead Sciences, ViiV Healthcare, Merck Sharpe & Dohme.

Claudin18.2 (CLDN18.2) has emerged as a clinically important therapeutic target in gastrointestinal malignancies following the success of zolbetuximab-based therapy. However, unlike conventional overexpression-driven biomarkers, the clinical relevance of CLDN18.2 is fundamentally shaped by target accessibility rather than expression alone. Physiologically concealed within gastric epithelial tight junctions, CLDN18.2 becomes exposed during malignant transformation, creating a spatially heterogeneous and dynamically accessible target landscape. This exposure-dependent biology challenges conventional tissue-based assessment and provides a strong rationale for molecular imaging. Recent advances in CLDN18.2-targeted molecular imaging across antibody, engineered-fragment, and nanobody platforms have enabled noninvasive whole-body assessment of target accessibility and lesion-level heterogeneity beyond localized biopsy. Importantly, imaging signals reflect not only antigen expression, but also probe-dependent pharmacokinetics, tissue penetration, and microenvironmental accessibility. In this review, we discuss the biological basis of CLDN18.2 accessibility, summarize emerging imaging and theranostic strategies, and examine the evolving role of molecular imaging in patient stratification and imaging-informed precision oncology.

RECQL5 is a member of the RecQ helicase family involved in DNA replication, homologous recombination, and maintenance of genomic stability. While germline pathogenic variants in other RecQ helicases cause established cancer predisposition syndromes, the role of RECQL5 in human cancer susceptibility remains uncertain. We report monozygotic adolescent twins with distinct tumors: dysembryoplastic neuroepithelial tumor in one twin and Burkitt lymphoma in the other. Clinical genome sequencing was initially nondiagnostic, but reanalysis identified a rare heterozygous nonsense variant in RECQL5 (NM_004259.7:c.2698C>T, p.(Gln900Ter)), present in both twins and their unaffected mother. The variant is predicted to undergo nonsense-mediated mRNA decay or produce a truncated protein lacking the C-terminal SRI (Set2-Rpb1 interacting) domain, which mediates interaction with RNA polymerase II. However, tumor sequencing data were not available to evaluate loss of heterozygosity or second somatic events. Given the unaffected carrier parent, lack of tumor molecular confirmation, and the biological heterogeneity of the tumors, a causal relationship for this variant cannot be established. This case highlights the challenges of interpreting rare germline variants in genes with emerging but incompletely characterized disease associations. Although the available evidence is insufficient to establish a definitive causal relationship, the identification of a shared loss-of-function RECQL5 variant in monozygotic twins with distinct tumors is noteworthy and adds to the limited clinical evidence suggesting a potential role for RECQL5 in cancer susceptibility. Additional functional studies, tumor-based analyses and the accumulation of well-characterized clinical cases will be essential to determine whether RECQL5 contributes to hereditary cancer predisposition.

Nearly 11 years after T-VEC became the first oncolytic virus approved in a major market, the closest candidate to follow it-Replimune's RP1, now facing an FDA advisory committee-remains a locally injected herpesvirus for melanoma, underscoring how little the field has advanced. Researchers blame a stack of unresolved obstacles-flawed trial designs, mistimed drug combinations, tumor defenses, such as hypoxia, and the inability to deliver most viruses intravenously-even as smarter sequencing, neoadjuvant approaches, and a bladder-cancer virus near approval hint at renewed momentum.

Timothy Yap, MBBS, PhD, vice president and head of clinical development in the Therapeutics Discovery Division and professor in the Department of Investigational Cancer Therapeutics (Phase I Program) at The University of Texas MD Anderson Cancer Center in Houston, spoke with Cancer Discovery's Suzanne Rose about how clinical trials have evolved, the role of AI, and the challenges facing today's drug developers. His research focuses on the first-in-human and combinatorial development of molecularly targeted agents, immunotherapies, antibody drug conjugates, and radioconjugates.

Although pesticide exposure occurs through multiple pathways and has been linked to diverse health outcomes, the overall evidence remains unclear. We systematically searched PubMed/MEDLINE, Embase, CINAHL, and Google Scholar up to November 20, 2025, for meta-analyses examining pesticide exposure and adverse health outcomes. We recalculated pooled estimates using random-effects models and converted all effect metrics to equivalent odds ratios with 95% confidence intervals. The quality of included reviews was assessed using AMSTAR 2, and the credibility of evidence was graded using predefined criteria (Class I to IV). A total of 53 meta-analyses covering 235 associations were included. Among the 77 main associations, 48.1% achieved statistical significance, of which 51.4% were supported by highly suggestive or suggestive evidence. Significant associations with pesticide exposure were observed across several health domains, particularly oncological, metabolic, neurological, and respiratory outcomes, with additional signals emerging in subgroup analyses. Particularly, exposures to organochlorine pesticides had increased odds of non-Hodgkin lymphoma. Exposure to dichlorodiphenyldichloroethylenes was associated with diabetes mellitus and non-Hodgkin lymphoma, while exposure to organochlorine insecticides was associated with hypothyroidism. Pesticide exposure was associated with childhood lower respiratory infections among respiratory outcomes. These findings identify priorities for prevention and highlight the need for better exposure assessment.

China has approved the first CAR T-cell therapy that targets solid tumors, satricabtagene autoleucel (satri-cel), for patients with claudin 18.2-positive, HER2-negative gastric or gastro-esophageal junction cancer. The approval stems from a phase II trial that found the cells increased progression-free survival and overall survival. Scientists are excited about the decision but caution that more research is necessary to improve the effectiveness of CAR T cells against solid tumors.

To present a summary of the 2026 version of the European Association of Urology (EAU)-European Association of Nuclear Medicine (EANM)-European Society for Radiotherapy and Oncology (ESTRO)-European Society of Urogenital Radiology (ESUR)-International Society of Urological Pathology (ISUP)-International Society of Geriatric Oncology (SIOG) guidelines on screening, diagnosis, and treatment of clinically localised prostate cancer (PCa). The Panel performed a literature review of all new data published in English, covering the time frame between May 2023 and 2025. The Guidelines were updated, and a strength rating for each recommendation was added based on a systematic review of the evidence. A risk-adapted strategy for identifying men who may develop PCa is advised, generally commencing at 50 yr of age and based on individualised life expectancy. The use of multiparametric magnetic resonance imaging to avoid unnecessary biopsies is recommended. When a biopsy is considered, a combination of targeted and regional biopsies should be performed. A new five-tier EAU classification has been introduced. Prostate-specific membrane antigen positron emission tomography imaging is the most sensitive technique for identifying metastatic spread. Active surveillance is the appropriate management for men with low-risk PCa, as well as for patients with selected favourable intermediate-risk ISUP grade group 2 lesions. Local therapies are addressed, as well as the management of persistent prostate-specific antigen after surgery. A recommendation to consider hypofractionated radiotherapy in intermediate-risk patients is provided. Patients with cN1 PCa should be offered radiotherapy to the primary tumour combined with long-term intensified hormonal treatment. The evidence in the field of diagnosis, staging, and treatment of localised PCa is evolving rapidly. These PCa Guidelines reflect the multidisciplinary nature of PCa management.

The nonprofit organization Blood Cancer United acquired the remaining supply of luveltamab tazevibulin and will continue the compassionate use program for children with a rare form of AML. The move points to the challenges of funding pediatric cancer drug development.

Kaposiform haemangioendothelioma (KHE) is a rare vascular neoplasm usually presenting in childhood. Its high morbidity and mortality is secondary to compression, invasion, and coagulopathy. The pathogenensis of KHE remains poorly understood although mTOR inhibitors have been used successfully in management. PTENHarmatoma Tumour Syndrome (PHTS) is a well characterised tumour predisposition syndrome with an ∼85% lifetime tumour risk. In addition, over 50% of patients have vascular anomalies. However, malignant vascular tumours are not a recognised association. A 28-year-old female with a KHE was enrolled in the IMAGINE study (Integrating Medically Actionable Genomics Into Early Phase Trials). A PTEN heterozygous c.277C>T p.(His93Tyr) pathogenic variant was detected initially in tumour tissue and subsequently saliva confirming a constitutional (germline) variant. Reverse phenotyping revealed the patient had a large head circumference, palmar keratosis and papillomas on the hand, feet, and gums, in keeping with PHTS. As malignant vascular neoplasms have not been well described in PHTS, this case further demonstrates the utility of genomic tumour profiling in early diagnosis of an Inherited Cancer Susceptibility Disorder (ICSD); particularly in young patients or in those where other features may be subtle. A plausible biological mechanism through upregulation of mTOR signalling also suggests malignant vascular tumours may be a rare tumour association with PHTS.

To evaluate a decentralized virtual Molecular Tumor Board (MTB) model in routine oncology practice in Russia. This retrospective analysis included 53 patients with advanced solid tumors discussed at a virtual MTB (December 2021-February 2025) after comprehensive genomic profiling. The primary endpoint was the rate of molecularly targeted therapy (MTT) recommendations, including biomarker-matched (BMT, ESCAT I-II) and biomarker-informed therapy (BIT, ESCAT III-V). Secondary endpoints included recommendation implementation, actionability, progression-free survival (PFS), and overall survival (OS). Potentially actionable biomarkers (ESCAT I-IV) were identified in 92,9% of patients. The MTB recommended MTT in 52,8%(28/53), including BMT in 10 (18,9%) and BIT in 18 (33,9%), while standard-of-care treatment was recommended in 25 patients (47,2%). Follow-up was available for 40 patients, among whom implementation of the primary MTB recommendation was observed in 27 cases (67,5%). In the evaluable MTT cohort, disease control rate was 59,3% (95% CI 40,7-75,5%) and objective response rate was 11,1% (95% CI 3,9-27,4%); outcomes were more favorable in the BMT subgroup than in the BIT subgroup. Implementation of MTB recommendations was associated with longer PFS (median 5 vs 2 months; HR 0,36, 95% CI 0,17-0,76; p = 0002) and longer OS (median 8 vs 5 months; HR 0,37, 95% CI 0,17-0,78; p = 0,01) than non-implementation. Our virtual MTB was feasible in routine practice and provided clinically relevant recommendations, with improved outcomes observed when these recommendations were implemented. However, the benefit appeared greatest in patients with clearly targetable alterations supported by higher levels of evidence, particularly ESCAT I-II.

ETV6::RUNX1 fusion is one of the most common genetic alterations among children with precursor B-cell acute lymphoblastic leukemia (B-ALL). This fusion generally results from a t(12;21), which fuses the 5' region of ETV6 to the 3' region of RUNX1. Here we report a novel atypical fusion in a 2-year-old male patient with B-ALL. Chromosome, FISH, and microarray were first performed to define a genetic subtype. Chromosome analysis showed three copies of chromosome 21. FISH results revealed ETV6::RUNX1 fusion. However, the signal pattern was atypical with the two fusion signals sitting on two different chromosome 21s, a RUNX1 signal on the third chromosome 21, and two ETV6 signals on two chromosome 12s. Microarray did not readily resolve the FISH signal pattern but confirmed gain of the entire chromosome 21 without any imbalances and showed two losses affecting exon 2 and exons 6-8 of ETV6. Subsequently, optical genome mapping (OGM) identified the fusion created by excision and insertion of ETV6 exons 3-5 into intron 2 of RUNX1. Long-read DNA and RNA sequencing confirmed the fusion identified by OGM and resolved the fusion breakpoints. The identified novel RUNX1::ETV6::RUNX1 fusion contains all important elements reported in the classical ETV6::RUNX1 fusion and is assumed to be oncogenic. Further, transcriptome profiling demonstrated that this case clusters with other typical ETV6::RUNX1 fusion B-ALL cases, suggesting similar functional behavior from this atypical fusion. The patient was treated using a standard risk ALL protocol and is in remission for two years. This report demonstrates how comprehensive genomic profiling with OGM and long-read sequencing can help resolve atypical findings from conventional methods, enable proper sub-classification and potentially inform risk assessment in relapsed or drug-resistant disease.

Cutaneous adverse drug reactions (cADRs) are unintended and harmful skin responses to medications that impose significant clinical and economic burdens worldwide. The increasing use of novel agents, particularly immune checkpoint inhibitors, has further compounded this challenge. While most cADRs are mild and resolve upon drug discontinuation, approximately 2-6.7% progress to severe, potentially fatal conditions. The most common severe forms include acute generalized exanthematous pustulosis, Stevens-Johnson syndrome/toxic epidermal necrolysis, drug reaction with eosinophilia and systemic symptoms/drug-induced hypersensitivity syndrome, and drug-associated bullous pemphigoid. Current therapeutic options for refractory cADRs remain limited, primarily relying on systemic corticosteroids, conventional immunosuppressants, and intravenous immunoglobulin. Growing insights into disease pathogenesis and the subsequent repurposing of novel targeted therapies offer promising solutions to the persistent challenges of cADRs. Emerging agents, such as biologics targeting tumor necrosis factor-α, interleukins (IL-4/IL-13, IL-5, IL-6, IL-17, IL-36), immunoglobulin E, and CD20, along with small molecule inhibitors of Janus kinases and phosphodiesterase 4, hold the potential to revolutionize management paradigms, though their long-term efficacy and safety profiles await robust clinical validation.

The vast majority of reported ALK rearrangements occur at exons 17-20. ALK-rearrangements occurring earlier in the ALK gene (eALK) are poorly characterized and rarely reported, potentially because most clinically available biomarker testing is optimized for detecting common ALK rearrangement regions. Two databases, the Tempus deidentified database and Cedars-Sinai database, were retrospectively queried to characterize eALK rearrangements, as defined by rearrangements involving ALK exons 1-16. Select specimens harboring eALK rearrangements were evaluated using available FDA-approved methods (immunohistochemistry [IHC] and fluorescent in-situ hybridization [FISH]); clinical management and outcomes were evaluated for these cases. 39 eALK rearrangements were detected in the Tempus database and 3 were identified in the Cedars-Sinai database. Across these 42 rearrangements, there were 35 unique partner genes; rearrangements were identified in all early exons except exons 8, 10, 13 and 15, with exons 2 and 4 being most common. Prostatic, breast, leiomyosarcoma, and ovarian serous carcinoma were the most common tumor types, which may reflect database bias. In three cases, IHC and FISH were unreliable at detection (0/ 3 IHC positive, 1/3 FISH atypical positive). Limited clinical information shows partial response to ALK targeted therapies. Novel eALK rearrangements are a distinct subclass of ALK rearrangements and were not reliably detected through FDA-approved testing modalities. Limited clinical data suggests these cancers could still respond to ALK targeted therapy.

Pediatric acute myeloid leukemia (pAML) has a poorer prognosis than acute lymphoblastic leukemia, and hematopoietic stem cell transplantation (HSCT) offers curative potential in high-risk or relapsed cases. Current models cannot accurately determine which individual patients will truly benefit from HSCT, leading to overtreatment or undertreatment. We developed HSCT-64, the first parallel transcriptomic risk framework for pediatric AML, conceptually analogous to a causal G-formula approach. It comprises two treatment-specific models, aHSCT-64 for allo-HSCT recipients and nHSCT-64 for non-HSCT patients, derived from a shared 64-gene signature identified from diagnostic RNA-sequencing data, enabling individualized survival prediction under both treatment scenarios at diagnosis. Trained on 1647 cases from four COG/TARGET cohorts and validated in 233 independent patients, HSCT-64 achieved a C-index of 0.791 and AUC of 0.794 for allo-HSCT overall survival, outperforming existing clinical, cytogenetic, and leukemia stem cell-based models. Comparing risk ranks between two models identified an HSCT-benefiting subgroup patients with a predicted risk rank reduction from HSCT who experienced a 5.88-fold mortality reduction post-transplant (Hazard Ratio, HR = 0.17, P = 0.0066), while no survival gain was seen in the nonbenefiting subgroup (HR = 0.94, P = 0.899). HSCT-64 enables precise, diagnosis-time identification of pAML patients most likely to benefit from transplantation, marking a shift from high-risk-based recommendations toward individualized, transcriptome-driven decision-making.

Genomic testing is now embedded in contemporary prostate cancer care, yet the clinical meaning of different genomic platforms varies substantially by disease state and clinical context. In localized disease, tissue-based genomic classifiers primarily serve prognostic functions by refining risk estimates beyond clinicopathologic variables, whereas in advanced disease, germline and somatic testing identify predictive biomarkers linked to therapy selection. This distinction is clinically consequential because the supporting evidence, endpoints, and implementation challenges differ across assays and across points on the disease continuum. In this review, we position tissue-based assays, germline testing, somatic sequencing, circulating tumor DNA (ctDNA), and artificial intelligence-enabled biomarkers within a unified clinical framework spanning localized disease, biochemical recurrence, and metastatic progression. We critically compare commercially available genomic assays with respect to methodology, specimen type, intended use, validation cohorts, and clinically relevant outcomes. We distinguish prognostic classifiers from predictive biomarkers such as homologous recombination repair deficiency and mismatch repair deficiency, and we evaluate emerging approaches, including liquid biopsy, multimodal integration with imaging, and digital pathology-based algorithms. We further address implementation barriers that may limit real-world impact, including reimbursement uncertainty, disparities in access to next-generation sequencing, limited provider familiarity with genomic interpretation, and the need for patient-centered communication and navigation in genomics-informed care. A clinically useful framework for prostate cancer genomics must therefore move beyond cataloging tests and instead clarify when genomic results change management, where evidence remains immature, and how implementation strategies can improve equity and actionability.

Philadelphia chromosome-positive acute lymphoblastic leukemia (-Ph+ ALL) is a high-risk leukemia often accompanied by additional genomic abnormalities beyond BCR::ABL1. We analyzed 74 newly diagnosed adult -Ph+ ALL patients. Additional chromosomal abnormalities (ACAs) were assessed by conventional cytogenetics in 51 patients and copy number variations (CNVs) by combined array comparative genomic hybridization plus single nucleotide polymorphism (array-CGH+SNP) in 43 patients. ACAs were detected in 31 cases (60.8%), including complex karyotypes in 11 (35.5%). CNVs were identified in 38 cases (88.4%) and were predominantly deletions (69.2%). Recurrent abnormalities included double Ph chromosome, monosomy 7, chromosomal gains at 8q, 9q34, 22q11, and losses at 7p, 9p, and 20q. The t(3;9;22) was the most frequent t(9;22) variant (12.9%), despite being rare in previous studies of adult Ph+ ALL. IKZF1 deletions were most frequent (91.7%), followed by CDKN2A/B (44.4%), PAX5 (38.9%) and RB1 (30.6%). IKZF1 co-deletions with CDKN2A/B and/or PAX5 occurred in 66.7%. IKZF1 and 22q11.22 co-deletions occurred in 34.2%, and VPREB1 deletions in 15.8%; both have been associated with adverse outcomes in ALL. Copy-neutral loss of heterozygosity >5 Mb was detected in 16.3%, frequently involving chromosome 9p and encompassing the CDKN2A/B loci. Adult Ph+ ALL in this multi-ethnic cohort exhibited marked genomic heterogeneity, with frequent submicroscopic alterations detectable only through integrated genomic profiling. Comprehensive genomic characterization is needed to better understand leukemogenesis and refine risk stratification.

The aim of this study was to determine whether overall and time-specific patterns of hyperglycaemia, particularly soon after diagnosis, are associated with incident cancer in adults with newly diagnosed type 2 diabetes. We retrospectively analysed a territory-wide cohort of 52,926 Hong Kong Chinese people with newly diagnosed type 2 diabetes. We examined cancer risk across groups of individuals classified according to their time-weighted mean HbA1c over the entire follow-up period (n=49,978) or during specific early exposure periods (n=39,185). A weighted cumulative exposure model was used to determine the role of historical HbA1c exposures in cancer development (n=49,966). Among 49,978 individuals with newly diagnosed type 2 diabetes, 1758 cancer events occurred. Each 11 mmol/mol (1%) increase in time-weighted mean HbA1c was associated with a 27% relative higher risk of cancer at any site (HR 1.27; 95% CI 1.20, 1.33). Within the first 2 years after diagnosis, a time-weighted mean HbA1c ≥53 mmol/mol (≥7.0%) vs <53 mmol/mol (<7.0%) was associated with a 30-75% relative higher risk of cancer at any site, depending on the specific HbA1c category, even after adjusting for subsequent HbA1c. Longer durations of early exposure were associated with higher risk, reaching 51-213% in the first 5 years of exposure. Earlier high HbA1c exposures contributed more strongly to cancer risk than later exposures. A 11 mmol/mol (1%) HbA1c reduction at 1-2 years was associated with a 6% relative lower cancer risk over a hypothetical 10 year window (HR 0.94; 95% CI 0.91, 0.98), whereas reductions after 5 years showed no significant risk differences. Overall, hyperglycaemic exposure was associated with an elevated long-term cancer risk in type 2 diabetes. Notably, individuals who showed better glycaemic management soon after diagnosis exhibited a lower cancer risk than those whose glycaemic management improved later, despite comparable overall glycaemic burdens.

Core-binding factor (CBF) acute myeloid leukemia (AML) with t(8;21)(q22;q22)/RUNX1::RUNX1T1 is typically considered as a favorable-risk AML in the context of cytarabine-based intensive chemotherapy. However, in some situations such as additional adverse-risk mutations or cytogenetics, the prognosis and disease course may be more uncertain. Here, we report the case of a young patient diagnosed with CBF-AML and RUNX1::RUNX1T1 fusion gene, carrying a rare and complex three-way t(8;11;21)(q22;q13;q22) translocation, with mutated KIT, ASXL1 and TET2 genes, transforming into an aggressive and multi-refractory mediastinal myeloid sarcoma. This case illustrates that this scarcely reported variant might negatively impact the favorable prognosis of CBF-AML.

Philadelphia-negative myeloproliferative neoplasms are chronic blood neoplasms. Treatments control blood counts, but disease can progress to myelofibrosis or acute myeloid leukemia. We performed longitudinal whole-genome and targeted sequencing in 30 patients, integrating clonal dynamics with 7,986 blood counts and clinical histories. Distinct evolutionary patterns distinguished stable from progressive disease, with leukemic transformation arising via TP53 loss, stepwise driver mutation acquisition within complex clones, or emergence of independent leukemic clones. In contrast, stable disease showed long-term clonal equilibrium without new drivers. Phylogenetic analysis using 203 whole-genomes of hematopoietic colonies revealed age-appropriate polyclonal hematopoiesis in triple-negative essential thrombocythemia and germline predisposition to thrombocytosis, supporting non-neoplastic origins. Therapy-associated mutagenesis was observed, including C > G mutations following azacitidine and characteristic T > A/T > G after hydroxycarbamide exposure in blood cells, although not in skin where UV damage predominated. These findings demonstrate that progression is genomically encoded years in advance and support serial monitoring and further study of treatment-related mutagenesis. Longitudinal whole-genome sequencing shows MPN progression is genomically encoded years before clinical transformation, with distinct evolutionary routes to leukemia and MF. It identifies DNA mutagenesis associated with HC and 5-azacitidine, suggests some triple-negative cases are nonclonal, and supports serial clinical genomic monitoring for improved risk stratification and long-term management. See related commentary by Agarwal and Sankaran, p. 1724.

The purpose of this study was to evaluate the contribution of radiomics features extracted from various pancreatic structures on computed tomography (CT) images, including the main pancreatic duct and cystic lesion, for predicting the pathological grade of intraductal papillary mucinous neoplasms (IPMNs) using machine learning models. A retrospective study using preoperative CT images obtained during the venous phase of enhancement in patients with pathologically confirmed IPMNs (2003-2024) was conducted. Main pancreatic ducts and cysts were manually segmented. Machine learning models were trained to classify IPMNs into high-grade/associated invasive carcinoma (HG/I) IPMNs or low-grade (LG) IPMNs using radiomics features from three structures (i.e., cysts, main pancreatic duct, and a combination of both structures). Model performance was evaluated using area under the receiver operating characteristic curve (AUC). SHapley Additive exPlanations (SHAP) values were used to interpret feature importance. A total of 274 patients with IPMNs were included. There were 149 patients with HG/I IPMNs (70 women [47 %]; median age, 71.0 years; age range: 29-92) and 125 patients with LG IPMNs (73 women [58.4 %]; median age, 68.0 years; range: 35-86). HG/I IPMNs were predominantly mixed-type IPMNs (51.7 %; 77/149). LG IPMNs were unspecified (51/125; 40.8 %), main/mixed (40/125; 32 %), or branch-duct type (34/125; 27.2 %). A support vector machine trained on combined features achieved the largest AUC (0.85; 95 % confidence interval [CI]: 0.85-0.87; P < 0.001), with 90 % sensitivity (95 % CI: 90-93), and 60 % specificity (95 % CI: 58-62). SHAP analysis identified main pancreatic duct radiomics features as having the largest contribution to model output. Integrating CT-based radiomics features from pancreatic ducts and cysts improves classification performance, with main pancreatic duct features being the most contributive predictor of IPMN grade.

It remains uncertain whether lower CD19 expression is clinically relevant for outcomes of CD19 CAR-T therapy of large B-cell lymphoma (LBCL). We conducted an integrative analysis of tumor CD19 levels with centralized quantitative assessment by flow cytometry (FC), immunohistochemistry and RNA-sequencing in a LBCL cohort (n=301). Pre-treatment CD19 by FC correlated with 1-year progression-free survival following axicabtagene ciloleucel or lisocabtagene maraleucel (16%, 53% and 64% for CD19low, CD19intermediate and CD19high, respectively; p=0.005). After CAR-T relapse, proportion of CD19low tumors increased (from 17% to 35%). Concordant associations were observed by immunohistochemistry and RNA-sequencing. Transcriptomic profiling linked lower CD19 to inflammatory pathways, validated in an external LBCL cohort (n=1,017). Genetic loss of the CD19 locus was observed in 8% of pre-CAR-T and 11% of post-CAR-T tumors (p=0.82), while no CD19 coding mutations were identified. Overall, CD19 expression is a clinically meaningful determinant of CAR-T outcomes, supporting quantitative assessment to improve risk stratification.

Cancer is associated with a significant short-term risk of venous thromboembolism (VTE). With substantial improvements in cancer survival, concern has been raised about an increased risk of VTE in long-term cancer survivors. We aimed to investigate the long-term risk of VTE in 5-year cancer survivors according to cancer subtype, age, and sex compared with a matched background population. Using Danish nationwide registries, we matched 5-year survivors of 21 cancers (≥18 years; diagnosed 2000-2017; included 2005-2022 [5 years after cancer diagnosis]) with individuals from the background population (1:1 matching for the overall population and 1:4 matching for each cancer subtype). Rates of VTE were examined using Cox regression. In the overall population, 222 297 five-year cancer survivors were matched with 222 297 individuals from the background population (median age 69.2 years; 43.6% males). During a median follow-up of 5.6 years, cancer survivors had a higher associated rate of VTE compared with the background population (adjusted hazard ratio [aHR] 1.85 [95%CI, 1.78-1.93]). Survivors of all 21 cancer subtypes had a higher associated rate of VTE compared with the background population, with the highest incidence rates found for multiple myeloma. HRs were higher in younger cancer survivors compared with older cancer survivors (<50 years: aHR, 2.85 [95%CI, 2.33-3.47]; 50-64 years: aHR, 2.26 [95%CI, 2.07-2.46]; ≥65 years: aHR, 1.69 [95%CI, 1.61-1.77]; Pinteraction<0.0001). Similar HRs were found in females and males (females: aHR 1.84 [95%CI, 1.75-1.95]; males: aHR 1.86 [95%CI, 1.75-1.98]; Pinteraction=0.94). Five-year cancer survivors had a higher associated rate of VTE compared with the background population.

Waldenström macroglobulinemia (WM) is a rare, indolent lymphoplasmacytic lymphoma characterized by bone marrow infiltration of IgM-secreting lymphoplasmacytic cells and associated with the MYD88(L265P) mutation in over 90% of cases. This mutation drives constitutive NF-κB signaling through aberrant MYD88 oligomerization, rendering WM dependent on BCR-proximal kinases including BTK for survival. Despite significant therapeutic advances over the past decade, including the established efficacy of BTK inhibitors, proteasome inhibitors, and anti-CD20 monoclonal antibodies in both frontline and relapsed settings, unmet needs persist-particularly regarding treatment durability, acquired resistance to covalent BTK inhibitors via C481S mutations, and the need for chemotherapy-free alternatives. The 2025 American Society of Hematology (ASH) Annual Meeting featured several pivotal clinical trials addressing these challenges, spanning optimized immunochemotherapy combinations in treatment-naive disease, next-generation BTK-targeted agents for relapsed/refractory cases including non-covalent inhibitors that bypass C481S-mediated resistance and protein degraders that eliminate both BTK kinase activity and scaffolding function, and emerging modalities including BCL2 inhibitors that exploit the apoptotic dependence of WM cells on BCL2 family proteins, MALT1 inhibitors that target downstream NF-κB signaling, and CD19-targeted antibody-drug conjugates and cellular therapies.

Post-endoscopy upper gastrointestinal cancer (PEUGIC) is an emerging quality metric, but varying definitions complicate benchmarking. We characterized PEUGIC rates and outcomes by anatomical site and coded high-risk condition (HRC) status in a large US cohort. Retrospective cancer-first cohort study of 14,814 patients with upper gastrointestinal cancer (UGIC) from a multicenter database (2016-2024). Detected UGIC was diagnosed within 6 months of EGD. PEUGIC was defined as UGIC diagnosed 6-36 months after a non-diagnostic EGD. The primary outcome was the PEUGIC rate by cancer type (esophageal [EC], gastric [GC], duodenal [DC]) and HRC status. Secondary outcomes included 3-year overall survival. The overall PEUGIC rate was 9.4% (EC 9.6%, GC 8.6%, DC 10.7%). Rates were higher with HRC (18.2%) versus without HRC (5.0%, P < .001). PEUGIC rate without HRC plateaued (4.1-5.1% in 2022-2024), while PEUGIC rate with HRC for EC (16.8%) and GC (14.4%) decreased over time. In exploratory analysis, PEUGIC was associated with better survival versus Detected UGIC for EC (adjusted hazard ratio [aHR] 0.82, 95% CI 0.67-0.99) and GC (aHR 0.77, 0.62-0.96), and PEUGIC with HRC was associated with lower mortality versus PEUGIC without HRC for EC (aHR 0.51, 0.36-0.74) and GC (aHR 0.50, 0.34-0.73). One in 10 upper GI cancer patients had a prior non-diagnostic endoscopy within 3 years. PEUGIC rates varied by anatomical site and HRC status, suggesting site- and HRC-stratified benchmarks may better capture endoscopic performance than aggregate metrics.

Biosimilars are highly similar to originator biologics and were introduced to increase market competition, lower the price of biologics, and expand patient access by improving affordability. By the end of 2024, three anticancer biologics have experienced biosimilar entry in the US (bevacizumab, rituximab, and trastuzumab), underscoring the need to evaluate their financial implications for payers and patients. To assess market dynamics and examine financial implications associated with biosimilar use patterns among patients who received bevacizumab, rituximab, or trastuzumab. This retrospective cohort examined price and market share trends and payer and patient costs after biosimilar entry. It used data from the Merative MarketScan commercial database and Medicare database from July 2019 to December 2024. Based on use patterns within 12 months of treatment initiation, patients were categorized into 4 groups: those who exclusively used biosimilars, exclusively used the reference product (ie, originator biologics), switched from the reference product to biosimilars, and switched from biosimilars to the reference product. Data were analyzed between September 2025 and March 2026. The primary study outcomes were average sales price (ASP), market share of reference products, and mean monthly payer and out-of-pocket costs within the study period. Among 14 655 patients in this study, the mean (SD) age was 57 (13) years, 9985 (68.1%) were female, and the most common cancers were breast cancer (5182 [35.4%]), lymphoma (4320 [29.5%]), and colorectal cancer (1998 [13.6%]). A total of 8710 patients (59.4%) exclusively used biosimilars, 4760 (32.5%) exclusively used the reference product, 1015 (6.9%) switched from the reference product to biosimilars, and 170 (1.2%) switched from biosimilars to the reference product. After biosimilar entry, the ASP of the 3 reference products declined by an average of 3.8% annually. Their market share decreased 30% annually in commercial insurance and 31.5% in the Medicare Part B market. Compared with patients who only used the reference product, those who only used biosimilars rendered a $3820 lower mean monthly payer cost and $39.50 lower mean monthly out-of-pocket cost. The results of this cohort study suggest that biosimilar entry was associated with lower ASP and reduced market share for reference biologics in oncology. Market competition introduced by biosimilars was also associated with lowered payer costs and reduced patient financial burden.

The hippocampus comprises discrete anatomic subfields subserving different components of memory. We evaluated longitudinal, dose-dependent volumetric changes in hippocampal subfields after fractionated RT and determined their associations with verbal and visuospatial memory performance. Eighty-nine adults with primary brain tumors received fractionated RT on a prospective clinical trial. High-resolution 3D volumetric MRI and memory tests were obtained at baseline and 3, 6, and 12-months post-RT. Bilateral hippocampi and their thirty-eight subfields were segmented using robust automated parcellation. Linear mixed-effects (LME) models analyzed (1) time-dependent atrophy, (2) dose-volume relationships, and (3) subfield-memory associations at the same timepoint, corrected for multiple comparisons. Multiple left and right-sided hippocampal subfields demonstrated significant atrophy at 3, 6, and 12-months post-RT (all p ≤ 0.05). Dose-dependent atrophy was significant at 12 months (p = .01) in the left hippocampus and across all time points in the right (all p ≤ 0.05). Eight right-sided and one left-sided subfield exhibited dose-dependent atrophy across all time points (all p ≤ 0.05). Greater left hippocampal tail and molecular layer volumes were associated with higher verbal memory scores, while greater volumes of multiple right-sided subfields predicted better visuospatial memory performance (all p ≤ 0.05, 𝛽 > 0). Fractionated RT induces progressive, dose-related atrophy in discrete hippocampal subfields, with earlier and steeper dose-response curves in right-sided subfields. Higher volumes within multiple left and right-sided subfields were associated with better verbal and visuospatial memory, respectively. Subfield-sparing planning objectives may optimize cognitive outcomes in primary brain tumor patients when hippocampal avoidance is not feasible.

Watch-and-wait is standard of care in asymptomatic early-stage chronic lymphocytic leukemia (CLL). The CLL12 trial investigated ibrutinib vs placebo for patients with early-stage CLL with intermediate to very high risk of progression, improving event-free survival (EFS) but not overall survival (OS). Building on these findings, our analysis examined whether a benefit could be identified within distinct genetic subgroups. After a median follow-up of 69.3 months, there were 166 EFS and 32 OS events in 515 trial patients. In the placebo arm, del(17p), del(11q), +12, unmutated immunoglobulin heavy variable chain (U-IGHV), and mutations in NOTCH1, ATM, NRAS/KRAS/BRAF, and NFKBIE correlated with shorter EFS. With ibrutinib, only del(17p) and TP53 and NFKBIE mutations significantly compromised EFS. Ibrutinib offered substantial EFS benefit in subgroups with U-IGHV, del(11q), +12, NOTCH1, ATM, and NFKBIE mutations. No EFS improvement was seen for patients with asymptomatic early-stage CLL with del(17p) or TP53 mutations. Ibrutinib provided no OS benefits in any genetic subgroup. Multivariable analysis revealed ibrutinib treatment as independent favorable factor for EFS, whereas U-IGHV, del(17p), POT1, RAS/RAF, and NFKBIE mutations were adverse prognostic factors. Results confirm watch-and-wait as standard of care for patients with early-stage CLL, especially in high-risk CLL characterized by del(17p) and/or mutated TP53. This trial was registered at the European Union Drug Regulating Authorities Clinical Trials Database (2013-003211-22).

Endoscopic papillectomy (EP) enables resection of ampullary lesions but is limited by piecemeal resection and high recurrence rates. To overcome these limitations, we developed endoscopic submucosal dissection including papilla (ESDIP). This study aimed to evaluate clinical outcomes and identify risk factors for local residual/recurrent tumor after ESDIP for laterally spreading ampullary tumors. This single-center retrospective study analyzed consecutive patients who underwent ESDIP between 2011 and 2024. The primary outcome was local residual/recurrent tumor after ESDIP. Risk factors for local residual/recurrent tumor were evaluated using Cox proportional hazards regression analysis. Of the 81 consecutive patients in whom ESDIP was attempted, three were converted to EP or surgery. En bloc resection was achieved in all 78 patients. After excluding 23 patients based on the predefined exclusion criteria, 55 patients were analyzed. The median tumor size was 38 mm (range, 15-128 mm). Negative horizontal and vertical margins, excluding the papillary ductal margin (PM), were achieved in 48 patients (87%), whereas negative PM and R0 resection were achieved in 34 patients (62%) and 29 patients (53%), respectively. Median follow-up was 31 months. The 3-year cumulative incidence of residual/recurrent tumor was 33%, and all residual/recurrent tumors were confined to the papilla. Positive/indeterminate PM was a significant risk factor for local residual/recurrent tumor (hazard ratio, 8.35; 95% confidence interval, 2.35-29.64). Although ESDIP enabled en bloc resection of laterally spreading ampullary tumors, the high rate of positive/indeterminate PM, resulting in frequent papilla-confined residual/recurrent tumors, remained its main limitation.

Aneuploidy is a hallmark of cancer and is associated with drug resistance and poor clinical outcomes across diverse cancer types. However, no therapies have been clinically established to target highly aneuploid tumors. By analyzing nearly half a million tumor samples subjected to comprehensive genomic profiling, we identified a striking mutual exclusivity between POLE exonuclease domain mutations and high aneuploidy burden. This observation was independently validated using data from The Cancer Genome Atlas (TCGA) and the Cancer Cell Line Encyclopedia (CCLE). Probabilistic modeling revealed that the elevated quantity and unique spectrum of mutations induced by POLE exonuclease deficiency increase the likelihood of inactivating essential genes on chromosome arms harboring losses, leading to a synthetic lethal phenotype in highly aneuploid cells. Functional experiments demonstrated that POLE exonuclease activity is essential for the viability of highly aneuploid cancer cell lines but dispensable in diploid cells. These findings suggest that selective inhibition of POLE exonuclease activity may represent a promising therapeutic strategy for targeting highly aneuploid tumors. An integrated approach using large-scale genomic analyses, probabilistic modeling and functional validation identified POLE exonuclease as a potential synthetic lethal target to overcome cancer aneuploidy.

A population of cells within a tumor can be described as antifragile (the opposite of fragile) if they derive a benefit from fluctuations or perturbations in environmental conditions induced by treatment. Treatment fluctuations could either promote (antifragile tumor) or inhibit (fragile tumor) the evolution of treatment resistance to targeted therapies. In this study, we showed that analysis of the convexity of dose-response curves provided a direct prediction of response to prescribed fluctuations in treatment. Convexity predicted that continuous treatment protocols (i.e., zero prescribed fluctuations in treatment) would outperform uneven protocols when dose response is convex. This theory was applied to predict in vivo response to targeted therapy, and the predictions were validated by experimentally testing high/low intermittent dosing (uneven dosing) and continuous dosing (even dosing) schedules. Convexity (and its inverse, concavity) explained 2 phenomena: Dose response is a convex (fragile) function, but the resistance onset rate is a concave (antifragile) function. Thus, design and validation of alternative treatment schedules maximized response while maintaining prolonged sensitivity to treatment. Together, these analyses provide supporting evidence that fluctuations alter the evolutionary trajectory of tumors in response to targeted therapy, even without altering the cumulative dose. By using this insight to design alternative treatment protocols to limit the evolution of resistance by careful analysis of the dose-response curvature, the study supports the potential of "evolutionary antifragile therapy" as a subtype in the broad class of evolution-based treatment strategies. The mathematical method using dose response curvature to determine appropriate treatment dosing that limits the evolution of resistance could facilitate designing dose schedules that optimize tumor response. See related commentary by Pomeroy and Palmer, p. 4195.

Cellular function depends on the spatial organization of cells and biomolecules within the tissue microenvironment. Advances in spatial omics have enabled profiling of molecular features such as transcriptome, proteome and epigenome, and there has been rapid progress of imaging-based approaches to study spatial three-dimensional (3D) genome organization. Here we present Spatial-ATAC-Hi-C, a microfluidic‑based platform for genome-wide, spatially resolved joint-profiling of 3D genome organization and chromatin accessibility on tissue slides. Applied to mouse and human brains, Spatial-ATAC-Hi-C revealed distinct chromatin architecture and gene regulatory programs in neuronal and non-neuronal populations in their native tissue context. In glioblastoma and astrocytoma samples, we detected spatially resolved 3D genome alterations, copy number variations and structural variations across tumor regions, revealing clinically relevant oncogenic events and clonal heterogeneity. By co-profiling of genome architecture and chromatin accessibility while preserving tissue architecture, Spatial-ATAC-Hi-C provides a powerful tool for studying spatial gene regulation in human biology and disease.

The insulin-like growth factor 2 mRNA-binding protein (IGF2BP) family is overexpressed in cancer and associated with poor prognosis. IGF2BP2 has been linked to single metabolic alterations by acting on its RNA targets. Here, we used a comprehensive approach to elucidate the effects of IGF2BP2 on primary and lipid metabolism. 13C-metabolic flux analysis (MFA) combined with RNA-Seq data revealed that IGF2BP2 affects mitochondrial fluxes by regulating the expression of several mitochondrial transporters, such as mitochondrial pyruvate carrier 1 (MPC1) and uncoupling protein 2 (UCP2). Methyl pyruvate reversed the gene expression patterns of UCP2 and CPT1A in HCT116 IGF2BP2 knockout (KO) cells by bypassing MPC1. Interestingly, an altered expression of the transporter UCP2 was also observed in a patient-derived tumor organoid (PDO), in which IGF2BP2 was knocked down. The altered glutamine metabolism seen in the 13C-MFA and the citrate label data derived from extracted mitochondria confirm a rerouting of glutamine almost exclusively into the mitochondria and a reduction of glycolytic carbon intake into the mitochondria. Due to changes in palmitate labeling patterns, lipid stainings were performed, suggesting lipid accumulation in KO cells. A lipidomic analysis revealed altered compositions across almost all lipid species. Further, lipogenic genes involved in fatty acid and cholesterol metabolism were differentially expressed. Most of the differentially expressed genes are potential direct targets of IGF2BP2 based on publicly available IGF2BP2 CLIP data. Overall, these results show the influence of IGF2BP2 on the central carbon metabolism of cancer cells, primarily through its effects on MPC1 and the resulting effects on UCP2. The complex interaction of IGF2BP2 with the metabolic network provides important insights into tumor metabolism, particularly relevant to tumor growth and resistance to therapy.

Milk-derived extracellular vesicles (EVs) are a promising source of molecules with therapeutic potential. Bovine colostrum is particularly enriched in EVs, which carry cargo of proteins involved in immune regulation, development and cellular signalling. Some studies have explored their role as bioactive anti-cancer agents, however, their mechanistic effects remain underexplored. Here, we show that colostrum-derived EVs (Col-EVs) exert anti-proliferative effects in gastrointestinal cancer models, including cell lines and patient-derived organoids, which is independent of apoptosis induction. Using a multi-modal approach combining proteomics, imaging and functional assays, we demonstrate that Col-EVs induce a reversible growth-arrest state, characterized by widespread transcriptional and RNA-processing dysregulation, chromatin compaction, nuclear reorganization and cytoskeletal remodelling. Proteomic analyses reveal that Col-EV treatment disrupts key components of the transcriptional machinery and cell cycle regulatory pathways, effects that are reversible upon EV withdrawal and can be rescued pharmacologically using an EZH2 inhibitor. Col-EVs enhance the sensitivity of cancer cells as well to DNA-targeting chemotherapies such as 5-fluorouracil, indicating their potential as modulatory adjuvants rather than cytotoxic agents. Overall, our findings reveal that Col-EVs can reversibly suppress cancer cell proliferation by reprogramming transcriptional and nuclear architecture, offering a natural, biocompatible strategy for modulating tumour growth and sensitizing cancer cells to conventional therapies.

RNA-binding motif protein 15 (RBM15) is a key regulator of hematopoiesis and leukemogenesis, linking RNA metabolism, epitranscriptomic regulation, and lineage specification. Identified through the RBM15::MKL1 fusion in acute megakaryoblastic leukemia (AMKL), RBM15 functions within the N6-methyladenosine (m6A) writer complex, where its RNA-recognition motifs and Spen paralog and ortholog C-terminal domain enable site-specific methylation and protein recruitment. In hematopoietic stem and progenitor cells, RBM15 supports stem cell quiescence, self-renewal, and balanced lineage output, with additional roles in B cell, myeloid, and megakaryocytic differentiation. Mechanistically, RBM15 links m6A deposition to chromatin regulation, RNA export, and splicing. This review summarizes RBM15 structure, function, and regulation and highlights its emerging roles in leukemia, including AMKL and myelodysplastic syndromes, while outlining key questions for future studies.

Cancer-associated cachexia (CAC) is a progressive, multiorgan wasting syndrome that critically limits treatment response and survival. In CAC, the nervous system senses, amplifies, and coordinates responses to a broad range of tumor-induced perturbations, including both direct tumor-derived factors and the resulting systemic disturbances in inflammation, metabolism, and immunity. By dysregulating central neural circuits and remodeling peripheral innervation, tumor-hijacked neural coordination drives both anorexia and multiorgan wasting, forming a self-reinforcing cachexia loop. Additionally, treatment-associated neurotoxicity further augments dysfunction across the central and peripheral nervous systems. In this review, we synthesize evidence from patient studies and animal models to examine how tumors co-opt the nervous system to systemically organize cachexia, thereby providing a pathophysiological framework for studying early disease detection, biomarker development, and mechanism-based intervention for CAC.

Proteogenomic cancer antigen discovery now extends beyond somatic mutations to include unmutated, aberrantly expressed, and non-canonical sources. We distinguish two orthogonal dimensions: molecular origin (canonical or non-canonical) and normal-tissue distribution (tumor-specific antigen, tumor-associated antigen, or lineage-specific antigen). Importantly, non-canonical origin does not imply tumor specificity. For human leukocyte antigen (HLA)-restricted immunotherapy, selectivity must be established at the level of naturally presented peptide-HLA (pHLA) complexes rather than inferred from RNA expression or total protein abundance. We therefore propose a normal-immunopeptidome denominator: a context-aware atlas of benign pHLA ligands that can be used to exclude unsafe candidates before therapeutic nomination. However, this denominator is necessary but not sufficient. Candidate nomination also requires evidence of analytical confidence, natural presentation on tumor cells, and, where available, peptide abundance and tumor heterogeneity. Additional considerations include whether the target is patient-specific or shared, its functional selectivity, the potential for immune escape, receptor cross-reactivity, HLA alloreactivity, and the modality-specific therapeutic index. We present a tiered framework that distinguishes core requirements, risk-triggered evaluation criteria, and recommended enhancements. These requirements escalate as uncertainty increases. Importantly, the absence of a peptide from existing databases should not be interpreted as evidence of its absence in humans.

Acute myeloid leukemia (AML) is an aggressive hematologic malignancy with dismal outcomes, especially in relapsed/refractory settings. Chimeric antigen receptor natural killer (CAR-NK) cell therapy holds promise but is constrained by the immunosuppressive tumor microenvironment (TME), where adenosine-mediated suppression is a key barrier. To develop a novel CAR-NK construct cotargeting AML cells and the adenosine-rich TME to enhance antileukemia efficacy. Ex vivo expanded primary NK cells were used to compare the effects of CD39 versus CD73 blockade on NK cell function via messenger RNA-electroporated antibodies. A CD33-CD73 dual-function CAR-NK construct (integrating CD33-specific lysis and anti-CD73scFv secretion for TME disruption) was designed and transduced into NK cells via retrovirus. Engineered NK cells were characterized for transduction efficiency, expansion, purity, viability, and CAR stability. In vitro cytotoxicity against AML cell lines and primary blasts was assessed, and in vivo efficacy was evaluated in a MOLM-13 xenograft mouse model. CD73 blockade more potently enhanced NK cell activity than CD39 blockade. Retroviral transduction achieved >50% efficiency, and expansion with K562-4-1BBL-mbIL-21/-15 feeder cells yielded NK cells with ≥6,000 fold expansion, >93% purity, >98% viability, and stable CAR expression. At an effector-to-target ratio of 0.5:1, CD33-CD73 CAR-NK cells mediated ~80% specific lysis, with superior cytotoxicity vs conventional CD33 CAR-NK cells. In xenografts, CD33-CD73 CAR-NK cells achieved robust tumor clearance, extended median survival by 24.5 days (59.5 vs 35 days) versus standard CD33 CAR-NK cells, and five out of six mice achieved long-term survival (>50 days). The CD33-CD73 dual-targeting CAR-NK platform synergistically targets AML cells and the adenosine-rich TME, exhibiting superior anti-leukemia efficacy. This strategy advances AML immunotherapy and provides a translational blueprint for TME-targeted therapies in other cancers.

Neoantigen vaccines have rekindled interest in therapeutic cancer vaccination, yet their clinical efficacy remains constrained by imperfect antigen prioritization, incomplete modeling of immunogenicity, tumor heterogeneity, and immune evasion mechanisms. Current computational pipelines are dominated by discriminative models that rank pre-existing mutant peptides based on features related to HLA binding and antigen presentation. Although these approaches have improved candidate prioritization, their ability to optimize antigen selection and vaccine constructs across multiple determinants, including presentation, recognition potential, and translational feasibility, remains limited. Generative artificial intelligence offers a complementary, design-oriented framework that can explore and iteratively optimize biological sequence space under explicit constraints, rather than merely scoring predefined candidates. In this review, we discuss how generative AI may broaden the computational scope of tumor-anchored antigen discovery, support tumor-context-integrated optimization of neoantigen candidates, facilitate the engineering of multi-epitope and mRNA vaccine constructs, and incorporate tumor-specific constraints such as antigen-presentation defects, clonal architecture, and the state of the tumor immune microenvironment. Within this framework, generative models are considered components of an AI-assisted integrated workflow rather than substitutes for tumor-derived evidence, established prediction tools, or experimental validation. We further discuss the role of immunopeptidomics-guided calibration and iterative validation in improving biological realism and translational relevance. A longer-term frontier is immunopeptidomics-anchored synthetic immunogen design, in which validated tumor-presented immunogenic peptides may serve as templates for designing neoepitope mimetics or heteroclitic peptide analogs with improved HLA compatibility, pHLA stability and T-cell priming capacity. Finally, we examine current bottlenecks, including limited functionally validated immunogenicity datasets, uncertain generalizability, experimental validation burden, and the emerging regulatory demand for interpretability and traceability. At present, generative AI should be viewed as a promising design-enabling biotechnology platform whose clinical value remains to be established through prospective comparison with standard neoantigen pipelines.

Hepatobiliary cancers are highly heterogeneous malignant tumors, and the drug resistance frequently emerges at advanced stages. Patient-derived organoids (PDOs) are three-dimensional ex vivo cultures. They can reproduce essential biological aspects of primary tumors, and play a key role in personalized oncology. With advances in methodology and technology, PDOs are increasingly being used in hepatobiliary cancer research at both the fundamental and translational levels. This review summarizes recent advances in patient-derived organoids, including culture matrices, microenvironment reconstruction, and organoid-on-chips. We also discuss the applications of PDOs in hepatobiliary cancer modeling and organoid biobank construction. Hepatobiliary cancer organoids hold promise for drug development and may serve as complementary tools for preclinical prediction. Integrating in vitro drug testing, resistance mechanism identification and combination regimen screening, organoid-based workflows can provide platforms for treatment stratification and personalized treatment optimization in hepatobiliary cancers. The incorporation of artificial intelligence may further expand the capabilities of organoid-based platforms. Overall, hepatobiliary cancer organoids represent clinically promising models for overcoming therapeutic resistance and developing personalized treatment strategies. They are powerful tools for precision oncology and clinical transformation.

Osteosarcoma (OS) is an aggressive bone malignancy with poor outcomes in recurrent and metastatic disease. Although ferroptosis represents as a promising therapeutic strategy, its regulation in OS remains incompletely understood. Here, we identified STK25 as a regulator of ferroptosis susceptibility and tumor growth in OS. Transcriptomic analysis of paired parental and multidrug-resistant OS cell lines identified lower STK25 expression in the resistant derivatives. In a retrospective tissue microarray, low tumor STK25 expression was associated with a lower histological tumor necrosis rate after neoadjuvant chemotherapy and shorter survival. Mechanistically, METTL3 overexpression reduced STK25 reporter activity and accelerated STK25 mRNA decay in an m6A-site-dependent manner. STK25 depletion was accompanied by reduced Hippo pathway activity, increased YAP nuclear localization, and increased GPX4 expression, whereas STK25 restoration increased LATS1 and YAP phosphorylation, reduced GPX4 expression and sensitized OS cells to ferroptosis. GPX4 re-expression partially rescued cell viability and oxidative-stress phenotypes induced by STK25 overexpression, supporting GPX4 as a functional downstream effector. In xenografts, STK25 overexpression significantly suppressed tumor growth and increased ferroptosis-associated changes. These findings support an METTL3-STK25-YAP-GPX4 regulatory model that links epitranscriptomic control to ferroptosis susceptibility in OS and warrants further preclinical and prospective clinical validation.

Intratumoral administration is widely used to achieve high local drug delivery while minimizing systemic toxicity. However, the fundamental assumption that locally injected agents remain within tumor tissue has rarely been quantitatively examined. Here, we show that drug leakage from the tumor surface can reduce leakage-adjusted local retention and limit therapeutic efficacy after intratumoral injection. Using multiple tumor models, conventional needle injection frequently resulted in drug escape from the tumor surface, leading to reduced leakage-adjusted retention and diminished therapeutic efficacy. Mechanical measurements revealed pronounced differences in tissue stiffness across tumor types, suggesting that tumor mechanical properties may contribute to differences in drug leakage and local retention. Histological analysis further showed that guided jet injection induces structural remodeling within tumor tissue, creating transient interstitial spaces that may facilitate intratumoral fluid propagation. Consistent with these structural observations, three-dimensional imaging demonstrated distinct cavity-formation patterns between conventional needle injection and guided jet delivery. Together, these findings indicate that drug leakage is an underrecognized physical limitation of intratumoral drug delivery and that tumor mechanical heterogeneity is associated with local retention. This study provides a rationale for improving local drug delivery through mechanically guided injection strategies.

Drug development is slow, costly, and prone to late-stage failure, in part because animal models poorly predict human responses. Two human-relevant technologies are maturing in parallel: biological avatars, defined as patient- or stem-cell-derived models such as organoids and organ-on-a-chip systems, and digital twins, defined as computational models that integrate a patient's molecular and clinical data to forecast treatment responses. We propose the ex vivo clinical trial concept, in which an avatar and a digital twin are coupled in an iterative loop so that laboratory measurements refine the computational prediction and the prediction guides the next experiment, allowing candidate therapies to be tested and prioritised before a patient is exposed. We review the platforms, their predictive performance in cancer, cystic fibrosis, and liver toxicity, the conditions under which they fail, and the qualification, turnaround, and standardisation requirements that must be met before such trials can inform drug development or clinical care.

Potent and sustained inhibition of epidermal growth factor receptor (EGFR) signaling is critical for suppressing colorectal cancer (CRC) growth, yet current EGFR therapies are often limited by incomplete EGFR blockade and on-target/off-tumor toxicities, particularly skin rash. Here, we co-targeted EGFR and cadherin-17 (CDH17) with a bispecific antibody (bsAb), overcoming these limitations by reducing skin-related toxicities and achieving superior tumor growth inhibition compared to EGFR- or c-MET/EGFR-targeting antibodies. Mechanistically, the high-affinity CDH17-binding arm anchored the EGFR/CDH17 bsAb to the tumor cell surface, facilitating rapid engagement of unbound or newly synthesized EGFR. This "anchor and capture" mechanism allowed the EGFR/CDH17 bsAb to achieve superior and more sustained suppression of the EGFR pathway than cetuximab and amivantamab. Further incorporation of an anti-CD16A nanobody transformed the EGFR/CDH17 bsAb into a trispecific natural killer (NK)-cell engager. Extensive format screening revealed that NK cell activation is heavily influenced by the spatial distance between the tumor antigen-binding Fab and the anti-CD16A nanobody, with longer distances impairing the bulky CD45 phosphatase exclusion from the immunological synapse and leading to significantly reduced cytotoxicity. Consequently, the widely adopted Morrison-type antibody consistently underperformed compared to architectures with shorter CD16A-TAA-Fab spacing. The final optimized molecule was IBI3019, a CDH17/EGFR/CD16A trispecific antibody that integrates potent CDH17-enhanced EGFR blockade with optimal architecture for efficient NK cell engagement. It demonstrated superior in vivo efficacy and a good safety profile in cynomolgus monkeys, with no observable skin toxicity. These promising pre-clinical findings warrant the clinical development of IBI3019. .

The use of anti-programmed cell death-1 (PD-1) antibody increases heart failure (HF) risk in patients with cancer with preexisting cardiovascular conditions. However, the underlying mechanism remains incompletely understood. To evaluate the effects of anti-PD-1 antibody on transverse aortic constriction (TAC)-induced cardiac remodeling and HF, anti-PD-1 antibody-treated mice; T cell-, myeloid-, and CD8+ T cell-specific Pdcd1 knockout; C-X-C motif chemokine receptor 3 (Cxcr3) knockout; and granzyme B (Gzmb) knockout mice combined with flow cytometry, Western blotting, immunofluorescence staining, pharmacological approaches, and bulk RNA-sequencing analyses were used. Administration of anti-PD-1 antibody, T cell-, or CD8+ T cell-specific Pdcd1 deletion, but not myeloid-specific Pdcd1 knockout, aggravated TAC-induced cardiomyopathy and HF in mice. Mechanistically, PD-1 blockade or deletion increased myocardial infiltration of CXCR3+ CD8+ T cells, leading to granzyme B/perforin-mediated impairment of cardiomyocyte mitochondrial complex I to exacerbate TAC-induced cardiac injury and HF. TAC-enhanced chemotaxis, between cardiac fibroblast-derived CXCL9/CXCL10 and CXCR3+ CD8+ T cells, was a driving force for recruiting CXCR3+ CD8+ T cells under the conditions of PD-1 blockade or deletion. The worsened TAC-induced cardiomyopathy caused by anti-PD-1 antibody or T cell-specific Pdcd1 deletion was rescued by genetic deletion or pharmacological blockade of granzyme B and CXCR3. Anti-PD-1 antibody enhances myocardial infiltration of CXCR3+ CD8+ T cells under TAC condition through CXCL9/CXCL10-mediated chemotaxis. The increased granzyme B and perforin likely derived from CD8+ T cells impair function of mitochondrial respiratory chain complexes to cause cardiomyocyte apoptosis, thereby exacerbating TAC-induced cardiomyopathy and HF. CXCL9/CXCL10-CXCR3+ CD8+ T cell axis may represent a promising target for combating anti-PD-1 antibody-associated cardiotoxicity.

Cancer nanomedicine offers a versatile platform for improving therapeutic index, but its clinical translation remains limited by unpredictable in vivo behavior, heterogeneous biological contexts, and inefficient design paradigms. Artificial intelligence (AI) is emerging as an integrative framework that links data-driven modeling with nanomedicine design, biological transport, and clinical decision-making. In this review, we discuss AI-guided strategies for material design, targeting, payload optimization, and in vivo delivery, with particular attention to protein corona-mediated biological identity, microenvironment-responsive activation, and biodistribution modeling. We further examine the translational requirements for AI-enabled nanomedicine, including data standardization, preclinical learning workflows, clinical stratification and risk-based governance. Finally, we outline future directions centered on transferable learning architectures, dynamic multiscale modeling and patient-aware therapeutic strategies. Together, these advances suggest that AI can move cancer nanomedicine beyond empirical formulation toward a more predictive, biologically informed, and clinically responsive discipline.

The immune response is a highly dynamic and precisely controlled process, and relies on an intricate network of protein interactions to maintain its homeostasis. Protein posttranslational modifications (PTMs), by covalent addition of chemical groups or peptide chains, or by proteolytic cleavage, directly alter the structure, activity, electrical charge, thermal stability, and function of immune-related proteins. Here, we enumerate the main PTMs in the immune system and highlight their global regulatory roles across innate immunity, T-cell-mediated cellular immunity, and B-cell-mediated humoral immunity, including phosphorylation, ubiquitination, SUMOylation, acetylation, methylation, glycosylation, and other new types. We focus on how these modifications regulate antigen presentation and recognition, immune signal transduction, antibody production and class switching, immune contraction, immune memory establishment, as well as immune tolerance and homeostasis. Critically, dysregulation of host protein PTMs disrupts immune homeostasis, which in turn induces numerous immune-related diseases, including cancer, autoimmune diseases, and infectious diseases. By integrating mechanistic insights with emerging therapeutic strategies, this review provides a comprehensive PTM network that elucidates the molecular basis of immune-related diseases and highlights promising immunotherapeutic strategies targeting these modifications.

As a key regulator of the immune system, regulatory T cells (Treg cells) suppress excessive immune responses and maintain self-tolerance through various immunosuppressive mechanisms, both contact-dependent and contact-independent. They have demonstrated significant therapeutic potential in immune-related diseases and some non-immune diseases. In this paper, we systematically review the induction strategies of Treg cells, covering various aspects including antigenic stimulation, cytokine modulation, metabolic pathways, epigenetic regulation, drug induction, and engineered technology. Although each strategy has its own advantages, no single approach can comprehensively address the complex issues surrounding Treg cell stability, target specificity, and large-scale production. Therefore, we propose that integrating multiple induction strategies during the Treg cell induction process is crucial for designing Treg cell therapies with superior functionality and broader applications, thereby overcoming the current limitations of Treg cell therapies. In addition, we introduce their therapeutic applications in immune diseases (such as autoimmune disease, organ transplantation, allergic asthma, and cancer) and non-immune diseases (such as insulin resistance, tissue repair, fibrotic diseases, and osteoporosis), and recent advancements. We also discuss current research bottlenecks of Treg cell-based therapeutic strategies and provide an outlook on their future development, aiming to deepen the understanding of Treg cells and promote the advancement of Treg cell-based cellular therapeutic strategies.

Pharmacological tools to selectively modulate extracellular vesicle (EV) secretion are scarce. Here, we identify the ALK5 (TGF-β receptor I) inhibitor SD-208 as a potent suppressor of small EV (sEV) secretion that acts independently of its canonical anti-fibrotic activity. SD-208 not only reversed myofibroblast activation but also markedly inhibited sEV secretion. Strikingly, this inhibitory effect persisted in non-activated cardiac fibroblasts and non-fibrotic HEK293 cells, demonstrating that SD-208 regulates EV secretion through mechanisms uncoupled from TGF-β/Smad signalling. Mechanistic analyses revealed that SD-208 disrupts vesicle trafficking rather than EV biogenesis. Reduced secretion of CD63+ EVs was accompanied by intracellular accumulation of CD63+ structures and their selective diversion into LAMP1+ lysosomes. Proteomic profiling of SD-208-treated and control HEK293 cells and cardiac fibroblasts revealed dysregulation of vesicle trafficking pathways, enrichment of ubiquitin ligase complexes, and enhanced endosome-to-lysosome transport. Together, these findings demonstrate that SD-208 diverts CD63+ multivesicular bodies (MVBs) from a secretory fate toward lysosomal degradation. This work identifies SD-208 as a small-molecule tool to interrogate the secretory-versus-degradative fate of MVBs and uncovers a new regulatory link between lysosomal pathways and EV trafficking. Beyond its established role as an anti-fibrotic agent, SD-208 provides mechanistic and therapeutic opportunities for the control of EV secretion in diseases such as fibrosis, cardiac remodelling, hypertrophic cardiomyopathy, and cancer.

Helminths modulate host immunity, and in this respect, the porcine whipworm Trichuris suis has been explored clinically as an immunotherapy for inflammatory diseases. T. suis secretes a complex mixture of excretory/secretory products (ESP), including extracellular vesicles (EVs), nanosized membranous particles with bioactive cargoes of lipids, proteins and nucleic acids that are implicated in intercellular communication within organisms. Here, we characterise the EV protein and miRNA cargo of T. suis to identify potential roles in host-parasite communication. In contrast to EVs from clade III/V helminth species characterised to date, we show that T. suis EVs did not suppress LPS-induced TNF production by human macrophages under the conditions tested, in contrast to non-vesicular ESP fractions which did. By generating the first extracellular miRNA complement for T. suis and quantifying EV-associated versus free miRNAs across larval and adult stages, we identify selective enrichment of two miRNAs in adult EVs with homology to human miR-22-3p and miR-29-3p, canonical anti-fibrotic and tumour-suppressor miRNAs. Functional assays in human colon cancer epithelial cells reveal that these helminth-derived orthologues downregulate conserved oncogenic targets, indicating evolutionary conservation of their regulatory capacity. Our findings uncover a previously unrecognised axis of whipworm biology in which selective EV-mediated export of anti-fibrotic miRNAs has the potential to modulate host tissue remodeling independently of classical immunomodulation, with implications for understanding helminth-driven epithelial homeostasis and cancer risk.

This study developed an integrated strategy combining surface plasmon resonance (SPR) for real-time biomolecular interaction analysis, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS) for compound profiling, and Global Natural Products Social Molecular Networking (GNPS)-based molecular networking for structural annotation in complex herbal systems. Using this platform, we identified hypermonol J (HPJ), a polycyclic polyprenylated acylphloroglucinol (PPAP), as a high-affinity small-molecule binder of tumor necrosis factor (TNF) (K D = 25.9 pM). SPR and orthogonal isothermal titration calorimetry (ITC) supported direct TNF binding, whereas mechanistic assays suggested that HPJ may preferentially stabilize TNF dimers, thereby impairing trimer assembly and downstream TNF receptor 1 (TNFR1)/nuclear factor kappa B (NF-κB) signaling. In a preclinical psoriasis model, topical 1% HPJ alleviated imiquimod (IMQ)-induced epidermal hyperplasia and suppressed IL-17/IL-23 pathways. In an acute lung injury (ALI) model, 10 mg/kg HPJ reduced neutrophil infiltration, lowered serum inflammatory cytokine levels, and restored pulmonary tight junction integrity. Biosafety assessments supported the preliminary tolerability of HPJ under the tested conditions. These findings identify HPJ as a promising small-molecule lead for further pharmacological optimization and underscore the value of SPR-guided natural product discovery for inflammatory storm intervention.

Apoptosis is a core program regulating organismal homeostasis and plays a pivotal role in the onset and progression of most diseases. Increasing evidence in recent years indicates that mitochondria are not only central to cellular metabolism but also play a pivotal role in regulating apoptosis. However, no systematic review elucidating how mitochondria finely regulate apoptotic processes through multidimensional mechanisms, including apoptosis-resistant diseases such as cancer. This paper systematically summarizes the molecular mechanisms by which mitochondria mediate apoptosis. We focus on the regulation of cytochrome c (Cyt c) release by the Bcl-2 protein family and the activation of downstream caspase cascades. Furthermore, we provide an in-depth analysis of intrinsic factors, including mitochondrial structural remodeling (membrane rupture, cristae remodeling, and membrane lipid redistribution), dynamics imbalance (fusion, fission, and mitophagy), and mitochondrial DNA abnormalities, as well as extrinsic factors involving interorganelle interactions with the endoplasmic reticulum, lysosomes, and other organelles. Additionally, we review clinical and preclinical advances in drugs targeting these pathways. This review aims to provide a comprehensive perspective on the complex network of mitochondrial regulation of apoptosis and offer valuable insights for developing novel clinical therapeutic strategies for cancer and other diseases.

Tumor metabolic reprogramming now extends far beyond the Warburg effect, spanning glycolysis, mitochondrial oxidative phosphorylation (OXPHOS), de novo lipogenesis, cholesterol biosynthesis, and fatty acid oxidation. This network is shaped by cell-intrinsic regulatory layers-transcriptional programs, histone lactylation and other posttranslational modifications, and m6A RNA modification-and by metabolic crosstalk with tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs) that drives immune evasion and drug resistance. How these diverse inputs converge remains undefined: no integrated framework connects regulatory layers to signaling networks such as PI3K-AKT-mTOR, HIF-1α, and cGAS-STING, or explains how their convergence generates metabolic plasticity. This review organizes tumor metabolic reprogramming into functional modules centered on three convergence hubs: the SREBP1/2-FASN-SCD1 lipogenic axis, the HIF-1α-GLUT1-LDHA glycolytic axis, and the LDLR-SCARB1-LXR cholesterol homeostasis system. The discussion traces how multilayered regulation sustains each hub, how metabolic competition between tumor cells and immune effectors creates immunometabolic checkpoints, and how compensatory rewiring undermines single-agent inhibitors. Therapeutic strategies span rational combination regimens that preempt compensatory rewiring to nanomedicine platforms enabling spatially controlled metabolism-immunity reprogramming. By identifying the hubs upon which diverse oncogenic signals converge, this framework reveals context-specific metabolic vulnerabilities and outlines priorities for biomarker-driven patient stratification and hub-targeted combination therapy.

RNA-binding proteins (RBPs) are central regulators of post‑transcriptional gene expression, controlling RNA stability, localization, translation, and alternative splicing. Their functions arise not only from intrinsic RNA-binding domains but also from dynamic interactions with noncoding RNAs, metabolites, cofactors, and other RBPs. Here, we summarize the structural diversity and core biological activities of canonical and noncanonical RBPs, and delineate how competitive and cooperative regulatory networks dictate RBP function in disease, with an emphasis on cancer. Competitive mechanisms, including lncRNA-mediated sequestration, antagonistic crosstalk between miRNAs and RBPs, and competition among RBPs for shared substrates, can redirect RNA fate. In contrast, cooperative mechanisms assemble multimolecular ribonucleoprotein complexes that reinforce oncogenic or tumor-suppressive programs. Dysregulation of these networks promotes proliferation, metastasis, immune evasion, and therapy resistance. We also review emerging therapeutic strategies that target RBP-centered regulatory circuits, including antisense oligonucleotides, small molecules, protein degraders, and natural products, and we evaluate representative preclinical studies and clinical trials. By integrating mechanistic principles with translational evidence, this review provides a network-based framework for exploiting RBPs as therapeutic vulnerabilities and for advancing next-generation precision oncology.

NADPH oxidases (NOXs) have emerged as central hubs that link environmental, metabolic, and immune cues through spatially organized redox signaling. However, their roles across tissues and disease states have not been comprehensively evaluated in an integrated manner. This review integrates recent advances in structural biology, immunometabolism, toxicology, and systems biology to provide an updated, comprehensive, and accessible view of NOX biology. Recent advances in high‑resolution cryo-EM, AlphaFold‑based modeling and molecular dynamics studies have provided new insights into NOX architecture, catalytic sites, post‑translational modifications and regulatory mechanisms, and docking interfaces for RAC1 and p47phox. Emerging evidence further indicates that cellular NOX-derived ROS can reprogram macrophage and T-cell metabolism, stabilize HIF-1α, and tune the balance between effector and regulatory states, thereby linking NOX activity to checkpoint control and tumor immune escape. A second focus is on how real‑world pollutants converge on NOX isoforms as proximal mediators of redox signaling across lung, vascular, hepatic, renal, and neural tissues. NOX activation during cellular injury may contribute to oxidative stress, mitochondrial dysfunction, inflammasome activation, and fibrotic signaling, through extracellular vesicles, lipid rafts, and noncoding RNAs. Finally, the review evaluates emerging therapeutic strategies, including isoform-selective/pan-NOX/peptide inhibitors, and nanozymes. It also discusses emerging approaches such as exosome-based biomarkers, network pharmacology, and machine learning for patient stratification and pharmacodynamic monitoring. By highlighting key mechanistic gaps and translational opportunities, this review establishes NOXs as actionable nodal regulators at the intersection of immunity, metabolism, environmental exposure, and human disease.

Ferroptosis is a novel, iron-driven form of cell death characterized by iron-dependent membrane-lipid peroxidation. Accumulating evidence has revealed that ferroptosis is heavily involved in multiple physiological and pathological conditions, resulting in remarkable progress in the treatment of multiple diseases. This review summarizes recent advances in and the fundamental features of ferroptosis and outlines the molecular mechanisms that regulate core metabolic pathways and their relevance to cellular physiology. In addition, the mechanisms and functional implications of ferroptosis in cancer, cardiovascular diseases, neurodegenerative disorders, and acute organ injury are discussed, and current diagnostic and therapeutic strategies that target ferroptosis are summarized. Finally, the review identifies persistent gaps in the mechanistic understanding, relevant biomarker development, and translational validity of ferroptosis to provide a comprehensive foundation for future research and clinical innovation in the era of precision medicine.

Neutrophil extracellular traps (NETs) released by C-X-C motif chemokine receptor 4-high (CXCR4hi) neutrophils are major contributors to skin inflammation in psoriasis, yet the upstream epitranscriptomic mechanisms that regulate NETosis remain poorly defined. Here, we investigated the role of methyltransferase-like protein 7A (METTL7A) in N 6-methyladenosine (m6A)-mediated neutrophil activation and psoriasis pathogenesis. Using transcriptomic profiling of CXCR4hi neutrophils from psoriasis patients, m6A RNA immunoprecipitation, in vitro functional assays, and in vivo murine models of skin inflammation, along with genetic silencing and adoptive transfer experiments, we demonstrated that METTL7A was preferentially upregulated in pathogenic and aged CXCR4hi neutrophils and was positively associated with disease severity. METTL7A expression also increased during human promyelocytic leukemia cells(HL-60 cell differentiation, maturation, and activation. Silencing METTL7A inhibited neutrophil maturation and aging, reduced NETosis, and alleviated psoriasiform inflammation in vivo. Adoptive transfer of METTL7A-deficient neutrophils further protected mice from skin inflammation. Mechanistically, METTL7A stabilized phospholipid scramblase 1 (PLSCR1) mRNA through m6A-associated regulation, thereby enhancing reactive oxygen species production and neutrophil extracellular traps (NETs) formation. These findings identify METTL7A as a novel epitranscriptomic regulator of pathogenic CXCR4hi neutrophils, acting through the METTL7A-PLSCR1 axis to drive NETosis and skin inflammation, thereby providing a rationale for developing m6A-targeted therapies in neutrophil-associated disorders.

Midkine (MDK) is a heparin-binding growth factor that is minimally expressed in most adult tissues but markedly upregulated in a wide range of malignancies. Accumulating evidence positions MDK as a multifunctional signaling hub that orchestrates tumor progression through its pleiotropic effects on cancer cells and the tumor microenvironment. MDK engages multiple cell surface receptors, including anaplastic lymphoma kinase, low-density lipoprotein receptor-related protein 1, Notch2, and integrins, thereby activating key downstream pathways such as PI3K/AKT, MAPK/ERK, JAK/STAT, and mTOR. Through these signaling networks, MDK promotes tumor cell proliferation, survival, invasion, angiogenesis, metabolic reprogramming, and resistance to therapy. In parallel, MDK exerts profound immunomodulatory effects by shaping the tumor microenvironment, facilitating the recruitment and polarization of immunosuppressive myeloid cells, impairing effector lymphocyte function, and regulating stromal and neural cell interactions via paracrine signaling mechanisms. Recent single-cell transcriptomic studies further highlight MDK's role in mediating intercellular communication across diverse pathological contexts, underscoring its function beyond tumor-intrinsic signaling. Given its extracellular accessibility, disease-associated overexpression, and central role in multiple oncogenic processes, MDK has emerged as an attractive therapeutic target. This review provides a comprehensive overview of the functional regulation of MDK in cancer, summarizing its molecular signaling pathways, contributions to tumor microenvironment remodeling, and involvement in therapeutic resistance. We also discuss current and emerging strategies for targeting MDK, including small-molecule inhibitors, nucleic acid-based approaches, and combination therapies, and highlight key challenges and future directions for translating MDK-targeted interventions into clinical practice.

Neurodegenerative diseases (NDs) are marked by selective neuronal vulnerability and progressive failure of neural circuits. Increasing evidence indicates that neuronal loss is not driven by a single terminal event, but emerges from interacting regulated cell death (RCD) programs. These programs are closely coupled to mitochondrial injury, proteostatic collapse, lysosomal stress, metabolic imbalance, glial state transitions, and chronic neuroinflammation. Yet, how distinct death pathways are organized across cell types, disease stages, and disease-specific microenvironments remains unresolved. This review examines RCD as an integrated pathogenic network in major NDs. Caspase and B-cell lymphoma 2 (BCL2) family signaling, receptor-interacting protein kinase 1 (RIPK1), RIPK3, and mixed lineage kinase domain-like protein activation, NOD-like receptor family pyrin domain containing 3 and gasdermin signaling, GPX4-linked lipid peroxidation control, and autophagy lysosomal failure are discussed as convergent stress response modules rather than isolated pathways. Across Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and Huntington's disease, these modules shape neuronal fate through disease-specific interactions with mitochondrial dysfunction, iron dyshomeostasis, inflammasome activation, and microglial metabolic remodeling. We further evaluate emerging therapeutic strategies that target cell death crosstalk, restore autophagy lysosomal competence, or improve delivery to the central nervous system, highlighting the importance of molecular selectivity, cellular context, disease stage, and translational feasibility.

Multiple myeloma (MM) is a severe plasma-cell malignancy that continues to cause substantial morbidity and mortality despite major therapeutic advances. Although current therapies target distinct molecular processes, many patients eventually relapse, raising the question of whether diverse treatments impose different pressures that nevertheless converge on shared transcriptional adaptation programs. While many studies have profiled transcriptional changes before and after treatment, these responses are often analyzed within individual drugs, leaving recurrent programs across therapies insufficiently explored. Here, we developed PRISM-MM (Perturbation Response Inference by Sparse integrative Modeling for Multiple Myeloma), an interpretable sparse integrative dictionary-learning framework that decomposes drug-control transcriptional shifts into signed response programs while accounting for drug identity, study background and cell-source state. By incorporating paired scRNA-seq as a calibration layer, PRISM-MM preserved cell-state-level interpretability and outperformed representative perturbation-prediction models in recovering reproducible response structure. Applied to a curated multi-study MM perturbation compendium, PRISM-MM identified conserved treatment-associated programs validated in held-out bulk and external paired scRNA-seq datasets. These programs revealed a recurrent remodeling pattern characterized by depletion of plasma-cell-like secretory activity and enrichment of inflammatory, adhesion-associated and stress-tolerant immune-interface states. We further nominated state-maintaining genes and highlighted a RFXAP-centered immune-regulatory axis that may drive the Program 8 inflammatory adaptation state and shape MM drug response. Together, PRISM-MM provides an interpretable framework for discovering conserved drug-response programs and generating mechanistic hypotheses about treatment adaptation in MM.

Aberrant activation of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway has been increasingly recognized as a key driver of autoimmune and inflammatory diseases. In recent years, accumulating evidence has highlighted the close association between cGAS-STING signaling and the pathogenesis of these disorders, suggesting that pharmacological targeting of this pathway may represent a promising therapeutic strategy. This review provides a comprehensive overview of the molecular mechanisms underlying cGAS-STING hyperactivation and its pathological roles across diverse diseases, including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), non-alcoholic steatohepatitis (NASH), STING-associated vasculopathy with onset in infancy (SAVI), Aicardi-Goutières syndrome (AGS), COPA syndrome, Niemann-Pick disease type C (NPC), neurodegenerative diseases and cancer. We critically evaluate current and emerging pharmacological strategies targeting the cGAS-STING pathway, encompassing direct cGAS and STING inhibitors, protein degradation technologies, epigenetic modulation, regulation of biomolecular phase separation, and artificial intelligence (AI)-enabled drug discovery approaches. By integrating disease-specific pathogenic mechanisms with therapeutic opportunities, this review highlights key challenges and future directions in the development of cGAS-STING-targeted therapies. Collectively, these insights provide a translational perspective for precision targeting of pathological cGAS-STING activation.

Restoring lipid droplet (LD) content has been reported to reverse hepatic stellate cell (HSC) activation during liver fibrosis. Although the mitochondrial citrate carrier SLC25A1 is known to drive metabolic reprogramming in cancer and steatohepatitis, its specific role in regulating LDs remains unclear. This study aimed to elucidate the role of SLC25A1 in controlling LD homeostasis during HSC activation and to determine whether therapeutic inhibition of SLC25A1 could ameliorate liver fibrosis by restoring peroxisome proliferator-activated receptor-γ (PPARγ)-dependent lipid storage. Knockdown of Slc25a1 was achieved using either an adeno‑associated viral vector expressing Slc25a1 short hairpin RNA or hyaluronic acid-modified, HSC membrane-biomimetic nanovesicles containing Slc25a1 small interfering RNA (HA@JMNVs/siSlc25a1). Both approaches effectively attenuated HSC activation in three distinct mouse models, induced by carbon tetrachloride, methionine- and choline-deficient diet, and bile duct ligation, respectively. Mechanistically, SLC25A1 deficiency reduced cytosolic acetyl-coenzyme A levels in activated the human immortalized hepatic stellate cell line LX-2, resulting in reduced overall acetylation of neural precursor cell expressed, developmentally downregulated 4 (NEDD4). This reduction subsequently suppressed the NEDD4-mediated ubiquitination and degradation of PPARγ at lysine 197 (K197). The metabolic-post-translational signaling cascade increased perilipin-2 (PLIN2) transcription, inhibited lipophagy, and ultimately restored LD accumulation, thereby reversing HSC activation. Collectively, these findings establish the SLC25A1-regulated metabolic axis as a promising therapeutic target and offer a robust preclinical proof-of-concept for targeted gene therapies against liver fibrosis.

Platelets have been increasingly recognized as versatile regulators of ageing, immunity, and cancer, yet their functional heterogeneity has remained poorly defined. We performed the first large-scale single-cell RNA sequencing of 28,192 platelets from healthy, aged, metastatic, and treated mice using the BD Rhapsody platform. Our analysis revealed four conserved and functionally distinct platelet transcriptional states: haemostatic platelet (HP), neural gene-enriched platelet (NEP), platelet-leukocyte aggregate (PLA) and platelet-erythrocyte aggregate (PEA). Among these states, Tpm2-high HP is linked to ageing-associated lung metastasis and is characterized by cytoskeletal remodelling gene signatures. The PLA state was predicted to be a signalling hub for immunothrombosis, with a PLA-Bridge subpopulation coordinating immune-adherent platelets via the Ppbp-Cxcr2 and Thbs1-Cd47 checkpoint axes. Strikingly, AAV-mPf4 gene therapy was associated with a NEP-associated transcriptional program, which mitigates age-related functional decline. This study provides a single-cell transcriptomic atlas of murine platelets under ageing and metastasis conditions and reveals transcriptional state-specific heterogeneity. This study also proposes PF4-based interventions and cytoskeletal candidates for diagnosis and therapy.

Optogenetics, a biotechnology that combines optical and genetic strategies to regulate cellular and tissue functions with high spatial and temporal precision, serves as a powerful tool-level regulatory technology widely employed to investigate cellular processes and elucidate disease mechanisms. Nanotechnology-driven optogenetics, which incorporates nanomaterials/nanostructures to improve the efficacy and broaden the applications of optogenetic systems, synergistically integrates the precision, tunability, and multifunctionality of nanotechnology with the spatiotemporal control inherent to optogenetics. The development not only enhances the flexibility and efficiency of optogenetic technology but also advances the field toward therapeutic-grade interventions. In this review, we summarize recent advances in nanotechnology-driven optogenetics, with a particular emphasis on three key areas: (1) nanostructured light sources for the precise activation of optogenetic systems, which include both externally light-stimulated systems and systems that operate independently of external light sources; (2) nanotechnology-enabled targeted delivery of light-sensitive proteins and genetic constructs to ensure efficient modulation of optogenetic pathways, which specifically involves the nanotechnology-assisted gene, protein, and recombinase enzyme delivery approaches; and (3) nanotechnology-driven therapeutic applications of optogenetics, including CAR T cell immunotherapy, cancer treatment, neurological interventions, and cardiac therapies. We further discuss the current challenges facing this emerging field and outline future research directions. This review aims not only to highlight recent breakthroughs but also to position nanotechnology-driven optogenetics as a promising tool for next-generation precision medicine.

Ferroptosis has long been regarded as a cell-autonomous form of regulated cell death driven by iron-dependent lipid peroxidation. Recent work, however, suggests that ferroptotic commitment can extend beyond the initiating cell, spreading to neighboring cells and, in some contexts, across tissue-scale distances. Multiple, non-mutually exclusive modes of contagion have now been described, including reactive oxygen species-triggered death waves, direct membrane contact-dependent transfer, and extracellular vesicle-mediated paracrine signaling. These findings redefine ferroptosis from a single-cell execution pathway into a spatially coordinated, multicellular process. In this perspective, we integrate mechanistic insights with experimental evidence on ferroptotic contagion and propose a unifying, multiscale framework in which distinct modes of transmission operate over different spatial scales, from short-range membrane transfer to longer-range oxidative and extracellular relay mechanisms. We discuss how nonlinear redox amplification, membrane biophysics, and tissue architecture together determine the dynamics, limits, and patterning of ferroptotic injury in vivo. This emerging framework has important implications for developmental tissue remodeling, the progression of organ injury, and ferroptosis-based cancer therapy. More broadly, defining how ferroptotic contagion is initiated, constrained, and manipulated therapeutically may help establish ferroptosis as a fundamental organizing principle for understanding tissue-level regulation and pathological escalation.

NOD-like receptor (NLR) family CARD domain containing 3 (NLRC3), an intracellular member of the NLR family, is a negative regulator of both immune cell modulation and tumor cell proliferation. However, the role of NLRC3 and the mechanisms underlying its effect on the tumor immune microenvironment remain unclear. In this study, we report that NLRC3 promotes antitumor immunity by specifically negatively regulating the infiltration and immunosuppressive function of monocytic myeloid-derived suppressor cells (M-MDSCs). Mechanistically, NLRC3 inhibits the stimulator of interferon genes (STING) signaling pathway, leading to reduced expression of programmed cell death ligand 1 (PD-L1) and CCR2 in M-MDSCs, thereby limiting the infiltration and immunosuppressive function of M-MDSCs. Notably, overexpression of NLRC3 in combination with the STING agonist c-GAMP significantly inhibited tumor growth. These findings reveal a critical role for NLRC3 in modulating the immune microenvironment and provide insights into the optimization of NLRC3-targeted therapeutics.

Hemorrhage is a leading cause of potentially preventable deaths in pre-hospital settings. Hence, there is a great need to develop a therapeutic that can be administered in the field to slow or stop hemorrhage before definitive treatment can be sought. The aim of this work was to develop a nanoscale delivery vehicle that could successfully localize to sites of active hemorrhage in the vasculature. We first synthesized four peptides derived from Factor VII as putative targeting epitopes for tissue factor (TF). The peptide sequence ERTFSERT was chosen after microscale thermophoresis revealed a micromolar dissociation constant for TF. The ERTFSERT peptide was covalently linked to a peptide amphiphile (PA) molecule and co-assembled to form supramolecular nanofibers, which were found to have a parallelepiped shape with a width of ∼12 nm and β-sheet secondary structure. Using a microfluidic channel with cultured human umbilical vein endothelial cells, we found that co-assemblies of the targeted nanofibers preferentially localized to endothelial cells activated with tumor necrosis factor alpha, validating the targeting capability of ERTFSERT in a physiologically relevant in vitro model. When administered intravenously in a rat model of liver hemorrhage, biodistribution assessed using ex vivo imaging revealed an increased concentration of targeted nanofibers near the site of liver injury with no adverse effect on mean arterial pressure or blood loss. Overall, these data suggest that TF-targeted PA nanofiber co-assemblies can be used in future studies to deliver therapeutic agents to the site of injury to treat hemorrhage. STATEMENT OF SIGNIFICANCE: We developed and validated a peptide amphiphile with a targeting moiety derived from Factor VII that can localize to tissue factor at the site of hemorrhage. This can be used in future studies to deliver hemostatic agents to the site of injury to treat hemorrhage or to treat other conditions where tissue factor is upregulated.

Excitability is a fundamental dynamical paradigm underlying both local and collective activity across a broad range of living systems, from neurons and cardiomyocytes to pancreatic β-cells, cancer cells, and the emerging field of network physiology. This review summarizes the current state of research on coherence-incoherence patterns in coupled excitable systems, covering theoretical advances and experimental evidence for their roles in physiological and pathological processes. Particular emphasis is placed on how excitable dynamics modifies the mechanisms of pattern formation relative to coupled oscillator networks, highlighting the importance of inhibitory/repulsive interactions and the constructive role of noise through phenomena such as coherence resonance, in generating pattern classes characteristic of excitable media, including bumps, patched patterns, and noise-facilitated chimera states. We further discuss how existing theoretical concepts can be extended to biological systems characterized by heterogeneous local dynamics, complex coupling architectures, and metastable behavior. In addition, we survey state-of-the-art electrophysiological and optical imaging techniques for observing coherence-incoherence patterns and assess current evidence linking them to sleep, cognition, spatial navigation, epilepsy, cardiac arrhythmia, pancreatic islet dynamics, and cancer progression. Finally, we outline major open challenges, including characterization of chaos, long-term dynamics and finite-size effects, experimental validation, control of pattern emergence/termination and switching, and the development of biologically realistic theoretical frameworks.

ATP-citrate lyase (ACLY) is a key metabolic enzyme that links mitochondrial citrate export to the generation of cytosolic acetyl-CoA, thereby supporting de novo lipogenesis, cholesterol biosynthesis, protein acetylation, chromatin remodelling, and transcriptional control. Interest in ACLY inhibition initially arose from its lipid-lowering properties and led to the clinical development of bempedoic acid, whose ability to reduce low-density lipoprotein cholesterol and improve cardiovascular outcomes now provides the strongest clinical proof of concept for targeting this pathway. Beyond dyslipidaemia, preclinical evidence suggests that targeting the ACLY pathway and related bempedoic acid-responsive metabolic programs may ameliorate metabolic dysfunction-associated steatotic liver disease (MASLD). ACLY inhibition is expected to reduce de novo fatty-acid and cholesterol synthesis by limiting cytosolic acetyl-CoA availability, whereas parent bempedoic acid can directly activate PPARα, and thereby enhance fatty-acid oxidation. However, unlike the cardiovascular setting, robust clinical data supporting ACLY inhibition in MASLD are still lacking. More recently, the identification of nuclear ACLY functions has substantially expanded its biological significance, establishing ACLY as a metabolic-epigenetic integrator that couples nutrient availability to chromatin remodelling, transcriptional programs and immune responses. In cancer, dysregulated ACLY activity contributes to tumour growth, metabolic plasticity, therapy resistance and immune evasion, and its inhibition has shown promising antitumour effects in preclinical models. This review summarizes ACLY biology and pharmacology, emphasizing established cardiovascular applications, emerging MASLD opportunities and exploratory oncologic indications, while highlighting unresolved translational questions.

Industrial benthic systems are major reservoirs of hydrophobic organic contaminants (HOCs) derived from industrial discharges, urban runoff, wastewater effluents, and maritime activities. This study quantified polycyclic aromatic hydrocarbons (PAHs), phthalate esters (PAEs), and alkylphenols (APs) in sediments and benthic crustaceans from Kaohsiung Harbor, Taiwan, and evaluated the factors controlling their distribution, bioaccumulation, and associated risks. Outside-harbor sediments exhibited ΣPAH, ΣPAE, and ΣAP concentrations ranging from 2.27 to 230, 25.4 to 340, and 1.40 to 72.8 ng g⁻¹ dw, respectively, whereas harbor sediments showed elevated concentrations of ΣPAHs (72.5-918 ng g⁻¹ dw), ΣPAEs (101-1406 ng g⁻¹ dw), and ΣAPs (34.5-603 ng g⁻¹ dw). PAHs were primarily controlled by total organic carbon (TOC), explaining up to 67% of the variance, while PAEs and APs were influenced by spatial variability and seasonal inputs. PAEs were enriched during the wet season due to runoff and wastewater transport, whereas APs accumulated during the dry season under reduced dilution and flushing conditions. Diagnostic ratios and Positive Matrix Factorization indicated predominantly pyrogenic PAH sources, with stronger wet-season petrogenic influence near harbor entrances and marine outfalls. Crustacean contaminant concentrations reflected sediment contamination patterns (PAEs > APs > PAHs), while compound-specific biota-sediment accumulation factor (BSAF) demonstrated that bioaccumulation was regulated by sediment bioavailability, contaminant partitioning, and species-specific exposure pathways. Risk assessments revealed episodic carcinogenic risks for PAHs, with incremental lifetime cancer risk (ILCR) values exceeding 10⁻⁵, whereas PAEs and APs posed persistent ecological risks. These findings highlight the importance of integrating source apportionment, bioaccumulation, and ecological risk assessment in harbor pollution management.

Whole Slide Images are central to modern pathology and computational histopathology. However, tissue processing and slide scanning can introduce artifacts that degrade image quality and hinder computer-aided diagnosis (CAD) systems. Therefore, automated artifact processing has drawn much attention in recent years. Nevertheless, current methods exhibit limitations, including reliance on annotated data and inadequate pixel-level localization capabilities, resulting in fragmented workflows. To address these challenges, we propose an unsupervised automatic pipeline for artifact detection, localization, and restoration. First, based on the features extracted by the histopathology foundation model, anomaly heatmaps are generated using Normalizing Flow to enable precise artifact detection. Subsequently, optimal artifact masks are identified via anomaly heatmaps and unsupervised semantic segmentation for accurate localization. Finally, a mask-guided artifact restoration is performed using a diffusion model. Experimental results demonstrate that our method effectively handles synthetic and real-world artifacts using only normal training data and improves downstream performance on both BCSS segmentation and TCGA-BLCA tumor staging classification, confirming our method's efficacy in bolstering the robustness of CAD systems, reducing manual verification, and providing an automated solution for artifacts in histopathology images.

Diffusion magnetic resonance imaging (dMRI) enables noninvasive mapping of tissue microstructure by probing water molecule diffusivity. While advanced multi-shell diffusion models offer improved sensitivity to cellular properties, their requirement for densely sampled q-space data leads to prohibitively long acquisition times. Current deep learning approaches for parameter estimation face three key limitations: (1) dependency on fixed acquisition protocols, (2) model-specific assumptions that constrain applicability, and (3) reliance on supervised learning paradigms that demand large labeled datasets and exhibit poor generalization to out-of-distribution cases. To address these challenges, we propose MINeR, a novel unsupervised subject-specific framework for reconstructing dense q-space data from highly undersampled acquisitions. Our method leverages direction-modulated implicit neural representation to flexibly sample diffusion signals across q-space, supporting the estimation of parameters for diverse diffusion models. Comprehensive evaluations demonstrate that MINeR maintains high fidelity in microstructural parameter estimation, particularly for advanced multi-shell diffusion models. The framework shows remarkable generalization capability, as evidenced by its robust performance on tumor data. Notably, MINeR effectively reconstructs high-quality diffusion signals by interpolating from 6 directions, significantly reducing acquisition time, while maintaining robust parameter estimation. This work presents a practical approach for enabling microstructural modeling from sparsely sampled q-space data, thereby improving the clinical applicability of diffusion MRI. The code is available at: https://github.com/AMRI-Lab/MINeR.

Early cancer initiation under chronic oxidative stress requires the survival of epithelial cells that should otherwise be eliminated by regulated cell death. However, the spatial and morphological identity of such stress-adapted cells remains poorly defined. Here, we integrated Xenium single-cell spatial transcriptomics, quantitative nuclear morphometrics, mitochondrial analyses, and human cancer datasets to investigate iron-driven renal carcinogenesis in Brca1(L63X/+) rats. Ferric nitrilotriacetate exposure induced dynamic remodeling of proximal tubular epithelium, characterized by loss of homeostatic transport and mitochondrial programs, and activation of Myc, Met, Lcn2, Anxa2 and iron-handling genes. BRCA1 haploinsufficiency expanded a karyomegalic proximal tubular state associated with mitochondrial dysfunction, chromatin hyperchromasia, nuclear remodeling, and ferroptosis-resistant transcriptional features. Subclassification of karyomegalic cells identified distinct stress-adaptive populations, including a BRCA1-associated K2 state enriched for Myc/Met/Lcn2 signaling and preferentially embedded within a pro-oncogenic epithelial-stromal microenvironment. Of note, neighboring non-karyomegalic proximal tubular cells acquired related iron-handling and survival-biased programs, indicating that karyomegaly marks a broader pre-neoplastic field rather than an isolated abnormality. Human TCGA renal cancer analyses and morphometric assessment of BRCA1-mutant breast tissue further supported the clinical relevance of karyomegaly-associated gene modules and nuclear remodeling. These findings define a spatially organized, ferroptosis-resistant karyomegalic niche as an early lesion in iron-driven carcinogenesis and establish quantitative nuclear morphology as a functional readout of exposome-related oxidative stress adaptation in genetically susceptible tissues.

The sharp rise in medical tomography examinations has created a demand for automated systems that can reliably extract informative features for downstream tasks such as tumor characterization. Although 3D volumes contain richer information than individual slices, effective 3D classification remains difficult: volumetric data encode complex spatial dependencies, and the scarcity of large-scale 3D datasets has constrained progress toward 3D foundation models. As a result, many recent approaches rely on 2D vision foundation models trained on natural images, repurposing them as feature extractors for medical scans with surprisingly strong performance. Despite their practical success, current methods that apply 2D foundation models to 3D scans via slice-based decomposition remain fundamentally limited. Standard slicing along axial, sagittal, and coronal planes often fails to capture the true spatial extent of a structure when its orientation does not align with these canonical views. More critically, most approaches aggregate slice features independently, ignoring the underlying 3D geometry and losing spatial coherence across slices. To overcome these limitations, we propose TomoGraphView, a novel framework that integrates omnidirectional volume slicing with spherical graph-based feature aggregation. Instead of restricting the model to axial, sagittal, or coronal planes, our method samples both canonical and non-canonical cross-sections generated from uniformly distributed points on a sphere enclosing the volume. Triangulating these viewpoints yields a spherical graph that captures spatial relationships among views, and we use a graph neural network to aggregate their features accordingly. Experiments across six oncology 3D medical image classification datasets demonstrate that omnidirectional volume slicing improves the average performance in Area Under the Receiver Operating Characteristic Curve (AUROC) from 0.7701 to 0.8154 compared with traditional slicing approaches relying on canonical view planes. Moreover, we can further improve AUROC performance from 0.8198 to 0.8372 by leveraging our proposed graph neural network-based feature aggregation. Notably, TomoGraphView also surpasses large-scale pretrained 3D medical imaging models across all datasets and tasks, underscoring its effectiveness as a powerful framework for volumetric analysis and therefore represents a key step toward bridging the gap until fully native 3D foundation models become available in medical image analysis. We provide a user-friendly library for omnidirectional volume slicing at https://pypi.org/project/OmniSlicer.

Spatial transcriptomics (ST) enables spatially resolved gene expression profiling within intact tissue sections. However, its widespread adoption is constrained by the high cost and low throughput of current sequencing-based protocols. This has motivated growing interest in computationally predicting gene expression directly from routinely acquired histology images. Existing methods are largely restricted to isolated 2D tissue slices and fail to capture richer spatial relationships or structured dependencies among spot-level gene expression profiles. In this paper, we propose STAG, a dual-branch framework for gene-aware expression prediction and spatial context modeling. A Query branch predicts ST expression for an individual target spot, while a Neighbor branch acts as an auxiliary branch to model structured relationships among multiple spots. By leveraging hypergraph learning, the Neighbor branch captures higher-order spatial and molecular dependencies, enabling unified modeling of both intra-slice and inter-slice relationships. This design supports standard 2D settings (a single slice) and naturally extends to 3D scenarios when adjacent tissue sections are available. Moreover, STAG leverages gene semantic information as biological guidance by encoding gene names with a foundation model, enabling coordinated gene-aware interactions beyond independent gene prediction. STAG achieves an average gain of 5.16% in PCC@250 across six datasets. Under highly variable gene selection, STAG maintains the lowest RMSE and highest PCC@50 across three datasets. The effectiveness of the learned representations is further demonstrated in pseudo-3D prediction and downstream cancer classification tasks. Code is available at https://github.com/MCPathology/STAG.

Methanethiol (MeSH) is a pivotal volatile sulfur compound in plant, human, and environmental chemistry, but its study is hindered by analytical challenges. Here, we present an accessible fluorometric two-component assay toolkit for quantitative assessment of its content and enzymatic degradation by methanethiol oxidase (MTO). The first assay sensitively detects MeSH with a limit of detection of 124 nM using a novel "tube-in-tube" design for efficient MeSH capture, circumventing the need for complex chromatography and expensive instrumentation. The second assay is a real-time kinetic MTO activity assay. Using this toolkit, we (i) established an inverse correlation between MTO activity and MeSH levels in a colon cancer cell line, (ii) quantified MeSH as a key scent marker in various types of cheese, (iii) evaluated MeSH release from the selected Brassica and Allium species and (iv) determined its production in aquatic systems. The MeSH metabolism toolkit presented here will provide researchers with simple, sensitive and specific assays to elucidate multifaceted roles of MeSH and MTO in human pathophysiology and to accelerate discoveries in a broad spectrum of scientific disciplines.

Testicular cancer is the most common solid tumor in young men aged 15-44 years, with seminoma being a major subtype. To elucidate its poorly understood molecular mechanisms, we performed integrated spatial transcriptomics (ST) and spatial metabolomics (SM) on fresh-frozen seminoma tissues from four patients. ST identified 16 distinct cell clusters and spatially resolved transcriptional programs in tumor, invasive, tubular, and stromal regions, revealing tumor-enriched pathways like glycolysis, oxidative phosphorylation, and ferroptosis. Pseudotime analysis suggested a branched trajectory of malignant progression. SM showed pronounced dysregulation of glutathione metabolism in tumor areas. Integrated analysis pinpointed glutathione metabolism and ferroptosis as convergent regulatory pathways in seminoma. Functional validation in TCam-2 cells and a xenograft mice demonstrated that pharmacologic inhibition of glutathione synthesis (using buthionine sulfoximine, BSO) or genetic knockdown of xCT induced ferroptosis and suppressed tumor cell growth both in vitro and in vivo. This study establishes glutathione metabolic reprogramming as a key adaptive mechanism in seminoma that maintains redox homeostasis and protects against ferroptosis, suggesting that targeting this pathway or promoting ferroptosis represents a novel therapeutic strategy.

Targeted protein degraders (TPDs), including proteolysis-targeting chimeras (PROTAC) and molecular glue degraders (MGD), are among the most promising small-molecule-based drug treatments in oncology. The May 2026 U.S. Food and Drug Administration (FDA) approval of vepdegestrant provides a regulatory milestone for heterobifunctional protein degradation and for PROTAC therapeutics. First-generation TPDs were developed for oral delivery; however, the intrinsic physicochemical properties of TPDs impose constraints on their oral bioavailability, systemic exposure, target-site accumulation, and therapeutic efficacy. As the field transitions toward a second wave of TPD development, nanoparticle-based targeted protein degraders (nano-TPD) are gaining momentum for broadening the therapeutic landscape of protein degradation. In this context, drug delivery systems offer opportunities to overcome key translational barriers by improving pharmacokinetics, tissue distribution, target site localization, cellular uptake, and therapeutic index. Here, we provide an overview of TPD discovery, from early laboratory to (pre-) clinical progress, discuss translational challenges, and suggest advanced drug delivery solutions to help realize the full potential of TPD therapies.

Tumor cells face constant environmental and therapeutic stresses, relying on interconnected survival mechanisms involving autophagy, metabolism, and cell death regulation. This review explores the triangular relationship between these processes and how their dynamic interplay supports tumor growth and drives resistance to various treatments. We discuss key molecular pathways regulating autophagy and metabolic reprogramming, and their impact on different cell death modalities. Finally, we address current challenges in translating this knowledge into clinical therapies, emphasizing the need for precise biomarkers and combination strategies to overcome resistance. Understanding this complex network offers promising avenues for developing more effective cancer treatments.

Glycosylated proteins and lipids regulate multiple cellular processes and play key roles in organ damage and regeneration. Recently, glycosylated non-coding small RNAs (glycoRNA) have been identified, however, their expression in the liver and their involvement in hepatic pathology has not yet been reported. We detected the presence of glycoRNAs by northern blot and an imaging approach called sialic acid aptamer and RNA in situ hybridization-mediated proximity ligation assay that enables direct visualization of glycoRNAs. Restoration of key mediators of glycoRNA biosynthesis was achieved in vivo, via adeno-associated viral vectors (AAV). Here, we show that glycoRNAs are synthesized in human liver tissue, primary hepatocytes and hepatic tumor cells. In tissues from patients with key features of metabolic dysfunction-associated steatotic liver diseases (MASLD), the most prevalent liver disease worldwide, expression of most glycoRNAs was reduced. Mechanistically, we found that reduced expression of SID1 transmembrane family member 1 (SIDT1) and DTW domain containing 2 (DTWD2), two key mediators of glycoRNA biosynthesis, contribute to loss of glycoRNAs in MASLD. Inhibition of SIDT1 and DTWD2 increased fatty acid load in primary human hepatocytes and enhanced inflammation signals upon co-culture with macrophages. Importantly, AAV-mediated in vivo restoration of SIDT1 and DTWD2 attenuated metabolic dysfunction-associated steatohepatitis in mice. Furthermore, sequencing of enriched glycoRNA samples identified eight glycoRNAs that were downregulated in steatotic human liver and hepatocytes. Collectively, our study demonstrates the presence of glycoRNAs in human hepatocytes and human liver tissues and their dysregulated expression in experimental steatosis models and in patients with MASLD. Recent estimates suggest that up to 24% of the world's population is affected by metabolic dysfunction-associated steatotic liver diseases (MASLD). Our study reports for the first time that glycosylated RNAs (glycoRNA), a recently discovered class of RNA, are present in the liver and their expression is dysregulated in fatty liver injury. Importantly, in vivo restoration of glycoRNA biogenesis, attenuated fatty liver injury in preclinical models of MASLD. Furthermore, our results suggest that glycoRNA expression is dysregulated in human livers with MASLD, indicating their potential as novel biomarkers of liver injury.

Dysregulated aerobic glycolysis represents a defining metabolic feature of renal cell carcinoma (RCC), yet the regulatory networks driving this metabolic shift remain incompletely elucidated. In the present study, we found that the deubiquitinase Josephin domain-containing protein 2 (JOSD2) is necessary for both glycolytic and proliferative activities in RCC. Furthermore, JOSD2 is significantly upregulated in RCC and its high expression correlates with poor clinical outcomes. Functional studies demonstrated that JOSD2 enhances RCC cell growth, metastatic capacity, and glycolytic activity in vitro and in vivo. Mechanistically, JOSD2 directly interacts with the transcription factor CREB1 and prevents its proteasomal degradation by removing K48-linked polyubiquitin chains, thereby stabilizing the CREB1 protein. Meanwhile, CREB1 transcriptionally upregulates JOSD2 by binding to its promoter, forming a reciprocal regulatory circuit between the two molecules. Reciprocal activation between JOSD2 and CREB1 leads to increased expression of key glycolytic regulators, including GLUT1, HK2, and PFKP, thereby driving metabolic reprogramming in RCC cells. Importantly, the tumor-promoting functions of JOSD2 were found to be largely dependent on CREB1 activity. Together, these results support a role for the JOSD2-CREB1 reciprocal axis in glycolysis-associated RCC progression and suggest its potential prognostic and therapeutic relevance.

Precisely delineating the transcriptomic profiles of glandular epithelial (GE) cells in prostate cancer (PCa) remains a significant challenge primarily due to their diffuse and multifocal distribution. To address this, we employed spatial transcriptomics (ST) to analyze 12 PCa tissue samples from 10 patients, aiming to identify PCa progression-associated genes by analyzing expression patterns across histologically distinct regions. Transcriptomic classification via principal component analysis (PCA), uniform manifold approximation and projection (UMAP), and Louvain clustering revealed spatially resolved histological structures within each tissue section. The malignancy status, progression stages, and developmental trajectories of GE clusters were further assessed using inferred copy number variation (inferCNV), diffusion pseudotime (DPT), and partition-based graph abstraction (PAGA) analyses. Based on the preliminary characterization of developmental trajectories, pairwise comparisons of GE clusters identified key oncogenes-including TFF3, OR51E2 (PSGR), FOLH1 (PSMA), AMACR (P504S), FOS (a subunit of AP-1), SLC4A4, EGR1, NDUFB9, and H2AFJ-that are positively associated with PCa progression. Immunohistochemistry (IHC) validation further confirmed the elevated expression of SLC4A4 and H2AFJ in advanced-stage PCa. Overall, this study establishes an ST-based framework for predicting PCa progression and provides valuable insight for the identification of progression-associated genes holding promise as clinical biomarkers.

Pullulan, a linear microbial α-glucan composed of maltotriose units linked by alternating α-(1 → 4) and α-(1 → 6) glycosidic bonds, has evolved from a food-grade excipient into a versatile platform for nanomedicine and regenerative biomaterials. This review synthesizes the structure-activity relationships (SAR) of pullulan and its derivatives, linking molecular design with macroscopic material performance and clinical potential across diverse biomedical applications. The presence of three hydroxyl groups per glucose unit (nine per maltotriose) enables diverse and site-specific chemical modifications, transforming an otherwise biologically inert backbone into highly functional and tunable biomaterials. Key strategies include cholesteryl grafting for self-assembling nanogels in cancer immunotherapy, periodate oxidation and methacrylation for dynamically crosslinked and 3D-printable hydrogels, and regioselective sulfation for heparin-mimetic anticoagulants. We also examine pullulan's intrinsic hepatotropism via the asialoglycoprotein receptor, enabling liver-targeted drug and gene delivery. Despite progress, including early-phase clinical evaluation of nanogel-based vaccines, broader pharmaceutical translation remains limited. Major challenges include poor mechanical strength of native networks (typically exhibiting a storage modulus (G') < 1 kPa), rapid enzymatic degradation, batch variability in molecular weight, and the lack of parenteral-grade regulatory standards. This review outlines future directions in precision modification, stimuli-responsive design, and standardization for clinical advancement and scalable biomedical applications.

Low-frequency, low intensity ultrasound (LIUS) has emerged as a promising physical modality capable of inducing selective apoptosis of cancer cells, while sparing healthy epithelial cells and fibroblasts. Hitherto, the mechanism underlying this selectivity has been unclear, but we now propose and develop a theoretical framework linking the distinct mechanical behaviours of cancer versus healthy cells to their differential responses to LIUS. We point out that cancer cells exhibit inhomogeneous ventral stress-fiber networks, which can produce irregular focal adhesion geometry and inward membrane curvature near focal adhesions under low-intensity ultrasound (LIUS). These curvature irregularities can favor loose packing of Piezo1 channels, thereby preserving their activity. In contrast, healthy epithelial cells and fibroblasts display more homogeneous cytoskeletal organization, which can result in more regular curvature profiles adjacent to focal adhesions. This leads to curvature-driven cholesterol redistribution, resulting in altered spatial organization of Piezo1 clusters and reduced coordinated channel activity and allowing cells to remain in their active, proliferative state when exposed to LIUS. Based on theoretical modeling and previous experimental findings, we propose that differences in cytoskeletal organization and membrane curvature can contribute to distinct Piezo1 activation patterns between healthy and cancerous cells. Our analysis identifies curvature-mediated Piezo1 redistribution as a potential physical basis for LIUS selectivity and provides a mechanistic foundation for designing ultrasound-based therapies to exploit the intrinsic cytoskeletal vulnerabilities of cancer cells.

Retinoblastoma is an aggressive intraocular tumor that originates from the developing retina in early childhood. Biallelic loss of RB1 has long been considered the main event for initiating RB development in most cases. Additional genetic events following RB1 loss, such as MYCN amplification, were found to be required for RB progression. Advancements in next-generation sequencing technologies have enabled a deeper understanding of the contributors to RB origin and development, revealing that secondary genetic alterations following RB1 inactivation are infrequent. In contrast, epigenetic changes were shown to be critical promoters of RB tumorigenesis. Several epigenetic regulators, including DNA methylation, histone modifications and noncoding RNAs, have been proven to be dysregulated in RB, contributing to its progression. Understanding the underlying mechanisms involved in RB and exploring new treatment strategies is a crucial step toward developing more effective and less invasive therapeutic approaches that can improve patient outcomes. This review summarizes the significant genetic and epigenetic alterations involved in RB tumorigenesis, current therapeutic strategies, and future treatment prospects for patients with RB.

Metabolic hijacking of tryptophan (Trp) via the IL4I1-AHR axis is a pivotal immune evasion mechanism in cancers, yet therapeutic strategies to disrupt this pathway remain unexplored. Here, we report the identification of ZY-MY-111, a selective small-molecule inhibitor of interleukin-4-induced-1 (IL4I1), through an in-house compound library screening and structural optimization. ZY-MY-111 exhibits potency (IC50 = 1.86 ± 0.13 μM) in blocking IL4I1-mediated oxidative deamination. Mechanistically, ZY-MY-111 acts as a mixed-type inhibitor, competitively occupying the catalytic pocket of IL4I1 and disrupting Trp-AHR signaling in cells. Functionally, ZY-MY-111 promotes T cell proliferation, enhancing immune responses against the tumor cells. In syngeneic tumor models, ZY-MY-111 achieved 49% tumor growth inhibition in CT26 colon carcinoma (P < 0.001) and 56% tumor growth inhibition in A20 lymphoma (P < 0.001) by remodeling the immunosuppressive microenvironment: increasing CD8+/CD4+ T cell ratios, reducing myeloid-derived suppressor cells (MDSCs, 59% decrease), and enhancing effector memory T cell infiltration. Our findings position IL4I1 inhibition as a potential strategy to restore anti-tumor immunity.

The emerging field of cancer neuroscience has fundamentally reshaped our understanding of malignancy, revealing tumors as complex ecosystems deeply integrated with and actively shaped by the nervous system. Malignancies recruit, reprogram, and exploit neural components, a phenomenon we conceptualize as "neural hijack" to fuel growth, metastasis, immunosuppression, and therapy resistance. Concurrently, drug repurposing, leveraging the established safety profiles of approved neuroactive agents, has emerged as a powerful strategy to therapeutically dismantle this tumor-neural axis. This review synthesizes the neural hijack paradigm, detailing how cancers co-opt neuropeptide, neurotransmitter, and synaptic signaling pathways. We provide a comprehensive evaluation of repurposed neuroactive drugs, including β-blockers, CGRP antagonists, NK1R antagonists, SSRIs, and anti-epileptics, framing their mechanisms within the disruption of specific hijacking nodes. We further propose "neural signatures", composite biomarkers integrating innervation density, receptor profiles, and neuro-immune contexture as critical tools for precision patient stratification. Finally, we discuss translational challenges and innovative solutions, charting a roadmap toward "neural disconnection", the therapeutic severing of the tumor's neural lifeline, as an emerging pillar of cancer therapy.

Piezo ion channels, notably Piezo1 and Piezo2, are key mechanosensors that transduce mechanical forces into intracellular signals, playing indispensable roles in digestive physiology. These channels regulate essential functions such as intestinal motility, epithelial barrier integrity, bile secretion, and host-microbiota balance. Emerging evidence links aberrant Piezo signaling to a wide range of gastrointestinal disorders, including functional bowel diseases, inflammatory conditions, and digestive cancers. However, translating these insights into therapeutic applications remains challenging. Most current findings are derived from animal models or in vitro studies, which do not fully recapitulate human tissue complexity. Advanced human-relevant platforms, such as organoids and organ-on-a-chip systems, are needed to bridge this translational gap. Furthermore, Piezo1 and Piezo2 play both overlapping and distinct roles in gastrointestinal pathophysiology, necessitating selective modulation strategies. While Piezo1 promotes processes such as epithelial remodeling and tumor invasion via pathways like RhoA/ROCK and YAP/TAZ, Piezo2 is more associated with sensory neuron activity, immune modulation, and tumor aggressiveness. The lack of specific agonists and inhibitors, especially for Piezo2, further limits its clinical translation. Lastly, Piezo channels are deeply integrated into complex molecular networks involving focal adhesions, cytoskeletal dynamics, and transcriptional regulation. This review synthesizes current advances in the mechanobiology of Piezo channels within the digestive system and highlights future directions for mechanistically-informed, Piezo-targeted therapies.

Glucocorticoids (GCs) are essential endocrine regulators coordinating stress responsiveness, metabolic flexibility, inflammatory resolution, and circadian physiology. While acute GC fluctuations are adaptive, sustained exposure (arising from psychosocial stress, circadian disruption, obesity, chronic inflammation, neoplasms, or steroid therapy) elicits pleiotropic effects that overlap with biological aging. Prolonged GC signaling intersects with multiple hallmarks of aging by altering nutrient sensing, suppressing autophagy, impairing mitochondrial quality control, and promoting cellular senescence. In this context, the GC-responsive polypeptide ACBP/DBI (acyl-coenzyme A [CoA]-binding protein/diazepam-binding inhibitor) has emerged as a stress-induced inhibitor of macroautophagy that amplifies several metabolic and immune consequences of GC excess linked to aging phenotypes. Clinically, chronic GC elevation is associated with earlier and more severe manifestations of age-related diseases, including metabolic syndrome, osteoporosis, sarcopenia, neurodegeneration, cardiovascular disease, immunosenescence, and cancer. Here, we review mechanistic links between GC signaling and systemic aging and discuss strategies to mitigate the age-accelerating consequences of persistent GC exposure.

Zinc ions (Zn2+) are crucial for cellular homeostasis, with their intracellular concentrations tightly regulated by multiple zinc transporters located at the plasma and organelle membranes. Zinc dysregulation has been implicated in esophageal squamous cell carcinoma (ESCC), yet the oncogenic roles of zinc transporters remain poorly understood. We herein identify SLC30A7 as a zinc transporter markedly elevated in ESCC and associated with poor prognosis. Genetic deletion of Slc30a7 significantly suppressed 4NQO-induced esophageal tumorigenesis, whereas SLC30A7 knockdown inhibited ESCC cell proliferation, migration, and tumor progression. Mechanistically, ERK1 specifically binds SLC30A7 and phosphorylates SLC30A7 at T297 site, driving the redistribution of zinc (Zn2+) from the cytosol into the Golgi lumen. SLC30A7 cooperates with the zinc metallochaperone ZNG1 to mobilize Golgi-resident zinc toward matrix metalloproteinases MMP2/3/9 activation, leading to E-cadherin degradation, β-catenin nuclear translocation, and MYC transcription. In ESCC PDX models, a tumor-targeted biomimetic ERK inhibitor nanoplatform (Q3ME@PBA-NPs) significantly restrained tumor growth and disrupted the ERK1-SLC30A7-MMP2/3/9-β-catenin-c-Myc axis. These findings reveal a previously unrecognized zinc-dependent oncogenic pathway in ESCC and highlight SLC30A7 as a valuable therapeutic target in cancer.

Single-cell RNA sequencing has become an essential approach for investigating cellular heterogeneity in cancer, enabling detailed characterization of malignant cells, tumor microenvironments, and genetically distinct subpopulations. However, accurate and reproducible cell type annotation remains a major analytical bottleneck, limiting the reliability of downstream genetic and molecular interpretation. In this study, we develop a robust retrieval-augmented generation framework for automated cell type annotation that improves stability, biological grounding, and reproducibility. The proposed approach integrates hybrid lexical-semantic information retrieval with ontology-aware evaluation, combining keyword-based search and biomedical domain-specific embeddings to enhance semantic relevance. Retrieval performance is further strengthened through query expansion, paraphrasing, and lightweight reranking, while biological validity is ensured using Cell Ontology-grounded similarity scoring. The framework is systematically evaluated across nine publicly available single-cell RNA sequencing datasets spanning diverse human tissues. Compared with baseline prompting, naïve retrieval-based pipelines, and established annotation tools, the optimized framework demonstrates consistently higher annotation accuracy, reduced performance variability, and improved resolution of ambiguous and rare cell populations. By strengthening the reliability of cellular annotation, this approach facilitates more accurate analysis of genetic and molecular alterations relevant to cancer biology and provides a scalable foundation for translational and diagnostic applications in cancer genetics.

Mass spectrometry imaging (MSI) has emerged as a transformative technology in pharmaceutical research, offering unprecedented capabilities to visualize drug distribution, metabolism, and target engagement in biological tissues. By combining the molecular specificity of mass spectrometry with spatial imaging resolution, this label-free approach enables simultaneous mapping of drugs, metabolites, and endogenous molecules across tissue sections, providing comprehensive insights into drug absorption, distribution, metabolism, excretion, and toxicity properties. Recent technical advances have dramatically enhanced MSI capabilities, achieving spatial resolutions down to the cellular level. The integration of tandem mass spectrometry, ion mobility separation, and dedicated data analysis tools powered by artificial intelligence has further expanded the analytical power of MSI, enabling robust molecular identification and pattern recognition in complex biological matrices. MSI applications span critical areas of drug development, from characterizing blood-brain barrier permeability and CNS drug distribution to mapping tumor microenvironment heterogeneity and evaluating anticancer drug penetration. This technology has proven invaluable for assessing drug disposition in the heart, liver, kidney, lung, as well as the gastrointestinal tract and skin. As drug research increasingly embraces multimodal approaches and single-cell analysis, MSI continues to evolve as an indispensable tool for understanding drug behavior in complex biological systems, ultimately accelerating drug development and reducing clinical failures.

The function of the interferon-stimulated gene ISG15 in cervical cancer (CC) is complex and remains unclear. We analyzed the expression and clinical significance of ISG15 in the TCGA-CESC cohort. Its function was investigated in Caski cells using RNA-seq, siRNA, and pharmacological inhibition methods, and validated in vivo in a xenograft model. ISG15 was significantly overexpressed in cervical cancer tissues (P < 0.001). However, high ISG15 expression was associated with better overall survival (HR=0.67, 95% CI: 0.48-0.94, p = 0.02). Gene set enrichment analysis (GSEA) indicated that high ISG15 expression was associated with the NOD-like receptor signaling pathway. Mechanistically, ISG15 binds to and stabilizes the NOD2 protein via ISGylation. This binding promotes cell proliferation and invasion. The RIPK2 inhibitor (GSK583) eliminated the oncogenic effects of ISG15. In vivo experiments showed that ISG15 knockdown or GSK583 treatment inhibited tumor growth by approximately 50%. Our study reveals the dual nature of ISG15: it is both a prognostic biomarker and a functional oncoprotein in cervical cancer cells. Its pro-tumorigenic effect is mediated by the overactivation of the NLR pathway through ISGylation. The ISG15-NOD2/RIPK2 axis is a targetable vulnerability, suggesting that treatment strategies should be decoupled from its prognostic association.

Male reproductive health is declining globally, reflected by reduced sperm counts, impaired semen quality, increasing infertility, and a rising incidence of testicular cancer. Endocrine‑disrupting chemicals include metabolic disruptors, within which obesogens represent a specific subgroup that dysregulate lipid metabolism and cellular energy homeostasis and are increasingly implicated in these adverse trends. Although obesogens act across multiple tissues, their lipid‑metabolism‑related effects on testicular cells remain comparatively understudied. We investigated male reprotoxicity of an environmentally and human‑relevant organochlorine mixture (OC‑MIX), consisting of 20 persistent contaminants, including polychlorinated biphenyls (PCBs), chlordane, and dichlorodiphenyltrichloroethane/dichlorodiphenyldichloroethane (DDT/DDE), originally identified in ringed seal blubber and representative of Arctic food‑chain exposures. Although OC‑MIX has shown male reproductive toxicity in animal models, its cellular mechanisms in Sertoli cells remain insufficiently defined. Using immature murine Sertoli TM4 cells, we demonstrated that OC‑MIX (5-50 µg/mL) disrupted lipid homeostasis and induced oxidative stress, culminating in lipotoxicity, processes known to compromise Sertoli cell support of spermatogenesis. These responses differed from those elicited by a fatty acid mixture or the lipotoxicant amiodarone and did not appear to involve androgen receptor, aryl hydrocarbon receptor, or peroxisome proliferator‑activated α signaling, suggesting distinct lipid‑centered mechanisms. Comparative profiling in immature murine Leydig TM3 cells revealed cell‑type‑specific responses, likely reflecting intrinsic differences in lipid composition. Notably, OC‑MIX selectively altered phospholipid and lysophospholipid species in Sertoli TM4 cells, indicating interference with membrane remodeling and lipid‑dependent signaling. Overall, these findings support the hypothesis that persistent organochlorines act as testicular obesogens, contributing to male reproductive dysfunction through selective, lipid‑disrupting mechanisms.

Intracerebral hemorrhage (ICH) is a neurological disorder characterized by a high mortality rate for which there is currently no definitive cure. Research has demonstrated that adipose-derived mesenchymal stem cells (ASCs) exhibit considerable potential in treating ICH. However, the advanced age of ICH patients and the necessary cell expansion before transplantation therapy could result in the senescence of ASCs, thereby compromising their viability and therapeutic efficacy. This study aims to investigate whether FGF21 (fibroblast growth factor 21) can rejuvenate aged ASCs by enhancing macroautophagy/autophagy flux and subsequently enhance the therapeutic efficacy of ICH. We demonstrated that the autophagy flux of aged ASCs was significantly decreased and FGF21 treatment significantly reversed the senescence phenotype and increased the viability of aged ASCs. Mechanistically, our findings suggested that FGF21 rejuvenates aged ASCs by augmenting autophagy flux, a process partly mediated by TFE3 (transcription factor E3) nuclear translocation. The FGF21-induced TFE3 nuclear translocation was partially facilitated potentially via the FGFR1-SIRT1-MTOR pathway. In addition, FGF21 enhanced the potential of senescent ASCs to differentiate into neurons. In the in vivo study, we further verified that FGF21 could enhance the therapeutic effect of ASCs on acute ICH rats. In conclusion, these results indicated that FGF21 could restore ASC viability by upregulating TFE3-mediated autophagy flux in part through the FGFR1-SIRT1-MTOR signaling pathway, enhanced the potential to improve the differentiation of ASCs into neural stem cells and enhanced the therapeutic effect of ASCs transplantation in acute ICH.Abbreviations: FGF21: fibroblast growth factor 21; TFE3: transcription factor E3; TFEB: transcription factor EB; DMEM: Dulbecco's modified Eagle medium; RAPA: rapamycin; 3-MA: 3-methyladenine; CQ: chloroquine; DMSO: dimethyl sulfoxide; RT-qPCR: quantitative real-time PCR; pAb: polyclonal antibody; mAb: monoclonal antibody; LAMP1: lysosomal associated membrane protein 1; SQSTM1/p62: sequestosome 1; MAP1lc3/LC3: microtubule associated protein 1 light chain 3; GFAP: glial fibrillary acidic protein; MAP2: microtubule associated protein 2; SOX2: SRY-box transcription factor 2; MOI: multiplicity of infection; FGFR1: fibroblast growth factor receptor 1; SIRT1: sirtuin 1; MTOR: mechanistic target of rapamycin kinase; ROS: reactive oxygen species; siRNA: small interfering RNA; OD: optical density; SASP: senescence-related secretion phenotype; IL6: interleukin 6; IL1B/IL-1β: interleukin 1 beta; TNF/TNF-α: tumor necrosis factor; CCL2/MCP-1: C-C motif chemokine ligand 2; BDNF: brain derived neurotrophic factor; VEGF: vascular endothelial growth factor; ICH: intracerebral hemorrhage; MLPT: modified limb placement test.

Rheumatoidarthritis (RA) is an autoimmune disease accompanied by joint swelling,stiffness, and pain, leading to a sharp decline in quality of life. However,the treatment of RA still faces numerous challenges. Clinical studies indicatethat specific hypoglycemic agents alleviate the symptoms of RA, while the potentialmolecular mechanism remains unknown. Herein, we initially assess the efficacyof various categories of anti-diabetic medications including biguanides, GLP1R(glucagon like peptide 1 receptor) agonists, SLC5A2/SGLT2 (solute carrierfamily 5 member 2) inhibitors, DPP4 (dipeptidyl peptidase 4) inhibitors,sulfonylureas, thiazolidinediones, and insulin analog in RA models ofcollagen-induced arthritis (CIA) and serum-transfer arthritis (STA). Resultsdemonstrate that solely thiazolidinediones (pioglitazone [PIOG]) confersuperior efficacy, whereas the other anti-diabetic agents provide minimal or notherapeutic benefits. Mechanistically, thiazolidinediones (PIOG) activatesPPARG/PPARγ (peroxisome proliferator activated receptor gamma) to promotemitophagic flux, thereby inhibiting aberrant NLRP3 inflammasome activation andreducing pro-inflammatory factors IL1B/IL1-BETA (interleukin 1 beta) and IL18 (interleukin18) release. Notably, loss of autophagy either genetically or pharmacologicallysubstantially diminishes the anti-inflammatory effects of PIOG both in vitroand in vivo. In summary, these results offer new mechanistic insight intodisease crosstalk and support the translational value of thiazolidinedionesPIOG as a candidate for precision therapy in RA or multimorbidity of RA and type2 diabetes mellitus (T2DM).Abbreviations: 3-MA: 3-methyladenine; ACP5/TRAP: acid phosphatase 5, tartrate resistant; AIM2: absent in melanoma 2; ALUM: aluminum hydroxide adjuvant; ANOVA: analysis of variance; PYCARD/ASC: PYD and CARD domain containing; ATP: adenosine triphosphate; BMDM: bone marrow-derived macrophage; BV:TV: bone volume:tissue volume; CIA: collagen-induced arthritis; DAPI: 4',6-diamidino-2-phenylindole; DNA: deoxyribonucleic acid; ELISA: enzyme-linked immunosorbent assay; FG: Fast Green; GFP: green fluorescent protein; GSDMD: gasdermin D; IL1B/IL1-BETA: interleukin 1 beta; IL18: interleukin 18; LDH: lactate dehydrogenase; LPS: lipopolysaccharide; Micro-CT: micro-computed tomography; MSU: monosodium urate; mtDNA: mitochondrial DNA; mtROS: mitochondrial reactive oxygen species; NAC: N-acetylcysteine; NLRP1B: NLR family, pyrin domain containing 1B; NLRP3: NLR family, pyrin domain containing 3; NLRC4: NLR family, CARD domain containing 4; OGTT: oral glucose tolerance test; PBS: phosphate-buffered saline; PINK1: PTEN induced putative kinase 1; PIOG: pioglitazone; PPARG/PPARγ: peroxisome proliferator activated receptor gamma; RA: rheumatoid arthritis; ROS: reactive oxygen species; STA: serum transfer arthritis; STZ: streptozotocin; T2DM: type 2 diabetes mellitus; Tb.N: trabecular number; Tb.Sp: trabecular separation; Tb.Th: trabecular thickness; THP-1: human monocytic leukemia cell line; TNF/TNF-α: tumor necrosis factor; TOMM20: translocase of outer mitochondrial membrane 20.

Ovarian cancer is the most prevalent and deadly gynecological malignancy worldwide, with a 5-year overall survival rate of only 10%-40% for patients with advanced disease. Copper chaperone for superoxide dismutase 1 (CCS) is a metallochaperone that plays a multifaceted role in the maturation of copper and displays aberrant expression levels and functions in cancer. Ferroptosis, a new form of cell death resulting from iron-dependent lipid peroxidation, is closely related to cancer. However, whether CCS regulates ferroptosis in ovarian cancer is unknown, and its underlying mechanisms have not been reported. Here, we report that highly expressed CCS contributes to ovarian cancer tumor growth. Moreover, suppressing CCS induced ferroptosis in ovarian cancer cells and increased their sensitivity to ferroptosis inducers. Mechanistically, high CCS expression was found to reduce intracellular copper ion levels and increase Solute Carrier Family 7 Member 11 (SLC7A11) or Glutathione Peroxidase 4 (GPX4) expression by increasing p53 ubiquitination, thus affecting ferroptosis. Additionally, DC_AC50, a small-molecule inhibitor of CCS that targets its copper transport interface, regulates ferroptosis and ovarian cancer growth. Analysis of clinical data revealed a positive correlation between high CCS expression and high SLC7A11 and GPX4 expression in ovarian cancer patients. In summary, our study reveals that CCS protects ovarian cancer cells from ferroptosis by promoting SLC7A11 and GPX4 expression in a p53-dependent manner.

Boron neutron capture therapy (BNCT) is a targeted radiotherapy that exploits the selective accumulation of the 10B isotope within tumors, followed by irradiation with low-energy neutrons to induce high linear energy transfer particles with short path lengths, resulting in localized tumor cell destruction while sparing surrounding healthy tissue. Achieving optimal therapeutic efficacy requires precise quantification of 10B and 11B in biological samples such as blood, tissue, and cells to inform treatment planning, dosimetry, and patient selection. This review provides a comprehensive assessment of analytical methods for boron (isotope) determination in BNCT, covering neutron-based techniques, mass spectrometry, nuclear imaging, and spectroscopic and magnetic approaches. Each method is discussed in terms of analytical principles, sample preparation, sensitivity, isotopic specificity, invasiveness, real-time capability, infrastructure complexity, and clinical applicability. Particular emphasis is placed on spatially resolved and in vivo techniques, as well as emerging theranostic strategies that integrate boron delivery with multimodal imaging. Despite significant technological progress, the lack of widely available, real-time, non-invasive, and 10B-specific dosimetry remains a key limitation for routine clinical implementation. In particular, while positron emission tomography-based approaches such as 4-borono-2-[18F]fluoro-L-phenylalanine provide essential information on boron biodistribution for treatment planning, they reflect tracer-level pharmacokinetics and therefore offer indirect rather than absolute quantification of therapeutic boron concentrations. This review critically assesses current methodologies in the context of clinical readiness and outlines future directions to support the translation of BNCT from bench to bedside.

o-Toluidine has been used for multiple industrial purposes, but causes bladder carcinogenesis in exposed workers. However, its underlying mechanism is still unclear. In this study, we examined oxidative and nitrative DNA damage caused by o-toluidine and its metabolites, 2-amino-m-cresol, 4-amino-m-cresol (4AC) and N-(4-hydroxy-2-methylphenyl) acetamide, using T24 human bladder epithelial cells. MTT assay showed that 4AC induced the most potent cytotoxicity among o-toluidine and its metabolites. In flow cytometry, 4AC showed the strongest ability of generating reactive oxygen species (ROS) and nitric oxide (NO) in exposed cells. Fluorescence immunocytochemistry revealed that 4AC significantly increased the staining intensities of an oxidative DNA lesion, 8-oxo-2'-deoxyguanosine, and a nitrative DNA lesion, 8-nitroguanine, at 5 nM. Their staining intensities were stronger than the other compounds. The comet assay showed that o-toluidine and its metabolites significantly increased the olive tail moment after the treatment with formamidopyrimidine DNA glycosylase. DNA damage was significantly reduced by transfection with small interfering RNA and antibodies against high-mobility group box-1 (HMGB1), receptor for advanced glycation end-products (RAGE) and Toll-like receptor (TLR) 9. These findings indicate that o-toluidine and its metabolites induce the release of HMGB1, which forms a complex with CpG DNA to bind to RAGE on neighboring cells and then is recognized by TLR9 in lysosomes, leading to ROS and NO production and DNA damage. In o-toluidine-induced carcinogenesis, TLR9-mediated inflammatory response and DNA damage induced by its metabolites, especially 4AC, may play a substantial role.

Histone deacetylases (HDACs) are key epigenetic regulators involved in a variety of cancers, rendering them attractive therapeutic targets. Although several HDAC inhibitors have achieved clinical success, challenges such as poor isoform selectivity, acquired resistance, and off-target toxicity limit their broader application. Proteolysis-targeting chimeras (PROTACs) represent an innovative therapeutic strategy that enables ubiquitin-proteasome-mediated degradation of HDACs. This approach enhances specificity, overcomes resistance mechanisms, including those resulting from point mutations or persistent target activity, and enables sustained suppression at low concentrations, owing to its catalytic and event-driven mode of action. This review summarizes the structural classification and biological functions of HDACs and surveys recent advances in the design of HDAC-directed PROTACs. Key emphasis is placed on rational warhead selection, linker optimization, and the strategic choice of E3 ligase recruiters to guide degradation efficiency and isoform specificity. Representative degraders are evaluated for their pharmacological characteristics and antitumor efficacy across diverse malignancies. Current challenges and future directions for the development of HDAC degraders as clinically viable agents are also discussed.

Lipophagy, a selective form of macroautophagy/autophagy, degrades lipid droplets (LDs) to provide energy and is implicated in metabolic disorders. The molecular mechanism underlying lipophagy induction remains incompletely understood. This study explored the role of SETDB1 in starvation-induced autophagy and lipophagy. We demonstrate that SETDB1 deficiency exacerbates starvation-induced hepatic lipid accumulation by inhibiting lipophagy. Mechanistically, starvation promotes ATM-mediated phosphorylation of SETDB1, which enhances its interaction with and methylation of the RNA helicase DDX5. In SETDB1-knockout hepatocytes, hypomethylation of DDX5 facilitates the formation of the DDX5-METTL3-METTL14 complex, increasing m6A modification of BECN1 and TFEB mRNAs. This modification promoted YTHDF2-mediated decay of these transcripts, thereby inhibiting starvation-induced autophagy and lipophagy. Furthermore, administration of the SETDB1 activator (R, R)-59 significantly enhances lipophagy and attenuates starvation-induced hepatic steatosis. Collectively, our findings reveal a novel pathway in which SETDB1 deficiency drives m6A-mediated mRNA degradation to suppress lipophagy, thereby contributing to hepatic steatosis.Abbreviations: AA free: amino acid deprivation; ATG14: autophagy related 14; ATG5: autophagy related 5; ATG7: autophagy related 7; ATM: ATM serine/threonine kinase; Baf A1: bafilomycin A1; DDX5: DEAD-box helicase 5; FASN: fatty acid synthase; LAMP1: lysosome associated membrane protein 1; LAMP2A: lysosome associated membrane protein 2A; LIPE/HSL: lipase E, hormone sensitive type; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MAP3K5/ASK1: mitogen-activated protein kinase kinase kinase 5; METTL3: methyltransferase 3, N6-adenosine-methyltransferase complex catalytic subunit; METTL14: methyltransferase 14, N6-adenosine-methyltransferase non-catalytic subunit; MGLL/MGL: monoglyceride lipase; OA: oleic acid; OSBPL8/ORP8: oxysterol binding protein like 8; PLIN2: perilipin 2; PNPLA2/ATGL: patatin like domain 2, triacylglycerol lipase; SETDB1: SET domain bifurcated histone lysine methyltransferase 1; TFEB: transcription factor EB; TP53/p53: tumor protein p53; ULK1: unc-51 like autophagy activating kinase 1; YTHDF2: YTH N6-methyladenosine RNA binding protein F2.

Traditional tumor detection relies on invasive biochemical analysis, which poses risks of biological toxicity and tissue damage. In contrast, label-free techniques based on biophysical properties(such as mechanical stiffness and electrical impedance)offer a promising non-invasive alternative. However, existing biophysical properties detection methods are largely limited to single-modal measurements, capable of assessing only either mechanical or electrical properties individually, which results in limited tumor detection rates and restricts their clinical applicability. Although combined electro-mechanical measurements exist, they are often limited to microscopic scales or fixed tissue specimens. To overcome these limitations, this study proposes a method for synchronously detecting the mechanical and electrical properties of fresh ex vivo tissues. Utilizing a piezoelectric-driven system, dynamic mechanical loading (1 Hz) was applied to tumor tissue from 4T1-bearing mice and normal mammary tissue from healthy BALB/c mice. The mechanical response and high-frequency electrical impedance (1 MHz) were acquired simultaneously via integrated force/displacement sensors and concentric electrodes. Experimental results show that tumor tissue exhibits distinct viscoelastic features, with significantly higher complex modulus and damping coefficient than normal tissue. Concurrently, the electrical impedance of tumor tissue (276.4 Ω) was significantly lower than that of normal tissue (587.2 Ω). This multi-parameter synchronous detection method effectively discriminates tumor from normal tissue and demonstrates strong potential as a rapid, label-free tool for intraoperative assessment and real-time monitoring.

Ferritins are vital macromolecules that have been widely used in a number of biotechnological fields. Ferritin-based hybrid nanoparticles, composed of different types of subunits and conjugates, represent a next generation of tools, which can significantly enhance their efficiency and expand the range of existing applications. This review outlines the application landscape of these hybrids in developing recombinant vaccines, drug delivery and imaging systems. We highlight the increasing trend towards the development of ferritin-based mosaic vaccines and some of them are already in the first or second phases of clinical studies. In comparison, drug delivery research, which is mostly focused on cancer theranostics, to our knowledge, has not progressed beyond the preclinical stage. Herein, we describe the key limitations and challenges of ferritin-based drug delivery systems development, suggest strategies that address these limitations and discuss promising future research directions. We conclude that engineered ferritin hybrids hold significant potential as useful tools for immunology, theranostics and other biomedical applications.

Emerging insights into the gut microbiome have sparked interest in exploring microbial therapeutics for treating inflammatory bowel diseases (IBD). However, no microbial therapeutics have yet shown clinical efficacy for IBD. Escherichia coli Nissle 1917 (EcN), although effective for maintenance of remission, is only marginally effective for treating active colitis. We postulated that EcN effectiveness is hindered by the inflamed intestine, which prevents colonization because EcN lacks stress-resistance mechanisms necessary to persist during colitis. To address this, we introduced a fitness advantage, the ttr operon, to EcN (EcN::ttr), enabling tetrathionate, a byproduct of intestinal inflammation, to be used as fuel. We hypothesized that EcN::ttr bioengineered to bloom during colitis would effectively treat colitis. We evaluated the efficacy of EcN::ttr in murine colitis including an acute dextran sodium sulfate model and a chronic mucin 2-deficient model. To determine the role of interleukin (IL) 10 in EcN::ttr protection, we tested its efficacy in IL10-deficient mice. Finally, we co-incubated EcN::ttr with human colonoids to understand its effect on barrier proteins. EcN::ttr ameliorated colitis more effectively than EcN and 5-aminosalicylate. EcN::ttr bloomed during inflammation and promoted immunoregulatory responses reliant on IL10 that limited leukocyte infiltration and decreased tumor necrosis factor-α+ myeloid resident cells. EcN::ttr induced functional changes in the gut microbiome related to mucosal healing, increased butyric acid, reduced bacterial translocation, and improved zonula occludens-1 organization. We provide a proof-of-concept study that bioengineering ttr into EcN unlocks a robust therapeutic effect during colitis. EcN::ttr may be a novel microbiome therapeutic for IBD due to its enhanced ability to successfully colonize the inflamed gut.

Tumor recurrence, metastasis, and therapeutic resistance remain major challenges in oncology, driving the need for advanced therapeutic strategies with improved precision and controllability. Optogenetics, which enables light-mediated regulation of cellular functions, has emerged as a promising modality for cancer therapy by offering unparalleled spatiotemporal precision. This capability allows dynamic control of intracellular signaling and transgene expression, enabling selective targeting of malignant cells while minimizing damage to surrounding tissues. However, clinical translation is hindered by key challenges, including inefficient in vivo delivery of optogenetic components, limited tissue penetration of activating light, and suboptimal performance of existing tools. Addressing these barriers requires a convergence of molecular engineering and materials science, wherein advanced biomaterials play a critical role in enabling gene delivery and overcoming tissue-penetration limitations in complex tumor environments. In this review, we provide a comprehensive oriented overview of optogenetics in oncology. We first analyze the molecular mechanisms and engineering principles of representative optogenetic tools, with a focus on LOV- and CRY2-based systems. We then highlight recent advances in biomaterial-assisted optogene delivery and light delivery strategies, emphasizing their material-dependent mechanisms that enable precise spatiotemporal control in vivo. Furthermore, we summarize emerging preclinical applications in cancer immunotherapy, gene regulation, and intracellular signaling control. Finally, we discuss key challenges in biosafety, kinetic optimization, and clinical scalability, and outline future directions that integrate optogenetics with functional materials and intelligent design to realize clinically viable platforms. This review aims to provide a framework for the development of clinically viable optogenetic platforms for next-generation cancer therapy.

The application of nanoparticles (NPs) in medicine as delivery platforms has experienced a significant expansion in the past decade and as a result, various imaging techniques have been applied to monitor cellular NP uptake. Although, high-resolution images of a NP contained within the cell can be generated allowing for quantification, live NP monitoring remains a challenge. Digital holographic microscopy (DHM) has emerged a convenient label-free method to study the cell surface dynamics in real-time by recording a hologram of an interference pattern generated by light which has been scattered from the cell sample, compared to the reference sample. Herein, we report that DHM can be used to monitor the cellular NP uptake in real-time by analysing the changing in surface roughness. As a proof-of-concept, we demonstrated an increase in cell roughness of MCF-7 human breat cancer cells when PLA-coated magnetite, Fe3O4 NPs interacted with the MCF-7 cell surface. By measuring cell roughness every minute, the entire NP internalisation process could be monitored. DHM also addressed challenges within existing imaging techniques, by enabling multi-cellular analysis as well as successfully monitor how changes in NP density and size impacted internalisation rate, highlighting how this method can be further applied in monitoring cellular dynamics and in the development of future nanoparticle-based therapeutics, as well as, tissue engineering.

Despite recent advances, refractory celiac disease (RCD) poses challenging questions. In type 2 RCD (RCD2), the lack of response to the gluten-free diet is attributed to an intestinal intraepithelial lymphoma-carrying driver JAK1 or STAT3 mutations. However, it remains unclear whether these can be safely targeted for therapy. In RCD1, pathogenic insights are still lacking. Duodenal biopsy specimens and peripheral blood mononuclear cells from patients with RCD1, RCD2, active celiac disease (CeD), CeD in remission, and controls were analyzed. Lymphocyte populations were characterized using single-cell transcriptomic, genomic, and T cell receptor (TCR) repertoire profiling. Functional and exome sequencing analyses were performed on patient-derived RCD2 cell lines exposed to JAK inhibitors. We show that clonal malignant RCD2 lymphocytes exhibit interpatient similarities but substantial intratumoral heterogeneity, and provide in vitro evidence that JAK inhibitors can select drug-resistant tumor cells, arguing against their use as monotherapy. In RCD1, we identified clonal T-cell expansions harboring mutations that enhance the JAK-STAT pathway. The detection of both RCD2 and a CD4+ lymphoproliferation in a patient initially diagnosed with RCD1 further illustrates the diversity of lymphoproliferative outcomes in CeD. These findings suggest that RCD subtypes may share underlying mechanisms driven by clonal evolution and JAK-STAT dysregulation. They also highlight the potential limitations of JAK inhibitor monotherapy and the importance of molecularly informed therapeutic strategies.

Aberrant O-linked glycosylation of the IgA hinge segment resulting in galactose-deficient IgA1 (Gd-IgA1) is frequently observed in patients with IgA nephropathy (IgAN), and it is hypothesized to be pathogenic. Here, we genetically disrupted the expression of galactosyltransferase 1 (C1galt1) to elevate Gd-IgA1 levels in mice and examine its role in glomerular deposition. We previously established a mouse model expressing the human IgA1 heavy chain via an IGHA1-knock-in allele. To investigate the role of aberrant glycosylation, we introduced a B cell-specific deletion of c1galt1 into this model. Circulating Gd-IgA1 levels were measured, and kidney phenotypes were evaluated under physiological conditions and following inflammatory stimulation. Furthermore, using a passive mouse model, we compared the glomerular deposition potential of IgA1 derived from various human sources, including serum, ileocecal mucus, as well as from patients with myeloma. B cell-specific deletion of c1galt1 resulted in markedly elevated circulating Gd-IgA1 levels under both physiological conditions and inflammatory stimulation (Lactobacillus casei cell wall extract and complete Freund's adjuvant), and these mice exhibited impaired B cell development and reduced IgA production in both systemic circulation and intestinal mucosa. Correspondingly, glomerular IgA deposition remained limited under physiological conditions and was not substantially enhanced following inflammatory stimulation. In contrast, mucosa-derived IgA1 from patients with IgAN induced significantly stronger mesangial deposition than serum- or myeloma-derived IgA1, despite having a similar or lower Gd-IgA1 content. Our findings support Gd-IgA1 as a correlate of mucosal immune activation rather than a direct pathogenic driver. Instead, tissue origin and immune context are likely key determinants of IgA1 deposition and pathogenic potential.

Chemical inducers of proximity (CIPs) stabilize biomolecular interactions, often causing an emergent rewiring of cellular biochemistry. While the discovery of heterobifunctional CIPs is expedited by rational design strategies, molecular glues have relied predominantly on serendipity. We hypothesized that preexisting ligands could be systematically decorated with chemical modifications to discover compounds that recruit proteins to a composite protein-ligand interface. Using sulfur(VI) fluoride exchange-based high-throughput chemistry (HTC) to install 3,163 structurally diverse building blocks onto ENL (eleven-nineteen leukemia) and BRD4 (bromodomain-containing protein 4) ligands, we screened each analog for degrader activity. This revealed dHTC1, an ENL degrader that recruits CRL4CRBN complex through an extended interface of protein-protein contacts and only engages CRBN after pre-forming the ENL:dHTC1 complex. We also identified dHTC3, a molecular glue that selectively dimerizes BRD4 bromodomain 1 to SCFFBXO3, an E3 ligase not previously accessible for chemical rewiring. Altogether, this study introduces HTC as a facile tool to discover new CIPs and new effectors for proximity pharmacology.

Gαs serves as the prototypical signal transducer for G-protein-coupled receptors (GPCRs) and is the heterotrimeric G protein most frequently mutated in cancer. The classical view of the plasma membrane as the only cellular location where GPCR signal transduction occurs has been challenged by evidence suggesting that Gs also signals from intracellular compartments. However, progress on this topic has stalled because of insufficient approaches with adequate spatiotemporal resolution. Here we describe genetically encoded probes and cell-penetrating compounds that block the effector-binding site of active Gαs in cells to prevent signal propagation at discrete subcellular locations, at user-specified times and across diverse experimental conditions. Using these tools, we show direct evidence of Gαs-mediated signaling on intracellular organelles, unique spatiotemporal features of signaling by Gαs oncomutants and specific regulation of physiologically relevant responses in cardiac or immune cells. These findings pave the way to harnessing the spatiotemporal modulation of Gs signaling and its untapped therapeutic potential.

The eukaryotic translation initiation factor 4E (eIF4E) has emerged as a compelling target for cancer therapeutics due to its pivotal role in regulating cap-dependent translation of oncogenic mRNAs and its implication in various malignancies. However, the clinical potential of current eIF4E inhibitors is limited by suboptimal potency and binding affinity. Based on an analysis of the eIF4E/eIF4G binding pocket and structural features of existing inhibitors, 75 compounds were designed, synthesized, and screened. The binding affinity, molecular mechanism and antitumor activity of the most potent compound b14 were evaluated in vitro and in vivo. Through structure-activity relationship analysis, 75 thiazole derivatives were synthesized and screened for binding affinity using fluorescence polarization (FP) and surface plasmon resonance (SPR). Hit compounds were evaluated for antitumor activity using the SRB assay. The most promising compound, b14, was further investigated for its antitumor activity and molecular mechanism via Western blotting (WB), quantitative real-time PCR (qRT-PCR), immunofluorescence, co-immunoprecipitation, and proteomics. The in vivo antitumor activity and safety of b14 were assessed using HeLa xenograft models and acute/subacute toxicity models, respectively. Compound b14 emerged as a lead molecule, exhibiting a 10-fold higher binding affinity to eIF4E than the reference inhibitor 4EGI-1. Mechanistic studies revealed that b14 disrupts eIF4F complex formation by inhibiting AKT-mTOR-4EBP1 and ERK-eIF4E phosphorylation, subsequently triggering mitochondrial dysfunction and apoptosis in tumor cells, with relatively low IC50 values. Moreover, proteomics analysis further demonstrated that b14 suppresses oncogenic lipogenesis by downregulating key enzymes involved in lipid metabolism. Finally, oral administration of b14 significantly inhibits HeLa xenograft growth in vivo without measurable side effects. Together, our results demonstrate that b14 is an excellent novel small-molecule inhibitor of eIF4E for future cancer therapy.

Ameloblastoma (AM) frequently develops resistance to the BRAF inhibitor vemurafenib, primarily mediated by ameloblastoma-associated fibroblasts (AMFs). However, the potential contribution of the circadian clock in this resistance has not been explored. This study aimed to elucidate the role of the BMAL1-LHX8 axis during tumor-stroma crosstalk, and to investigate the therapeutic potential of targeting this axis to augment vemurafenib sensitivity for AM. Patient-derived AM cells and AMFs were utilized to reconstruct the stroma-rich AM tumoroid in-vitro for recapitulating the tumor-stroma interplay and assessing the pharmacological effect of vemurafenib. Time-series RNA-sequencing, luciferase assays, and CRISPR/Cas9 gene editing were used to define the transcriptional landscape and the BMAL1-LHX8 regulatory network. The efficacy of combining the clock modulator GSK4112 with vemurafenib was assessed in stroma-rich AM tumoroids and cell line-based xenograft models. Patient-derived AMFs exhibit enhanced secretory activity, and elevated metabolic and contractile functions contribute to increased stromal stiffness through ECM remodeling in AM. Tumor-stroma crosstalk was recapitulated in stroma-rich AM tumoroid, which is evidenced by the dynamic alteration of normal fibroblasts (NFs) to AMF-like state. Transcriptomic profiling of stroma-rich tumoroids revealed severe disruption of the molecular clock and LHX8 in AMFs. Functional studies demonstrated that BMAL1-driven LHX8 expression promotes pro-tumorigenic AMF activities, including growth factor secretion and ECM remodeling. Critically, pharmacological inhibition of the BMAL1-LHX8 axis with GSK4112 potently sensitized AM to vemurafenib in both stroma-rich tumoroid and xenograft models. Our findings reveal the role of the BMAL1-LHX8 axis in underlying AMF-mediated drug resistance in AM, and propose that the molecular clock modulation in tumor-stroma crosstalk represents a potential therapeutic avenue for ameloblastoma.

Membrane proteins, including small four-transmembrane proteins, play roles in membrane organization and signal transduction. Generally, they share capacities for protein-protein association and subcellular translocation to regulate cellular functions. Despite known commonalities and differences in structure and function, the relationships among the tetraspanin, CD20-like, golgi 4-transmembrane spanning transporter (Mtp), and transmembrane 4 L six (L6) families remain unclear. Here, we mostly compared TM4SF5, a transmembrane 4 L6 family member, to the genuine tetraspanins and the CD20-like family, exploring unique aspects that evolved from the small four-transmembrane proteins. TM4SF5 and other L6 membrane proteins have two conserved cysteines, affecting N-glycosylation in the large extracellular loop (LEL) and favoring a dimeric state, whereas the tetraspanins maintained their cysteine residues in addition to the CCG motif. However, TM4SF5 and other L6 membrane proteins have more sequence similarity to proteins in the CD20-like family, which lack these features in the LEL. Uniquely, TM4SF5 and other L6 membrane proteins have a β-sheet configuration in the LEL, unlike other small four-transmembrane proteins with α-helices. Therefore, the unique features in the LEL and the N-glycosylation of TM4SF5 may contribute to dimer formation and cholesterol binding, potentially influencing regulatory roles in liver malignancy development.

Pyridoxal-5'-phosphate (PLP), the ubiquitous and ancient cofactor, plays important roles in enzymatic elimination, transamination and other reactions. The catalytic efficiency of PLP-dependent enzymes is significantly higher than that of free PLP. The recruitment of solution PLP by the enzymes, particularly through interactions outside the active-site entrance, is the key step determining the occurrence of PLP-mediated catalysis. However, the precise mechanism by which enzymes recruit solution PLP remains elusive. This study aims to elucidate the mechanism by which enzymes, specifically methionine γ-lyase (MGL) that suppresses cancer cell proliferation through serum or dietary methionine depletion, recruit solution PLP and to investigate the role of the C-terminal domain in this process. The study also seeks to explore the relationship between PLP recruitment and the efficient L-methionine catabolism in the host organism. We report the crystal structure of yMGL, which belongs to a newly identified subgroup of cystathionine γ-lyases, in complex with L-methionine and PLP. To investigate the functional role of the C-terminal domain in PLP recruitment and enzyme activity, we performed C-terminal domain truncations and mutations and assessed their effects both in vitro and in vivo, analyzing changes in PLP binding affinity and L-methionine catabolism, respectively. Through structural, biochemical, bioinformatic and metabolic analyses, we demonstrate that the C-terminal domain of yMGL, outside the canonical PLP-binding domain, is essential for the specific interaction between yMGL and PLP, and L-methionine catabolism. A conserved Ser residue within this domain, located outside of active-site entrance, determines PLP recruitment. These findings elucidate a previously uncharacterized mechanism of PLP recruitment by MGLs and enable rational design of MGLs with enhanced PLP binding and catalytic performance.

Splicing dysregulation frequently occurs in cancers, yet the functional implications of most mis-spliced genes remain elusive. Through a comprehensive pan-cancer analysis of splicing dysregulation, we identified universally mis-spliced genes in cancers and were surprised to find their significant functional enrichment in cell migration. Interestingly, the STE20-like protein kinase (SLK) gene encoding a scaffold protein showed a significant splicing shift from SLK-S to SLK-L isoforms across various cancer types. Detailed studies demonstrated this splicing shift promoted cancer metastasis in cellular and animal models. Mechanistically, RNA-binding Fox-1 Homolog 2 (RBFOX2) protein was identified as a splicing factor that regulates SLK splicing. The two isoforms interact with occludin with different affinities, contributing to their unique activities in cancer metastasis. Notably, the antisense oligonucleotides designed to suppress SLK-L splicing effectively inhibited cancer migration and invasion. Collectively, this study shows a new splicing switch with a key role in controlling cancer metastasis, shedding light on new cancer therapy via splicing manipulation.

Genomic integrity in transcriptionally active regions is pivotal for suppressing oncogenic mutations, yet the mechanisms that govern precise homologous recombination (HR) repair within these regions remain elusive. Here, we report that the IRAK1-spliceosome axis operates with small nuclear RNA (snRNA) as a central hub, potently promoting accurate repair at DNA double-strand break (DSB) sites within active chromatin in human cancer cells. Mechanistically, IRAK1 phosphorylates spliceosomal serine/arginine (SR)-rich proteins to recruit snRNA to DSBs, inducing robust condensation of the MRE11-RAD50-NBS1 (MRN) complex near transcriptionally active regions to create an ATM activation platform. Collectively, our findings define a prevalent mechanism governing region-specific precise repair in transcriptionally active domains, where snRNA acts as a "transcription repair bridge" to link transcriptional processes to HR repair and ultimately preserves genomic stability. Inhibiting IRAK1 axis impairs HR repair in transcriptionally active regions, causing a marked increase in mutation rates specific to these regions and cancer-cell chemosensitivity.

Oncogenic KRAS and NRAS mutations are common in hematologic malignancies, but their signaling in this context remains less well characterized than in carcinomas. Using multi-omics screens in multiple myeloma, we sought to identify regulators of RAS activity. We found that the phosphatase PP1C dephosphorylated conserved RAS residue T148, permitting LZTR1-dependent proteasomal degradation. LZTR1 was ineffective against KRAS A146 gain-of-function mutations, which lie adjacent to T148 and are enriched in hematologic cancers, such as diffuse large B cell lymphoma and acute myeloid leukemia. Remarkably, KRAS protein stability was four-fold lower in hematologic versus carcinoma cells, revealing a unique therapeutic opportunity targeting RAS protein stability. PAK1 and PAK2 shielded RAS from LZTR1-dependent degradation by phosphorylating T148, and inhibiting PAK1/2 activity improved RAS-directed therapy. Collectively, these findings reveal a regulatory circuit governing RAS stability that is preferentially active in blood cancers and potentially druggable.

The PI3K/AKT signaling pathway serves as a pivotal regulatory hub that governs essential cellular processes, including growth, proliferation, metabolism, survival, and migration. In the context of tumorigenesis and cancer development, aberrant activation of this pathway drives malignant transformation, enhances invasive and metastatic potential, and confers resistance to therapeutic interventions, ultimately resulting in tumor progression and poorer clinical outcomes. This study comprehensively investigates PI3K/AKT signaling, elucidating its involvement across diverse human systemic tumors and analyzing the interplay between this pathway and other signaling cascades implicated in tumorigenesis. Emphasis is placed on mechanistic insights into the role of the PI3K/AKT pathway in tumor initiation and progression across various systems, alongside an overview of therapeutic strategies targeting PI3K/AKT signaling. Notably, advances in research on the PI3K/AKT pathway have identified a wide array of potential therapeutic targets, facilitating significant progress in related treatment modalities. Given the extensive involvement of PI3K/AKT signaling in diverse biological processes, optimizing the therapeutic efficacy of PI3K/AKT-targeted interventions while minimizing associated adverse effects remains a critical challenge, necessitating innovative approaches for substantial therapeutic breakthroughs. Therefore, a systematic and nuanced understanding of the pathway's role in tumor biology is essential for advancing foundational research and informing future clinical developments in this field.

Durable treatment of leukemia remains challenging because most therapeutic regimens rely on a single cytotoxic mechanism. Here, we report a rationally loaded cuprous oxide ferritin nanoplatform (Cu2O@Fn) that enables targeted induction of dual cell death in leukemia. By exploiting the intrinsic binding of ferritin to CD71, which is broadly expressed across leukemia types, Cu2O@Fn achieves efficient and selective intracellular delivery in diverse leukemia cell lines and patient-derived bone marrow samples. Upon cellular uptake, the controlled release of copper ions engages dual death pathways: Cu+ mediated Fenton-like reactions drive ferroptosis, and subsequent Cu2+ overload triggers cuproptosis, collectively resulting in irreversible mitochondrial damage. Comprehensive evaluation across multiple in vitro systems, cell-line-derived xenografts, and a CD71-humanized leukemia model demonstrates robust and broad-spectrum anti-leukemic efficacy, including performance exceeding established clinical treatment regimens. Together, this work establishes ferritin-mediated copper delivery as a neotype therapeutic strategy against leukemia.

Small ubiquitin-like modifier (SUMO) conjugation, or SUMOylation, is a dynamic post-translational modification that regulates protein stability, localization, transcriptional activity, DNA damage response, and cellular stress adaptation. Increasing evidence indicates that dysregulated SUMOylation contributes to tumor progression, immune disorders, and neurodegenerative diseases, highlighting the SUMO pathway as a promising therapeutic target. However, despite extensive advances in SUMO biology, a comprehensive pharmacological assessment of SUMO pathway inhibitors, including their mechanisms of action, evidence quality, target engagement, and translational challenges, remains insufficient. This review presents an inhibitor-centered analysis of pharmacological strategies targeting the SUMOylation machinery. This review describes the molecular architecture of the SUMO conjugation cycle and identifies major druggable nodes, including the SUMO-activating enzyme (SAE), the SUMO-conjugating enzyme UBC9, SUMO-specific proteases (SENPs), and SUMO-dependent protein-protein interactions. It also examines the biological rationale for SUMO pathway inhibition in major disease contexts, with an emphasis on cancer, autoimmune disorders, and neurological diseases. Furthermore, SUMO pathway modulators are systematically classified according to their molecular targets and evidence maturity, including SAE inhibitors, UBC9-directed compounds, SENP inhibitors, direct SUMO binders, and natural product-derived modulators. Their mechanisms of action, biochemical potency, structural validation, cellular target engagement, in vivo efficacy, PK/PD properties, and developmental status are critically evaluated. Overall, this review provides a comprehensive pharmacological framework for understanding SUMO pathway inhibition and highlights key considerations for the development of next-generation SUMO-directed therapeutics.

Atopic dermatitis (AD) is a heterogeneous inflammatory skin disorder characterized by recurrent eczematous lesions and persistent pruritus. Its pathogenesis involves epidermal barrier dysfunction, immune dysregulation, microbial dysbiosis, and altered neuroimmune signaling. Genetic defects in structural proteins, particularly filaggrin, and altered epidermal lipids increase allergen and microbial penetration, perpetuating itch, scratching, tissue injury, and inflammation. Molecular evidence further reveals AD endotypes beyond conventional clinical phenotypes, with heterogeneity in immune polarization, barrier dysfunction, microbial colonization, and pruritic pathways. This heterogeneity may underlie differences in clinical presentation and treatment response. Conventional treatments, including skin hydration, antihistamines, and topical anti-inflammatory agents, often provide incomplete or transient control. Mechanism-based therapies targeting IL-4/IL-13, OX40/OX40L, JAK, and PDE4, together with AhR agonists, have broadened treatment options. However, efficacy must be balanced against treatment-specific risks. Biologics require monitoring for ocular and injection-site reactions, whereas systemic JAK inhibitors require monitoring for infections and laboratory abnormalities and carry regulatory class warnings regarding major adverse cardiovascular events, venous thromboembolism, and malignancy; however, these warnings are largely extrapolated from studies in older patients with rheumatoid arthritis, and the magnitude of these risks in patients with AD remains uncertain. Long-term safety and real-world evidence remain limited for several emerging therapies. This review synthesizes current knowledge of barrier dysfunction, immune heterogeneity, molecular endotypes, microbiome imbalance, neuroimmune signaling, and mechanism-based therapies. Integrating clinical phenotypes with molecular endotypes, predictive biomarkers, efficacy, and individualized safety assessment may enable more precise treatment selection and durable disease control.

Engineering transgene expression to respond specifically to intracellular cues holds transformative potential for monitoring cell states and modulating cellular functions. Here we introduce TRADER (Target-induced Transgene Activators via Reprogramming of dCas6-barnase Degrader), a versatile and modular platform that enables transgene activation in response to intracellular molecules at the post-transcriptional level. TRADER leverages a proximity-dependent RNA degrader system, in which a catalytically inactive Cas6 (dCas6) is fused to an engineered RNase, barnase, to suppress exogenous gene expression through targeted mRNA degradation. This repression can be relieved through two distinct mechanisms: (1) incorporating microRNA (miRNA) target sites into the dCas6-barnase transcript allows for endogenous miRNA-mediated downregulation of the degrader, and (2) inserting protease-cleavable linkers enables protease-triggered degrader disassembly, and further engineering a positive-feedback loop achieves signal amplification. As a proof of concept, we demonstrate the utility of TRADER in two applications: selective sensing of intracellular miRNA and ATP levels in target cells using fluorescent proteins, and conditional elimination of tumor cells overexpressing cathepsin B protease (CaBp) through induced expression of a cytotoxic protein. Collectively, TRADER offers a flexible and expandable toolkit for cell-state-specific control of transgene expression, with broad implications for synthetic biology, cell-based diagnostics, and precision therapeutics.

Metastasis is an inefficient cellular process in which most disseminated tumor cells fail to form secondary lesions in distant tissues due to hostile conditions, such as protective immune surveillance. The few cells that survive these threats can seed subclinical metastatic lesions, known as micrometastases, which are the least-known stage of the metastatic cascade. In this study, we review micrometastasis immunobiology, which differs from that of larger, clinically manifested metastasis. Key mechanisms such as epithelial-to-mesenchymal transition, stemness, dormancy, and immune evasion shape this bottleneck of metastasis, determining long-term disease evolution, therapy responses, and patient outcomes. Understanding micrometastasis immunology may reveal therapeutic opportunities to fully eradicate disseminated cells. Thus, we discuss emerging time-tailored immunopreventive strategies to intercept the progression to overt metastasis.

The human transcriptome contains millions of A-to-I editing sites arising from an unclear number of poorly characterized dsRNAs. Editing sites reveal the presence of dsRNA, but this method is limited by transcription levels, read depth, and ADAR expression and cannot identify unedited dsRNA. To address these limitations, we developed dsRNAscan. Applying dsRNAscan to the human genome predicted 5 million dsRNAs, mostly in repetitive and intergenic regions. Machine learning models trained on A-to-I editing and RNA structure-probing data identified ∼2.4 million high-confidence predictions, which were enriched at dsRNA-binding protein binding sites. Additionally, we predicted hundreds of dsRNAs conserved across vertebrates and observed thousands of editing-enriched regions suspected to arise from intermolecular dsRNAs formed with sense-antisense transcripts. Quantifying expression of intramolecular and intermolecular dsRNAs accessible to cytoplasmic immune sensors revealed that their ratio correlated with ADAR dependency across cancer cell lines. The human dsRNAome is available as a resource at https://dsrna.chpc.utah.edu/.

The Fc region of therapeutic IgG antibodies is often engineered to remove or "silence" Fc effector functions, but it remains unclear whether these mutations eliminate all Fc-mediated effector activity. Human Fc receptor-like 5 (FCRL5/FcRH5) is a low-affinity IgG Fc receptor expressed on B cells and is an actively pursued antibody target in multiple myeloma. Here, we show that common Fc function-silencing mutations do not impair FCRL5-mediated activity and therefore attenuate, rather than eliminate, Fc effector function. The crystal structure of the FCRL5-IgG1 Fc complex, solved at 3.4 Å resolution, revealed that FCRL5 binds IgG1 Fc in a 1:1 complex through a binding mode distinct from that of classical Fcγ receptors, explaining why mutations that attenuate Fc effector function spare FCRL5 binding. Fc-engineered antibodies that selectively engage FCRL5 inhibited B cell receptor-induced Ca2+ flux in FCRL5-expressing B cells. These findings demonstrate that Fc-attenuated therapeutic IgG retains the ability to engage FCRL5, identifying an unappreciated pathway for B cell modulation.

Nanoparticle carriers are widely used for nucleic acid delivery, with poly(ethylene glycol) (PEG)-conjugated nanoparticles emerging as a frontrunner in the field. Growing concerns surrounding PEG immunogenicity have generated interest in zwitterionic polymers as alternative biocompatible surface grafts. At present, there exists a paucity of data regarding zwitterion-conjugated nanoparticle performance in primary immune cells and across biological sexes. Here, we synthesized pH-responsive PEG- and zwitterion-grafted cationic nanoparticles, including PEG methyl ether methacrylate (PEGMMA), carboxybetaine methacrylate (CBMA), sulfobetaine methacrylate (SBMA), and polymeric SBMA, and assessed their cytotoxicity and small interfering RNA (siRNA) delivery in primary male and female murine macrophages. All formulations enabled intracellular siRNA uptake and suppressed tumor necrosis factor-α expression. Sex-dependent effects varied by nanoparticle formulation and biological end point. Zwitterionic nanoparticles exhibited sex-specific effects in cytotoxicity and gene knockdown despite similar uptake, whereas PEGMMA showed higher uptake in females but comparable knockdown across sexes. These findings demonstrate that uptake alone does not predict functional efficacy and highlight the importance of sex-informed evaluation when developing nanomedicines.

Manual palpation is a cornerstone of medical assessment, yet its subjective nature limits its ability to provide quantitative data of characteristics like tissue hardness. Here, we introduce DigiPalp, a wearable smart glove designed to enable real-time 4D tactile scanning, combining 3D surface mapping with a tissue hardness measurement at each point. This quick (typically <0.5 seconds) and noninvasive measurement is achieved through a fusion of custom piezoresistive pressure sensors and magnetic position sensors embedded into the glove's fingertips. By featuring silicone-encapsulated stretchable wiring, the hand's natural range of motion is maintained to support a workflow like conventional palpation. We show that the system can reliably differentiate six hardness levels across the soft tissue range, identify small, harder nodules (down to 5-millimeter radius) embedded in silicone phantoms, mimicking tumor detection, and demonstrate the system's capability on complex tissue through a full 4D scan of the torso of a living person.

Solitary fibrous tumor (SFT) is a rare and aggressive sarcoma driven by NAB2::STAT6 gene fusions, yet effective targeted therapies remain unavailable. Here, we report that the NAB2ex4::STAT6ex2 fusion variant forms nuclear condensates via liquid-liquid phase separation (LLPS) in engineered fibroblast models and primary SFT cells. These condensates co-localize with BRD4S and EGR1, key transcriptional regulators, and are functionally active, driving widespread transcriptional reprogramming. Treatment with Mithramycin A, a compound that disrupts EGR1-DNA interactions, dissolves NAB2::STAT6 condensates and reverses their aberrant gene expression and chromatin binding signatures. Our findings uncover a previously unrecognized role for NAB2::STAT6 in condensate-mediated oncogenic signaling and provide a mechanistic rationale for condensate-targeted therapy in SFT.

PIM kinases are overexpressed in castration resistant prostate cancer (CRPC) and many small molecule PIM kinase inhibitors (smPIM inhibitors) have been designed to block the catalytic activity of PIM. However, smPIM inhibitors have shown limited efficacy in solid tumors. Notably, all these inhibitors share the common property that they increase total PIM protein levels, which limits their efficacy because PIM1 has kinase-independent pro-survival effects. Here, we identify high mobility box group 1 (HMGB1) as a novel PIM1 binding partner. Stabilization of PIM1 by smPIM inhibitors increases the cytosolic accumulation of HMGB1, which leads to activation of mitophagy and suppresses oxidative-stress induced cell death. Knockdown of PIM1/2/3 and/or HMGB1 sensitizes cancer cells to smPIM inhibitors. In contrast, treatment with a PIM PROTAC (PIMTAC) that we developed overcomes the kinase-independent pro-survival effects of PIM1 and is more effective than smPIM inhibitors in vitro and in vivo. These results uncover a mechanism of resistance that has limited the success of smPIM inhibitors and provides compelling evidence that targeted degradation of PIM is needed to realize its potential as an anti-cancer target.

Epidermal growth factor receptor (EGFR) is an oncogenic driver in multiple cancers and a therapeutic target of tyrosine kinase inhibitors and neutralizing monoclonal antibodies. However, resistance to EGFR-targeted therapies, particularly the anti-EGFR antibody cetuximab, remains a clinical challenge in colorectal (CRC) and head and neck (HNSCC) cancers. Cetuximab exerts its antitumor activity by blocking ligand-dependent EGFR signaling and by engaging immune effector mechanisms. To investigate cetuximab resistance mechanisms, we cultured the cetuximab-sensitive CRC cell line DiFi in 3D with cetuximab, generating the cetuximab-resistant derivative (DiFi-CR). Genomic and transcriptomic profiling revealed that DiFi-CR cells harbor a mutation of the S442 residue within the EGFR ectodomain. Patient samples revealed recurrent EGFR S442 mutations following anti-EGFR therapy, suggesting S442 as a potential resistance hotspot. For mechanistic analyses, we reconstituted the EGFR S442I mutation, using a doxycycline-inducible system, and showed that it was necessary and sufficient to induce cetuximab resistance in CRC and HNSCC cells using in vitro cultures and in vivo mouse experiments. In silico studies, live-cell binding assays, and antibody enrichment in nude mice xenografts revealed that the S442I mutation leads to weaker EGFR-cetuximab binding. Weaker cetuximab binding was also predicted in silico for other S442 patient mutations. We found that mutant EGFR-driven resistance could be overcome by targeting the EGFR family member, ERBB2, with trastuzumab-deruxtecan. This combinatorial response required a physical interaction between EGFR and ERBB2, determined by co-immunoprecipitation. Our study supports EGFR S442 mutations as cetuximab resistance drivers and highlights co-targeting ERBB2 as a therapeutic strategy to restore anti-EGFR efficacy.

Resistance to temozolomide (TMZ) is a major driver of treatment failure and high mortality in patients with glioblastoma (GBM). However, the mechanisms underlying TMZ resistance, especially intrinsic resistance, remain incompletely elucidated. Through integrative multi-omics analyses, we identified a novel super-enhancer-driven circular RNA, circMLB, which is highly overexpressed in TMZ-R GBM. Both in vitro and in vivo studies robustly demonstrate that circMLB significantly enhances TMZ resistance in GBM. Mechanistically, the transcription factor PAF1 forms a protein complex with BRD4 that is enriched in the super-enhancer region, thereby upregulating circMLB expression. Furthermore, highly expressed circMLB directly binds to PKR, triggering stress granule assembly and ultimately reinforcing TMZ resistance in GBM. Importantly, we identified a novel PKR inhibitor, lurasidone, which synergizes with TMZ to exert potent anti-tumor activity against TMZ-R GBM in preclinical models. Our findings uncover a previously unrecognized SE-circMLB-PKR axis mediating intrinsic TMZ resistance and highlight the lurasidone-TMZ combination as a promising therapeutic strategy to surmount TMZ resistance in GBM.

It remains unclear whether super-enhancers (SEs) can drive expression of circular RNAs (circRNA) in gastrointestinal cancers (GI cancers). We first identified circRREB1 as a novel SE-driven circRNA in GI cancers and HNRNPL as the RNA binding protein controlling circRREB1 biogenesis. Indeed, SE-driven HNRNPL leads to high circRREB1 expression in GI cancers, which is regulated by transcription factor MAZ. circRREB1 could promote invasion and metastasis of cancer cells in vitro and in vivo. Mechanistically, circRREB1 binds to the SRSF3-XRN1-YTHDC2 complex in cytoplasm, destabilizes the m6A-modified CD44 mRNAs, and, thus, enhances metastasis of cancer cells. Consistently, high circRREB1 levels in malignant tissues are associated with poor patient prognosis. Although showing no impacts on tumor growth in nude mice, circRreb1 accelerates tumor proliferation in immunocompetent mice. Interestingly, circRreb1 elevates Ets1 expression through a competing endogenous RNA mechanism, up-regulates Cxcl2 expression and secretion, drives myeloid-derived suppressor cells (MDSCs) infiltration to tumors and contributes to formation of immunosuppressive tumor microenvironments. In line with this, depletion of MDSCs abrogates the oncogenic effects of circRreb1 in vivo. Notably, circRREB1 silencing synergizes with anti-PD1 therapy to activate antitumor immunity, offering novel insights into circRNA-mediated cancer epigenetics and promising therapeutic targets.

DNA methylation is an epigenetic modification that influences gene expression and cellular signaling. Abnormalities in the DNA methylation landscape have been associated with diverse pathologies and diseases, making accurate and accessible detection methods essential for both research and clinical diagnostics. Conventional methylation analysis techniques, such as bisulfite conversion-based assays, are the gold standard; however, they can be labor-intensive, chemically harsh, and poorly suited for real-time or point-of-care applications. Additionally, the potential for incomplete or over-conversion may affect diagnostic performance and has been a longstanding topic of discussion around bisulfite conversion. Alternative strategies, such as methylation-sensitive restriction enzymes (MSREs) and catalytic approaches, have addressed some limitations but still face challenges in specificity and sensitivity, throughput, and operational complexity. Recent advances in bioengineering and nanotechnology have leveraged the development of molecular elements, broadly referred to as "methylation readers", capable of specifically recognizing methylated DNA. Methylation readers are naturally occurring proteins, such as methyl-CpG binding domain proteins (MBDs), but also synthetically engineered proteins, such as transcription activator-like effectors (TALEs), as well as abiotic analogs, such as molecularly imprinted polymers (MIPs) and methylation-sensitive intercalators. The recent integration of these methylation readers into various biosensor platforms has enabled a significant advancement in epigenetic analysis, offering rapid, sensitive, and conversion-free DNA methylation detection without the need for relatively complex sample preparation. Such capabilities hold significant potential for early disease detection, personalized diagnostics, and fast monitoring of methylation-associated diseases. This review aims to provide a comprehensive and focused overview of the recent developments in the integration of methylation readers into biosensing technologies. We highlight the potential of these integrated approaches to transform epigenetic analysis in both research and clinical settings, discuss existing limitations, and outline future directions for advancing the field.

Electrochemical (EC) and photoelectrochemical (PEC) biosensors offer accuracy, convenience, and affordability, making them ideal for early cancer screening. However, in conventional EC and PEC biosensors, signal crosstalk between the photocurrent and redox currents poses a significant challenge for the simultaneous detection of multiple biomarkers, as it prevents their independent readout. Here, we present an interference-free EC-PEC biosensor that simultaneously detects the early-stage cancer biomarkers miR-155 and miR-451 using a direct hybridization method, and enhances the EC-mode signal via a hybridization chain reaction. Through ingenious design, the photocurrent generated by the poly(o-phenylenediamine) (PoPD)/WSe2 heterojunction is decoupled from the redox peak current of methylene blue, effectively avoiding signal interference and enabling separate quantification of miR-155 and miR-451. Moreover, the PEC mode employs a photocurrent polarity-switching strategy to effectively eliminate false-positive results. This current-decoupled biosensor exhibits excellent analytical performance for miR-155 and miR-451 over the range of 1.0×10°-1.0 × 105 fM, with detection limits as low as 9.0 × 10-2 fM and 3.7 × 10-1 fM, respectively. This work not only provides a new approach for early cancer screening but also offers an easy-to-use electrochemical platform for multiplexed detection of coexisting targets.

Metabolism is fundamental to cell function, yet its activities vary across tissue environments. Resolving these processes in situ at single-cell resolution is crucial for understanding physiology in health and disease. However, existing methods lack biochemical specificity or direct linkage to cell identity. Here we report a method, Raman Enhanced Delineation of Cell Atlases in Tissues (REDCAT), an all-optical platform integrating Raman scattering microscopy and high-plex immunofluorescence to co-map metabolism and cell types. REDCAT achieves subcellular profiling of protein, lipid, nuclear metabolites and redox metabolism in human tissues. In lymph nodes, it revealed cell-type-specific metabolic specialization. In lymphoma, REDCAT uncovered profound reprogramming and transitional states during tumor transformation. In the liver, it resolved zonation-dependent metabolic gradients. By linking cell identity to spatial metabolic states, REDCAT provides a framework for studying immunity and cancer, offering a path to deciphering the metabolic basis of disease.

RAS, a frequently mutated cancer-driver gene, has been the focus of intensive research, with numerous inhibitors developed to target RAS and its signalling molecules. With the advent of approved subtype-specific RAS inhibitors targeting KRASG12C mutation, the development of RAS inhibitors that are effective regardless of RAS mutation status is the next major challenge to address clinically prevalent RAS mutations and further overcome RAS-driven acquired resistance to currently available drugs. With the goal of obtaining broad-spectrum aberrant RAS-signalling inhibitors, we conduct multimodule drug screening for small-molecule compounds capable of RAS/RAF-binding inhibition. Structural studies of the hit compounds demonstrate that they covalently bind to a distinct site in the RAS-binding domain of RAF and allosterically disrupt RAF conformation, thereby preventing RAS/RAF interaction and downstream signalling. The compounds exhibit antitumour efficacy against multiple cancers with varying RAS mutations, including KRAS, NRAS and HRAS, and wild-type RAS-driven cancers in preclinical models. Furthermore, they effectively suppress tumour growth of BRAFV600E-melanoma with acquired BRAF inhibitor resistance by preventing RAS signal reactivation. These findings provide a possible approach for designing RAF-targeting compounds, and the resulting molecules may serve as the basis for developing RAS/RAF-signalling inhibitors with distinct mechanisms to overcome multiple RAS-driven cancers.

Chimeric antigen receptor (CAR) T cell therapy for pancreatic ductal adenocarcinoma (PDAC) has been challenging, in part due to the paucity of tumor-specific cell-surface targets. Here, we explored the design feasibility and anti-tumor efficacy of CAR T cells recognizing the CA19-9 glycan, a tumor antigen commonly associated with PDAC and other gastrointestinal tumors. We designed and screened 14 CA19-9 CAR constructs harboring different single-chain variable fragments (scFvs) and intracellular signaling domains. After narrowing down to one optimal CAR configuration, we further evaluated its anti-tumor efficacy in various in vitro and in vivo models. Out of 14 CAR constructs tested, we identified one design - AbLIFT15.28z - that enabled robust T cell expansion and efficient tumor cell-killing properties. Experiments with human PDAC cell lines and patient-derived organoids (PDOs) revealed that AbLIFT15.28z CAR T cell-mediated cytotoxicity was confined to cancer cells expressing CA19-9, both in vitro and in vivo. We then tested the efficacy of AbLIFT15.28z CAR T cells in immunocompetent murine models of PDAC, revealing significant effects against both primary and metastatic tumors. Efficacy was not limited to PDAC, as robust and specific anti-tumor effects were observed against CA19-9-expressing tumors of the colon, stomach, esophagus, and biliary tract. Our findings demonstrate the effectiveness of CA19-9 CAR T cells in GI cancers using various pre-clinical tumor models, providing a rationale for further clinical development.

Data-driven protein engineering has enabled biosensors that directly couple molecular recognition with measurable signal outputs, providing a powerful strategy for rapid protein analysis. However, their integration into automated and portable analytical systems remains limited. Here, we developed an integrated photoelectrochemical and fluorescence digital microfluidic (PF-DMF) platform for automated biomarker detection by combining a programmable "lock-and-cage" protein sensor (LucCage/LucKey) with precise digital microfluidic manipulation. The platform enabled automated detection of human tumor necrosis factor-α (hTNF-α) with a detection limit of 0.1 μM over a linear range of 0.1-10 μM, while exhibiting high specificity, good reproducibility, and minimal sample consumption. Direct analysis of hTNF-α in simulated body fluids was achieved without complex sample pretreatment. The platform was further extended with a CRISPR-based nucleic acid sensing module, enabling nucleic acid detection of Salmonella typhimurium (S. typhimurium) within the same automated workflow. In addition, a standardized reagent kit maintained stable analytical performance for at least 30 days under appropriate storage conditions. Overall, the proposed PF-DMF platform provides a versatile and scalable strategy for automated multimodal biomarker analysis, offering significant potential for portable diagnostics in healthcare and food safety.

Clonal acquisition of multiple chromosomal abnormalities in hematopoietic stem and progenitor cells (HSPCs) is a hallmark of high-risk acute myeloid leukemia with complex karyotype (AML-CK). AML-CK is associated with TP53 mutations and chromosome 5q deletions [del(5q)]; however, the drivers and clonal trajectories of aneuploid evolution in HSPCs remain unknown. We developed a patient-derived induced pluripotent stem cell (iPSC) model in which preleukemic HSPCs clonally evolve to distinct, highly aneuploid states after transient mitotic inhibition. By tracking chromosome evolution at single-cell resolution, we show that TP53-mutant HSPCs with del(5q), but not TP53 mutation alone, evolve complex chromosomal changes. Clonal evolution was marked by the stepwise acquisition of numerical and structural chromosome changes seen in patients with AML-CK, with individual abnormalities conferring a fitness advantage. iPSC-derived aneuploid HSPCs and primary samples from patients with AML-CK exhibited a conserved gene expression signature marked by upregulation of PTEN, cohesins, and the antiapoptotic factor BCL2, indicative of a shared aneuploid cell state in HSPCs. The clinical BCL2 inhibitor venetoclax eradicated BCL2-dependent aneuploid clones, with resistant clones undergoing a lineage switch to upregulate alternative BCL2 factors. In summary, we demonstrate that mutant TP53 and del(5q) drive chromosome evolution marked by stepwise acquisition of individual abnormalities. Moreover, aneuploid HSPCs exhibit a shared gene expression state that confers unique, targetable therapeutic vulnerabilities in AML-CK.

T-cell exhaustion has been shown to be a key resistance mechanism to efficacy of T-cell engagers (TCE) in multiple myeloma (MM). Mezigdomide, a potent cereblon E3 ligase modulator that targets IKZF1 and IKZF3 simultaneously for proteasomal degradation, has been shown to modulate T-cell activity in patients with MM. We explored the possibility that targeting IKZF1/IKZF3 could address T-cell exhaustion and restore functionality. We conducted extensive transcriptomic and epigenetic profiling on ex vivo generated exhausted T cells (Tex), using their autologous activated T cells as a comparison. Our study revealed that IKZF1 and IKZF3 are critical regulators that contribute to the development and maintenance of T-cell exhaustion. They regulate transcription by directly binding to promoters and enhancers, both proximal and distal, thereby altering transcriptional potential. Increased IKZF1 binding to exhaustion genes after multiple T-cell stimulations results in the enhancement of transcription, whereas binding to cytokine genes results in transcription repression. Mezigdomide treatment in Tex results in decreased expression of exhaustion-related markers, increased proinflammatory cytokine expression, and enhanced target cell killing with alnuctamab, a B-cell maturation antigen targeting TCE. This study provides crucial mechanistic insights into the roles of IKZF1/IKZF3 in T-cell exhaustion, supporting the rationale for combining mezigdomide with TCE to enhance therapeutic outcomes in MM.

Natural products (NPs) are foundational to drug discovery, owing to their diverse molecular scaffolds and novel bioactivities. However, optimizing NPs for optimal bioactivity and improved drug properties is hindered by material scarcity and labor-intensive purification processes. Here, we present an automated ultrahigh-throughput experimentation (ultraHTE) and direct-to-biology (D2B) platform that enables rapid structure-activity relationship (SAR) mapping from miniaturized libraries of synthetic NP analogues, unlocking NPs as a viable starting point for late-stage diversification in drug-lead optimization. Starting from leelamine, a conifer resin-derived tricyclic diterpene with prostate cancer antiproliferative activity, we synthesized leelamine analogues in a 1536-well format, achieving a 73% success rate while consuming only 0.03 mg of NP per reaction. Key to success was the development of a platinum-catalyzed amine-acid reductive amination reaction, which performed well on both amine and complementary acid-derived NPs, in a so-called Janus strategy which more than doubled the accessible chemical space. Direct phenotypic screening in LNCaP cells enabled rapid identification of potent, cell-permeable analogues with robust quality control achieved through iterative resynthesis on preparative scale. SAR insights revealed two key substitution patterns driving potency based on 4-methoxyphenyl and 3-indolyl derivatives. Mechanistic studies─including androgen receptor binding, chemoproteomics, and a lysosomal cholesterol colocalization assay─demonstrated distinct modes of action for optimized analogues compared to leelamine. Collectively, this amine-acid amination-based D2B platform showcases accelerated SAR elucidation from minimal material and cellular profiling, unlocking a framework for mechanism-driven lead discovery in NP chemical space.

Methionine (Met) plays a pivotal role in numerous cellular functions. Methionine restriction has been demonstrated to provide metabolic benefits in aging, obesity, diabetes and as an adjunct to cancer therapy. However, the methionine-sensing proteins and how cells directly sense the methionine level have remained elusive. In this study, we developed a photoaffinity analogue of methionine to capture proteins that specifically recognize and sense methionine in living cells. Using chemoproteomic profiling and biochemical validation, we found that PKM2 is a specific methionine sensor that transduces methionine availability signals through the interaction with the GATOR2 complex, which, in turn, modulates the downstream response of the mTORC1 pathway through a novel methionine-recognition pocket on PKM2. As our findings indicate that the sensing of methionine by PKM2 is independent of its enzymatic activity, we envision that disrupting the binding of methionine to PKM2 or stabilizing the PKM2-GATOR2 interaction would create a methionine pseudostarvation state in living cells, which holds promise as a novel therapeutic avenue that could emulate the physiological benefits of a methionine-restricted diet and circumvent the drawbacks of dietary methionine restriction.

Polyethylene glycol (PEG) can stabilize nanoparticles and facilitate transport across biological barriers, yet the same PEG layer suppresses cellular uptake─a trade-off known as the PEG dilemma. Current strategies seek to overcome this limitation by enabling PEG shedding in response to pathological biochemical cues, but their dependence on heterogeneous microenvironments limits predictability and cross-route applicability. Here, we report a thermoresponsive PEG shedding strategy based on PEG-lipid conjugates with oxanorbornadiene linkers that cleave at physiological temperature. By varying the linker structure, PEG shedding kinetics can be modulated over pharmacologically relevant time scales, enabling mRNA-lipid nanoparticles to undergo surface transitions from sterically shielded to cell-interactive states. In inhaled delivery, rapid PEG shedding after mucus traversal restored cellular uptake and markedly enhanced pulmonary mRNA expression, resulting in inhibition of pulmonary metastases. In systemic administration, intermediate PEG shedding preserved the circulation benefit of dense PEGylation while enabling subsequent intracellular delivery, leading to improved tumor expression and antitumor efficacy. These results establish kinetic programming of PEG shedding as a design principle for dynamically regulating nanoparticle biointerfaces across distinct delivery routes.

Efficient and stimulus-modulated intersystem crossing (ISC) in photodynamic therapy (PDT) is usually designed via molecular engineering of π-conjugated core structures. Herein, we report that nonconjugated protonatable imide side chains of perylene diimide (PDI) photosensitizers can function as through-space electrostatic regulators of ISC, enabling acid-responsive reactive oxygen species (ROS) generation. Incorporation of a conformationally flexible dimethylamine chain at the imide position enables acid-triggered protonation, inducing a through-space cation-carbonyl interaction between protonated dimethylamine and the neighboring imide carbonyl group. This localized electrostatic field redistributes electrons within the PDI-DMA core, markedly reduces singlet-triplet energy gaps, and promotes ISC. Consequently, PDI-DMA exhibits acid-enhanced ROS generation, achieving over an order-of-magnitude enhancement relative to its alkyl-substituted analogue PDI-Bt. Systematic molecular controls, host-guest shielding experiments, and theoretical calculations collectively identify this nonconjugated side-chain-induced electrostatic interaction as the dominant origin of the enhanced ROS generation, overturning the conventional assumption that nonconjugated imide substituents are photophysically inert. When formulated into biocompatible nanoparticles, they preferably accumulate at lysosomes and retain acid-enhanced ROS production, delivering potent photodynamic antitumor efficacy following intratumoral administration in tumor-bearing mice. This work establishes nonconjugated side-chain electrostatic modulation as an unexplored design paradigm for reprogramming excited-state processes in stimuli-responsive organic photosensitizers.

Aberrant O-glycosylation is a prominent feature of cancer and contributes directly to multiple hallmarks of malignant transformation. Cancer-associated O-glycans include both truncated structures that are unique to mucin-type O-glycosylation and more elongated terminal motifs that function as ligands for glycan-binding proteins. Instead of representing a simple binary change from elongated to truncated structures, it is increasingly understood that these glycans decorate the cancer cell surface with a heterogeneous pattern of truncated, elongated, capped, and partially processed glycoforms, which coexist even on individual proteins and within the tumor glycocalyx. Functionally, truncated aberrant O-GalNAc glycans, such as Tn, STn, and T perturb epithelial differentiation, barrier integrity, signaling, and immunosuppression through interactions with glycan binding receptors, such as Siglecs and MGL. In addition, more elongated O-glycans, including Lewis antigens, interact with selectins promoting metastasis and cellular trafficking. Moreover, individual GalNAc-transferase isoenzymes, governing site-specific O-glycosylation, are emerging as context-dependent regulators of oncogenic processes through glycosylation of selected proteins. These observations establish cancer-associated O-glycans as context-dependent molecular drivers of tumor progression and promising therapeutic entry points. In this review, we discuss the expression, regulation, and functions of cancer-associated O-glycans, their roles in glycan-binding protein biology, and emerging therapeutic strategies that may enable a new generation of glycan-directed precision treatments in oncology.

Protein arginine deiminase-4 (PAD4) catalyzes hydrolysis of arginine to citrulline in proteins that promote widespread cellular changes that can induce innate immunity and promote cancer. Hyperactivity of PAD4 leads to a form of cell death called NETosis, releasing PAD4 to the extracellular space to promote various autoimmune diseases through the generation of anticitrulline protein antibodies (ACPAs). Little is known about the specific citrullinated substrates that lead to autoimmunity, but there is growing evidence that PAD4 is localized to the cell surface in response to inflammation. Here, we characterize the cellular consequences of exogenous PAD4, showing that it induces morphological changes that increase cell migration, a hallmark of cancer. We then devised a robust proteomics approach to identify PAD4 substrates. We identified ∼ 3000 citrullinated peptides from 1300 proteins upon exogenous addition of PAD4 both inside and outside of cells. This extracellular set can be further augmented by targeting PAD4 to cancer cells using a HER2 binding protein conjugate. Finally, we studied how citrullinated cells can induce a humoral response in vivo to produce ACPAs. We believe these studies further our understanding of cellular consequences of extracellular PAD4 and identify new PAD4 substrates that are potential neoepitopes for ACPA generation.

Ligand dimerization represents a powerful strategy to enhance avidity, potency, and selectivity. Leveraging the natural-product molecular glue rocaglamide (RocA), we identified BisRoc, a dimeric rocaglate ligand that potently and durably suppresses translation and exhibits greater specificity across a cancer cell line panel than the monomeric RocA. CRISPRi screening revealed that BisRoc activity is influenced by cellular context, including IFITM-mediated uptake, ABC-type efflux transporters, and the translation initiation factor eIF4A2. Mechanistic studies showed that the paralogs eIF4A1 and eIF4A2 are differentially sensitive to BisRoc-induced dimerization. Owing to the presence of multiple binding sites on RNAs, BisRoc-bridged eIF4A-RNA motifs assemble into higher-order complexes that promote stress-granule formation more efficiently than monomeric RocA. Given the widespread multivalency of RNA-RBP interactions, this ligand dimerization strategy may be extended to modulate the higher-order assembly of other RNA-binding proteins.

The genus Myotis is one of the largest clades of bats, and it exhibits some of the most extreme variation in lifespans among mammals, alongside unique adaptations to viral tolerance and immune defence1-3. Here, to study the evolution of these phenotypes, we generated cell lines and near-complete genome assemblies for eight closely related Myotis species. Using genome-wide screens of positive selection, analyses of structural variation and functional experiments in primary cells, we identify patterns of adaptation contributing to longevity, cancer resistance and viral interactions. We demonstrate distinct modes of adaptation to DNA and RNA viruses compared with all other mammals, with bats exhibiting genome-wide over-representation of positive selection for DNA-virus-interacting proteins and elevated rates of copy-number variation for RNA-virus-interacting proteins. Characterization of Myotis-specific duplications of the key immune factor EIF2AK2 (also known as PKR) reveals multiple ancient segregating trans-species copy-number polymorphisms. We show that the recurrent evolution of longevity seen in Myotis is associated with positive selection in cancer pathways, and demonstrate a unique response to DNA damage in primary cells of the long-lived Myotis lucifugus. Together, our results suggest that bats' remarkable longevity and immunity are linked through pleiotropic adaptations to viruses and ageing-related disease.

Fluorescence Molecular Tomography is a promising technique for non-invasive 3D visualization of fluorescent probes, but its reconstruction remains challenging due to the inherent ill-posedness and the necessity of adopting presumed tissue optical properties, whose exact ground-truth values are arduous to acquire. Emerging deep learning approaches have shown promising reconstruction results with higher spatial resolution, but mostly rely on supervised training with a large scenario-specific paired dataset and often degrade under optical-parameter mismatch. To address these problems, we propose μNeuFMT, a self-supervised neural-field reconstruction framework that integrates a differentiable finite-element-method solver in the loss function. A core advantage of μNeuFMT lies in its dual capability: it reconstructs fluorophore spatial distributions while adaptively optimizing paired background optical parameters, thereby lowering the dependence on prior information regarding background absorption and scattering coefficients. Extensive numerical, phantom, and in vivo lymph node and tumor imaging validation studies demonstrate consistent gains in accuracy and robustness across varying acquisition settings, even with moderately biased optical parameter initializations. We envision μNeuFMT as a meaningful step toward more reliable fluorescence molecular imaging in complex biological tissues. Our code and example data are publicly available at: https://github.com/Star6azeR/NeuFMT.

Brain metastases are a major clinical challenge with poor prognosis, particularly in patients with a history of smoking. While neurons are abundant in the brain and capable of synaptic interaction with tumor cells, their role in brain metastasis remains poorly understood. Emerging evidence suggests that neuronal activity may contribute to metastatic progression, yet the mechanisms by which smoking alters the brain microenvironment to promote tumor growth are unclear. We examined the impact of nicotine exposure on microglia-neuron-tumor interactions in brain metastasis models using in vivo and ex vivo approaches. We analyzed patient-derived brain metastatic tissues, performed molecular and metabolic profiling, and utilized pharmacologic inhibition strategies to dissect the underlying mechanisms of tumor-neuron communication. Our findings reveal a significant association between smoking history and perineural invasion in brain metastases. Nicotine exposure stimulates microglia to release exosomal miR-32-3p, which activates GABAergic neurons and enhances GABA release. GABA acts as a metabolic substrate in the tumor microenvironment, fueling tumor growth through the GABA shunt pathway. Pharmacological inhibition of GABA uptake with FDA-approved GABA transporter (GAT) inhibitors Tiagabine and NO-711 significantly suppress brain metastatic progression in vivo. This study identifies a novel GABAergic neuron-microglia-tumor signaling axis by which smoking promotes brain metastasis via GABAergic metabolic reprogramming. Targeting GABA uptake on tumor cells using repurposed anticonvulsant drugs offers a promising therapeutic strategy, particularly for patients with smoking-associated brain metastases.

Cells employ lysosomal degradation to eliminate extracellular and membrane proteins, which is a process that can be exploited for therapeutic purposes. Current lysosome-targeting chimeras (LYTACs) that hijack endogenous lysosome-associated receptors (LARs) to degrade pathogenic proteins are constrained by their reliance on endogenous receptor trafficking. Here we introduce synthetic chimeric antigen lysosome-associated receptors (CALARs) that reprogram cells to effectively sort extracellular proteins to lysosomes, operating independently of native ligand competition. Encoded within circular RNA, CALARs enable sustained suppression of immune checkpoints and growth factor receptors for over 1 week. This prolonged suppression enhances immune activation and substantially reduces the tumor burden in vivo. This study introduces a modular, genetically encodable platform for modulating protein expression and advancing cell-based therapeutics.

Immune checkpoint inhibitors (ICIs) have transformed cancer therapy, yet the basis of variable patient responses remains unclear. We assemble a longitudinal single-cell RNA sequencing atlas of 441 samples from 241 patients across ten cancer entities to map treatment-associated remodeling of the tumor immune microenvironment (TIME). With a hierarchical reference-guided deep-phenotyping framework, we define 77 immune and stromal subtypes and resolve four conserved TIME subtypes. Approximately 40% of tumors shift between states during therapy, and the transition is more predictive of outcome than the baseline state. Favorable transitions toward inflamed or B cell-enriched subtype track with improved response and survival, while persistence in or shifts toward myeloid dominance indicate resistance. We derive a 59-gene signature that predicted response and survival for 1,383 baseline tumors across 19 independent cohorts. These findings establish immunotype transitions as a central determinant of ICI efficacy, offering new avenues for response prediction and rational immunotherapy design.

The abscopal effect is a systemic antitumor response to localized radiotherapy (RT) that has emerged as a central focus in the era of immunotherapy. Clinical evidence demonstrates that local RT can induce regression of distant, nonirradiated metastases, including cases of complete remission. However, the effect remains unpredictable, with marked variation in clinical outcomes ranging from complete regression to paradoxical progression. The mechanistic basis for this heterogeneity and optimal strategies to enhance abscopal effects remain incompletely understood. This review synthesizes current evidence into a mechanistic framework organized around a three-stage immune cascade: ignition, orchestration, and execution. We examine how tumor-intrinsic factors, RT parameters, and biomarkers shape this cascade. We then appraise the clinical trial landscape and discuss how these mechanistic insights may inform patient selection, treatment timing, and radioimmunotherapy combination strategies. Critically, we establish a bidirectional framework distinguishing the favorable abscopal response from the pro-tumorigenic badscopal effect, in which RT paradoxically promotes distant progression through immunosuppressive pathways. By clarifying these mechanisms and variables, this review offers a framework for improving future clinical trial design and advancing the abscopal effect from a sporadically observed phenomenon to a predictable therapeutic strategy.

Liver metastases (LM) in small cell lung cancer (SCLC) are associated with a limited response to immune checkpoint inhibitors (ICIs). In this trial, we evaluated the efficacy of first-line envafolimab plus chemotherapy in SCLC patients with LM and explored potential predictive biomarkers. Patients received envafolimab in combination with chemotherapy for four cycles, followed by envafolimab monotherapy until disease progression or unacceptable toxicity. The primary endpoint was progression-free survival (PFS). Secondary endpoints included safety, objective response, overall survival, and identification of predictive biomarkers. Whole-exome sequencing (WES), transcriptome sequencing, DNA methylation sequencing, circulating tumor DNA (ctDNA) analysis, and cell-free DNA (cfDNA) methylation sequencing were performed on tumor tissues and peripheral blood samples. The objective response rate (ORR) was 75.9%, the disease control rate (DCR) was 100%, and the median PFS was 4.3 months. Granulocyte turnover and TNFRSF9 expression emerged as key indicators across transcriptomic and epigenomic analyses, and were associated with prognosis and liver dysfunction. cfDNA-based assessment of granulocyte turnover reflected both liver damage and tumor dynamics, suggesting its potential for real-time monitoring. These findings suggest that granulocyte activity may contribute to an immunosuppressive tumor microenvironment and serve as a predictive marker for treatment stratification in SCLC with LM.

Lung cancer remains a leading cause of cancer-related mortality worldwide, and the clinical translation of chimeric antigen receptor (CAR)-based immunotherapy into solid tumors continues to be constrained by antigen heterogeneity, an immunosuppressive microenvironment, and stromal barriers that impede immune cell trafficking and curtail efficacy. Here, we present a comprehensive bibliometric analysis of CAR-based immunotherapy for lung cancer, drawing on publications indexed in the Web of Science Core Collection and Scopus between 2016 and 2025. Bibliometric and network analyses elucidate global research trajectories, collaboration patterns, and emergent therapeutic themes, while clinical trial data from ClinicalTrials.gov further appraise translational progress. This landscape has expanded rapidly, with growing emphasis on CAR engineering, microenvironment modulation, precision targeting, and combinatorial strategies. Clinical evidence, though drawn from a still-limited number of trials, indicates generally tolerable toxicity in most reported cases; nevertheless, severe and occasionally fatal adverse events have been documented, underscoring the need for continued vigilance in safety evaluation. Antitumor efficacy remains modest, reflecting persistent challenges in target selection, tumor accessibility, cellular persistence, and functional maintenance. Emerging strategies, including multi-target CAR architectures, rational drug combinations, and biomarker-guided patient stratification, may offer avenues for overcoming these limitations. This study furnishes a data-driven pharmacological atlas of CAR-based immunotherapy in lung cancer, illuminating key therapeutic targets, evolving treatment paradigms, and prospective opportunities for clinical translation.

Small-cell lung cancer (SCLC) is widely considered one of the most aggressive human malignancies, characterized by development of rapid metastases and eventual resistance to platinum chemotherapy and immunotherapy. For decades, despite rigorous scientific investigation and well-established mouse models providing a basis for mechanistic understanding, there were limited therapeutic advances for SCLC. The past decade has seen reinvigoration of drug development for SCLC as well as US Food and Drug Administration (FDA) approval of tarlatamab, a bispecific T-cell engager with unprecedented survival advantage in this recalcitrant disease. Multiple other promising clinical trials in the first and later-line settings are underway that seek to challenge standards of first-line chemoimmunotherapy, investigate combination treatments in the later line, and introduce T-cell engagers earlier in treatment. This review provides a comprehensive primer on the biology and molecular landscape of SCLC, in the context of how both shape ongoing investigational strategies. A broad overview of established multidisciplinary management of both limited-stage and extensive-stage SCLC is discussed to provide understanding of how the next decade is likely to bring significant clinical gains as emerging therapeutics seek to redefine the management of SCLC and build upon novel advances.

Inhibition of androgen receptor (AR) signaling remains the cornerstone of systemic therapy for advanced prostate cancer (PC). However, a subset of aggressive tumors either arises de novo with neuroendocrine features or emerges under treatment pressure through lineage plasticity and AR independence. These lethal states are encompassed within the spectrum of aggressive-variant prostate cancer (AVPC), an umbrella term that includes both histologically confirmed neuroendocrine prostate cancer (NEPC)-comprising de novo NEPC and treatment-emergent NEPC (t-NEPC)-and clinically or molecularly defined AVPC lacking histologic confirmation but sharing neuroendocrine-like, AR-indifferent, or small-cell features. These phenotypes are characterized by rapid progression, visceral dissemination, low or discordant prostate-specific antigen (PSA) levels relative to tumor burden, and poor prognosis. Treatment options for NEPC/AVPC remain limited and largely rely on platinum-based chemotherapy, which usually provides only modest and transient benefit. This unmet need has intensified interest in lineage-associated vulnerabilities. Delta-like ligand 3 (DLL3), an inhibitory Notch ligand with restricted expression in normal adult tissues, is aberrantly upregulated in several neuroendocrine malignancies and has emerged as a clinically actionable target. In prostate cancer, DLL3 expression is enriched in neuroendocrine tumor cells, being detected in approximately 76.6% of castration-resistant NEPC compared with only 12.5% of castration-resistant adenocarcinoma, supporting its development as both a biomarker and therapeutic vulnerability. Clinical success of DLL3-targeted therapies in small-cell lung cancer further supports evaluation of DLL3-directed strategies in NEPC and related AVPC states. This review summarizes the biological rationale, translational evidence, and emerging clinical data supporting DLL3-targeted therapies in prostate cancer. Investigational platforms include antibody-drug conjugates, bispecific and trispecific T-cell engagers, and DLL3-directed radiopharmaceuticals. Early clinical studies suggest that activity is largely confined to DLL3-expressing neuroendocrine tumors, highlighting the importance of biomarker-guided patient selection. Delta-like ligand 3-directed therapies may reshape the management of DLL3-expressing prostate cancer if ongoing efforts to refine biomarkers improve patient enrichment and optimize trial design are successfully translated into clinical practice.

The management of endometrial cancer (EC) has transitioned from a histopathological model to a framework based on molecular precision. This change is centered on the integration of immune checkpoint inhibitors, which has altered clinical standards for different genomic subsets. For patients with mismatch repair deficient (dMMR) or microsatellite instability-high (MSI-H) tumors, PD-1 and PD-L1 blockade has moved from late-line therapy to a frontline standard. Results from the RUBY and NRG-GY018 trials show that combining dostarlimab or pembrolizumab with platinum-based chemotherapy improves survival in patients with untreated advanced disease. Mismatch repair proficient (pMMR) tumors, which make up the majority of cases, remain difficult to treat due to an immune-cold microenvironment. Overcoming this resistance requires combination strategies, such as using anti-angiogenic agents like lenvatinib with immune checkpoint inhibitors, as shown in the KEYNOTE-775 trial. Data from the DUO-E study also suggest that including DNA damage response inhibitors may improve results in pMMR populations. As molecular monitoring via liquid biopsy and the use of antibody-drug conjugates (ADCs) evolve, the personalization of immunotherapy continues to progress. This review examines current clinical evidence and the mechanisms of resistance to define the future of precision medicine in endometrial cancer.

Cell-based immunotherapies have emerged as a transformative modality in modern cancer treatment, complementing conventional approaches such as surgery, chemotherapy, radiotherapy, and molecularly targeted therapies. This review provides an integrated and up-to-date synthesis of the rapidly evolving landscape of cellular immunotherapy, encompassing chimeric antigen receptor (CAR) T cells, T cell receptor (TCR)-engineered T cells, tumor-infiltrating lymphocytes (TILs), dendritic cell (DC) vaccines, natural killer (NK) cell-based therapies, and macrophage-directed strategies. We delineate the mechanistic foundations underlying each modality, summarize clinical outcomes across both hematologic malignancies and solid tumors, and critically evaluate therapeutic performance in the context of treatment-associated toxicities, resistance mechanisms, and barriers to durable response. Furthermore, we highlight emerging next-generation strategies designed to mitigate antigen escape, overcome immunosuppressive tumor microenvironments, and address challenges related to manufacturing, scalability, and accessibility. Collectively, these advances establish cell-based immunotherapies as a central component of precision oncology, with expanding potential to deliver durable and broadly accessible clinical benefit across diverse cancer types.

In the phase II trial SWOG S1616 (NCT03033576), patients with advanced melanoma with primary resistance to anti-programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 (PD-L1) therapies had improved outcomes with the combination of the anti-cytotoxic T lymphocyte-associated protein 4 (CTLA-4) antibody ipilimumab with continued anti-PD-1 therapy with nivolumab, compared with ipilimumab alone. Baseline biopsies from patients responsive to combination therapy showed increased transcriptomic expression of complement by myeloid cells, interferon pathways by endothelial cells, and oxidative phosphorylation and lipid metabolism by melanoma cells. Using spatial proteomics, some on-therapy biopsies from patients responding to combination therapy exhibited networks of activated CD8 T cells near melanoma cells, whereas others had T and myeloid cells, reflecting different time points in a dynamic antitumor response. Conversely, biopsies from patients progressing on combination immunotherapy displayed impaired T-cell infiltration adjacent to plasma cells. Our results define cellular neighborhoods and transcriptomes in melanoma biopsies when reversing resistance to anti-PD-1 with the addition of anti-CTLA-4, and plasma cell sheets in nonresponding biopsies. Patients with melanoma with primary resistance to anti-PD-1/PD-L1 therapies are treated with anti-PD-1 in combination with anti-CTLA-4. Melanoma biopsies responding to combination immunotherapy in the second line of care have distinct tumor microenvironments, with antitumor immune responses, compared with those that progress, which have impaired T-cell infiltration and plasma cell networks.

Immune checkpoint blockers (ICB) improve outcomes in metastatic melanoma (MM), but resistance limits benefit. This phase I/II (NCT02706353) study evaluated intratumoral sotigalimab (anti-CD40 agonist) with pembrolizumab in 32 patients with ICB-naïve MM. Primary endpoints were safety and objective response rate (ORR). Sotigalimab was well tolerated. At the recommended phase II dose, the ORR was 50%, and the disease control rate was 92%, with ORRs of 67% in injected and 50% in non-injected tumors. Multiomic analyses of tumor and blood showed that sotigalimab effectively engaged the CD40 pathway, boosting infiltration and activation of myeloid cells, including CD11c+DC-LAMP+ dendritic cells and macrophages. The combination therapy activated innate and adaptive immunity in injected tumors and cytotoxic responses in non-injected tumors. T-cell receptor sequencing showed increased T-cell clonality with expanded new clones shared across tumors. Clinical responses correlated with these immunologic changes but not with baseline features associated with response to anti-PD-1 monotherapy. In this study, we provide compelling data that intratumoral sotigalimab combined with pembrolizumab is safe, activates antigen-presenting cells, and elicits broad innate and adaptive immune responses in both injected and non-injected tumors, supporting further randomized phase II trials to evaluate sotigalimab's potential to enhance anti-PD-1 therapy through "in situ" immunization.

Breast cancer is the most frequently diagnosed cancer, with metastasis accounting for the majority of cancer-related deaths. The mechanisms of early-stage breast cancer metastasis to regional immune sites like lymph nodes remain elusive. Here, we performed an in-depth proteomic and phosphoproteomic analyses of a substantial series of breast cancer samples, alongside genomic and transcriptomic evaluations. This cohort encompasses 195 specimens: 65 primary breast tumors, their corresponding normal tissues, and metastatic axillary lymph nodes. We offer an overview of the molecular alterations at the transcriptomic, proteomic, and phosphoproteomic levels during lymph node metastasis. Notably, the findings indicate that regional lymph node metastasis is primarily influenced by proteomic and phosphoproteomic alterations, rather than genomic or transcriptomic changes. We found that ANGPTL4 and HMGB1 could serve as biomarkers of lymph node metastasis. Data analysis and cell experiments involving silencing of the alternative splicing (AS) factor HNRNPU demonstrated that AS plays a significant role in modulating protein expression, phosphorylation profiles, and cell proliferation. The key phosphorylation sites, including MARCKSL1-S104 and FKBP15-S320, as well as the upstream kinase PRKCB, were identified as playing crucial roles in breast cancer lymph node metastasis. Targeted intervention of the kinase PRKCB resulted in effectively suppressing the proliferation and metastasis of breast cancer tumor cells. Immune profiling analysis and experimental validation of breast cancer cell co-cultured with CD8+ T cell reveals correlations between phosphorylation of MARCKSL1-S104 and FKBP15-S320 with immune checkpoint PD-L1 expression, and their impact on tumor cell apoptosis, suggesting a potential mechanism of immune evasion in metastasis. This study systematically characterizes the molecular landscape and features of primary breast tumors and their matched metastatic lymph nodes. These insights enhance our understanding of early-stage breast cancer metastasis and may pave the way for improved diagnostic tools and targeted therapeutic strategies.

To elucidate the cellular and molecular mechanisms underlying therapeutic resistance (refractoriness) following transarterial chemoembolization (TACE) in hepatocellular carcinoma (HCC) by comprehensively characterizing the post-treatment tumor microenvironment (TME). We employed an integrative spatial multi-omics strategy, combining bulk and single-cell RNA sequencing, subcellular-resolution spatial transcriptomics, and spatial proteomics on tissues from TACE-treated and treatment-naïve HCC patients. Public datasets were used for prognostic and predictive validation, and key findings were confirmed with multiplex immunofluorescence and in vitro experiments. TACE induced a profoundly hypoxic TME, which drove the selective enrichment of a pro-fibrotic tumor-associated macrophage (TAM) population characterized by high SPP1 expression (SPP1 + TAMs). Spatial mapping demonstrated that these SPP1 + TAMs localize to hypoxic tumor cores, where they remodel the extracellular matrix by producing fibronectin (FN1), establish fibrotic niches, and restrict T-cell activation, thereby orchestrating an immune-excluded microenvironment. The abundance of this macrophage subtype was a robust predictor of TACE resistance. TACE-induced hypoxia promotes a macrophage-driven fibrotic program that is a key determinant of immune evasion and treatment failure in HCC. Targeting this SPP1 + TAM-mediated fibrotic niche presents a potential therapeutic strategy to overcome TACE resistance and improve clinical outcomes.

Targeted and biologic therapies are increasingly being used for treatment of dermatological conditions. Certain agents are associated with cardiometabolic toxicities, including major adverse cardiovascular events and altered lipid profiles, necessitating baseline and on-treatment monitoring. Close cardiometabolic monitoring is needed throughout treatment with co-management by cardiology, endocrinology, rheumatology, oncology, and primary care for symptomatic or high-risk patients. Conversely, some therapies may reduce systemic inflammation and atherogenic factors, possibly conferring reduced risk of cardiometabolic events. Randomized, controlled trials in dermatological populations are needed to corroborate these observations. In part II of this two-part continuing medical education series, we describe cardiometabolic effects of targeted and biologic therapies, including immune checkpoint inhibitors, antitumor monoclonal antibodies, intravenous immunoglobulins, Janus kinase inhibitors, tumor necrosis factor inhibitors, interleukin inhibitors, and apremilast. In addition, we outline recent advances in pathophysiology and supplement with practical recommendations for baseline assessment and monitoring in dermatological patients.

Neuroendocrine carcinomas (NECs) comprise a highly heterogeneous group of lethal malignancies arising from diverse anatomical sites. We aimed to provide a functional blueprint for bridging the translational gap between fundamental discovery and clinical application in NEC. Building on our previously established cross-tissue transcriptional framework, we here performed comprehensive proteomic and phosphoproteomic profiling of 267 NECs spanning 26 anatomical sites. This integrated analysis recapitulated the five ANHPY transcriptional subtypes driven by master transcription factors (ASCL1, NEUROD1, HNF4A, POU2F3 and YAP1). These subtypes hijacked distinct lineage developmental programmes, which converged with specific RB1 status to dictate their molecular identities. Synergistic profiling of tissue and plasma proteomes, alongside cell secretomes, nominated subtype-specific diagnostic and therapeutic biomarkers. Specifically, we uncovered NAD+ dependency as a metabolic vulnerability unique to subtype H, which is primarily distributed across gastroenteropancreatic NEC. Pharmacological inhibition of the NAD+ salvage enzyme NAMPT achieved complete tumour regression in vivo by triggering synthetic lethality in the context of an inherent deficiency in NAD+ biosynthesis. We also defined four tumour microenvironment (TME) subtypes that capture the extrinsic diversity of NEC. Notably, the 'inflamed' TME subtype was associated with superior immunotherapy responses, yielding an inflammatory signature that robustly stratified patients across diverse tissue origins. This study delineated conserved molecular identities, multidimensional biomarkers, the immune landscape and subtype H-specific therapeutic strategies, paving the way for precision medicine in NEC.

Neoadjuvant immunotherapy has emerged as a promising strategy for patients with head and neck squamous cell carcinoma. However, the systemic immune mechanisms underlying treatment response remain insufficiently defined. This study evaluated the clinical feasibility and immunological effects of a single neoadjuvant dose of a programmed death-ligand 1 (PD-L1) blocking antibody in patients with resectable head and neck squamous cell carcinoma, with a particular focus on immune cell dynamics. In this phase II, open-label, non-randomized window-of-opportunity study (PIONEER trial, NCT04939480), patients received one dose of the PD-L1-targeting antibody atezolizumab before surgery. Clinical safety, operability, and pathological response were assessed. A translational research program incorporated intensive immune monitoring based on immediate processing of freshly collected peripheral blood samples, enabling reliable quantification of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC), which are highly sensitive to freeze-thaw procedures. Additional immune profiling was performed to track treatment-induced systemic immune activation and cytokine dynamics. All patients proceeded to surgery without delay, and neoadjuvant atezolizumab was well tolerated. Pathological tumor regression exceeding 50% was observed in 7 of 20 patients (35%), including 5 patients with major pathological response. Treatment induced a transient peak in type II interferon-associated cytokines and defined signatures of cell-mediated immunity approximately 15 days after antibody administration. Patients who achieved robust pathological regression displayed higher baseline frequencies of activated natural killer T cells and exhausted T cells, with further expansion after treatment. Importantly, immediate blood processing revealed that pronounced tumor regression was consistently associated with an early and marked reduction in mature immunosuppressive PMN-MDSC-a finding that would have been obscured by conventional frozen sample workflows. A single neoadjuvant dose of atezolizumab was feasible, safe, and elicited measurable systemic immune activation in patients with resectable head and neck squamous cell carcinoma. Direct analysis of fresh blood uncovered dynamic PMN-MDSC changes as a potential novel pharmacodynamic biomarker of treatment response, emphasizing the advantages of real-time, non-frozen immune cell profiling for complete assessment of immune cell dynamics in neoadjuvant immunotherapy studies. NCT04939480.

As cancer survival rates improve, the long-term burden of treatment has become increasingly evident. Cancer survivors face a markedly higher risk of cardiovascular-related complications and premature, non-cancer-related mortality. In particular, cardiovascular disease (CVD) is disproportionately prevalent, with survivors approximately 40% more likely to develop and die from CVD compared to the general population. Although this increased morbidity reflects both acute cardiotoxic events and the later development and progression of more chronic CVDs, traditionally viewed as direct consequences of cancer therapies, the underlying mechanisms, especially those that are feasible to modify, are poorly understood. Emerging evidence positions the gut-heart axis as a central regulator of cardiovascular risk in cardio-oncology. This avenue is especially compelling to explore as the gut microbiome is well documented to be altered by cancer therapies, including chemotherapy, immune checkpoint inhibitors, targeted therapies, and radiation, with profound and persistent changes in diversity, composition and function widely reported across clinical cohorts. Comparable microbial changes have been observed in non-cancer cohorts with cardiovascular disease. For example, specific microbial metabolites have been reported to exert cardiovascular protective benefits in hypertension. Thus, there is a compelling opportunity to explore the gut microbiome to advance our understanding and ability to prevent cardiovascular disease in cancer survivors. This review synthesizes current evidence linking the gut microbiome to cancer therapy-related cardiac dysfunction (CTRCD), evaluates microbial metabolites as predictive biomarkers of cardiotoxicity, and discusses microbiome-targeted modulation as an emerging strategy for improving cardiovascular outcomes in cancer survivors.

Renal cell carcinoma (RCC) is a highly heterogeneous malignancy characterized by diverse molecular alterations and dynamic evolutionary trajectories. Advances in genomics and molecular profiling have shifted RCC management from morphology-based classification toward molecularly informed diagnosis and therapy. In this review, we integrate recent multi-omics and clinical advances to define the evolving biological and therapeutic landscape of RCC. We highlight how the 2022 WHO classification incorporates molecular features to refine pathological subtyping and summarize key driver genetic alterations underlying major RCC entities. We then discuss insights derived from multi-region sequencing studies, including TRACERx Renal, which demonstrate how intratumoral heterogeneity, branched evolution, and clonal selection shape disease progression and therapeutic resistance. We further examine subtype-specific metabolic programs, with particular emphasis on dysregulation of the AMPK-mTORC1 signaling axis and distinct metabolic dependencies across clear cell and non-clear-cell RCC. We additionally highlight cell death regulation and differentiation plasticity as integrative themes that connect molecular pathogenesis to therapeutic vulnerability, with sarcomatoid and rhabdoid dedifferentiation as a key example. Finally, we review the evolution of systemic therapies, from VEGF-targeted tyrosine kinase inhibitors to immune checkpoint-based combinations, and outline emerging strategies, including HIF-2α inhibition, PD-1/VEGF bispecific antibodies, and subtype-specific metabolic and epigenetic dependencies. Collectively, this review provides an integrated framework linking molecular evolution, metabolic reprogramming, and therapeutic development, with the goal of advancing biomarker-driven precision management in RCC.

The global prevalence of metabolic diseases, notably obesity, diabetes mellitus, and thyroid disorders, has risen dramatically in recent decades, posing a significant threat to public health. While conventional pharmacological interventions remain the cornerstone of management, they face considerable limitations, including adverse effects and the development of drug resistance, underscoring the urgent need for alternative therapeutic strategies. Intriguingly, immune checkpoint molecules, originally recognized for their transformative role in cancer immunotherapy, are now gaining attention as pivotal regulators of metabolic homeostasis. Growing research demonstrates that immune checkpoint pathways, particularly the PD-1/PD-L1 axis, play dual roles in immune regulation and metabolic modulation by orchestrating inflammatory responses and energy metabolism. For instance, in obesity, PD-1/PD-L1 signaling promotes the polarization of adipose tissue macrophages from a proinflammatory M1 phenotype toward an anti-inflammatory M2 state, thereby mitigating chronic low-grade inflammation and associated metabolic dysfunction. These findings highlight the potential of immune checkpoint modulation as a novel therapeutic strategy for metabolic disorders. However, despite these advances, the role of immune checkpoints in metabolic diseases remains underexplored, with limited comprehensive reviews on the subject. This review comprehensively elucidates the pathophysiological mechanisms and translational potential of immune checkpoint regulation in obesity, diabetes, and thyroid disease while proposing novel directions for clinical intervention.

Mouse models remain the premier preclinical model organisms for cancer immunotherapy, yet how they faithfully recapitulate the human tumor microenvironment has remained poorly defined. In a recent Nature Immunology study, Courau and colleagues addressed this gap through a systematic, cross-species immune profiling spanning 15 widely used mouse models and corresponding human cohorts. Their analysis revealed that commonly used mouse tumors only capture a portion of human disease-predominantly macrophage-rich, T cell-poor microenvironments-but largely miss immune-rich, CXCL13-organized human tumor types, which are more responsive to immune checkpoint blockade in the clinic. Beyond cellular composition, the authors also uncovered species-specific chemokine networks and cell-cell interactions that may account for the unique immune profiles in mouse and human tumors. Despite the differences, conserved transcriptional modules were detected by consensus gene expression profile analysis. Notably, a strong association between interferon-responsive myeloid cells and T-cell cytotoxicity marks a key transcriptional program in which mouse biology and human biology converge, which is able to predict patient survival. Taken together, this work serves as a cautionary guide to the limitations of mouse models; more importantly, it offers a queryable atlas for precisely aligning preclinical models with the human conditions they credibly emulate.

Chemotherapy induces cancer cell apoptosis and the release of apoptotic bodies (AB) that are poorly immunogenic or immunosuppressive, creating a major barrier to the success of coadministered or second-line immunotherapies. In this study, we found reduced circulating levels of thymosin alpha-1 (Tα-1), a key endogenous peptide hormone with immunomodulatory activity, after chemotherapy treatment in patients with multiple types of cancer and in mice bearing established tumors. Tα-1 bound to tumor ABs and interacted with AB-borne microRNAs, including miR-146a-5p, following the phagocytosis of ABs into the endolysosomal compartment of dendritic cells (DC). The interaction with Tα-1 protected miR-146a-5p from lysosomal RNase A-mediated degradation, allowing miR-146a-5p-mediated activation of Toll-like receptor 7 (TLR7) that licenses DC maturation, migration to tumor-draining lymph nodes, and presentation of tumor antigens to activate tumor-specific CD8+ T cells. Therapeutic Tα-1 supplementation produced strong synergy with chemotherapy to control established tumors in mice with high miR-146a-5p expression in a TLR7-dependent manner. These findings establish Tα-1 as a pivotal endogenous microRNA chaperone that unlocks a critical limiting step of DC licensing, empowering robust antitumor immunity after chemotherapy. Tα-1 promotes miR146a-5p-mediated TLR7 signaling in dendritic cells to enhance chemotherapy-induced immunity, offering a foundation for developing Tα-1-based approaches to elicit or amplify immune responses against diverse tumors.

Epigenetic silencing of interferon (IFN) signaling contributes to the resistance of tumors to programmed cell death protein 1 (PD-1)/programmed cell death ligand 1 (PD-L1) immune checkpoint blockade. In this study, we conducted a fluorescence-activated cell sorting (FACS)-based CRISPR-Cas9 screen to identify tumor-intrinsic regulators of PD-L1 surface expression and identified the histone-lysine methyltransferases EHMT1 and EHMT2 as key suppressors of IFN signaling. TNG917 was developed as a histone substrate-competitive dual-inhibitor of EHMT1/2 with low nanomolar potency in cells and high selectivity over other methyltransferases. In cancer cell lines, TNG917 relieved H3K9-mediated repression, restored IFN-stimulated gene expression, and triggered secretion of T-cell chemoattractant cytokines, including CXCL10. When dosed orally in both syngeneic and humanized mouse models, TNG917 monotherapy led to marked tumor growth inhibition, whereas combination with anti-PD-1 therapy produced complete, durable regressions and established protective immune memory. Early pharmacokinetic and toxicology assessments revealed favorable exposure profiles and a wide safety margin. These findings establish EHMT1/2 inhibition by TNG917 as a strategy to convert immune-cold tumors into T cell-inflamed lesions and potentiate checkpoint blockade efficacy, supporting its advancement into clinical development in combination with immunotherapy. By reversing EHMT1/2-dependent repression of interferon signaling, TNG917 enhances tumor immunogenicity and T-cell infiltration, identifying EHMT1/2 as key epigenetic drivers of immune evasion and actionable targets to restore immune checkpoint blockade responsiveness.

The clinical benefit of therapies targeting the KRAS G12C mutation is substantially limited by the development of resistance through multiple mechanisms. Therefore, to achieve lasting benefit with these therapies, effective strategies to tackle the evolution of drug resistance are required. To investigate this, we developed a preclinical model that mimics the development of resistance to KRAS G12C inhibitors (G12Ci), such as adagrasib and RMC-4998. Treatment of tumors containing a minor subpopulation of resistant cancer cells with G12Ci led to their rapid outgrowth, replacing drug-sensitive cells within a few weeks. However, combining G12Ci with therapies that enhance immune responses, such as anti-PD-1 or SHP2 inhibitors, resulted in the elimination of resistant cells and durable complete responses, even though anti-PD-1 or SHP2 inhibitor monotherapy did not induce regression of resistant tumors in the absence of drug-sensitive cells. This bystander killing of resistant cells following targeting of drug-sensitive cells was dependent on an intact adaptive immune system. Mechanistically, these combination therapies led to profound remodeling of the tumor immune microenvironment, making it less immunosuppressive, and promoted cancer cell death that primed an immune response, with an influx of cytotoxic T lymphocytes recognizing tumor-associated antigens shared between G12Ci-resistant and -sensitive cancer cells. Promotion of immune-mediated bystander elimination of drug-resistant cells may provide a paradigm for tackling the problem of drug resistance in cancer more broadly. Combination therapies that stimulate antitumor immunity promote immune-mediated bystander killing of KRAS-G12C inhibitor resistant subpopulations, providing a strategy to overcome drug resistance that might have relevance in broader therapeutic settings.

Lactylation, an emerging posttranslational modification, plays a crucial role in epigenetic regulation and tumorigenesis. Characterization of the key enzymes governing lactylation in tumors, including writers, erasers, and readers, could help identify potential therapeutic targets. In this study, we used lactylome analysis and immunoprecipitation mass spectrometry to identify a role for PARK7 as a delactylase. PARK7 catalyzed the delactylation of the RNA splicing-related protein SF3B2 at lysine 280 (K280), limiting tumor growth in an immune-dependent manner. RNA immunoprecipitation sequencing revealed that the lactylation modification of SF3B drives abnormal splicing of serpin family RNA, resulting in excessive serpin secretion into the extracellular matrix and subsequent tumor fibrosis. Clinically, elevated PARK7 expression correlated with longer survival among patients with cancer. These findings not only identify a delactylation modification enzyme, PARK7, with tumor-suppressive effects but also reveal a connection between lactylation and RNA splicing regulation in cancer biology. PARK7 functions as a delactylase that modifies SF3B2 and inhibits aberrant RNA splicing to suppress fibrosis in an immune-dependent manner, supporting the potential of harnessing this axis to reprogram the tumor microenvironment.

The transition of endothelial cells (EC) to tumor-associated ECs (TEC) is a critical process in tumor progression. The underlying mechanisms driving this angiogenic state could represent potential therapeutic targets. In this study, we revealed that a co-opted gene regulatory program from EC progenitors promotes the angiogenic phenotypes of TECs. Specifically, the TEC-specific gene regulatory program, derived from large-scale single-cell transcriptomic and epigenomic atlases across multiple solid tumors, was projected onto multiomic maps of human embryonic stem cell-derived EC differentiation. TECs reactivated a gene regulatory program governing extracellular matrix remodeling originally confined to the late EC progenitor state. Notably, integrin αv emerged as a mediator associated with endothelial plasticity in the tumor microenvironment (TME). Stimulation of primary ECs with predicted TME ligands reactivated these progenitor-associated regulatory programs, whereas inhibition of integrin αv attenuated this effect, suppressing endothelial migration, invasion, and tumor vascularization in vivo. Collectively, these findings reveal that tumor endothelial plasticity is governed by the co-option of a developmental cis-regulatory program, highlighting a regulatory mechanism underlying TEC adaptation in tumors and providing a rational therapeutic strategy to counteract angiogenic reprogramming. Tumors exploit intrinsic endothelial differentiation programs to drive vascular evolution, providing a conceptual framework for targeting endothelial plasticity as an anti-angiogenic strategy.

Despite the presence of oxygen, tumors frequently preferentially perform fermentative glycolysis, producing lactate and acidifying the tumor microenvironment (TME). Although studies have observed that high concentrations of lactate in the TME help tumors gain a proliferative advantage, a detailed understanding of the molecular mechanisms is needed to uncover strategies to overcome lactate-mediated growth. In this study, we investigated how lactate exerts progrowth effects in clear cell renal cell carcinoma (ccRCC), a highly glycolytic tumor primarily caused by alterations in the von Hippel-Lindau tumor suppressor and constitutive activation of HIF signaling. High lactate concentrations activated GPR132, a lactate sensor highly expressed by ccRCC, which conferred protumor growth signaling by elevating mitochondrial respiration through the ERK/STAT3/JAK2 pathway. Furthermore, GPR132 facilitated the uptake of lactate through elevation of HIF signaling downstream of AKT/mTOR to fuel mitochondrial respiration in a feed-forward manner. Treatment with a small-molecule GPR132 antagonist demonstrated the essentiality of GPR132 to support ccRCC growth in vivo. Together, these findings reveal that GPR132 signaling promotes ccRCC by sustaining mitochondrial integrity and elevating lactate import. The cross-talk between lactate and tumor cells is a metabolic vulnerability that can be disrupted by targeting GPR132, providing a potential treatment strategy for ccRCC. Lactate sensing through GPR132 represents a tumor dependency mechanism that reprograms metabolic signaling to support clear cell renal cell carcinoma growth, suggesting GPR132 could represent a potential target for developing cancer therapies.

Distinguishing indolent from aggressive tumors remains a key challenge in the clinical management of prostate cancer (PC), highlighting the need for better tools for accurate risk stratification. A defining feature of aggressive PC is its propensity for perineural invasion (PNI), a pathological finding that is associated with poor prognosis. Despite its clinical significance, very little is known about the spatial and molecular determinants of PNI in PC. To address this, we used high-definition spatial transcriptomics (Visium HD) to profile the PNI-associated tumor microenvironment (TME) of a representative PC patient at near single-cell resolution. Spatial mapping of the expressed genes and inferred cell types revealed transcriptionally divergent malignant cell states spatially linked to PNI within this patient. Nerve-invasive PC cells were organized within a distinct spatially localized niche that exhibited altered TME characteristics, including increased proportions of macrophages, CD4 T-cells, and endothelial cells, suggesting coordinated changes in the tumor- and immune microenvironments. The PNI-associated niche in this patient further displayed enrichment of pathways involved in immune regulation and extracellular matrix remodeling, consistent with PNI-associated niche remodeling. APP-CD74 was identified as a potential signaling axis associated with tumor-nerve, nerve-macrophage, and nerve-endothelial cell interactions, suggesting PNI in this patient may be associated with distinct microenvironmental signaling programs. We further revealed a PNI-associated PC signature that held biomarker potential at the early-localized and advanced-metastatic disease stages. Although based on a single patient, these results contribute to our understanding of the spatial and molecular features of PNI in PC and may help guide personalized treatment choices for PC patients in the future.

Recent advances in immunotherapy have significantly revolutionized cancer treatment landscape and prompted extensive research into novel vaccine-based anti-tumor therapy. As the primary defense against external infections, the mucosal immune response plays a pivotal role in maintaining normal physiological functions and preventing pathogen invasion. Mucosal tissues are among the most common sites of tumor initiation, and therefore harnessing mucosal immune to fight against malignant cells represents a feasible option for optimizing and improving current antitumor therapies. Over the past few decades, numerous efforts have been devoted to designing ideal and effective mucosal vaccine. Some clinical trials using mucosal vaccine for cancer therapy have been also launched continually. In this review, we introduce the mechanisms of mucosal immunity, with a focus on the respiratory, gastrointestinal, and urogenital tracts. We also summarize recent research and clinical trials on the application of mucosal vaccines in anti-tumor therapy, aiming to provide new perspectives and directions for future studies.

Cancer remains a major therapeutic challenge due to drug resistance and metastasis, processes driven by oxidative stress and redox imbalance. Targeting this vulnerability through ferroptosis (iron-dependent lipid peroxidation) and cuproptosis (copper-driven mitochondrial dysfunction), two ROS-mediated cell death pathways, offers a promising therapeutic strategy. However, clinical translation is hindered by incomplete understanding of their redox regulation and limited immunogenicity. A genome-wide CRISPR knockout screen was performed to identify key regulators of ferroptosis. Genetic depletion or pharmacological inhibition of candidate genes was evaluated across multiple cancer cell lines for sensitivity to ferroptosis inducer RSL3 and the cuproptosis inducer elesclomol (Es). Antitumor efficacy was assessed in xenograft, orthotopic, metastatic, and syngeneic mouse models, alone or combined with immune checkpoint inhibitors. Mechanistic studies also examined ROS production, mitochondrial stress, mitochondrial DNA release, cGAS-STING activation, and immune responses within the tumor microenvironment. Glutathione reductase (GSR), a central enzyme maintaining reduced glutathione (GSH) homeostasis, was identified as the top suppressor of ferroptosis. GSR knockout or pharmacological inhibition markedly sensitized diverse cancer cell lines to RSL3-induced ferroptosis, while GSR overexpression conferred resistance. Strikingly, GSR depletion also enhanced sensitivity to cuproptosis triggered by the copper ionophore Es. In multiple in vivo tumor models, GSR inhibition synergizes with RSL3 or Es to suppress tumor growth, inhibit lung metastasis, and prolong survival. Mechanistically, GSR deficiency amplified ROS production, induced mitochondrial stress, and triggered the cytosolic mitochondrial DNA release under ferroptotic or cuproptotic stress, activating the cGAS-STING pathway in vitro and in vivo. This increased inflammatory cytokine production, promoted immunogenic cell death, and enhanced the release of damage-associated molecular patterns (DAMPs), including HMGB1. Together, GSR inhibition combined with a ferroptosis or cuproptosis inducer transformed the tumor microenvironment into a highly immune stimulatory state, thereby enhancing the efficacy of immune checkpoint blockade through increased dendritic cell activation and T-cell infiltration and activation. GSR represents a key molecular node connecting and modulating ferroptosis and cuproptosis through redox regulation. Targeting GSR amplifies ROS-mediated immunogenic cell death, triggers cGAS-STING activation in cancer cells, and enhances the efficacy of cancer immunotherapy, providing a promising redox-based therapeutic strategy.

Prostate cancer (PCa) is the most prevalent malignancy in men, often progressing to a more refractory form castration-resistant prostate cancer (CRPC) after primary androgen deprivation therapy. Immunotherapy has brought new hope to PCa patients, but the immunosuppressive characteristics of PCa cells limit its efficacy. Cuproptosis, which can affect tumorigenesis, therapeutic resistance, and immune modulation has received significant attention recently. However, it is limited by insufficient copper ion (Cu2+) concentration, hypoxia, and overexpression of glutathione (GSH) in tumors. To overcome these limitations, a targeted nanoplatform (CuS@MYC-PEG-FA) is fabricated to achieve a synergistic cuproptosis and immunotherapy. CuS@MYC-PEG-FA can not only promote reactive oxygen species (ROS) generation and Cu2+ release, but also elicit a powerful antitumor immunity response. Meanwhile, the loaded c-MYC inhibitor (MYCMI-6) can reduce the expression of glutaminase and subsequently inhibit the generation of GSH, further sensitizing cells to cuproptosis effectively. Notably, cuproptosis combined with photothermal therapy (PTT) can turn the "cold" tumor properties of PCa into "hot" ones, leading to its sensitivity to anti-programmed death ligand 1 (anti-PD-L1) immunotherapy. The results show that the combination of CuS@MYC-PEG-FA and PD-L1 inhibitor treatment can significantly inhibit the development of PCa, providing a new strategy for the treatment of PCa.

Cell-surface sialoglycans overexpressed on cancer cells suppress activation of tumor-infiltrating immune cells (TIICs) and thus represent promising targets for cancer immunotherapy. However, the functional roles of sialylation in TIICs remain incompletely understood. Here we developed a strategy for single-cell correlative analysis of α2,3- and α2,6-linked sialoglycans and transcriptomes to determine the correlations of cell-surface sialylation with gene expression of the same cells in a linkage-specific manner by single-cell RNA sequencing. We found that α2,3- and α2,6-sialoglycans were distinctively regulated across specific TIIC subpopulations in murine tumors. Notably, downregulation of α2,6-sialoglycans was identified as a cell-surface marker for intratumoral tumor-specific antigen-reactive CD8+ T cells with high avidity. Mechanistically, high-avidity (but not low-avidity) activation of murine CD8+ T cells within the tumor suppressed β-galactoside α2,6-sialyltransferase 1 expression, which downregulates α2,6-sialylation and exposes galectin-1 ligands. Functionally, galectin-1 binding promoted T cell apoptosis and immune evasion in cancer. This work demonstrates the functional significance and therapeutic potential of specific sialoglycans on TIICs.

Antibody-drug conjugates (ADCs) and bispecific antibodies represent a rapidly advancing frontier in oncology, yet the abnormal tumor microenvironment (TME) hinders their delivery and reduces efficacy. Emerging immunomodulatory ADCs (IM-ADCs) demand mechanistic mathematical models that couple drug transport with immune dynamics. Here, we present a mechanistic framework for the delivery of HE-S2 ADC, an anti-programmed cell death ligand 1 (PD-L1) antibody bearing the bifunctional immunomodulator D18. Our model integrates cancer-immune cells interactions, TME properties, such as dysfunctional vessels, elevated interstitial fluid pressure, tissue hydraulic conductivity, and vascular permeability, spatiotemporal distributions across growing tumor and adjacent host tissue, convective-diffusive transport, ADCs binding and internalization kinetics and tumor-draining lymph node biology governing antigen presentation and the generation of effector CD8+ T cells. Parameters were calibrated simultaneously with the murine MC38 and B16 tumor growth data and effector CD8+T cell data following treatment with D18, anti-PD-L1, and ADC. Our mechanistic spatiotemporal model captures the superior antitumor efficacy of the HE-S2 ADC relative to its individual components and provides mechanistic predictions for unmeasured variables, such as spatiotemporal dynamics of drug/immune-cell distributions. It explains reduced intratumoral D18 exposure via rapid clearance, while antibody/ADC achieves higher tumor retention through leaky tumor vasculature. The model suggests a reinforcing loop in which improved ADC exposure enhances CD8+T cell infiltration, driving tumor shrinkage that lowers fluid pressure and improves drug delivery. Parametric analyses findings support TME normalization strategies that increase functional vessel density prior to ADC administration; however, such approaches should preserve sufficient vascular permeability by maintaining vessel pore radius >~40 nm, ensuring pores remain large enough for ADC extravasation and effective intratumoral delivery. The proposed mechanistic model successfully captures how TME properties regulate the delivery and efficacy of IM-ADCs while suggesting TME normalization as a potential strategy to improve treatment outcomes.

Patient-centric Biologics-Device Combination Products (BDCP) improve patient experience and compliance, while enhancing therapeutic outcomes by enabling novel routes of delivery, tackling frequent dosing and large-volume delivery requirements, and/or by simplifying administration in a healthcare setting as well as in out-patient dosing (e.g. self-administration). They also present promising solutions to overcome vaccine immunization challenges, particularly in developing nations. The BDCP opportunities, however, are often tempered by intrinsic properties of biologics (e.g. stability, viscosity etc.) and devices (e.g. design control, human factor engineering etc.) which can be further compounded by regulatory complexity across multiple jurisdictions. This comprehensive review examines the current landscape of combination product spanning across the complexities of modality (e.g. proteins, vaccines, oligonucleotide, mRNA/lipid nanoparticle (LNP) and to a limited extent cell & gene therapy) and delivery (parenteral, oral, ocular etc.) through the lens of business drivers, phase-appropriate technical development, regulatory frameworks, and patient-centric design principles. Selected case studies and commercially approved products for each modality are also presented here. Continued investments in new and improved devices such as prefilled syringes, wearable pumps, pen devices, jet devices and autoinjectors for liquid and lyophilized products highlight the recent paradigm shift for integration of delivery devices from self-administration to personalized medicine. Persistent innovation in the field, for example, has enabled device integration beyond chronic conditions such as diabetes and autoimmune diseases and prophylactic vaccination such as flu to cancer immunotherapy. Furthermore, technical, clinical and regulatory successes in achieving novel routes of delivery (e.g. oral biologics, inhaled insulin, inhaled vaccines, intradermal vaccination etc.), smart delivery systems and digital technologies have further advanced the boundaries of BDCP to not only increase market penetration but create new markets. This not only benefits patients worldwide but also paves the way for precision medicine in the future.

Biomarkers for immune checkpoint inhibitors have largely treated the tumor immune microenvironment as a fixed property. In this issue of Cancer Cell, Lin et al. assemble a longitudinal pan-cancer single-cell atlas and show that the direction of immune-state remodeling during therapy is more informative than baseline composition.

Combination approaches using systemic immunotherapy agents are now standard of care for patients with advanced-stage hepatocellular carcinoma (HCC), resulting in improved overall survival. However, even with optimal systemic regimens, fewer than 40% of cases respond to treatment, presumably due to resistance mechanisms, including antidrug antibodies and acquired resistance related to alterations in the tumor immune microenvironment (TIME). As a result, new strategies are needed to improve immunotherapeutic efficacy in this setting. Early investigations into the local and systemic effects of yttrium 90 (90Y) radioembolization using resin and glass microspheres demonstrated activation of both innate and adaptive immune systems, leading to sustained therapeutic efficacy in a subgroup of patients with HCC undergoing curative surgical resection after downstaging procedures. Preliminary prospective and retrospective studies have confirmed the safety of combining liver-directed interventions with immunotherapy. Based on these findings, clinical trials are being designed to evaluate the efficacy of different therapeutic strategies combining 90Y radioembolization and immune checkpoint inhibitor therapy. The Society of Interventional Oncology is committed to advancing research on how local-regional therapies influence the TIME and systemic inflammatory response. This review aims to inform the interventional and medical oncology community about essential considerations and strategies for implementing these combination therapies involving 90Y radioembolization and immunotherapy.

Interferon (IFN) signaling plays a pivotal role in orchestrating antitumor immunity and shaping the response to immune checkpoint blockade (ICB). Although genetic alterations that impair the IFN pathway have been reported, such events are relatively rare, suggesting a potential contribution of epigenetic dysregulation. Here, we identified a RUNX2-mediated epigenetic mechanism that disrupts the type I interferon (IFN-I) signaling pathway in osteosarcoma (OS), thereby limiting the efficacy of ICB. Development of an algorithm to assess the association of 1,425 transcription factors with IFN pathway activation in human OS tumors enabled identification of RUNX2 as a potential negative regulator of IFN signaling. RUNX2 depletion in OS cells activated the IFNB1-driven IFN-I response. Mechanistically, RUNX2 formed a transcriptional repressor complex with NCOR1 and HDAC3 that reduced H3K9 acetylation at the enhancers of key IFN-I genes, leading to their downregulation. Inhibition of the RUNX2-NCOR1-HDAC3 complex enhanced IFN-I signaling, with cGAS, STING, and IFNB1 being required for the induction of interferon-stimulated genes and tumor suppression. Paradoxically, reactivation of IFN-I signaling also upregulated immune checkpoint molecules PD-L1 and PD-L2. Combination treatment with a selective HDAC3 inhibitor and anti-PD-1 antibody led to durable tumor regression in syngeneic OS mouse models, accompanied by increased cytotoxic T cell infiltration. These findings reveal a mechanistic link between RUNX2-driven epigenetic repression and impaired antitumor immunity via the cGAS-STING-IFN-I axis and suggest a rational combinatorial strategy to overcome OS resistance to ICB.

Current advances in tumour immunotherapy remain constrained by inadequate cytotoxic T lymphocyte activation and the immunosuppressive tumour microenvironment. Innovative strategies are urgently required to enhance anti-tumour immune potency for clinical improvements in immunotherapy-resistant malignancies. Herein, we developed pyroptosis-amplified decoy nanovesicles (PADVs) to synergistically dismantle tumour resistance mechanisms and rejuvenate endogenous anti-tumour immunity. PADVs integrate checkpoint-neutralizing capability with precise regulation of the pyroptosis molecular switch in tumour cells, thereby initiating a sequential activation cascade that enables efficient presentation of tumour-specific antigens to the reshaped immune system. Systemically infused PADVs demonstrated dual-targeting priority towards lymph node and deep tumour tissues, reversing immunosuppression through competitive blockade of PD-1/PD-L1 and SIRPα/CD47 axes while providing co-stimulatory signals for immune reactivation. Furthermore, PADVs alleviate the epigenetic suppression of pyroptosis in tumours via cytosolic delivery of decitabine and LPS, enabling gasdermin D (GSDMD) upregulation and cleavage. This coordinated strategy reverses T cell exhaustion and triggers explosive antigen release via GSDMD-mediated membrane perforation, initiating a self-amplifying immune cascade involving dendritic cell activation, effector and memory T cell formation. Consequently, PADVs demonstrate excellent efficacy in suppressing tumour progression, recurrence, and metastasis in melanoma, breast cancer, and colon cancer models. This study pioneers a versatile nanoplatform that not only counteracts tumour immune evasion but also establishes durable anti-tumour immunity, offering a transformative approach to tumour immunotherapy.

Proteasome inhibitors show limited efficacy against solid tumors. We previously show the combination of Bortezomib with ammonium tetrathiomolybdate (TM) or AMD3100 could inhibit breast cancer growth rely on intact immune system. However, it remains to be explored the broad application of these drug combinations in cancer treatment. We show drug combinations inhibit the growth of multiple tumor models but fails against B16F10. We identify GSDME-mediated pyroptosis triggered by drugs as the critical switch: GSDME mediated pyroptosis drives CCL5 release, recruiting dendritic cells (DCs) and CD8⁺ T cells to initiate adaptive immunity. B16F10 cells express minimal GSDME and therefore undergo apoptosis to retain CCL5 intracellular and abrogating antitumor immunity. Overexpression of GSDME in B16F10 restores pyroptosis, CCL5 secretion, and treatment sensitivity, while GSDME loss abolishes efficacy in responsive models. Notably, pyroptosis in GSDME-proficient subsets propagates systemic immunity, controlling distant GSDME-deficient lesions. In human cancers, GSDME expression correlates with CD8⁺ T cell and DC infiltration. High GSDME predicts superior survival in BTZ-treated multiple myeloma patients. These findings establish GSDME mediated pyroptosis as the primary route for CCL5 secretion, explaining differential efficacy across tumor models and suggesting patients with high GSDME expression may benefits more from proteasome inhibitor-based therapies.

Radiotherapy elicits dual immunomodulatory effects in cancer, activating antitumor immunity while paradoxically inducing immunosuppression, which limits therapeutic efficacy. The molecular pathways mediating postradiation immune escape in hepatocellular carcinoma (HCC) remain poorly defined. Here, we elucidate a previously uncharacterized mechanism whereby radiotherapy drives the accumulation of CD200+ tumor-associated macrophages (TAMs) that suppress eosinophil-mediated antitumor immunity in patients with HCC. Through single-cell RNA sequencing of postradiotherapy HCC specimens, we demonstrated that radiation-induced DNA damage activated the cytosolic DNA-sensing STING pathway in TAMs, triggering NF-κB-dependent CD200 upregulation independent of canonical type I interferon signaling. These radiation-induced CD200+ TAMs exhibited an immunosuppressive phenotype and correlated with adverse clinical outcomes in HCC patients. Mechanistically, CD200+ TAMs established an immunosuppressive axis by recruiting CCR1+ eosinophils through CCL3-mediated chemotaxis, subsequently inhibiting their antitumor functions via CD200-CD200R engagement. This interaction comprehensively suppressed NF-κB activation in eosinophils, impaired their antigen-presenting capacity and Th2 cytokine secretion and abrogated their ability to support CD8+ T-cell-mediated cytotoxicity. Therapeutic blockade of CD200R following radiotherapy restored eosinophil effector functions, promoted central memory T-cell formation, and significantly enhanced tumor control across multiple preclinical HCC models. Remarkably, CD200R antagonism sensitized PD-1-refractory "cold" tumors to radioimmunotherapy combinations, overcoming primary resistance. Our findings establish STING-driven CD200+ TAM accumulation and subsequent eosinophil dysfunction as critical determinants of radioresistance, positioning CD200R blockade as a promising therapeutic strategy to potentiate radioimmunotherapy responses in patients with HCC. Graphical abstract of the study findings. Radiotherapy-induced STING signaling activation promotes the accumulation of CD200+ TAMs. These CD200+ TAMs facilitate the recruitment of eosinophils via the CCL3-CCR1 chemotaxis axis while simultaneously suppressing eosinophil-mediated antitumor activity through CD200-CD200R engagement (left panel). Targeted blockade of CD200R following radiotherapy unleashed the antitumor potential of eosinophils, leading to enhanced infiltration and effector function of CTLs (right panel).

Radiotherapy is frequently combined with chemotherapy, targeted therapy and immunotherapy to improve the efficacy of cancer treatment. Nevertheless, some radioresistant cancer cells seem to possess higher drug resistance, leading to failure of cancer treatment. Thus, it is crucial to explore the mechanisms of drug resistance in cancer RT. This review outlines molecular, cellular, and microenvironmental adaptations in cancer RT. These adaptations confer enhanced chemotherapy resistance in cancer cells surviving from ionizing radiation (IR). In addition, RT also activates oncogene signaling pathways, induces epigenetic remodeling and alters post-translational modifications, collectively driving resistance to targeted therapy. RT alters tumor intrinsic properties, promotes immunosuppressive effects and remodels TME, thus inducing immunotherapy resistance. Various emerging strategies including phytochemicals, small molecules, synthetic compounds, macromolecules, nanoparticles, photodynamic therapy (PDT), photothermal therapy (PTT), MicroRNA (miRNA) therapy, PROTACs, adoptive cell therapies, and engineered Salmonella, have been developed to overcome radioresistance and drug resistance in cancer RT. We highlight the importance of evaluating therapeutic effects and side effects of these emerging strategies. Through summarizing mechanisms and emerging strategies for drug resistance in cancer RT, this review aims to clarify obstacles in combined cancer treatment and guide future direction of research on cancer therapy.

Hypoxia and increased tissue stiffness are well established hallmarks of the lung tumor microenvironment and have each been independently implicated in promoting cancer progression, therapy resistance, and metastatic potential. Despite this recognition, how these biophysical and biochemical cues interact to jointly regulate lung cancer behavior remains poorly understood. This review synthesizes current literature examining the individual and combined roles of hypoxia and extracellular matrix (ECM) stiffness in shaping lung tumor progression and argues for the advancement of experimental models that can capture their dynamic crosstalk. We discuss the cellular and molecular mechanisms through which hypoxic signaling and matrix stiffening arise in lung tumors, including disrupted vascularization, rapid proliferation, aberrant ECM remodeling, and increased matrix deposition, and highlight how these processes converge to influence cancer cell signaling, invasion, and treatment response. In addition, we evaluate existing in vitro lung tumor microenvironment models used to study hypoxia and stiffness, with particular attention to their strengths, limitations, and capability to recapitulate combinatorial microenvironmental cues. Finally, we outline emerging strategies and design considerations for next generation model systems capable of integrating oxygen gradients with physiologically relevant mechanical properties. By emphasizing the need for combinatory approaches, this review aims to guide researchers toward more predictive in vitro platforms and to identify critical gaps that must be addressed to better understand lung cancer mechanobiology and inform future therapeutic strategies. STATEMENT OF SIGNIFICANCE: Tumor progression is strongly shaped by the physical and chemical properties of the tumor microenvironment, yet these factors are often oversimplified in experimental models. This review focuses on how ECM mechanics and oxygen availability regulate cell-matrix interactions and tumor mechanobiology in lung cancer. By highlighting current limitations within the field, this review aims to guide the development of more physiologically relevant platforms for studying lung cancer progression and therapeutic response.

Incomplete radiofrequency ablation (iRFA) of hepatocellular carcinoma frequently results in tumor recurrence driven by residual tumor cells and an immunosuppressive microenvironment dominated by M2 macrophages. Although RFA transiently stimulates antitumor immunity through tumor antigen release, post-ablation efferocytosis of apoptotic tumor cells reinforces immune tolerance and limits durable responses. Here, we report a multifunctional liposomal nano-vaccine (R/B@Lipo-I, composed of the MerTK inhibitor BMS777607, the TLR7/8 agonist R848, and surface-anchored interferon-γ) to overcome post-RFA immunosuppression by inhibiting efferocytosis while promoting macrophage M1 polarization. Mechanistically, BMS777607 suppressed efferocytosis, converting apoptotic tumor cells into immunogenic signals, whereas R848 and interferon-γ activated NF-κB and JAK-STAT pathways to drive pro-inflammatory macrophage reprogramming. In vitro, R/B@Lipo-I induced M1 polarization, enhanced phagocytic and antigen-presenting functions, and promoted dendritic cell maturation and CD8+ T-cell activation. In vivo, it significantly potentiated RFA efficacy, suppressed tumor growth, and remodeled the tumor immune microenvironment by increasing M1 macrophages, mature dendritic cells, and cytotoxic T cells while reducing M2 macrophages and regulatory T cells. Transcriptomic analyses further confirmed activation of interferon-γ-responsive, TNF-related, and innate immune pathways. Collectively, this work establishes macrophage reprogramming via efferocytosis inhibition as an effective strategy to overcome RFA-induced immunosuppression and provides a rational nanotherapeutic approach to reduce tumor recurrence. STATEMENT OF SIGNIFICANCE: Incomplete radiofrequency ablation leaves behind an immunosuppressive tumor niche that conventional therapies fail to overcome. We demonstrate a rationally engineered liposomal nano-vaccine that hijacks apoptotic tumor signals, rewires macrophages toward a pro-inflammatory state, and orchestrates a systemic anti-tumor immune response. This strategy transforms post-ablation immune tolerance into durable immunity, offering a mechanistically guided approach to prevent tumor recurrence and advance nanomedicine-based cancer immunotherapy.

Innovative biomaterial-based drug delivery systems have shown notable promise in the clinical cancer treatment. However, the therapeutic effect for patients remains highly variable, and new strategies urgently need to be developed. Heat shock protein 90 (HSP90) is multifunctional molecular chaperones that are associated with various human diseases, including cancer, inflammation, and protein misfolding disorders. Notably, when combined with biomaterials, they can enable targeted and regulated tumor microenvironments, making progress in cancer treatment in recent years. Herein, the design concepts, therapeutic mechanisms, and synergistic applications of biomaterials combined with HSP90 inhibitors for cancer therapy will be comprehensively introduced. Additionally, this review will focus on the foundational concepts and structural type classification of HSP90 inhibitors, and summarize the application of biomaterials as inhibitor delivery carriers. Further, we systematically analyze the progress of HSP90 nanoinhibitors combined with other therapies for cancer therapy. Finally, we reasonably discuss the challenges inherent to the combined strategy of biomaterials and HSP90 nanoinhibitors, and the development prospects of new technologies in the engineered biomaterials for HSP90, aiming to provide a new paradigm for improving cancer treatment. STATEMENT OF SIGNIFICANCE: Accumulating evidence has demonstrated that heat shock protein 90 (HSP90)-based nanomedicines are emerging as promising platforms for tumor treatment. This review first categorizes the structural types and therapeutic mechanisms of HSP90 inhibitors, then summarizes the design and preparation strategies of various biomaterials conjugated with these inhibitors. Moreover, the applications of HSP90 nanoinhibitors in both monotherapy and rational combination strategies, as well as the present challenges and future directions for biomaterial-HSP90 inhibitor systems, are systematically discussed, providing new insights for nanotechnology-enabled cancer therapy.

Intratumoral bacteria have attracted considerable attention as critical components of the tumor microenvironment (TME) that influence tumor initiation, progression, therapeutic efficacy, and immune responses. Conventional antibiotic approaches for eradicating intratumoral bacteria are limited by severe side effects, disruption of microbial homeostasis, and the emergence of multidrug resistance, while offering no direct tumoricidal effects. Here, we developed bimetallic Copper-Tin nanozymes (CuSn nanozymes) with a galvanic cell effect to overcome bacteria-associated challenges, overcome drug resistance, and enhance cancer therapy. The nanozymes can generate highly active electrons and metal ions at tumor sites without external stimulation, achieving massive reactive oxygen species (ROS) production and effective tumor eradication. The released electrons disrupt bacterial electron transport chains (ETC), thereby inhibiting energy synthesis and achieving potent antibacterial effects. The release of copper ions further enhances ROS-mediated antibacterial activity. Moreover, bacterial eradication promotes antitumor immune activation, further enhancing therapeutic outcomes. Using breast, endometrial, and liver cancer models, we demonstrated the broad applicability of this approach. This work not only offers a new paradigm for designing innovative bimetallic nanozymes with enhanced catalytic activity but also provides a potential therapeutic approach for simultaneously eliminating bacteria and tumors, overcoming chemoresistance, and offering effective treatment without reliance on conventional antibiotics or chemotherapeutics for microbiota-enriched tumors.

Circadian rhythm and oxidative stress are fundamental biological regulators whose intricate interplay plays a pivotal role in cancer progression and immunosuppression. These two factors engage in a bidirectional, self-reinforcing pathological loop that drives tumor progression and fosters an immunosuppressive tumor microenvironment. In the context of cancer, circadian rhythm disruption promotes immunosuppression by reducing cytotoxic CD8+ T cell infiltration while expanding populations of immunosuppressive cells such as regulatory T cells and myeloid-derived suppressor cells. Oxidative stress acts as a pivotal effector with dual roles, it promotes immunosuppression by polarizing macrophages toward an M2 phenotype, yet it is also strategically harnessed to disrupt immunosuppressive networks. Critically, oxidative stress serves as a central link, wherein circadian rhythm disruption, through the accumulation of ROS, drives the recruitment of immunosuppressive cells and upregulates immune checkpoints such as PD-L1, thereby suppressing anti-tumor immunity. Furthermore, emerging evidence highlights the potential of circadian rhythm as a prognostic biomarker and underscores the promise of chrono-immunotherapy, which strategically times interventions to align with the circadian rhythm to optimize anti-tumor immune function. Collectively, the circadian rhythm and redox balance represent novel therapeutic targets, with pharmacological agents that restore circadian rhythm and enable cell-selective modulation of oxidative stress. Thus, integrating circadian rhythm and oxidative stress into oncology offers a promising strategy to improve cancer prevention, prognosis, and treatment outcomes.

Recent studies increasingly highlight the crucial role of macrophages in tissue development, homeostasis and inflammation. Consequently, macrophages have become an attractive therapeutic target in the rapidly evolving field of immunotherapy, ranging from alleviating autoimmune diseases to modulating the tumor microenvironment. However, enhancing the functions of these cells typically requires ex or in vivo intracellular delivery of cell-impermeable functional molecules, such as nucleic acids. Traditionally, this is achieved using viral and non-viral carriers or methods involving membrane-disruption strategies. As a result, several techniques have been established for the intracellular delivery of functional molecules, resulting in e.g., chimeric antigen receptor-expressing macrophages. Nonetheless, it is becoming increasingly clear that the transfection technology itself perturbs cell homeostasis. Such off-target effects could significantly impact the therapeutic potential of the final cell product. In this review, we first discuss aspects of macrophage biology that are essential to understand macrophage activation and why these cells are of therapeutic interest. This is followed by a summary of the approaches that facilitate the intracellular delivery of functional payloads into macrophages and a brief discussion on how these approaches may potentially affect macrophage phenotype. The aim of this work is to allow informed selection of intracellular delivery methods towards more efficacious and safer macrophage-based therapies.

Chimeric antigen receptor T (CAR-T) cell therapy has demonstrated remarkable success in hematologic malignancies but faces persistent challenges in solid tumors and broader clinical translation, including limited target specificity, heterogeneous tumor antigen expression, unpredictable functional outcomes, and complex manufacturing processes. Recent advances in artificial intelligence (AI) are beginning to transform how these challenges are addressed by enabling data-driven design, prediction, and optimization across the entire CAR-T development pipeline. In this review, we examine how modern AI approaches such as machine learning, deep learning, and generative models are reshaping key stages of CAR-T engineering. We first discuss AI-assisted antigen discovery strategies that integrate multi-omics and clinical datasets to identify tumor-specific targets. We then examine AI-enabled engineering of antigen-recognition modules, including computational design and optimization of antibody- and TCR-derived binding domains. Next, we highlight emerging efforts to program CAR architectures and synthetic signaling circuits using AI models. We further review AI-assisted prediction of CAR-T functional performance, therapeutic efficacy, and clinical outcomes. Finally, we discuss the growing role of AI in manufacturing, quality control, and process optimization, including image-based cellular phenotyping and digital monitoring of production pipelines. Together, these advances suggest a shift from empirical CAR-T engineering toward programmable, predictive, and increasingly autonomous design frameworks that may accelerate the development of safer and more effective cellular immunotherapies.

T cell exhaustion is a state of T cell dysfunction resulting from persistent antigenic stimulation, characterized primarily by the high expression of inhibitory receptors, metabolic reprogramming, and epigenetic remodeling. T cell exhaustion is closely associated with immune responses in chronic infections, tumor escape, and organ transplantation. In transplantation immunology, T cell exhaustion plays a dual role: moderate exhaustion can promote immune tolerance and reduce graft rejection, while excessive exhaustion may weaken the defensive capabilities of the immune system, increasing the risk of infection and tumorigenesis. Therefore, effective regulation of T-cell exhaustion has become a crucial issue in the field of immunotherapy. Epigenetic or metabolic interventions may offer novel insights for achieving graft-specific tolerance. Further studies can focus on precise modulation of T-cell exhaustion through metabolic reprogramming, epigenetic regulation, and immune checkpoint inhibition, ultimately enhancing the efficacy of transplantation immunology and immunotherapy. This review focuses on the molecular phenotype, metabolic patterns, and mechanisms of epigenetic changes in exhausted T cells. It also explores the research progress of T cell exhaustion in organ transplantation. Furthermore, the review introduces strategies to induce T cell exhaustion, discussing how these strategies can effectively reduce the side effects of immunosuppressive therapy and promote graft tolerance.

Castration-resistant prostate cancer represents a critical clinical challenge due to its propensity for resistance to conventional therapies and limited treatment efficacy. Ferroptosis is an iron-dependent form of programmed cell death driven by lipid peroxidation. It holds therapeutic potential and can be induced by glutathione peroxidase 4 (GPX4) inhibition, glutathione depletion, or iron overload using compounds such as RSL3 and Erastin. These approaches show promise in overcoming drug resistance and enabling synergistic effects with anti-androgen therapy, chemotherapy, and immunotherapy. This review systematically summarizes the core regulatory networks of ferroptosis in prostate cancer (such as the PI3K-AKT-mTOR, Hippo/YAP, PGE2, and their downstream pathways), summarizes combination treatment strategies and clinical trial progress, proposes a three-pronged translational framework of "ferroptosis regulatory network-biomarkers-precision therapy", and discusses the challenges it faces in terms of drug resistance, targeting accuracy, and clinical translation. These insights aim to accelerate biomarker discovery, optimization of multimodal combination regimens, and the translation of ferroptosis from fundamental research into transformative therapeutic interventions.

Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide, largely due to aggressive tumor behavior, immune evasion, and limited responsiveness to immunotherapy. Circular RNAs (circRNAs) have emerged as important regulators of cancer progression; however, the roles of exosomal circRNAs in tumor-immune communication in GC remain poorly understood. Here, circRNA profiling of exosomes derived from GC tissues and matched adjacent normal tissues identified circFXR1 as an upregulated circRNA associated with poor survival and adverse clinicopathological features. Functional analyses using GC cell lines, patient-derived organoid, and multiple mouse models demonstrated that circFXR1 promotes tumor proliferation, migration, and growth both in vitro and in vivo. Mechanistically, circFXR1 interacts with miR-497-5p, thereby upregulating PD-L1 expression, which in turn impairs CD8+ T cell-mediated cytotoxicity. In parallel, circFXR1 activates tumor-intrinsic mTOR signaling in a miR-497-5p-dependent manner, further enhancing malignant phenotypes. Importantly, circFXR1 is enriched in tumor-derived exosomes and transferred to recipient tumor cells and CD8+ T cells, resulting in increased PD-L1 expression, impaired T cell effector function, and enhanced tumor progression. Clinically, circFXR1 expression was elevated in non-responders compared with responders to anti-PD-1 therapy. Consistently, in an NSG xenograft model with adoptive transfer of human CD8+ T cells, circFXR1 overexpression attenuated the therapeutic efficacy of anti-PD-1 treatment. Collectively, these findings identify exosomal circFXR1 as a regulator linking tumor-intrinsic signaling and CD8+ T cell-associated immune evasion in GC. CircFXR1 may be associated with anti-PD-1 treatment response in GC, although further validation in molecularly stratified, multi-center cohorts is required.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by profound tumor microenvironment (TME) heterogeneity and therapeutic resistance. However, the cellular dynamics, spatial architecture, and molecular networks driving progression from primary tumors (PTs) to hepatic metastases (HMs) remain incompletely understood. Here, we performed high-definition single-cell spatial transcriptomics using Xenium In Situ (5K) on formalin-fixed paraffin-embedded (FFPE) matched PDAC PTs and HMs, generating a single-cell spatial atlas encompassing more than 900,000 resolved cells. Integrated analysis with single-cell RNA sequencing revealed distinct cellular composition, spatial organization, and cell-cell communication patterns between PTs and HMs. Hypoxia and genomic instability were identified as core drivers of tumor cell plasticity toward a basal-like phenotype, while cancer-associated fibroblasts (CAFs) further shaped malignant cell fate. Compared with stroma-rich PTs, HMs displayed spatial immune-stromal remodeling and formed an immunogenic invasive border with actionable therapeutic vulnerabilities. We further identified functionally specialized macrophage subsets that spatially orchestrate pro-metastatic programs: TREM2+ macrophages drive tumor vascular remodeling, whereas MMP12+ macrophages promote CAF-mediated extracellular matrix (ECM) reorganization to facilitate metastasis. Moreover, CXCL9/10+ cells recruit and activate CD8+ T cells, and ectopic expression of Cxcl9/10 significantly sensitizes PDAC to immune checkpoint blockade in immunocompetent mouse models. Collectively, our study delineates the spatially coordinated tumor-stromal-immune ecosystem that sustains PDAC progression and metastasis, unveils novel immune-modulating and pro-metastatic mechanisms, and provides targetable vulnerabilities to improve immunotherapy for metastatic PDAC.

Advanced and recurrent cervical cancer (CC) remains a clinical challenge due to limited therapeutic options and a poor 5-year survival rate (<20%). While immunotherapy has reshaped the treatment landscape, primary resistance driven by immune exclusion often limits its efficacy. This study aims to identify the key molecular determinants governing T-cell infiltration and to develop a robust prognostic framework by systematically analyzing the cancer immunoediting process. We integrated transcriptomic data from TCGA and multiple GEO cohorts (n = 7) with CIC scores via the Tumor Immunotherapy Gene Expression Resource (TIP). Weighted Gene Co-expression Network Analysis (WGCNA) was utilized to identify gene modules negatively correlated with T-cell infiltration (Step 5 of CIC). A refined prognostic signature was constructed using an ensemble of machine learning algorithms, including LASSO, Random Forest, and XGBoost. The clinical relevance, immune microenvironment landscape, and single-cell expression patterns of the lead candidate, TFRC, were validated using multi-omics integration. Finally, the oncogenic role of TFRC was characterized through in vitro functional assays and an in vivo xenograft model. WGCNA identified a key module of 973 genes significantly associated with impaired T-cell recruitment. A 4-gene prognostic model (FOXRED2, RAB5IF, SHC1, TFRC) was developed, demonstrating high predictive accuracy for 1-, 3-, and 5-year overall survival (AUC up to 0.74), which was further validated in independent cohorts. Among these, TFRC emerged as a critical biomarker, showing significant overexpression in CC tissues and correlating with disease progression. Bioinformatic analysis linked high TFRC expression to a "cold" tumor microenvironment characterized by reduced CD8+ T-cell infiltration and elevated TIDE exclusion scores. Functionally, TFRC silencing significantly suppressed CC cell proliferation, migration, and invasion, and markedly attenuated tumor growth in nude mice. Our study provides a novel CIC-based stratification tool for CC patients and identifies TFRC as a pivotal regulator of immune exclusion. These findings suggest that targeting TFRC not only inhibits intrinsic tumor progression but also potentially sensitizes CC to immunotherapy by remodeling the immune microenvironment.

Prostate cancer (PCa) remains a leading cause of cancer-related mortality among men particularly in metastatic castration-resistant disease, which is refractory to immune checkpoint blockade. Prostate-specific membrane antigen (PSMA) is highly overexpressed and correlated with PCa aggressiveness and invasiveness. In this study, we evaluated the prophylactic and therapeutic potential of a next generation Nanoplasmid-based DNA vaccine encoding human PSMA (N-hPSMA). We demonstrated that Nanoplasmid resulted in faster, higher, and more sustained transgene expression compared with a conventional plasmid vector, which led to a significant increase in hPSMA-specific cellular and humoral immune response. Prophylactic vaccination conferred tumor eradication and prevented lung metastasis. N-hPSMA vaccination increased systemic immune activation with abated regulatory T cell population and augmentation of effector-memory CD8+ T cells. In the therapeutic setting, N-hPSMA slowed tumor growth and potentiated PD-1 blockade, resulting in enhanced tumor regression and survival, while increasing activation of myeloid antigen presenting cells. In a humanized orthotopic PCa model, N-hPSMA vaccination plus PD-1 blockade significantly increased functional human CD8+ T cells and suppressed the growth of human PCa LNCaP tumors. These findings established N-hPSMA as a preclinical proof-of-concept DNA vaccine that enhances antitumor immunity and sensitizes PCa to immune checkpoint blockade.

The highly immunosuppressive tumor microenvironment in pancreatic ductal adenocarcinoma (PDAC) severely limits the efficacy of current immunotherapies. Identifying key molecular drivers of this immunosuppressive niche is therefore essential for developing new treatment strategies. In this study, through comprehensive bioinformatic analysis of PDAC clinical datasets and systematic experimental validation, we demonstrate that PGK1 is upregulated in PDAC and is associated with poor immune infiltration. Genetic knockdown of PGK1 upregulated STING expression, promoted the recruitment of antitumor immune cells into the tumor microenvironment, and significantly enhanced the in vivo efficacy of STING agonist treatment. Mechanistically, PGK1 stabilizes DNMT1 by blocking its ubiquitination-mediated degradation, which in turn promotes methylation of the STING promoter and suppresses its transcription. Our findings reveal a non-metabolic role of PGK1 in promoting an immunosuppressive microenvironment and propose a promising combination strategy targeting the PGK1-STING axis for the treatment of PDAC.

Dysregulated immune activity is a cornerstone of numerous diseases, where excessive suppression enables cancer progression and uncontrolled activation drives autoimmunity. Immunotherapy aims to restore this balance, however, conventional approaches are often limited by suboptimal precision, efficacy, or long-term safety. The emergence of the CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9) genome-editing system has opened transformative avenues for overcoming these hurdles. This review traces the development of the CRISPR-Cas9 toolkit, encompassing gene knock out (CRISPR-KO), knock in (CRISPR-KI), transcriptional activation (CRISPRa), interference (CRISPRi), base editing and prime editing-and examines their application in reprogramming immune responses. We focus on how these six strategies enhance anti-tumor immunity by engineering immune cells to bypass inhibitory checkpoints and the tumor microenvironment, and suppress autoimmunity by restoring immune tolerance through precise genomic and transcriptional interventions. Finally, we discuss translational challenges and future directions, including the adoption of next-generation editors and smart delivery systems, which are poised to maximize the therapeutic potential of CRISPR-based cellular interventions.

Targeting the transforming growth factor-β (TGF-β) pathway to reverse the immunologically "cold" tumor microenvironment of pancreatic ductal adenocarcinoma (PDAC) remains clinically unsuccessful, warranting novel therapeutic strategies. We performed multiplex immunohistochemistry on human PDAC samples to correlate cell-type-specific TGF-β pathway activation and CD8+ T-cell abundance and developed a tumor and T-cell coculture to interrogate the TGF-β pathways that promote T-cell-mediated cytotoxicity. We employed newly generated genetically engineered mouse models and a specific pathway inhibitor and confirmed our findings using single-cell RNA sequencing, flow cytometry, and multiplex immunohistochemistry. We performed proteomics and various in vitro and in vivo assays to establish the mechanisms. We found TGF-β-activated kinase 1 (TAK1, Map3k7) to be aberrantly activated in PDAC cells and correlates with T-cell dysfunction. Pharmacological inhibition with Takinib, or genetic deletion of tumor Map3k7 in an autochthonous p48-Cre/Trp53f/f/LSL-KrasG12D genetically engineered mouse model, enhances CD4+ and CD8+ effector T-cell infiltration and renders immune checkpoint blockade effective. Mechanistically, TAK1 inhibition induces DNA damage and cytoplasmic DNA leakage, which activates the cyclic GMP-AMP synthase-Stimulator of Interferon Genes DNA sensing pathway, triggering inflammatory responses that promote adaptive immune cell infiltration. At the molecular level, TAK1 phosphorylates Ephrin Receptor A2 at Serine 897, which in turn phosphorylates RAD51 at Tyrosine 315, a key DNA repair protein involved in homologous recombination. We uncover TAK1 as a critical mediator in maintaining genomic integrity and highlight its potential as a therapeutic target to induce an inflamed tumor microenvironment that sensitizes PDAC to immune checkpoint blockade.

The therapeutic efficacy of systemic treatments in cancer therapy is invariably limited by the biophysical barriers, including vascular endothelial barrier, extracellular matrix, and elevated interstitial fluid pressure. Ultrasonic cavitation, as a non-invasive modality, can induce a series of biological effects that leverage mechanical forces to breach these biophysical barriers and remodel the tumor microenvironment. This review traces the paradigm shift from thermal coagulation to mechanochemical modulation, where acoustic forces are transduced into profound biological responses via mechanosensitive ion channels and immunogenic signaling pathways. We summarize recent advances in the intelligent engineering of ultrasound-active materials, from vascular-targeted microbubbles and phase-change nanodroplets to oxygen-independent piezocatalysts. Meanwhile, we clinically evaluate the utility of cavitation in enhancing drug delivery and remodeling immune environment, and highlight the milestone approval of histotripsy for non-thermal ablation. Finally, we discuss critical challenges regarding stochasticity and biosafety, proposing a roadmap toward artificial intelligence-guided, closed-loop dosimetry. We predict that by integrating physical mechanics with biological engineering, ultrasonic cavitation may alleviate multidrug resistance and immunosuppression in cancer therapy.

L-arginine (L-Arg) deprivation in the tumor microenvironment (TME) drives effector T cell dysfunction and immunotherapy resistance. However, simply supplementing L-Arg can be counterproductive, as tumor cells and immunosuppressive myeloid cells act as dominant consumers, co-opting the nutrient to promote tumor progression. To break this detrimental cycle without fueling protumoral networks, we develop a near-infrared (NIR)-triggered nanoregulator (hPFL@Lipo) to simultaneously alleviate intratumoral L-Arg deficiency and redirect its metabolism to support antitumor immunity. This nanoregulator was constructed through coordination-driven self-assembly to co-load and stabilize L-Arg and Fe3+ within hollow Prussian blue (hPB) nanoparticles, followed by lipid membrane encapsulation for enhanced systemic stability. Under NIR irradiation, hPFL@Lipo releases Fe3+ and L-Arg while generating localized hyperthermia. Fe3+ repolarizes M2-like macrophages toward an M1 phenotype, thereby increasing the intratumoral M1-to-M2 ratio. The photothermal effect induces immunogenic tumor cell death, which promotes the infiltration of cytotoxic CD8+ T cells. Concurrently, the released L-Arg supplements the local pool, while thermal ablation reduces the overall cellular burden within the tumor, thereby alleviating arginine local depletion. Together, this strategy resolves the tumor-immune conflict over L-Arg by remodeling the intratumoral landscape of L-Arg consumers in favor of antitumor effector cells, thereby reprogramming the net metabolism of the tumor from a tumor-promoting to a tumor-suppressing state and achieving potent synergy with αPD-1 therapy.

Osteosarcoma (OS), the most common malignant bone tumor, remains challenging to treat because of poor drug delivery to bone tissue and limited response to immunotherapy. To overcome these hurdles, we developed a bone-targeted, glutathione (GSH)-responsive polymeric nanoparticle (NPALN/Mn-AP) that chelates manganese (Mn) and delivers an ATM inhibitor (AZD0156) and a PRMT5 inhibitor (GSK3326595). By functionalizing this nanoplatform with alendronate (ALN) into NPALN/Mn-AP, we achieve preferential accumulation in bone tumors. Upon cellular uptake, elevated intracellular GSH levels in OS cells trigger the controlled release of both inhibitors. Inhibiting ATM and PRMT5 amplifies DNA damage and activates the cGAS-STING pathway, while Mn ions further enhance this innate immune signaling by promoting cytosolic DNA sensing. Together, these effects reshape the tumor microenvironment toward a more immune-responsive state and promote antitumor immunity in osteosarcoma. In vivo studies demonstrate that NPALN/Mn-AP significantly inhibits OS progression and boosts systemic immune responses. This dual-action, bone-specific nanotherapeutic platform synchronized DNA-repair inhibition and Mn-enhanced immune-stimulation, offering a promising new approach for effective osteosarcoma treatment.

When tumor suppressor genes are lost through chromosomal deletion, the deletion of adjacent genes can generate therapeutic vulnerabilities. MTAP is frequently co-deleted with the chr9p21 tumor suppressor gene CDKN2A, creating a synthetic lethal dependency on protein arginine methyltransferase 5 (PRMT5). Telomeric to MTAP lies focadhesin (FOCAD), whose loss induces dependency on the HBS1-like translational GTPase (HBS1L)-protein pelota homolog (PELO) ribosome rescue complex for translational maintenance. FOCAD is deleted in ∼1 out of 3 MTAP-deleted cancers. We screened an immunomodulatory imide drug (IMiD)-focused diversity library and identified a weak hit that bound cereblon (CRBN), promoted HBS1L-CRBN-compound complex formation, and induced E3-ligase-dependent HBS1L ubiquitination and degradation. Guided by cryo-EM structures and proteome selectivity, we developed TNG961, a potent, selective HBS1L degrader that disrupts the HBS1L-PELO complex, inducing translational arrest, unfolded protein response activation, and growth inhibition in FOCAD-negative models. Oral administration of TNG961 regresses FOCAD-negative xenografts, including PRMT5 inhibitor-refractory models, establishing HBS1L degradation as a strategy to exploit FOCAD loss and supporting the clinical evaluation of TNG961 as a first-in-class precision oncology therapeutic. FOCAD deletion, frequently co-occurring with MTAP/CDKN2A loss, creates a synthetic lethal dependency on the HBS1L-PELO ribosome rescue complex. TNG961, a first-in-class molecular glue degrader of HBS1L, enforces translational arrest and drives tumor regressions in FOCAD-negative models, including PRMT5 inhibitor-refractory tumors, establishing a novel precision oncology strategy for chromosome 9p21 co-deletion contexts.

Gene fusions generated by chromosomal rearrangements function as oncogenic drivers in human cancers. We previously showed that EWSR1-ETS oncofusions of Ewing sarcoma directly induce surface expression of IL1 receptor accessory protein (IL1RAP), which along with limited expression in healthy tissues except in the placenta nominate IL1RAP as a promising Ewing sarcoma immunotherapy target. We therefore engineered antibody-drug conjugates (ADC) with different cytotoxic payloads to target IL1RAP. ADCs potently blocked tumor growth and induced durable regression of Ewing sarcoma xenografts in mice and diminished metastatic dissemination in vivo. Moreover, we show that other oncofusions also induce IL1RAP expression in diverse cancers, including NPM-ALK in anaplastic large cell lymphoma and ETV6-NTRK3 in multiple tumor types. IL1RAP expression rendered these malignancies similarly vulnerable to IL1RAP-targeting ADCs, which effectively blocked the growth of ALCL xenografts and syngeneic ETV6-NTRK3+ sarcomas. Lack of detectable normal tissue toxicity, including in nonhuman primates, supports the further clinical translation of IL1RAP-targeting ADCs. The IL1RAP surface protein is induced by multiple oncogenic fusions, including EWSR1-ETS, NPM-ALK, and EN in distinct malignancies, rendering these cancers vulnerable to anti-IL1RAP ADCs. We demonstrate that IL1RAP-targeting ADCs have potent efficacy in both primary and metastatic diseases and are well tolerated in nonhuman primates, warranting their further clinical translation.

Metabolic reprogramming within the tumor microenvironment (TME) impairs antitumor immunity and compromises the efficacy of immunotherapy. Through multiomics-based metabolic subtyping in intrahepatic cholangiocarcinoma, we identified a subgroup with the worst prognosis that demonstrates significant enrichment in both cyclooxygenase/arachidonic acid (COX/AA) metabolism and KRAS mutations. Mechanistically, KRAS mutation-mediated NF-κB pathway activation upregulates CXCL5 expression, thereby recruiting CXCR2+ polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) into the TME. Concurrently, KRAS mutation drives prostaglandin E2 (PGE2) production in tumor cells, and PGE2, in turn, enhances AA uptake and COX-2 expression in PMN-MDSCs, establishing an amplifying loop between tumor cells and PMN-MDSCs that exacerbates PGE2 production. PGE2 accumulation potently suppresses the antitumor activity of CD8+ T cells via the prostaglandin E receptor 4 (EP4). Therapeutic targeting of the COX-2-PGE2-EP4 axis, combined with anti-PD-1 immunotherapy, demonstrates profound synergistic efficacy in both KRAS-mutant murine models and patient-derived tumor fragments harboring KRAS mutations. This work identifies a PGE2-COX-2 positive feedback loop between tumor cells and PMN-MDSCs that may serve as a therapeutic vulnerability for KRAS-mutant cholangiocarcinoma.

Bacterial therapy represents a promising strategy for reshaping the immunosuppressive microenvironment of "cold" tumors, owing to the inherent tumor-homing and immunomodulatory capabilities of bacteria. Bacteria can be genetically modified to precisely and continuously release immune activators at tumor sites via synchronous lysis circuits or external physical triggers (e.g., photothermal and magnetothermal methods). However, tumor microenvironment (hypoxia and acidity) interferes with gene expression efficiency, and bacterial metabolites (butyric acid and lactic acid) have potential tumor-promoting risks. In this study, we developed bacteria-nanomaterial hybrid systems (IE-PPCs) to achieve stable and controllable delivery of immune factors while enhancing treatment safety. E. coli Nissle 1917 (EcN) was engineered to express IFN-γ and anchored onto PPCs composed of antimicrobial peptides, MMP-cleavable peptides and hydrophilic PEG. Prior to administration, the expression of IFN-γ in IE-PPCs was precisely regulated by IPTG induction. Upon intravenous injection, IE-PPCs selectively colonized the tumor microenvironment, then MMP-responsive PPCs spontaneously transformed into fibrous nanostructures, exposing antimicrobial peptide residues that lyse the bacteria, thereby releasing IFN-γ and bacterial lysates. IFN-γ directly inhibited tumor proliferation, while combined with bacterial lysates induced dendritic cell maturation and promoted M1 macrophage polarization. In 4T1 mouse breast cancer models, IE-PPCs combined with anti-PD-L1 therapy achieved 89.7% anti-tumor rate, and bacterial loads within tumors decreased by 98.9%. The IE-PPCs system provides a breakthrough strategy for reshaping the immunosuppressive microenvironment and offers a reliable and attractive therapeutic option for cancer patients.

Zinc ions (Zn2+) overload is considered an exceptionally safe and ideal approach for pyroptosis-mediated tumor treatment. However, achieving efficient and safe induction of tumor-specific Zn2+ overload remains a significant challenge. This study pioneers the concept of "sono-activated Zn2+ storm", developing a sono-activated and tumor microenvironment (TME)-responsive nanoplatform, Zn-TCPP@CaCO3, which enables a precise and spatiotemporally controllable induction of Zn2+ overload at the tumor site. Specifically, Zn-TCPP@CaCO3 selectively releases meso-tetra-(4-carboxyphenyl) porphine (TCPP) and delivers the "exogenous Zn2+" within the acidic TME. Upon tumor-site localized ultrasound irradiation, the sonosensitizer TCPP generates abundant reactive oxygen species (ROS) in situ, which oxidatively attack Zn-metallothionein complexes and effectively unlock the "endogenous Zn2+" reservoir. The synergistic influx of exogenous and endogenous Zn2+ drastically disrupts intracellular zinc homeostasis, further exacerbating oxidative stress by impairing mitochondrial function. This disruption initiates a self-amplifying positive feedback loop, ultimately inducing a potent "Zn2+ storm" that finally triggers tumor cell pyroptosis. The results demonstrate that this strategy not only effectively induces powerful antitumor effects, but also elicits a strong immunogenic response and significantly enhances the therapeutic efficacy of immune checkpoint inhibitors. Overall, this work provides valuable insights into the development of novel tumor specific ion interference therapies.

Tryptophan (Trp) catabolism has been recognized as a key immunosuppressive axis in cancer. However, this largely centered on indoleamine-2,3-dioxygenase 1 (IDO1). The clinical failure of IDO1 inhibitors has exposed the limitations of this reductionist view. To re-synthesize current knowledge into a further understanding of Trp metabolism, and propose biomarker-guided, multi-node intervention strategies that can resurrect Trp metabolism as a precision immuno-oncology target. Key Scientific Concepts of Review: This review comprehensively describes the pathways of Trp metabolism in the human body and the key enzymes that can serve as therapeutic targets, thus proposing the possibility of multi enzyme combined inhibition. Second, we synthesize how Trp metabolites can modulate the functionality of immune cells, mainly T cells, within the tumor microenvironment, thereby affecting tumor immune surveillance and the efficacy of immunotherapy. Then we discuss how tumor cells manipulate Trp metabolic pathways to enhance their survival and metastasis. We also propose a new framework for targeting Trp metabolism, such as combining enzymes inhibitors or Aryl hydrocarbon receptor (AhR) antagonists with immune checkpoint blockade. By shifting from "IDO1-focus" paradigms to comprehensive metabolic interventions, we may release more potential of Trp modulation in cancer immunotherapy.

Colorectal cancer (CRC) is a multifactorial disease characterized by disruptions in cellular and immune functions, influenced by genetic mutations, environmental factors, and infections. Recent studies have highlighted the intratumoral microbiota as a critical component of the tumor microenvironment (TME), with a significant role in CRC initiation, progression, and therapeutic response. While the gut microbiota's influence on CRC is well-established, the specific contribution of intratumoral microbiota remains inadequately explored. Emerging evidence suggests that intratumoral microbiota may promote cancer progression through inflammatory pathways, metabolic alterations, and resistance to chemotherapy. Conversely, certain microbial communities exhibit tumor-suppressive properties by modulating immune responses and inducing apoptosis in tumor cells. This review aims to highlight the dual role of the intratumoral microbiota in CRC and explore the potential of microbial interventions such as probiotics, phage therapy, and fecal microbiota transplantation (FMT) in enhancing therapeutic outcomes. Furthermore, the review examines the potential of microbiota-targeted therapies to optimize cancer treatment strategies and stresses the need for personalized approaches based on microbial biomarkers. Intratumoral microbiota, as emerging tumor components, has been identified in various solid tumors. The review emphasizes the mechanisms by which intratumoral microbiota mediate inflammation, metabolic alterations, and immune modulation in CRC. It highlights how certain intratumoral microbiota are associated with resistance or sensitivity to treatments, and how manipulating the microbiota could enhance immunotherapy efficacy. By integrating advancements in multi-omics and clinical research, targeting the intratumoral microbiota represents a promising avenue for improving CRC therapies and overcoming treatment resistance. The clinical application of the intratumoral microbiota has the potential to revolutionize the treatment of CRC, paving the way for novel therapeutic strategies in oncology.

Cancer immunotherapy exhibits limited efficacy in immunologically "cold" tumors due to insufficient T-cell infiltration and immunosuppressive tumor microenvironment. The efficacy of photodynamic therapy (PDT) is restricted by inadequate tumor-specific aggregation of photosensitizers and therapeutic resistance regulated by epigenetics. Herein, we engineered a light-driven, ROS-responsive hydrogel (PACL@Gel) for loading peptide-lipid scaffold nanoparticles encapsulating proteolysis targeting chimera (PROTAC) molecule and photosensitizer to enhance cancer immunotherapy. The system utilizes both endogenous and photodynamically amplified ROS to drive controlled hydrogel degradation and nanoparticle release, establishing a self-sustaining ROS feedback loop to overcome the limitations of insufficient endogenous ROS. The released PACL nanoparticles synergistically combine PDT with epigenetic BRD4 protein degradation to robustly amplify Caspase-3/GSDME dependent pyroptosis. Pyroptotic tumor cells would trigger immunogenic cell death, releasing inflammatory mediators and damage-associated molecular patterns that promote dendritic cells maturation and T-cell activation and infiltration to effectively remodel the immunosuppressive tumor niche and promote antitumor immunity. Experimental results validated that under laser irradiation, PACL@Gel demonstrated potent local tumor suppression and distant abscopal effect, prevented postoperative recurrence and established long-term immune memory. Collectively, this integrated PROTAC-PDT hydrogel platform effectively reprograms the immunosuppressive tumor microenvironment, providing a robust and actionable approach to enhance PDT therapy in immunologically cold tumors combined with epigenetic strategies.

Photothermal therapy (PTT) as a localized cancer treatment modality has gained considerable momentum recently. While conventional PTT relies on photothermal agents (PTAs) to produce abundant heat (above 50°C) via near-infrared (NIR) laser irradiation for tumor ablation, this high-temperature approach often inflicts collateral thermal injury on surrounding healthy tissues, which severely constrains the therapeutic feasibility of PTT near vital organs. Therefore, mild-temperature PTT (below 45°C) is of clinical interest. Herein, we construct an injectable hydrogel (IVH@gel) via the coordination between calcium ions (Ca2 +) and sodium alginate to encapsulate the PTA Hu Kaiwen Ink, the free radical initiator 2,2'-azobis[2-(2-imidazolin-2-yl)propane]dihydrochloride (VA-044), and the autophagy inhibitor hydroxychloroquine (HCQ) for realizing synergistic PTT and immunotherapy. Specifically, when irradiated under an NIR laser, the ink in the hydrogel generates heat, which leads to the degradation of VA-044 to produce free radicals for killing cancer cells; meanwhile, HCQ is released for realizing autophagy-inhibited mild-temperature PTT of tumor. More importantly, the IVH@gel-mediated PTT elicits strong immunogenic cell death (ICD), which activates a potent antitumor immune response, leading to significant tumor inhibition. This work provides a promising method for the development of multifunctional mild-temperature PTT drugs.

Combining efficient photocatalytic activity with precise targeting is crucial for covalent organic framework (COF)-based nanomedicines to remodel the immunosuppressive microenvironment and sensitize "cold" tumors to immunotherapy. Herein, we constructed two isomorphic frameworks featuring distinct electronic topologies: a zwitterionic squaric acid (SA) COF with inherent electrostatic asymmetry, and a charge-neutral terephthaldehyde (TA) COF. The spatial divergence of ion centers in SA COF triggers intense molecular-level ionic displacement polarization, creating a robust internal electric field. Such unique field-effect regulation accelerates photogenerated charge separation for efficient superoxide anion production, enhances intersystem crossing for singlet oxygen generation, and promotes nonradiative energy relaxation for high-performance photothermal conversion under dual-wavelength irradiation. After PEGylation and iRGD modification, the formulated SA@PEG-iRGD exhibits excellent physiological stability and active deep tumor penetration. With near-infrared absorption, it enables high-contrast photoacoustic imaging for guided dual-wavelength (660/808 nm) phototherapy. This spatiotemporally controlled therapeutic cascade triggers lethal ferroptosis and precise photothermal ablation, potently activating immunogenic cell death. Combined with aPD-L1 therapy, it eliminates primary tumors, produces abscopal effects, and establishes immune memory against recurrence and metastasis. This study highlights the value of electronic structure engineering in COF-based precision nanomedicine.

Personalized neoantigen cancer vaccine is a promising strategy for precision immunotherapy by targeting patient-specific and mutation-derived tumor antigens. Early clinical studies have demonstrated the feasibility, safety, and immunogenicity of these vaccines across multiple solid tumors, with encouraging outcomes particularly when combined with immune checkpoint blockade. However, broader clinical translation remains limited by sequential bottlenecks across the vaccine development pipeline, including false-positive neoantigen selection,  imperfect modeling of antigen processing and HLA presentation, limited prediction of T-cell receptor recognition, and challenges in formulation, delivery, and manufacturing. Artificial intelligence and advanced computational workflows are increasingly integrated into this pipeline to improve candidate prioritization and support more reproducible decision-making. In this review, we summarize clinical progress and key translational barriers in personalized neoantigen vaccination, and discuss how AI-enabled approaches may contribute across four major stages: multi-omics integration for neoantigen discovery, processing-aware HLA presentation prediction, structure-aware and TCR-informed immunogenicity modeling, and data-driven formulation optimization, particularly for lipid nanoparticle-based delivery systems. These approaches are able to help narrow biological and chemical search spaces, improve prioritization, and provide mechanistic insights into antigen presentation and immune recognition rather than replacing experimental validation. This articlefurther addresses future implementation challenges, including dataset diversity, model interpretability, prospective benchmarking, manufacturing traceability, and evolving regulatory frameworks for individualized mRNA cancer immunotherapies. Integrating computational innovation with rigorous immunological validation, scalable manufacturing, and regulatory oversight will be essential for advancing personalized neoantigen vaccines toward broader clinical implementation.

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive solid tumor, closely associated with its unique tumor microenvironment (TME), which is characterized by a dense desmoplastic stroma. Abundant stromal cells, primarily fibroblasts, constitute the majority of cells in the tumor mass and exhibit pronounced spatial heterogeneity. Importantly, the spatial distribution of tumors and fibroblasts is vital for shaping the TME and critically influencing therapeutic responses. Here, we present a facile microarray chip for generating architecturally defined 3D PDAC heterospheroids. This platform enables us to mimic the dynamic interactions between tumor and stromal cells and to investigate how spatial organization influences stroma heterogeneity, tumor invasion and chemoresistance. The chip incorporates square concave microstructure array allowing controllable and reproducible production of uniform-sized spheroids. By simply altering the cell seeding sequence, we successfully constructed heterospheroids with distinct spatial distributions of cancer cells and fibroblasts. We further demonstrated that these organizational patterns modulate tumor-stroma crosstalk and ultimately regulate tumor invasive behavior. Furthermore, the heterospheroids with defined patterns exhibited distinct drug responses, and the potential for combination therapy evaluation was also verified. Beyond providing a robust platform for engineering heterospheroids with controllable tumor-stroma architectures, this system offers a robust 3D co-cultured model for advancing cancer research and drug screening.

Peptide-drug conjugates (PDCs) are emerging as a next-generation class of targeted therapeutics designed to overcome key limitations associated with conventional chemotherapy and antibody-drug conjugates (ADCs). By integrating a tumor-homing peptide, a cleavable or stimuli-responsive linker, and a potent cytotoxic payload, PDCs offer enhanced tumor selectivity while maintaining structural simplicity and synthetic flexibility. Compared to bulky monoclonal antibody-based systems, PDCs possess significantly smaller molecular size, enabling improved tumor penetration, rapid tissue diffusion, and reduced immunogenicity. Recent advances in peptide engineering have facilitated the development of ligands targeting integrins, G protein-coupled receptors, and other tumor-overexpressed biomarkers, promoting receptor-mediated internalization and intracellular drug release. Linker chemistry plays a pivotal role in therapeutic performance, with enzyme-sensitive, redox-responsive, and pH-cleavable linkers enabling site-specific drug activation within the tumor microenvironment. Despite their promise, PDCs face challenges including rapid renal clearance, proteolytic degradation, and limited circulation half-life. Strategies such as cyclization, PEGylation, and albumin-binding modification have been explored to enhance stability and pharmacokinetics. Furthermore, emerging theranostic PDC platforms incorporate imaging moieties or radiolabels, enabling real-time visualization of tumor targeting, biodistribution, and treatment response. Such dual-functional systems facilitate biomarker-guided patient stratification and image-guided precision therapy. This review comprehensively discusses the structural design principles, delivery barriers, pharmacokinetic considerations, applications, imaging advancements, and current clinical landscape of PDCs, highlighting their advantages over ADCs and outlining future directions for precision oncology. Collectively, PDCs represent a promising and versatile platform poised to redefine targeted cytotoxic delivery in cancer therapy.

Selecting effective chemotherapy after immunotherapy remains a major clinical challenge in advanced gastric cancer because patient responses are highly heterogeneous. A major obstacle is the lack of personalized preclinical models that faithfully recapitulate the immune-activated tumor microenvironment (TME) following immunotherapy and enable evaluation of subsequent chemotherapy responses while preserving immune-tumor interactions. Here, we developed an immune-activated co-culture model comprising patient-derived gastric cancer organoids and autologous peripheral blood mononuclear cells (PBMCs). Immune activation was achieved using anti-PD-1-loaded gold nanocages (aPD-1@Au NCs), which integrate near-infrared-triggered photothermal tumor ablation with sustained anti-PD-1-mediated immune modulation. The resulting model recapitulates key features of the post-immunotherapy TME, including enhanced immune-cell infiltration, increased tumor-cell apoptosis, and CD8+ T-cell activation. We then applied this model to evaluate five clinically relevant chemotherapeutic agents, including oxaliplatin, irinotecan, fluorouracil, doxorubicin, and docetaxel, for personalized chemotherapy screening after immunotherapy. Compared with monoculture organoids or non-activated co-cultures, the immune-activated model revealed drug-specific differences in tumor-killing efficacy, immune-mediated chemosensitization, and PBMC toxicity. Among the tested agents, oxaliplatin showed the strongest immune-mediated chemosensitization, whereas docetaxel achieved favorable tumor cell killing with relatively low immunotoxicity. These findings demonstrate that chemotherapy responses after immune activation are highly drug-specific, underscoring the importance of personalized chemotherapy selection following immunotherapy. As a proof-of-concept study, this work establishes an immune-activated patient-derived gastric cancer organoid-PBMC co-culture model that partially recapitulates the post-immunotherapy TME. This model provides a promising framework for developing personalized chemotherapy screening strategies after immunotherapy. STATEMENT OF SIGNIFICANCE: Selecting effective chemotherapy after immunotherapy remains a major challenge in advanced gastric cancer because of substantial interpatient heterogeneity. Progress in personalized treatment is limited by the lack of preclinical models that faithfully recapitulate the post-immunotherapy tumor microenvironment (TME). Here, we developed an immune-activated co-culture model integrating patient-derived gastric cancer organoids with autologous peripheral blood mononuclear cells (PBMCs). Immune activation was achieved using anti-PD-1-loaded gold nanocages (aPD-1@Au NCs), which combine near-infrared-triggered photothermal tumor ablation with sustained aPD-1-mediated immune modulation. This platform preserves patient-specific tumor characteristics while capturing key immune-tumor interactions following immunotherapy. Using this model, we evaluated five clinically relevant chemotherapeutic agents and identified distinct drug-specific differences in tumor-killing efficacy, immune-mediated chemosensitization, and immunotoxicity. As a proof-of-concept study, this work establishes a promising framework for personalized chemotherapy screening after immunotherapy and supports the development of precision treatment strategies for gastric cancer.

Glioblastoma therapy is severely limited by poor blood-brain barrier (BBB) penetration and systemic toxicity of chemotherapeutics. Here, we engineered polyphenol-stabilized selenium nanoparticles (nGPSe NPs, <60 nm) via spatial confinement synthesis as a redox-dual nanocatalytic carrier for afatinib (AFA). These AFA@nGPSe NPs utilize their distinct physicochemical properties to facilitate efficient nose-to-brain delivery, achieving high tumor accumulation while bypassing the BBB. The platform exhibits unique tumor-selective redox duality by generating cytotoxic reactive oxygen species and depleting glutathione within the tumors, yet activating antioxidant defense pathways in normal neural tissues to prevent neurotoxicity. This dual mechanism synergizes with AFA-induced tumor cell death. In orthotopic glioblastoma models, intranasal administration achieved a 60% long-term survival rate, driven by a chemoimmunotherapeutic response involving robust CD8+ T cell and macrophage infiltration. This study presents a multifunctional nanoplatform that integrates tumor microenvironment-responsive catalysis, non-invasive delivery, and immune reprogramming for precise and safe glioblastoma therapy.

The central nervous system (CNS) harbors a distinct immune memory programming system, wherein immunogenic cell death (ICD) acts as a pivotal signaling hub. A spectrum of insults, from systemic metabolic dysfunction to local protein aggregation and ionic dyshomeostasis, can provoke ICD in neurons, glia, and resident immune cells. This process orchestrates the release of damage-associated molecular patterns (DAMPs) from distinct subcellular compartments. These DAMPs synergistically initiate both innate trained immunity (TI), characterized by profound metabolic-epigenetic reprogramming, and antigen-specific adaptive immune responses that traverse the blood-brain barrier. Together, these pathways constitute an integral network of central immune surveillance. Crucially, this ICD-driven immune programming exhibits a striking functional dichotomy depending on the pathological context. In non-neoplastic conditions such as neural injury and neurodegenerative diseases, uncontrolled ICD signaling can establish a pathological trained immune memory, driving a self-perpetuating cycle of chronic neuroinflammation and tissue damage. Conversely, within the tumor microenvironment of malignancies like glioma, the adaptive immune responses elicited by ICD are frequently subverted by potent immunosuppressive mechanisms, culminating in tumor immune escape. This review dissects the differential regulatory mechanisms of ICD-mediated immune memory in CNS tumors versus non-tumor diseases. We aim to elucidate the molecular switches that govern the transition of this immune program from a beneficial, compensatory state to a pathological, detrimental phenotype. By exploring emerging therapeutic strategies, including gene editing, nanomaterials, and bioactive phytochemicals that precisely target ICD pathways, we provide a theoretical framework for understanding CNS immune homeostasis and for the rational design of precision immunotherapies.

Tumor cells exhibit a hyper-glycolytic phenotype, resulting in massive lactic acid (LA) production that acidifies the tumor microenvironment (TME) and fosters immunosuppression. Current lactate-targeted therapies often lack synergistic dual-directional regulation. Herein, we engineer an exosomal nanoplatform, PpIX/siRNA@EXO-LOD, to simultaneously disrupt intra- and extracellular LA homeostasis, thereby enabling synergistic metabolic and photodynamic therapy (PDT). The system integrates three key components: protoporphyrin IX (PpIX) for PDT, siRNA targeting monocarboxylate transporter 4 (MCT4) for intracellular metabolic interference, and surface-displayed lactate oxidase (LOD) for extracellular catalytic starvation. Upon epithelial cell adhesion molecule (EpCAM)-mediated targeting, the nanovesicle triggers a cascade of synergistic effects. Crucially, siRNA-mediated silencing of MCT4 induces lethal intracellular acidosis, leading to significant intracellular H2O2 accumulation. This elevated H2O2 level acts as a booster for PpIX-generated reactive oxygen species (ROS) upon laser irradiation, creating an amplified oxidative stress burst that overwhelms tumor cell defenses. Concurrently, surface-anchored LOD consumes extracellular LA, alleviates lactate-induced immunosuppression. In vivo studies demonstrate that this dual-regulation strategy effectively inhibits tumor growth, downregulates metastasis-related factors (amphiregulin (AREG), ATP-binding cassette sub-family B member 1 (ABCB1)), and reprograms the TME from an immunosuppressive state to an immunologically active state. This work presents a precision nanomedicine strategy that leverages the interplay between metabolic modulation and photodynamic amplification for enhanced cancer treatment.

A biodegradable biomimetic nanoplatform (HMCDL@TK-M) was constructed by combining hydrogen-doped HxMoO3 nanoparticles, dual-drug loading, and a hybrid spinach-cancer cell membrane coating. The system features pH-responsive biodegradability, tumor-homing capability, and high NIR-II photothermal conversion. An oxygen-lactate cascade, formed via thylakoid membrane-mediated H2O2 decomposition and lactate oxidase-driven lactate oxidation, alleviates hypoxia and depletes lactate in the tumor microenvironment. This dual metabolic modulation reprograms M2 macrophages to M1, promotes dendritic cell maturation, and reduces Treg infiltration. In 4T1 tumor-bearing mice, HMCDL@TK-M achieves strong tumor accumulation, effective photothermal ablation, and combined with lactate depletion, complete tumor eradication without systemic toxicity. The treatment also induces robust CD4+/CD8+ effector memory T-cell responses, providing durable antitumor immunity. This work demonstrates a synergistic metabolic-photothermal immunotherapy strategy for efficient and long-lasting cancer treatment.

Cancer progression is systemically influenced by distant organ dysfunction induced by primary tumors, yet how long-distance tumor-organ crosstalk regulates antitumor immunity remains unclear. Here, we identify host metadherin (MTDH) as a critical regulator of tumor-induced immunosuppression and metabolic reprogramming via tumor-liver interactions. Using Mtdh knockout mouse models, we show that concurrent MTDH loss in hepatocytes and CD8+ T cells enhances effector T cell function and suppresses tumor growth and metastasis. Mechanistically, tumor-derived extracellular vesicles and particles (EVPs) activate Kupffer cells to secrete tumor necrosis factor α (TNF-α) and TGF-β, which suppress hepatic PPARα-mediated lipid oxidation via nuclear factor κB (NF-κB) signaling. MTDH loss restores hepatic lipid catabolism, reduces systemic lipid levels, and promotes mitochondrial metabolic reprogramming in CD8+ T cells under lipid-reduced conditions, thereby boosting antitumor immunity. Genetic or pharmacological targeting of MTDH synergizes with anti-PD-1 therapy. These findings establish host MTDH as a key mediator of tumor-liver crosstalk through metabolic and immune interactions, driving systemic cancer progression.

The clinical translation of cellular immunotherapies for solid tumors remains limited by the lack of adequate tools to evaluate therapeutic function within the native tumor microenvironment. Here, we demonstrate a miniaturized implantable microdevice (IMD) capable of spatially resolving and multiplexing the delivery and screening of multiple T cell therapies directly within live tumors. This customizable IMD is comprised of discrete fibrin-filled reservoirs that can be independently loaded with distinct immune cell formulations, which enables spatially confined release and region-specific immune-tumor interactions within the same tumor mass. Using this platform in glioblastoma xenografts, we simultaneously delivered EGFR-targeting CAR-T cells and control T cells and analyzed adjacent tissues via IHC-based quantitative image analysis and multiplexed immunofluorescence. The perireservoir region adjacent to CAR-T cell reservoirs exhibited dense CD8+ infiltration, cleaved caspase-3-mediated apoptosis, and suppressed Ki67 expression, in contrast to minimal activity in control regions. We believe our IMD platform facilitates the parallel in vivo evaluation of multiple immune cell therapies, providing a translational tool to accelerate the development of successful personalized adoptive cell therapies.

Immune checkpoint inhibitor (ICI) myocarditis is a rare but frequently fatal immune-related adverse event of cancer immunotherapy. Understanding the mechanisms of this toxicity is critical to balancing treatment with maintenance of antitumor immunity. Using integrated spatial and single-cell analyses in a pharmacological murine model, we identified regional infiltration of Ly6C+ monocytes and PD-1+ CD8+ T cells in the heart that organize into fibroblast-rich immune structures, which we term tertiary T cell niches (TTCNs). TTCNs serve as hubs for T cell activation, sharing features of tertiary lymphoid structures. A TTCN gene signature was strongly enriched in cardiac tissue from patients with ICI myocarditis. Complementary T cell receptor analyses revealed clonal expansion of cardiac T cells following ICI treatment. We further identified TTCN-associated cytokines and structural proteins as candidate therapeutic targets to reduce myocardial inflammation while considering tumor control. Together, these findings suggest that cardiac tertiary immune structures play a central role in ICI myocarditis and highlight pathways that could mitigate ICI cardiotoxicity.

Intercellular communication within the tumor microenvironment (TME) is highly active in gastrointestinal (GI) cancers, and exosomes have an important function in tumor development, progression, metastasis, angiogenesis, and drug resistance. Exosomes secreted by tumor cells actively remodel their local environment by delivering oncogenic proteins, genetic material, and integrins, contributing to processes such as invasion, immune escape, and metastatic niche formation. Cancer-associated fibroblasts (CAFs) are a main source of exosomes in the stroma, and their presence boosts tumor progression by orchestrating changes in the extracellular matrix composition, metabolism, and epithelial to mesenchymal transition. Similarly, exosomes produced by endothelial cells facilitate angiogenesis and increase vascular permeability, providing a mechanism for tumor spread. One of the key factors recently discovered in exosomes is the integrin β3, which plays a major role in organotropic metastasis, cell adhesion, migration, and activation of signaling pathways in GI cancers. In this review, the biological roles of exosomes isolated from tumors, CAFs, and endothelial cells have been highlighted, especially with reference to the functional significance of integrin β3 in GI cancers. This review also provides insight into the application of integrin β3 in exosomes as a biomarker for early detection, diagnosis, and treatment. The knowledge gained through exosome-based signaling could lead to new approaches for interfering with TME-associated tumor progression.

Pancreatic ductal adenocarcinoma (PDAC) remains among the most lethal solid malignancies, and its therapeutic failure reflects both aggressive tumor-cell biology and the highly restrictive tumor microenvironment (TME). A defining hallmark of PDAC is desmoplasia, an extensive, extracellular matrix (ECM)-rich fibroinflammatory reaction that frequently exceeds the tumor cell compartment itself. Evidence from genetically engineered mouse models and human specimens identifies pancreatic stellate cells (PSCs) as the dominant architects of this stroma. Upon activation, PSCs differentiate into matrix-producing fibroblasts that drive collagen and hyaluronan (HA) accumulation, tissue stiffening, and vascular compression. ECM remodeling elevates interstitial pressure, collapses perfused vessels, and establishes profound hypoxia, which in turn reinforces fibroblast activation and matrix deposition through feed-forward signaling loops. These hypoxic, high-stress conditions severely restrict the delivery of cytotoxic agents, biologics, and nanomedicines, while simultaneously activating mechanotransduction pathways that enhance tumor cell survival under therapy. Clinical attempts to ablate stromal components validated the barrier function of desmoplasia but also revealed its tumor-restraining roles, exposing the limitations of indiscriminate depletion. This review synthesizes PSC-driven stromal initiation, matrix biomechanics, spatial zonation, and formulation-aware delivery into a single framework to explain why stromal targeting has repeatedly failed clinically and which normalization strategies are most likely to improve therapeutically effective exposure in PDAC.

Metabolic reprogramming enables tumor cells to evade immune surveillance, yet the mechanisms linking specific metabolic alterations to immune evasion remain incompletely understood. We found that in metabolic dysfunction-associated steatohepatitis (MASH)-related hepatocellular carcinoma (HCC) cells, aberrant accumulation of bile acids impairs the NOD-like receptor family CARD domain containing 5-mediated major histocompatibility complex class I antigen presentation pathway, thereby leading to loss of antigenicity in tumor cells. Bile acid retention in tumor cells results primarily from the downregulation of the efflux pump ABCB11, driven by G protein-coupled receptor 120-mediated FXR suppression under lipid-rich conditions. Furthermore, genetic inactivation of NLRC5 abolishes the recovery of MHC-I expression and antitumor immunity upon bile acid reduction. Treatment with Tropifexor, a potent selective FXR agonist, synergizes with immune checkpoint blockade, significantly reducing tumor burden and potentiating the intratumoral adaptive immune response in mice. Thus, we reveal that intracellular bile acid accumulation suppresses both tumor antigenicity and intrinsic antitumor immunity. This insight provides a therapeutic target for improving immunotherapy outcomes in MASH-HCC.

Drug resistance remains a principal barrier to effective cancer therapy, with the tumor microenvironment (TME) serving as a central modulator. Among TME components, cancer-associated fibroblasts (CAFs) are the most abundant stromal cells and pivotal orchestrators of tumor initiation, progression, and therapeutic resistance. CAFs promote drug resistance through diverse mechanisms, including secretion of cytokines and chemokines, extracellular matrix remodeling, and direct tumor cell interactions. Accumulating evidence further reveals their multifaceted role in resistance via TME remodeling and dynamic crosstalk with immune and endothelial cells. This review provides a comprehensive synthesis of CAF-mediated drug resistance across multiple malignancies-breast, pancreatic, lung, ovarian, prostate, gastric, and hepatobiliary cancers-and examines how CAFs modulate tumor cell behavior, drug metabolism, efflux pathways, and the broader TME. Emerging CAF-targeted strategies and future therapeutic directions are also discussed, highlighting opportunities to overcome drug resistance and improve clinical outcomes.

Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality globally, with many patients presenting with advanced disease that does not respond to standard treatments. While immune checkpoint inhibitors (ICIs) have improved outcomes, their effectiveness is limited in immunologically "cold" tumors, highlighting the urgent need to elucidate the complexity of the tumor immune microenvironment (TIME). Tertiary lymphoid structures (TLSs), ectopic and organized lymphoid aggregates that form at sites of chronic inflammation, have become key players in antitumor immunity. This review thoroughly covers the composition, multistage formation, and functional maturation of TLSs in HCC. We emphasize that the prognostic and predictive value of TLSs depends not only on their presence but also on their spatial localization, structural maturity, and specific immune cell composition. A high density of intratumoral TLSs, especially those with mature germinal centers containing B cells, T follicular helper (Tfh) cells, and dendritic cells (DCs), is strongly linked to better survival and lower recurrence rates. Additionally, TLSs serve as biomarkers for predicting favorable responses to ICIs in both neoadjuvant and adjuvant settings. We also explore how locoregional therapies and specific chemokine axes induce or modulate TLS formation. Finally, we discuss the emerging field of non-invasive TLS evaluation using radiomics and advanced imaging techniques, which promises to guide patient stratification for immunotherapy. TLSs therefore represent promising therapeutic targets with the potential to reprogram the immune milieu of HCC and enable more effective, personalized treatment approaches.

Lactate and lactate-mediated protein lactylation are no longer viewed merely as accompanying phenomena of enhanced glycolysis, hypoxic responses, or tissue acidification. They are now recognized as an important regulatory axis that links local metabolic stress to chromatin regulation and altered protein function. With the rapid development of research on histone and non-histone lactylation, lactate-related signals have been implicated in inflammatory injury and repair, fibrotic remodeling, tumor immune escape, and therapy resistance. However, current studies often conflate elevated lactate levels, global increases in lactylation, site-specific lactylation events, and disease-dependent functional consequences, which can lead to overinterpretation of both the biological impact and therapeutic value of lactylation. This review first summarizes lactate production, transport, and local homeostatic regulation, then discusses the biochemical basis and detection strategies of protein lactylation. It further examines how histone lactylation reshapes transcriptional programs and how non-histone lactylation influences immune regulation by altering protein fate and signaling execution. Considering the distinct features of inflammatory and tumor microenvironments, this review compares the functional outputs of the lactate-lactylation axis during stage-specific inflammatory responses and persistent tumor-associated stress, with particular emphasis on its translational significance in immune checkpoint regulation, impaired antigen presentation, and therapeutic resistance. We propose a stratified framework for interpreting lactate-related events, distinguishing metabolic stress readouts, functional regulatory events, and disease-dependent nodes. This framework may support patient stratification, lesion-selective delivery, dynamic monitoring, and the design of precise combination therapies.

T-cell dysfunction is an important contributor to both multiple myeloma (MM) disease progression and failure of antimyeloma chimeric antigen receptor T-cell (CAR-T) and bispecific T-cell engager (TCE) therapies. Overcoming T-cell dysfunction is therefore key to improving outcomes of patients with MM. Immunomodulatory drugs and cereblon E3 ligase modulatory drugs have been observed to activate T cells and, more recently, reduce T-cell dysfunction; however, the underlying mechanisms behind this are incompletely understood. Here, using bone marrow samples from patients with MM, we demonstrate a significant reduction in dysfunctional T-cell populations expressing exhaustion markers, such as TIGIT, upon treatment with mezigdomide. We further demonstrate the ability of mezigdomide to improve T-cell function and cytotoxicity in primary T-cell models of T-cell dysfunction and bispecific TCE therapy in vitro. Using concurrent assay for transposase-accessible chromatin using sequencing, chromatin immunoprecipitation sequencing, high-throughput chromatin conformation capture, and RNA sequencing in primary T cells treated with mezigdomide, we demonstrate the novel role of transcription factor Ikaros in regulating an important T-cell exhaustion gene TIGIT. Finally, we demonstrate the ability of mezigdomide to enhance survival outcomes of anti-B-cell maturation antigen CAR-T therapy in vivo. Overall, our data show that mezigdomide treatment improves antimyeloma T-cell therapy efficacy and reduces T-cell dysfunction by abrogating Ikaros-mediated upregulation of exhaustion genes.

Interleukin-2 (IL-2) is a potent mediator of T-cell activation with significant potential for cancer immunotherapy, yet its clinical utility is severely constrained by its narrow therapeutic window. Here we report a chemically masked IL-2 prodrug (Cm-proIL2) that enables tumor-microenvironment-responsive cytokine activation through tumor-associated protease cleavage. Site-specific conjugation of a poly-(ethylene glycol) (PEG) moiety selectively masks peripheral receptor engagement, while PEG removal within the tumor microenvironment restores IL-2 receptor binding and reduces molecular size, thereby enhancing intratumoral lymphocyte penetration and effector T cell functionality. Building on this modular platform, we further engineered a PD-1-targeted nanobody fusion, PD1-Cm-proIL2, to enable the cis delivery of IL-2 activity to PD-1+ tumor-infiltrating T cells. PD1-Cm-proIL2 induces robust antitumor immunity, resulting in complete tumor regression and durable protection upon tumor rechallenge in a murine colorectal cancer model. Together, these findings demonstrate the feasibility of chemically masked, tumor-responsive cytokine activation as a strategy for improving the therapeutic index of IL-2-based immunotherapies.

Ligand-receptor interactions play a crucial role in disease therapy. However, enhancing ligand-receptor selectivity and precisely controlling subsequent cellular behavior remain significant challenges. In this study, we developed a receptor-mediated cascade amplification strategy via glycopeptides (GPs) to regulate the dynamic self-assembly of artificial ligands, enhancing ligand-receptor selectivity. By designing the assembly sequences, we control the antagonistic relationship between dynamic nanostructures and receptor-mediated internalization. Our findings show that in fast cascade amplification, rapid fiber network formation inhibits internalization, while in slow amplification, receptor aggregation enhances internalization, promoting cellular uptake. This precise regulation further impacts the spatial distribution and function of receptor, ultimately controlling cellular phenotypes. Based on real-world needs, we applied the cascade amplification strategy to enhance the efficacy of PD-L1 tumor immunotherapy. These findings provide new insights into the regulation of ligand-receptor selectivity and receptor activity, offering promising potential for targeted enhancement and precise intervention of cellular activities.

The clinical efficacy of melanoma immunotherapy is frequently compromised by an immunosuppressive, "cold" tumor microenvironment (TME) driven by redox dysregulation and the metabolic polarization of tumor-associated macrophages (TAMs). To overcome these intertwined barriers, a macrophage-targeted, biomimetic nanoplatform (MOF-808-Ro-G/N) is developed. This system integrates a catalytic metal-organic framework (MOF-808) core loaded with a PERK inhibitor and a CD47 blocking agent, encapsulated within a Rhizopus oryzae (Ro)-derived fungal shell. The biomimetic shell leverages Dectin-1 interactions to enable precise TAM targeting and accumulation. Upon endocytosis, the acidic endosomal microenvironment activates the MOF core to deplete glutathione and trigger a surge of reactive oxygen species, thereby inducing immunogenic cell death. Synergistically, the released payloads disrupt the PERK-ATF4 metabolic stress axis, reprogramming pro-tumorigenic M2 TAMs toward an anti-tumor M1 phenotype, while concurrently blocking the CD47-SIRPα checkpoint to restore macrophage phagocytosis. Through this orchestrated modulation, MOF-808-Ro-G/N effectively remodels the TME into an immunologically "hot" niche, promoting robust cytotoxic T lymphocyte infiltration and achieving substantial tumor growth suppression. Overall, this "metabolic-immune dual-reprogramming" strategy provides a promising materials-driven paradigm to overcome immunotherapy resistance in solid tumors.

High-grade serous ovarian cancer (HGSOC) is commonly accompanied by malignant ascites, a clinically relevant tumor niche that promotes immune evasion, metastasis, and treatment resistance. Although natural killer (NK)-cell dysfunction has been described in ovarian cancer, the broader innate lymphoid landscape of ascites and the mechanisms linking ascites-derived signals to innate immune suppression remain insufficiently resolved. We performed single-cell RNA sequencing of NK/innate lymphoid cells from ovarian cancer ascites to define cellular heterogeneity and differentiation states. Functional assays assessed NK-cell cytotoxicity, degranulation, and receptor expression following exposure to patient-derived ascites, with or without transforming growth factor-β (TGF-β) receptor inhibition. Proteomic profiling was used to characterize the soluble ascites milieu, and clinical associations were examined for innate lymphoid subsets. Single-cell analysis identified eight transcriptionally distinct NK/innate lymphoid states, including cytotoxic, precursor, early-like, tolerant/immunoregulatory, regulatory, proinflammatory, and innate lymphoid populations. Ovarian cancer ascites was characterized by depletion of cytotoxic and precursor NK-cell states together with enrichment of early-like, tolerant, regulatory, pro-inflammatory, and innate lymphoid cell (ILC) populations. Trajectory analysis indicated impaired maturation toward terminally differentiated cytotoxic NK cells. Notably, ascites contained an expanded population of programmed cell death protein 1 (PD-1)+ ILC2s, which were more abundant in patients with shorter progression-free survival. In functional assays, short-term exposure of healthy donor NK cells to ascites suppressed degranulation and tumor-cell killing, reduced expression of activating receptors including NKp30 and DNAM-1, increased inhibitory receptor expression, and shifted NK cells toward a CD56highCD16low phenotype. Proteomic profiling supported a soluble milieu consistent with type 2 immune skewing and NK-cell suppression. Importantly, TGF-β receptor inhibition partially restored NK-cell activation and function in the presence of ascites. HGSOC ascites establishes a type 2-skewed immunoregulatory niche that coordinately drives NK cell dysfunction and PD-1+ ILC2 accumulation. The findings identify TGF-β-linked suppression and ascites-associated immune regulators as candidate immunotherapeutic vulnerabilities for restoring antitumor immunity in ovarian cancer.

Tumor microenvironment (TME)-induced immunosuppression is a major driver of cancer treatment failure and resistance to immunotherapy. Myeloid-derived suppressor cells (MDSCs) are not only pivotal suppressors of effector immune responses but also central organizers of tumor-supportive metabolic, stromal, and cellular crosstalk. This review systematically summarizes the origin and phenotypic characteristics of MDSCs and the mechanisms governing their recruitment, expansion, and activation within the TME. This review integrates current knowledge of MDSC biology through two complementary frameworks. One framework highlights the bidirectional interactions between MDSCs and immune, tumor, stromal, endothelial, and adipocyte compartments. The other classifies therapeutic approaches according to their mechanisms and translational relevance, including combinations with immunotherapy. We further evaluate clinical translation, including lessons from unsuccessful or inconclusive trials, biomarker gaps, and rational combination strategies. Together, current evidence indicates that MDSCs represent context-dependent therapeutic nodes, while functional reprogramming, spatially resolved profiling, and patient stratification may improve immunotherapy outcomes.

The tumor microenvironment is crucial for cancer progression, but the mechanisms underlying the tumor-immune cell interactions in it remain poorly understood. Here, we identified latent transforming growth factor-β (TGFβ) binding protein 4 (LTBP4) deficiency in colorectal cancer (CRC) as a critical driver that reprogrammed tumor-associated macrophages (TAMs) and induced a distinct subset, which promoted tumor progression by coordinating immune evasion and extracellular matrix (ECM) remodeling. Clinically, LTBP4 deficiency correlated with CRC progression and poor patient survival. Ltbp4 knockout markedly promoted tumor growth and metastasis in immunocompetent mice, an effect attenuated in immunodeficient hosts, establishing the essential role of host immunity in mediating the effects of LTBP4 deficiency. Single-cell RNA sequencing revealed that LTBP4 deficiency induced a mannose receptor C-type 1-positive (MRC1+)/CD44+ TAM subset and correlated with reduced CD8+ T cell infiltration. Mechanistically, LTBP4 deficiency increased active TGFβ1 levels, which acted in a paracrine manner to up-regulate MRC1 in TAMs, whereas autocrine signaling induced HAS2 (hyaluronan synthase 2) expression and hyaluronan production to increase CD44. CD44 signaling in TAMs up-regulated matrix metalloproteinases for collagen degradation, whereas MRC1 mediated collagen internalization, cooperatively remodeling the ECM to facilitate tumor invasion. The TGFβ1-driven MRC1+/CD44+ TAMs further suppressed CD8+ T cell function by diminishing the C-X-C motif chemokine ligand 16-C-X-C motif chemokine receptor 6 (CXCL16-CXCR6) axis. Therapeutically, targeted depleting MRC1+/CD44+ TAMs enhanced the efficacy of PD-1 (programmed cell death-1) blockade in LTBP4-deficient tumors. Our study positions LTBP4 as a key modulator of tumor progression and reveals a therapeutic strategy for LTBP4-deficient CRC.

Poly(ADP-ribose) polymerase (PARP) inhibitors are highly effective in BRCA-deficient tumors, yet their therapeutic benefit in BRCA-proficient cancers remains limited. Here, we demonstrate that YCH1899, a next-generation, resistance-overcoming PARP inhibitor, elicits significantly stronger type I interferon signaling than currently approved PARP inhibitors in BRCA-proficient tumor cells. Mechanistically, YCH1899 facilitates the accumulation of cytosolic DNA, thereby robustly activating the cGAS-STING-IRF3 pathway. Consequently, YCH1899 exhibits potent inhibitory effects on the growth of various BRCA-proficient tumors in a STING-dependent manner and enhances the infiltration of CD8⁺ T cells within the tumor microenvironment. To identify tumor-intrinsic regulators of this immune response, we conducted in vivo CRISPR screens, which revealed USP20 as a prominent regulator. The genetic ablation of USP20 markedly diminished the antitumor immunity induced by YCH1899 in vivo, suggesting that USP20 may serve as a potential predictive biomarker. Furthermore, combining YCH1899 with STING agonists synergistically amplifies interferon pathway activation, driven by enhanced phosphorylation of IRF3 at Ser173, resulting in superior tumor control and profound immune cell infiltration and activation within the tumor microenvironment. Collectively, our findings establish YCH1899 as a potent PARP inhibitor capable of extending clinical benefit to BRCA-proficient cancers and provide a rationale for its clinical combination with STING agonists to elicit robust antitumor immunity, even in patients resistant to PARP inhibitors.

Terminal exhaustion of CD8⁺ T cells limits the efficacy of immune checkpoint blockade (ICB), particularly in glioblastoma, which is characterized by an immunosuppressive tumor microenvironment. Therefore, a better understanding of the regulators of terminal exhaustion could overcome intrinsic resistance to ICB and improve the efficacy of immunotherapy. In this study, we report that all-trans retinoic acid (ATRA) suppresses CD8⁺ T-cell terminal exhaustion. The administration of ATRA during CD8⁺ T-cell activation in vitro conferred resistance to terminal exhaustion and preserved effector cytokine production and effector function. Mechanistically, ATRA selectively induced the expression of the long isoform of T-cell factor 1 (TCF-1βBD) through activation of the canonical WNT/β-catenin pathway. In a mouse glioma model, adoptively transferred ATRA-conditioned CD8⁺ T cells exhibited increased TCF-1βBD and β-catenin expression and resistance to exhaustion, which led to superior tumor-infiltrating CD8+ T-cell polyfunctionality and enhanced glioma suppression. Importantly, oral administration of ATRA also suppressed terminal exhaustion of tumor-infiltrating CD8+ T cells and synergized with anti-PD-1 therapy, overcoming resistance to immune checkpoint blockade (ICB) in two mouse models of glioma. Combined ATRA and anti-PD-1 treatment also displayed synergistic therapeutic potential by significantly reducing recurrence in a mouse model of glioma that had undergone surgical resection. Single-cell transcriptomic analysis of glioblastoma patients receiving anti-PD-1 therapy revealed that CD8⁺ T cells from responders were enriched in genes that respond to retinoic acid and WNT-associated genes, which correlated with improved survival. These findings establish ATRA as a modulator of CD8⁺ T-cell exhaustion through the induction of WNT/β-catenin-dependent TCF-1βBD expression, suggesting that ATRA has therapeutic potential for overcoming resistance to ICB in glioma.

We performed an integrated clinical and genomic analysis of over 7,000 consecutively sequenced colorectal cancer (CRC) samples to comprehensively characterize genetic drivers and metastatic tropisms of CRC. We find that genomic evolutionary changes, such as clonal mutations and oncogenic mutant allelic imbalance, selectively enhance the impact of recurrent oncogenic alterations. We identify the relative timing of organ-specific metastasis, showing sequential metastatic progression in microsatellite stable CRC with brain and adrenal metastases as late events; metastatic sites that cluster together, such as lung, bone, and brain metastases; and genomic events that enhance or decrease risk for each metastatic site, with WNT pathway activation as overall protective while RAS pathway activation increased risk for spread to all metastatic sites. Our data suggest that despite the heterogeneity in CRC, genomic evolution increases the impact of recurrent alterations, and integrating information about tumor primary location and genomics can be used to predict organ-specific metastasis risk.

In this issue of Cancer Cell, Manca et al. investigate the genomic evolution and metastatic tropism of over 7,000 colorectal cancers. Integrating primary tumor location, genomic alterations, and clinical variables, they reveal how distinct patterns of tumor evolution shape metastatic competence and impact organ tropism, offering a framework to refine surveillance.

GIM/FP/GP: [Formula: see text] Gastroenterology: [Formula: see text] Public Health: [Formula: see text].

Associations between various genetic variants and the risk of hepatocellular carcinoma (HCC) have been extensively explored but produced contradictory results. The aim of the present systematic meta-analysis was to determine and validate genetic variants that are associated with HCC risk. Two-step literature searches of PubMed, Embase, Web of Science, and Google Scholar databases and various meta-analyses were performed, and a comprehensive field synopsis and epidemiological evidence were provided. A total of 20,081 publications were identified, of which 830 were deemed eligible for inclusion. Eventually, 36 variants in 27 genes were identified to be associated with HCC risk. Moreover, cumulative epidemiological evidence of an association was graded as moderate for nine variants in eight genes (ESR1 rs2234693, GRP78 rs430397, HLA-DP rs3077, HLA-DQ rs2856718, MnSOD rs4880, TNFα rs361525, HFE rs1800562 and rs1799945, and UGT1A7 High/Low) and strong for three variants in three genes (IL-1B rs1143627, COL18A1 rs7499, and NQO1 rs1800566); HFE rs1800562 was deemed to have a false-positive association. Thus, 11 variants in 11 genes were identified to be associated with HCC risk. This synopsis helps elucidate the mechanisms of carcinogenesis of HCC and provides insights into the early diagnosis and novel treatments of HCC by targeting those potential genes.

Recurrent hepatocellular carcinoma (HCC) can arise with variable molecular similarity to the primary tumor, but practical approaches for quantifying methylome divergence in paired tumors remain limited. We performed reduced representation bisulfite sequencing on paired primary and recurrent HCC lesions and matched normal margins from ten patients, quantifying clustered CpG methylation across 77,790 MspI-defined loci. Regression-based methylation shift patterns (MSPs) captured tumor-specific curve shapes, and Euclidean distances between regression-coefficient vectors grouped tumors into four MSP classes. We further introduced tH, a triangle-area metric summarizing within-patient methylome divergence between primary and recurrent tumor-associated MSPs. In this exploratory cohort, tH distinguished lower- and higher-divergence recurrence patterns that showed concordance with serum marker patterns, mutation-based tumor relatedness, and recurrence-free survival differences. Interval-based analyses revealed prominent hypermethylation at low-methylation loci, hypomethylation at high-methylation loci, and heterogeneous changes in intermediate methylation tiers. These findings establish an MSP/tH framework for quantifying methylome divergence between paired primary and recurrent HCCs and for stratifying tumors by shared epigenetic shift patterns, providing a basis for future validation in larger paired-tumor cohorts.

Hepatoblastoma (HB) is the most common primary liver malignancy in childhood, yet its molecular determinants, functional dependencies, and therapeutic vulnerabilities remain incompletely characterized. Integrative analyses combining transcriptomic profiling with functional genomic datasets provide a strategy to identify essential genes, biomarkers predictive of tumor behavior and treatment response. Differential expression analysis comparing HB tumors with normal liver was processed on training cohort. These genes were integrated with DepMap CRISPR-Cas9 dependency scores to prioritize HB-essential candidates. Elastic Net regression was used to derive a 16-gene predictive signature, which was validated in an external cohort. Single-cell RNA-seq datasets were analyzed to assess expression patterns across hepatic and tumor-associated cell populations. A supervised deep-learning classifier was trained on single-cell profiles to distinguish tumor cells from hepatocytes, and SHAP values were computed to interpret gene contributions. Drug-gene interactions were queried using curated repressive compounds from DGIdb, and approved drugs were screened for relevance in pediatric cancer clinical trials. A total of 789 genes were found overexpressed in HB tumors from the training transcriptome cohort. Chronos DepMap analysis identified 73 HB-essential genes that were not essential in adult liver cancer cell lines (hepatocellular carcinoma and cholangiocarcinoma). Elastic-net tuning based on the expression of 16 HB-essential genes in the split training cohort enabled robust tumor-normal discrimination, with AUC = 0.88, specificity = 0.90, and sensitivity = 0.90 in internal validation. This performance was confirmed in an independent external cohort, achieving AUC = 0.99, specificity = 1.00, and sensitivity = 0.98. Single-cell validation further demonstrated tumor-specific enrichment of the signature. The deep-learning classifier (tumor cells vs. normal hepatocytes) reached high accuracy (AUC = 0.99; F1-score = 0.97), with SHAP analysis highlighting PEG10, GREB1, PLCB4, RHOBTB1, CRIM1, FSD1L, CORO2A, KIT, ANKRD50, HDAC11, ZNF233, SEMA7A, and FABP4 as major contributors. Six of these genes were confirmed to be absent or lowly expressed in the background liver microenvironment. Drug-gene interaction analysis identified HDAC11 as a potential therapeutic target of approved drugs used in pediatric oncology. This integrative framework combining transcriptomics, CRISPR dependency mapping, machine learning, and pharmacogenomic annotation identifies clinically relevant HB-essential genes and predictive molecular signatures for tumor identity. The derived expression-based scores provide tools for patient stratification, while drug-gene mapping highlights actionable vulnerabilities on HDAC11 with pediatric approved drugs that support rational drug repurposing strategies in hepatoblastoma.

Pancreatic ductal adenocarcinoma (PDAC) is a highly mortal cancer whose only potentially curative treatment is surgical resection. Intraoperative assessment of its surgical margins is vital for patient survival. Frozen section biopsy is routinely performed for this purpose. However, its heavy reliance on pathologists' expertise often leads to diagnostic discrepancies. The inherent invasiveness of PDAC also leads to sampling errors. This study developed an intelligent molecular cytology approach that improves diagnostic objectivity and broadens sampling coverage. Our method, Multi-Instance Cytology with LEArned Raman-embedding (MICLEAR), leverages compositional information from label-free Raman imaging. First, 4085 cells were brushed off from the pancreases of 41 patients and imaged using stimulated Raman scattering microscopy. Then, a contrastive learning-based cell embedding model was developed to compress each cell's morphological and compositional information into a compact cell vector. Finally, a multi-instance learning-based diagnostic model using cell vectors was employed to predict the likelihood that a patient's margin is positive. MICLEAR achieved 80% sensitivity, 100% specificity, and an area under the receiver operating characteristic curve of 0.86 in 27 patients for validation, comprising 10 with positive margins and 17 with negative margins, in approximately 8 minutes per patient. It may hold promise for more efficient and accurate intraoperative assessment of PDAC surgical margins.

Hepatocellular carcinoma is characterized by considerable molecular heterogeneity, which complicates prognostic predictions and contributes to therapeutic resistance. This study aimed to develop a molecular classification framework grounded in lipid droplet-associated genes (LDAGs) and to comprehensively elucidate their biological significance and clinical applicability in guiding personalized treatment approaches. By leveraging multi-cohort datasets, we defined LDAG-based molecular subtypes and systematically characterized their genomic alterations, metabolic features, pathway activation patterns, and therapeutic vulnerabilities. Three distinct subtypes (C1-C3) were identified according to LDAG expression patterns, each demonstrating unique clinical outcomes, mutational profiles, and metabolic reprogramming. The C1 subtype correlated with the poorest overall survival, more advanced tumor stages, and activation of pro-proliferative signaling pathways. Therapeutic vulnerabilities were subtype-dependent, with C1 showing heightened sensitivity to sorafenib. Five pivotal LDAGs (PLIN3, SET, CKAP4, RAP1B, and PISD) were implicated in the aggressive phenotype of C1, among which PLIN3 exhibited the strongest prognostic value. Functional assays confirmed that PLIN3 knockdown reduced lipid accumulation, suppressed cell proliferation and migration, and impaired tumorigenesis, whereas its overexpression promoted aggressive tumor behavior. In conclusion, our LDAG-based classification system stratifies hepatocellular carcinoma into three clinically relevant subtypes. PLIN3 emerges as a promising prognostic biomarker and therapeutic target, thereby mechanistically linking lipid metabolism to hepatocellular carcinoma progression.

Ischemic stroke in patients with active cancer is heterogeneous and frequently classified as cryptogenic under the TOAST classification (Trial of ORG 10172 in Acute Stroke Treatment). The American Heart Association recently proposed an etiological classification for cancer-related ischemic stroke (CRIS). This study aimed to evaluate its clinical and prognostic implications. We analyzed data from the prospective SCAN study (Ischemic Stroke in Patients With Cancer and Neoplasia), which enrolled patients with acute ischemic stroke and active cancer in Japan. Among the registered patients, those with available D-dimer data were included in the analysis. Stroke subtypes initially classified according to the TOAST criteria were reclassified using the CRIS framework. Kaplan-Meier survival curves were constructed, and differences were assessed using the log-rank test. Of 135 enrolled patients, 132 (median age, 75; 37.9% female) were included. Under the TOAST classification, 9 patients had small vessel occlusion, 20 large artery atherosclerosis, 28 cardioembolism, 10 other determined cause, and 65 cryptogenic strokes. After reclassification, 2 patients originally categorized as other determined etiology due to disseminated intravascular coagulation, and 46 patients previously classified as cryptogenic stroke were reclassified as CRIS. Patients classified as CRIS demonstrated significantly worse 1-year survival than those classified as conventional etiologies or cryptogenic stroke (global log-rank, P<0.001). The 3-month survival rate was 37.5% in the CRIS group and 89.2% in the reclassified cryptogenic stroke group. The newly proposed CRIS classification reduced the proportion of cryptogenic strokes under the TOAST system and enabled prognostic stratification by identifying a subgroup with markedly worse outcomes.

Observational studies have long suggested an association between gut microbiota dysbiosis and gastric cancer, yet the causal relevance and underlying biological pathways remain poorly defined. Here, we applied a comprehensive Mendelian randomization (MR) framework to gut microbiota and gastric cancer GWAS summary statistics to identify 14 microbial taxa significantly associated with gastric cancer risk. Sensitivity analyses, including tests for horizontal pleiotropy, heterogeneity, and genetic co-localization, revealed no strong evidence of shared causal variants (H4 < 2%); however, convergent results across five complementary MR models support overall causal inference. Mediation analysis further identified a partial pathway from the taxon ebi-a-GCST90027718 to gastric cancer via the metabolite met-c-926 (sdLDL), accounting for 4.31% of the total effect. Preliminary 16S rRNA sequencing in a pilot cohort (n = 5 per group) revealed divergent shifts in Ruminococcus subgroups without disrupting overall community stability, though these observations require replication. Collectively, this study establishes a genetically anchored evidence chain linking host genetics, gut microbiota, and gastric cancer pathogenesis, providing insights for microbiota-targeted prevention strategies.

Cutaneous melanoma with regional or distant metastases (late-stage) is associated with worse survival than localized disease. Access to dermatologic care varies widely across the United States and may influence the stage at which melanoma is diagnosed. To examine the association between regional dermatologist density and the proportion of melanoma cases diagnosed at late-stage. This national ecological study used the State Cancer Profiles and Area Health Resource File data to assess proportions of late-stage melanoma and dermatologist counts per 100,000 population across National Cancer Institute Health Service Areas (HSAs) from 2017 to 2021. Five hundred fifty-seven HSAs met inclusion criteria. In multivariable analysis, higher dermatologist density was associated with significantly lower proportions of late-stage melanoma diagnosis. Compared with HSAs with no dermatologists, the odds of late-stage melanoma were reduced by 11% for >0-1 dermatologists per 100,000 population (OR = 0.89, P = .034), 11% for >1-2 (OR = 0.89, P = .020), 16% for >2-4 (OR = 0.84, P < .001), and 20% for >4 (OR = 0.80, P < .001). The ecological design limits causal inference and HSAs with low case counts were censored. Greater dermatologist density is associated with a lower proportion of melanoma cases diagnosed at a late-stage, underscoring the importance of improving access to specialty care.

Mohs surgery has excellent outcomes but is time-consuming due to the use of intraoperative frozen sections. Two-photon fluorescence microscopy (TPFM) is a slide-free technique that generates H&E-like histology images in fresh tissue specimens in real-time which could accelerate treatment. To determine the accuracy of TPFM with H&E coloring for the evaluation of basal cell carcinoma on Mohs specimens. Frozen section analysis and TPFM images were acquired from 133 specimens following completion of Mohs surgery for basal cell carcinoma. Two Mohs surgeons who had not previously seen TPFM histology evaluated the TPFM images. Their evaluations (TPFM read) were compared to a consensus frozen section analysis (frozen section analysis read) graded by a dermatopathologist and separate Mohs surgeon. The average sensitivity and specificity were 93.8% and 82.2%, respectively. If specimens that were discordant due to section plane differences or tissue fragments from frozen sectioning are excluded, average sensitivity and specificity improve to 95.0% and 94.3%, respectively, which is a truer measure of the diagnostic accuracy. This study evaluated agreement between histology after conclusion of surgery. Mohs surgeons can interpret H&E colored TPFM images with comparable accuracy to frozen sections on Mohs margins taken for basal cell carcinoma with minimal training.

Nail squamous cell carcinoma (NSCC) is the most common nail malignancy, yet it is under-recognized because of its variable presentation. Characterize demographic and clinical characteristics associated with in situ (NSCCis) and invasive (iNSCC) NSCC and evaluate treatment outcomes in a multinational cohort. Two-hundred-sixty-nine NSCCs (261 patients) were retrospectively reviewed. NSCCis and iNSCC characteristics were compared using t-tests/chi-square and multinomial logistic regression. NSCC mainly affected the right index and middle fingers and bilateral thumbs (62%), predominantly in men (72.4%) in their sixth decade. Koilocytosis, used to determine human papilloma virus (HPV) status, was detected in 32.2% of tumors (15 polymerase chain reaction-confirmed), and this was more common in NSCCis (P = .015). Subungual ooze independently predicted iNSCC (odds ratio = 3.98; 95% CI = 1.19-13.2; P = .025), while pain (P = .022) and nail plate loss (P = .004) were associated with iNSCC on univariate analysis. Mohs micrographic surgery was the most common treatment (39.5%). Overall recurrence was 9.7%, with no difference among treatments (P > .05). Retrospective design and histopathology-based HPV status. NSCC presents as a periungual papule/plaque with horizontal growth and a warty appearance, or as an oozing subungual ulcerating tumor, dermally invasive. Periungual NSCC is associated with HPV, often sexually transmitted. HPV-related cases are typically NSCCis. Digit-sparing surgery is first-line treatment, with comparable efficacy across modalities.

Patients at high risk for hepatocellular carcinoma (HCC) are recommended to undergo biannual abdominal ultrasound surveillance; however, ultrasound has low sensitivity for small HCC nodules and is associated with poor adherence. To address these limitations, the multianalyte HelioLiver Dx blood test was developed to aid in the detection of HCC in patients with cirrhosis who are at high risk for HCC. The performance of the HelioLiver Dx test and ultrasound for the detection of HCC in adults with cirrhosis was evaluated in a cross-sectional, prospective, blinded, multicenter validation study. All participants provided blood specimens for the HelioLiver Dx test and underwent ultrasound. All participants also underwent multiphasic MRI, which served as the reference standard for determining HCC status. Of 1,268 evaluable participants, 46 (3.6%) were considered to have HCC as determined by MRI, with many (46%) having small HCC lesions ≤2 cm in diameter. The HelioLiver Dx test had a sensitivity of 47.8% (95% CI 32.9-63.1) for all HCC lesions and 28.6% (95% CI 11.3-52.2) for HCC lesions ≤2 cm. In contrast, ultrasound demonstrated a lower sensitivity of 28.3% (95% CI 16.0-43.5) for all HCC lesions and failed to detect any (0%; 95% CI 0.0-16.1) HCC lesions ≤2 cm. The specificity of HelioLiver Dx and ultrasound were 87.6% (95% CI 85.6-89.4) and 93.9% (95% CI 92.5-95.2), respectively. The HelioLiver Dx test met prespecified co-primary endpoints for superior sensitivity and non-inferior specificity compared to ultrasound. Compared with ultrasound and alpha-fetoprotein, the HelioLiver Dx test identified more HCC lesions overall, including more small lesions. A convenient and accurate blood-based test may improve HCC surveillance by facilitating earlier detection and reducing patient barriers to testing. NCT03694600 IMPACT AND IMPLICATIONS: There is a significant, unmet clinical need for more sensitive and accessible methods for the early detection of hepatocellular carcinoma (HCC) in high-risk patient populations. The current study is the first blinded, multicenter, prospective study to evaluate the performance of a multianalyte blood test compared to abdominal ultrasound for the detection of HCC among patients with cirrhosis. The multianalyte HelioLiver Dx test met prespecified co-primary endpoints for superior sensitivity and non-inferior specificity compared to ultrasound for detection of HCC lesions. The availability of a more accessible, convenient and sensitive blood test to aid in the detection of HCC may improve utilization and consequently clinical outcomes for high-risk patients via reduction of care barriers and improved early HCC detection. gov identifier: NCT03694600.

Persistent infection with high-risk human papillomavirus (HPV) is a major cause of cervical cancer, and improved point-of-care (POC) detection is critical for early intervention. Although PCR-based assays are highly sensitive, their reliance on centralized laboratory infrastructure limits accessibility in decentralized settings. CRISPR-Cas diagnostics combined with lateral flow assays (LFA) offer a rapid alternative; however, visual interpretation of faint test bands remains subjective and variable. Here, we developed a smartphone-based CRISPR-Cas12a LFA platform integrated with an interpretable machine learning (ML) framework for quantitative detection of circulating HPV DNA in plasma. Standardized image acquisition was implemented using a light-controlled enclosure, and radiomics-inspired features were analyzed using a multivariable logistic regression model. The system was trained on 150 plasma samples and validated in an independent cohort of 60 samples. The optimized model achieved 96.7% sensitivity and 100% specificity, outperforming visual interpretation, particularly for low-signal samples. Performance remained stable across different smartphone models, lighting conditions, and operators, with rapid on-device inference enabling consistent and reliable operation. This integrated CRISPR-LFA platform demonstrates accurate and reproducible detection of circulating HPV DNA and supports feasibility for POC applications, pending further validation in broader clinical settings.

Achalasia appears to increase the risk of esophageal squamous cell carcinoma, but existing evidence relies on small studies that did not adjust for this cancer's main risk factors (ie, smoking and alcohol overconsumption). Whether achalasia increases the risk of esophageal adenocarcinoma, whose main risk factor is gastroesophageal reflux disease (GERD), is even more uncertain. This study aimed to clarify these associations. A population-based case-control study was conducted in 5 Nordic countries from 1987 to 2023. Esophageal cancer cases (n = 35,604) were matched to 10 times as many background population controls by age, sex, calendar year, and country (n = 355,869). Conditional logistic regression provided odds ratios (ORs) with 95% confidence intervals (CIs), adjusted for smoking and alcohol, and additionally for GERD in a mechanistic model. All variables came from national health data registries. Achalasia was present in 137 cases (96 squamous cell carcinomas and 41 adenocarcinomas) and 230 controls (99 squamous cell carcinomas and 131 adenocarcinomas). Achalasia was associated with a strongly increased risk of esophageal squamous cell carcinoma (OR, 9.46; 95% CI, 7.07-12.65). A moderate association was found for esophageal adenocarcinoma (OR, 3.05; 95% CI, 2.14-4.35), which attenuated after adjustment for GERD (OR, 1.44; 95% CI, 0.99-2.09). Achalasia treated with myotomy or dilation was associated with an even stronger association with esophageal squamous cell carcinoma (OR, 16.33; 95% CI, 11.26-23.67), but not with esophageal adenocarcinoma after adjustment for GERD (OR, 1.32; 95% CI, 0.76-2.30). Achalasia appears strongly and independently associated with esophageal squamous cell carcinoma. The association with esophageal adenocarcinoma is less strong and seems to be largely explained by GERD.

Endoscopic retrograde cholangiopancreatography-guided transpapillary biliary drainage (ERCP-BD) is the standard for primary palliation of malignant distal biliary obstruction (MDBO), but endoscopic ultrasound-guided choledochoduodenostomy (EUS-CDS) has demonstrated improved technical success, efficiency, and safety in randomized trials. However, cost-effectiveness data are lacking. In this modeling study, we analyzed the cost-effectiveness of EUS-CDS with lumen-apposing metal stent (LAMS) and ERCP-BD with self-expandable metal stent (SEMS) for primary MDBO palliation. A state-transition Markov model compared EUS-CDS and ERCP-BD over a 1-year time horizon from a US healthcare perspective. The base case was a 70-year-old with locally advanced, unresectable pancreatic cancer, common bile duct dilation >15 mm, and MDBO. Probabilities were derived from meta-analyses of randomized trials. Outcomes were incremental cost-effectiveness ratios (ICERs), with a willingness-to-pay (WTP) threshold of $100 000/quality-adjusted life year (QALY). Extensive sensitivity analyses were performed. EUS-CDS with LAMS was cost effective versus ERCP-BD with SEMS for primary treatment of MDBO at an ICER of $47 711/QALY. In one-way sensitivity analyses, EUS-CDS remained cost effective if it cost <$15 502 or if ERCP-BD cost >$11 174. ERCP-BD would become cost effective if technical success was >91%, reintervention <11%, or postprocedural pancreatitis <4%. Probabilistic sensitivity analysis showed EUS-CDS remained cost effective in 74.1% of iterations at a $100 000/QALY WTP threshold. In patients with MDBO and biliary dilation >15 mm, EUS-CDS with LAMS may be not only a clinically preferred option but also an economically viable primary approach. Continued efforts to minimize LAMS costs, decrease stent dysfunction, and identify optimal anatomic indications are warranted to facilitate wider adoption.

Traditional CRISPR-Cas12a mutation detection systems are limited by poor single-base specificity, target-specific crRNA redesign, and insufficient sensitivity for low-abundance mutations, restricting their clinical liquid biopsy applications. Herein, we developed a crRNA-universal, sensitive and specific CRISPR-Cas12a detection platform, termed DESIC (double-end blocker and split-input mediated CRISPR-Cas12a system), for single-base mutation detection. The DESIC system adopts two key structural designs: double-end blocker (DEB) and duplicated split-input (SIN). The DEB spatially isolates crRNA recognition and target-binding regions, enabling universal detection of various mutation sites without crRNA redesign. The SIN strategy amplifies thermodynamic differences from single-base mismatches, greatly improving single-nucleotide discrimination. We targeted four prevalent pancreatic cancer KRAS mutations (G12D, G12R, G12V, Q61H) and optimized the system to achieve optimal discrimination. The optimized DESIC system exhibited ultra-low limits of detection down to 0.01% mutant allele fraction with reliable linear quantitative performance. Clinical validation using 15 pairs of pancreatic cancer tissue and peripheral blood samples confirmed that DESIC results were highly consistent with gold-standard NGS data. With a flexible modular design, this low-cost, easy-operated platform can be readily extended to multiple tumor mutations, holding great potential for tumor liquid biopsy and early molecular diagnosis.

Exogenous L-glutamine has preclinical antitumor activity although formal clinical translation has not been attempted. We conducted a single-arm phase 1 trial to assess the safety and preliminary efficacy of clinical-grade, US Food and Drug Administration-approved L-glutamine therapy with gemcitabine and nab-paclitaxel (GA) in participants with treatment-naive, advanced pancreatic cancer (n = 16). The primary endpoint was to determine the recommended phase 2 dose (RP2D) by adaptive Bayesian design across standard doses of GA and a dose range of 0.1-0.3 g kg-1 twice-daily oral L-glutamine. Secondary endpoints included safety and preliminary efficacy of the study combination. The primary endpoint was met with the RP2D reached at maximum doses of L-glutamine and GA. The grade ≥3 treatment-related adverse event rate was 66.7%, primarily from GA. Addition of L-glutamine to GA induced tumor shrinkage in 94% of subjects with a best overall response rate (ORR) of 44% (12.5% complete response). Median progression-free survival and overall survival (OS) were 8.5 months (95% confidence interval (CI) 6-not reached (NR)) and 22 months (95% CI 11-NR), respectively. L-Glutamine induced distinct metagenomic and metabolomic signatures on exploratory analyses in glutamine-treated subjects as a single agent, while the combination of L-glutamine and GA nearly doubled the ORR and tripled the OS compared to historical GA alone (ClinicalTrials.gov registration: NCT04634539 ).

Gastric cancer (GC) ranks as the fifth most common cancer worldwide, however, accurate and non-invasive diagnostic modalities for GC remain limited. Cell-free DNA (cfDNA) fragmentomics has emerged as a promising tool for cancer cell detection. Here we develop a gastric cancer detection model, named GaFraD model. The GaFraD model uses four cfDNA fragmentomics features, including fragment size ratio (FSR), copy number variation (CNV), 9-bp end motif (Motif), and fragment size at transcription start sites (TF). This model achieves an area under the receiver-operating characteristic curve (AUC) of 0.970 (95% CI: 0.944 - 0.990), a sensitivity of 95.0% and a specificity of 80.9%. By combining the GaFraD model and conventional protein biomarkers CA19-9 and PG-I/PG-II, the CONFIRM model was generated. The CONFIRM model attained an AUC of 0.986 (95% CI: 0.966 - 1.000), a sensitivity of 95.0% and a specificity of 95.6% in detecting GC. Moreover, the CONFIRM model achieved remarkable performance (AUC = 0.983, sensitivity 95.6%, specificity 94.2%) in distinguishing patients with early-stage GC from controls. Our work showed the high discriminatory power in distinguishing GC patients from controls, indicating the clinical potential of using cfDNA fragmentomics combined with protein biomarkers for non-invasive GC detection. The results of the study provide a new avenue for early, accurate, and non-invasive clinical diagnosis of GC.

Colonoscopy quality and diagnostic outcomes in Lynch syndrome have historically been heterogeneous. Following the introduction of the Lynch syndrome Bowel Cancer Screening Programme (BCSP), we assessed colonoscopy quality and diagnostic yield in Lynch syndrome using the National Endoscopy Database (NED), comparing outcomes between BCSP and non-BCSP providers. We performed a cross-sectional analysis of UK Lynch syndrome colonoscopies from NED between June 2024 and June 2025. Colonoscopy quality indicators and endoscopic detection rates for polyp (PDR) and advanced neoplasia (eANDR; polyps ≥10 mm or cancer; histopathology linkage was unavailable) were calculated and compared between BCSP and non-BCSP providers. Factors associated with neoplasia detection were identified using logistic regression. Among 3196 colonoscopies, most were performed in England (90.2%), with 34.1% undertaken within BCSP. Overall colonoscopy quality was high: 97.6% were complete, 89.0% had adequate bowel preparation, and 96.3% had adequate tolerance. BCSP procedures demonstrated higher rates of completion (98.5% vs. 97.1%; P = 0.02) and adequate bowel preparation (92.7% vs. 86.7%; P < 0.001) than non-BCSP colonoscopies. PDR and eANDR were 46.8% and 4.7%, respectively, and were higher in BCSP than non-BCSP procedures (PDR 53.7% vs. 43.3%; eANDR 5.7% vs. 3.9%). Higher polyp detection was associated with increasing age (odds ratio [OR] 1.03, 95%CI 1.03-1.04), male gender (OR 1.48, 95%CI 1.26-1.73), BCSP status (OR 1.50, 95%CI 1.28-1.77), adequate bowel preparation (OR 1.81, 95%CI 1.40-2.34), and England nation (OR 1.99, 95%CI 1.22-3.36). Our NED evaluation demonstrated high colonoscopy performance in Lynch syndrome, with superior quality and endoscopic yield in BCSP procedures, supporting national integration of surveillance to establish Lynch syndrome-specific quality benchmarks.

The cardiovascular-kidney-metabolic (CKM) syndrome framework integrates the shared pathophysiology of cardiovascular disease, excess adiposity, diabetes, and chronic kidney disease into a unified staging framework. Clonal haematopoiesis of indeterminate potential (CHIP), the age-related expansion of hematopoietic stem cells harbouring somatic mutations, has been linked to several CKM components, but its relationship with CKM syndrome is unknown. UK Biobank participants without a history of hematologic malignancy were included. Exposures included any CHIP, large CHIP, and major gene-specific CHIP subtypes. Outcomes were CKM stage and progression to stage 4 CKM syndrome during follow-up. Associations of CHIP with CKM stage and progression were tested using multivariable-adjusted logistic and Cox regression, respectively. Proteomic mediation analyses prioritized circulating proteins associated with CKM progression. Among 451,460 participants (mean [SD] age, 56.5 [8.1] years; 245,055 females [54.3%]), 15,486 (3.4%) had CHIP. CHIP was associated with higher CKM stage, driven by associations with non-DNMT3A CHIP, including TET2 (adjusted OR [aOR], 1.10; 95% CI, 1.00-1.20; false discovery rate [FDR]-corrected P = 0.045) and JAK2 CHIP (aOR, 1.68; 95% CI, 1.20-2.35; FDR-corrected P = 0.003). Over a median (IQR) 13.5 (12.6-14.3) years of follow-up, CHIP was independently associated with progression to stage 4 CKM, again driven by associations with non-DNMT3A CHIP subtypes (aHR, 1.24; 95% CI, 1.17-1.31; FDR-corrected P < 0.001). Proteomic mediators of progression to stage 4 CKM in non-DNMT3A CHIP were enriched for immune and inflammatory signalling pathways. These findings identify non-DNMT3A CHIP as a marker of CKM progression and support inflammatory mechanisms linking CHIP to CKM syndrome.

The profound phenotypic heterogeneity of circulating tumor cells (CTCs) presents a major analytical challenge, demanding technologies capable of high-plex, quantitative single-cell profiling. Here, we introduce PRISM (Phenotypic Resolution via Immuno-SERS Mapping), a biosensing platform engineered to meet this challenge. The platform integrates a high-efficiency dual-antibody (anti-EpCAM/anti-CSV) capture substrate with a suite of seven spectrally orthogonal SERS nanoprobes, enabling crosstalk-free, multiplexed quantification of epithelial, mesenchymal, and stem-like (E-M-S) markers. We demonstrate the platform's robust analytical performance, including high capture efficiency for heterogeneous cell lines and excellent linearity. Its superior phenotypic resolving power was validated by quantitatively distinguishing canonical cell line archetypes and tracking dynamic protein expression shifts during induced epithelial-mesenchymal transition. Applying PRISM to CTCs from pancreatic cancer patients, we introduce a novel data analysis framework, including a 'Metastasis Potential Score' (MPS), to translate high-dimensional spectral data into a clinically relevant metric for risk stratification. Furthermore, longitudinal analysis of patient samples demonstrates the platform's utility as a dynamic monitoring tool, capable of tracking therapy-induced phenotypic shifts. PRISM establishes a powerful analytical methodology for high-dimensional single-cell analysis, providing a robust tool for both fundamental cancer biology research and translational clinical applications.

The role of preoperative radiotherapy after neoadjuvant chemotherapy for pancreatic cancer remains controversial. To evaluate whether the addition of radiotherapy to chemotherapy is associated with improved oncologic outcomes in patients undergoing resection for pancreatic cancer. This was a retrospective propensity score-matched cohort study of patients undergoing pancreatic cancer resection between 2015 and 2023, with a median follow-up of 44 months. Patients were recruited from 2 high-volume referral centers. Of patients undergoing pancreatectomy after neoadjuvant treatment for pancreatic ductal adenocarcinoma, those treated with fluorouracil, leucovorin, irinotecan, and oxaliplatin or gemcitabine/nab-paclitaxel with or without preoperative radiation met eligibility criteria. Patients with metastatic disease, surgery-related mortality, or follow-up less than 12 months were excluded. Treatment groups were matched 1:1 on age, sex, comorbidity, tumor characteristics, baseline carbohydrate antigen 19-9 level, resectability status, and chemotherapy exposure. Neoadjuvant chemotherapy with or without radiotherapy. Event-free survival, pattern of recurrence, and overall survival. Of 1201 consecutive patients undergoing pancreatectomy after neoadjuvant treatment for pancreatic ductal adenocarcinoma, 869 met eligibility criteria. Among 800 included patients (median [IQR] age, 65 [58-71] years; 402 men [50.2%]), 438 received neoadjuvant chemotherapy and radiotherapy and 362 chemotherapy alone. After matching, 226 patients per group were analyzed; most had resectable (117 of 226 [51.1%]) or borderline-resectable (104 of 226 [46.5%]) disease. Radiotherapy was associated with higher rates of pathologic complete response (15 of 226 [6.6%] vs 10 of 226 [4.4%]), node-negative disease (139 of 226 [61.5%] vs 76 of 226 [33.6%]), and R0 resection (192 of 226 [85.0%] vs 118 of 226 [52.2%]). Recurrence patterns were similar, with distant recurrence as the predominant first site of failure in both groups (116 of 226 [51.3%] vs 109 of 226 [48.3%]). In competing risk analysis, the cumulative incidence of locoregional recurrence did not differ (subdistribution hazard ratio [HR], 0.90; 95% CI, 0.53-1.54; P = .71). Radiotherapy was not associated with improved event-free survival (HR, 1.10; 95% CI, 0.88-1.37; P = .38) or overall survival (HR, 1.02; 95% CI, 0.79-1.32; P = .84). Results of this cohort study suggest that although the addition of radiotherapy to neoadjuvant chemotherapy was associated with an improved pathologic response, it was not associated with survival benefit or reduced recurrence in resectable and borderline-resectable pancreatic cancer. These findings highlight the limited impact of locoregional control in a disease driven by systemic progression.

Beneficial effects of the gut commensal Agathobacter rectalis (Ar) are reported in diseases, yet its role in colorectal cancer (CRC) remains unclear. Here, metagenomic analysis revealed consistent fecal Ar depletion across CRC cohorts. In Apc min/+ mice, Ar inhibited colon tumorigenesis, reducing tumor number and volume versus E. coli and PBS controls. LC-MS/MS metabolomics showed decreased fecal cholesterol and altered lipid/cholesterol pathways after Ar treatment. In vitro, Ar-conditioned medium suppressed CRC cell growth, clonogenicity, migration, and cell cycle progression. LC-MS/MS identified (-)-epigallocatechin (EGC) as an Ar-derived metabolite absent in control bacteria. EGC recapitulated Ar-mediated anti-CRC effects in vitro and in vivo, with metabolic changes linked to lipid/cholesterol pathways. Transcriptomics showed that EGC suppressed SREBP signaling, cholesterol metabolism, and MAPK pathways. Mechanistically, EGC reduced nuclear SREBF2 and its transcriptional activity, downregulated cholesterol synthesis/metabolism genes, including FDPS and PCSK9, and suppressed MAPK signaling. Molecular docking suggested that EGC may bind pSREBF2 or SCAP. Cellular thermal shift assay revealed that EGC interacts with and stabilizes pSREBF2, but not SCAP. Co-immunoprecipitation demonstrated that EGC reduces pSREBF2-SCAP interaction, thereby inhibiting SCAP-mediated SREBF2 cleavage activation. Together, these findings define an Ar-EGC microbe-metabolite axis and support Ar/EGC-based interventions targeting cholesterol metabolism in CRC.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by high resistance to anticancer therapies, which is caused in part by hypovascularization and dysfunctional vessels that inefficiently deliver chemotherapy. Exercise has been shown to improve tumor vascular function and chemotherapy efficacy in preclinical models. Uncovering the mechanism by which exercise modifies tumor vasculature could help identify potential strategies to improve drug delivery. In this study, we evaluated surgically resected PDAC from patients who exercised during neoadjuvant chemotherapy to demonstrate that exercise remodeled PDAC vasculature and improved tumor vascular function in patients. In mice bearing orthotopic PDAC and treadmill exercise, tumor vascular remodeling was dependent on sphingosine-1-phosphate receptor 1 (S1PR1) signaling in endothelial cells, and S1PR1 was necessary for improved chemotherapy efficacy by exercise. Exercise activated S1PR1 in tumor endothelium, improved tumor vascular function, and increased gemcitabine (Gem) delivery and efficacy. Mice with an endothelial cell-specific S1PR1 deletion did not display exercise-induced improvements in tumor vascular function or Gem efficacy. These findings demonstrate that exercise increases chemotherapy delivery and efficacy by improving vascular function, defining S1PR1 as a necessary mediator of exercise-induced vascular remodeling. Aerobic exercise induces tumor vasculature remodeling via endothelial S1PR1 to enhance drug delivery and suppress pancreatic cancer progression, supporting discovery of exercise efficacy biomarkers and therapeutic strategies for patients unable to exercise.

Recovering high-quality microbial genomes from metagenomic sequencing data is essential for accurate profiling and understanding microbial variation. However, existing clustering methods often suffer from limited accuracy and scalability. Here we present MetaCAT (Metagenome Clustering and Association Tool), a framework that combines recovery of microbial genomes from metagenomic data and analysis of their associations with host traits. MetaCAT incorporates a Sparse Weighted Dirichlet Process Gaussian Mixture Model (SWDPGMM) to accurately and efficiently decompose complex datasets and combines k-mer frequency with read coverage to improve genome reconstruction. It also provides a dedicated workflow for microbial single-nucleotide polymorphism identification and metagenome-wide association studies with the host. MetaCAT outperforms existing methods in both clustering accuracy and computational efficiency across diverse datasets. Using metagenomic data from colorectal cancer cohorts, it revealed previously unrecognized marker species and microbial single-nucleotide polymorphisms associated with colorectal cancer. MetaCAT provides a scalable framework for microbial community profiling and advances our understanding of host-microbe interactions.

Extracellular matrix (ECM) stiffness is a critical biomechanical factor that plays a key role in tumor progression, influencing tumor behavior and driving its malignancy. Transcriptional co-activator yes-associated transcriptional regulator (YAP) is a well-established mechanosensitive regulator; however, the molecular mechanisms by which matrix stiffness-mediated YAP activation promotes tumor progression through transcriptional regulation remain incompletely defined. Here, using cross-omics analysis, in vitro, and in vivo experiments, we identified ATPase Na+/K+ transporting subunit alpha 1 (ATP1A1) as a novel target of YAP/TEA domain transcription factor 4 (TEAD4) mechano-responsive transcription factor complex, whose increased expression in stiff matrix promotes tumor cell proliferation. Mechanistically, we revealed that stiff matrix promotes YAP/TEAD4 binding to the ATP1A1 promoter, thereby upregulating its expression. Increased ATP1A1 in turn promotes intracellular calcium signaling and activates the nuclear factor-kappa B (NF-κB) pathway, thereby promoting C-X-C motif chemokine ligand 1 (CXCL1) expression and secretion, and subsequently enhances tumorigenic potential. These findings demonstrate that increased matrix stiffness activates YAP/TEAD4-dependent transcriptional program and promotes ATP1A1 expression, thereby converting mechanical cues into chemical signaling output and finally promoting malignant phenotypes in the tumor. Furthermore, these findings suggest that CXCL1 may contribute to stiffness-associated tumor progression and warrant further investigation as a potential therapeutic target.

Colibactin is a bacterial genotoxin that damages DNA and is linked to colorectal cancer. It is implicated in interbacterial competition but its role in natural microbiomes is unclear. Here we show that Frischella perrara, a colibactin-producing symbiont restricted to honeybee guts, lowers bee lifespan but increases survivorship following challenge with the opportunistic bee pathogen Serratia marcescens. F. perrara mutants lacking colibactin production fail to improve survivorship following challenge. The symbiont reduces pathogen loads by causing colibactin-dependent DNA damage and prophage induction. Colibactin is toxic to the pathogen under anoxic conditions, reflecting the bee gut. clbS, a gene conferring protection from colibactin, is ubiquitous among bacteria restricted to bee guts but absent from opportunistic colonizers, including pathogens. Indeed, F. perrara colonization representing colibactin pressure had limited impact on the bee gut microbiome where strains encode clbS. Our findings suggest that colibactin functions in interbacterial competition and protects against pathogens while permitting co-evolved symbionts to persist.

Hepatocellular carcinoma (HCC), an aggressive type of liver cancer, has limited treatment options, and chemotherapy remains an important clinical approach. This study investigates the role of dual-specificity phosphatase 12 (DUSP12) and its interaction with the nucleolar protein N-acetyltransferase 10 (NAT10) in HCC models under genotoxic stress. We demonstrate that doxorubicin (DX) induces greater cytotoxicity than cisplatin, correlating with a stronger DNA damage response (DDR), nucleolar stress, and NAT10 relocalization. CRISPR-Cas9-mediated DUSP12 knockout sensitized cells to DX, increasing markers of DNA damage (γH2AX, p53) and delaying DNA break repair. This was accompanied by redistribution of nucleolar proteins NAT10 and TCOF1. Remodelin treatment enhanced DX sensitivity in DUSP12 knockout cells, suggesting a context-dependent interaction between DUSP12 loss and the NAT10 inhibition. Protein interaction assays confirmed that DUSP12 binds NAT10, corroborating a regulatory interaction. DUSP12 deletion increased NAT10 phosphotyrosine levels and reduced N4-acetylcytidine (ac4C) RNA modification, consistent with altered NAT10-dependent RNA acetylation. Analysis of patient data revealed frequent DUSP12 amplification in HCC, whereas elevated DUSP12 expression was associated with poor survival and enrichment of DDR- and ribosome biogenesis-related transcriptional programs. Our findings identify the DUSP12-NAT10-ac4C axis as a novel molecular link between DDR and nucleolar stress, unveiling a potential therapeutic vulnerability in HCC.

Given the limited efficacy of existing therapies for metastatic colorectal cancer (mCRC), there is an urgent need for novel strategies. Prohibitin 1 (PHB1) is significantly upregulated in CRC, where it plays a critical role in oxidative phosphorylation (OXPHOS) to meet the heightened energy demands of rapid tumor growth and metastasis. Here, we discover that PHB1 interacts with NADH: ubiquinone oxidoreductase core subunit S1 (NDUFS1), a subunit of the OXPHOS complex, thereby modulating mitochondrial respiratory function. Based on these insights, we developed TD6, a highly potent and selective small-molecule inhibitor of PHB1. In a mouse model of colorectal cancer pulmonary metastasis (CRPM), TD6 treatment significantly prolonged animal survival. Mechanistically, TD6 binds to PHB1 and induces a conformational change in the PHB complex, which reduces PHB-mediated stabilization of NDUFS1. Disruption of the PHB-NDUFS1 interaction promotes NDUFS1 degradation via the lysosomal pathway, leading to impaired activity of mitochondrial complex I (MCI) and reduced OXPHOS function. In summary, PHB1 maintains MCI stability and OXPHOS activity through its interaction with NDUFS1. By targeting PHB1, TD6 effectively disrupts this regulatory axis, demonstrating therapeutic potential against CRPM. This study presents a novel drug candidate and a theoretical foundation for PHB1-based interventions targeting cancer energy metabolism.

The technical implementation of volumetric magnetic resonance imaging (MRI) has the potential to substantially advance tumor tracking in MRI-guided radiotherapy (MRIgRT). However, existing three-dimensional cine MRI techniques remain constrained by tradeoffs between spatial and temporal resolution, with spatial resolution often limited to a voxel spacing of 5-6 mm. In this study, we proposed a respiratory motion augmentation (RMA) method for personalized super-resolution (pSR) reconstruction to achieve sufficiently high spatial and temporal resolutions for real-time volumetric MRI. The proposed method enhances the motion robustness of pSR networks by synthesizing high-resolution MR images across multiple respiratory phases using deformable image registration (DIR). A conventional SR network was initially trained on a public dataset of 78 patients and subsequently personalized using 1.5-T MR-LINAC data from 12 cancer patients (10 abdominal and 2 prostate cancer cases). For network personalization, the proposed RMA approach was applied by deforming a breath-hold high-resolution image to match free-breathing low-resolution images using DIR, thereby generating multiple paired datasets for each patient. The RMApSR network substantially improved both image quality and segmentation accuracy. The network achieved increases of 1.7% in the peak signal-to-noise ratio, 6.2% in the structural similarity index measure, and 10.7% in the Dice similarity coefficient compared with those of the conventional pSR while maintaining consistent performance across respiratory phases. Personalized training, which requires 13.2 min, can be performed in parallel with standard MRIgRT procedures. Combined with an inference time of 72 ms per volume, our approach demonstrates high clinical feasibility. This motion-aware personalization strategy represents a significant advancement toward achieving high spatiotemporal resolution requirements for volumetric MRIgRT implementation.

Enterotoxigenic Bacteroides fragilis (ETBF), which secretes B. fragilis toxin (BFT) and has been associated with colorectal cancer, can be enriched in patients with gall bladder cancer (GBC). Whether and how ETBF contributes to GBC is unclear. Here we confirm, through analysis of patient samples, that ETBF is enriched in GBC tumours, while experiments in mice show that ETBF colonizes the gall bladder. In vitro and in patient-derived organoids, ETBF promoted GBC proliferation. ETBF also promoted tumour growth in a BFT-dependent manner in a mouse GBC allograft model. Mechanistically, the tumorigenic activity of BFT was dependent on the ETBF surface protein, membrane-bound lytic murein transglycosylase D (MltD), which interacts with the host receptor protein, transmembrane serine protease 13 (TMPRSS13). This interaction activated JAK2-STAT3 signalling to promote tumorigenesis. BFT also activated NF-κB signalling, which increased CXCL1 secretion, leading to myeloid-derived suppressor cell recruitment and angiogenesis in the tumour microenvironment. Taken together, these findings uncover mechanisms through which ETBF facilitates GBC development, with potential promise as therapeutic targets to limit GBC progression.

Accurate delineation of pancreatic tumors on Magnetic Resonance Imaging (MRI) is important for diagnosis, radiotherapy treatment planning, and outcome assessment, but remains challenging due to complex anatomy and subtle tumor appearance. In routine practice, tumor contours on MRI are produced manually, which is time-consuming and subject to inter-observer variability. Radiotherapy on MRI-Linear Accelerator (MRI-Linac) systems further requires fast and consistent Gross Tumor Volume (GTV) contours for online adaptation, yet most public pancreas tumor segmentation benchmarks focus on Computed Tomography (CT). The Pancreatic Tumor Segmentation in Therapeutic and Diagnostic MRI (PANTHER) challenge addresses this gap by benchmarking automatic pancreatic tumor segmentation on MRI. The dataset includes contrast-enhanced T1-weighted diagnostic MRI and T2-weighted MRI-Linac scans with expert pancreas and tumor annotations, organized into two tasks: (1) tumor segmentation on diagnostic MRI and (2) tumor segmentation on MRI-Linac images. Performance was evaluated using overlap metrics, distance-based metrics, and tumor volume error. The challenge attracted 285 registered participants, with 12 and 9 final submissions for Tasks 1 and 2, respectively. On diagnostic MRI, top methods achieved performance close to inter-reader agreement. Multi-reader analysis suggested that models often reproduced the contouring style of the training annotator, highlighting the importance of annotation quality and consensus. In contrast, performance on MRI-Linac images was lower and more heterogeneous, including cases of complete localization failure. PANTHER provides the first public benchmark for pancreatic tumor segmentation on MRI, showing that clinically useful automation is feasible on diagnostic MRI, while robust MRI-Linac GTV segmentation remains an open challenge.

Hepatitis B virus (HBV) infection remains a severe global public health challenge, with hepatocellular carcinoma being a primary cause of HBV-related mortality. Occult HBV infection (OBI) represents a distinct type of HBV infection that has been increasingly linked to hepatocellular carcinoma development, yet the precise molecular mechanisms underlying this association remain poorly elucidated. Although HBV pre-S deletion mutations have been shown to enhance cell proliferation and contribute to hepatocarcinogenesis, the biological functions of other types of pre-S mutations, particularly point mutations, are still mostly unexplored. In our prior studies, we identified several high-frequency pre-S point mutations from OBI blood donors. Within this research, we systematically explored the effects of these OBI-associated pre-S mutations on host cell proliferation and assessed their potential oncogenic properties. Cell proliferation assays revealed that several pre-S mutations significantly enhanced the proliferative capacity of host cells. Mechanistically, five pre-S mutations (E39K, D44N, N98T, H128R, and I161T) activated the Akt/mTOR signaling cascade, up-regulated Cyclin D1 expression, and induced G1-to-S phase cell cycle progression. Further analyses suggested that the large HBV surface protein (LHBs) likely acts as the key mediator linking pre-S mutations to signaling activation and cellular proliferation. These findings provide novel mechanistic understandings of the oncogenic potential of pre-S point mutations in hepatocarcinogenesis and may facilitate the identification of high-risk individuals within OBI populations as well as the development of treatment strategies for hepatocellular carcinoma linked to HBV.

The invasive and metastatic potential of hepatocellular carcinoma (HCC) is tightly linked to lipid metabolic reprogramming. However, existing knowledge of the molecular regulatory network governing lipid metabolism in HCC remains incomplete. This study reveals for the first time that ADAR1 promotes HCC progression via direct binding to PPARγ mRNA to regulate lipid metabolism. Through multi-omics approaches (publicly available single-cell sequencing databases, clinical sample analysis, and cellular models), we showed that ADAR1 was aberrantly up-regulated in HCC and promoted tumor cell proliferation, migration, and invasion. The adenosine analog 8-chloroadenosine (8-Cl-Ado) dose- and time-dependently down-regulated ADAR1 expression. Transcriptomic analysis revealed that 8-Cl-Ado significantly suppressed key genes associated with cholesterol synthesis and fatty acid metabolism. Mechanistically, ADAR1 binds to PPARγ mRNA, thereby activating the PPAR signaling axis, while PPARγ knockdown significantly abrogates malignant phenotypes in HCC. Functional rescue experiments confirmed that overexpression of the ADAR1 p150 isoform rescued the tumor-suppressive phenotype induced by 8-Cl-Ado. Collectively, these findings demonstrate that 8-Cl-Ado inhibits hepatocarcinogenesis and progression by suppressing ADAR1 and subsequently regulating PPARγ-mediated lipid metabolic processes, providing novel therapeutic targets and potential intervention strategies for HCC.

Our previous preclinical study determined artesunate as a candidate drug for hepatocellular carcinoma (HCC) and identified glucosylceramidase (GBA) as one of its direct targets. This research aimed to identify the binding sites of GBA with artesunate and the potential anti-HCC mechanisms, which remain unclear. Artesunate effectively suppressed cell viability and proliferation, and enhanced apoptosis of HCC cell lines with more sensitivity in HepG2 than MHCC-97H cells. Network calculation and a series of in vivo and in vitro experimental data demonstrated that the apoptosis-related GBA-ceramide-CTSD-BID-BAX signaling was one of the key putative target pathways by which artesunate may inhibit the malignant progression of HCC. Furthermore, through integrated computational and experimental approaches, we identified Y313, E340, and N396 as critical binding residues within the GBA active site. Mutagenesis studies revealed that these residues were indispensable for the interaction, with E340R and N396R mutations exhibiting the most pronounced impairment in binding affinity and enzymatic activity, respectively. Crucially, disrupting this binding interface abolished artesunate's ability to modulate the downstream apoptotic pathway. Our findings provide the first structural and mechanistic elucidation of artesunate's target engagement with GBA, unveiling a specific signaling cascade for its anti-HCC activity and establishing a foundational framework for developing novel GBA-targeted therapies.

Non-rigid registration is essential for augmented reality-guided laparoscopic liver surgery, as it enables the fusion of preoperative information such as tumor location and vascular structures into the limited intraoperative view, thereby enhancing surgical navigation. A prerequisite is the accurate prediction of intraoperative liver deformation, which remains highly challenging due to factors such as large deformation caused by pneumoperitoneum, respiration and tool interaction as well as noisy intraoperative data, and limited field of view due to occlusion and constrained camera movement. To address these challenges, we introduce PIVOTS, a Preoperative to Intraoperative VOlume-To-Surface registration neural network that directly takes point clouds as input for deformation prediction. The geometric feature extraction encoder allows multi-resolution feature extraction, and the decoder, comprising inter-modality cross attention modules, enables information exchange between pre- and intraoperative features and accurate multi-level displacement prediction. We train the neural network on a large synthetic dataset created using a biomechanical simulation pipeline that explicitly targets the mentioned intraoperative challenges and validate its performance on both synthetic and real datasets. Results demonstrate superior registration performance of our method compared to baseline methods, exhibiting strong robustness against high amounts of noise, large deformation, and various levels of intraoperative visibility. The network is fast enough to run multiple times per second and directly generalizes to new patients without retraining. We publish training and test sets as evaluation benchmarks in an effort to contribute to the development of more robust liver registration methods based on volume-to-surface data. Code, docker container and datasets are available athttps://github.com/pengliu-nct/PIVOTS.

ADAR1 is overexpressed in hepatocellular carcinoma (HCC) and has been linked to poor prognosis, metastasis, and recurrence; however, its precise functions and underlying mechanisms, particularly in the context of metastasis, remain inadequately elucidated. This study seeks to elucidate the functions and underlying mechanisms of the most abundantly expressed isoform of ADAR1, ADAR1p110, in the setting of HCC metastasis. In this study, hepatocyte-specific ADAR1p110 knock-in mice and engineered HCC cell lines were utilized to investigate the function of ADAR1p110 in vivo and in vitro. Genome sequencing, transcriptome sequencing, microRNA sequencing, RNA immunoprecipitation qPCR (RIP-qPCR), and RNA pull-down assays were performed to elucidate the mechanism of ADAR1p110 in HCC. We demonstrated that ADAR1p110 overexpression promotes HCC metastasis by improving the motility of HCC cells. Mechanistically, ADAR1p110 overexpression increases TUBA1A expression, which plays a crucial role in regulating HCC cell motility. At the molecular level, ADAR1p110 suppresses miR-451a biogenesis by competitively binding to pri-miR-451a, thereby preventing its cleavage by the Drosha/DGCR8 complex. Furthermore, we confirmed that TUBA1A is a direct downstream target of miR-451a.

Advanced gastric cancer (GC) poses a significant threat to public health, leading to substantial consumption of healthcare resources due to its aggressive nature and poor prognosis. Hairy and enhancer of split 6 (Hes6), a member of the mammalian homologs of Drosophila's hairy and enhancer of split (HES) family, functions as a transcriptional cofactor owing to its unique structural features. Previous studies have indicated that Hes6 expression is elevated in several malignant tumors, contributing to the enhanced proliferation and invasion of tumor cells. However, the precise role and underlying mechanisms of Hes6 in gastric cancer initiation and progression remain poorly understood. In this study, we demonstrate that Hes6 expression is significantly up-regulated in gastric cancer tissues, with elevated levels of Hes6 correlating with poor prognosis in certain patient cohorts. Functionally, Hes6 mediates gastric cancer cells to obtain stronger proliferation, migration, and invasion abilities by activating the PI3K/AKT signaling pathway, thereby accelerating tumor progression. Moreover, Hes6 interacts with the Twist1 protein, stabilizing it and facilitating the epithelial-mesenchymal transition (EMT). Collectively, these findings offer valuable insights into the regulatory mechanisms underlying gastric cancer progression, highlighting the potential of transcriptional cofactors, such as Hes6, in PI3K/AKT signaling as promising targets for therapeutic intervention in gastric cancer.

Increasing evidence indicates that the galectin family played a significant role in tumor progression and is closely related to the hypoxic microenvironment within tumor tissues. However, the regulatory mechanisms behind this process are unexplored. In this study, we found that Gal-1 expression was significantly up-regulated in hepatocellular carcinoma (HCC) tissues and was closely associated with poor prognosis of patients. Intervention of Gal-1 at the cellular level significantly inhibited the malignant phenotype of hepatoma cells. For the first time, we discovered that Gal-1 was regulated by m6A modification, ALKBH5 mediated the demethylation of Gal-1 mRNA, and YTHDF2 recognized Gal-1 mRNA and altered its stability. This regulatory process was altered under hypoxic conditions, and hypoxia-inducible factors (HIFs) mediated the regulation of m6A modification in hepatoma cells by hypoxia. HIF-1α bound to the promoter region of ALKBH5 and up-regulated ALKBH5 expression, while HIF-2α bound to the promoter region of YTHDF2 and generated negative regulation. In vivo, after intervention of Gal-1, reduction of proliferative markers and inhibition of epithelial-mesenchymal transition occurred in the subcutaneous tumor. The use of LNP-siGal-1 also inhibited epithelial-mesenchymal transition.

The incidence of esophageal adenocarcinoma (EAC) is rapidly increasing in Western countries. Gastroesophageal reflux, containing acidic bile salts (ABS), is the main risk factor for EAC. Cancer cells develop adaptive abilities to recalibrate redox balance via hijacking the antioxidant systems to maintain reactive oxygen species (ROS) below lethal levels. We investigated the role of PRDX2 and its regulation in EAC chemoresistance under reflux conditions. We analyzed public databases to identify PRDX2's aberrant overexpression and potential role in chemoresistance in EAC. To model acute and chronic GERD in vitro, we applied transient and repeated ABS exposures. Using 2D and 3D organotypic culture models of EAC cells, we investigated PRDX2's anti-ferroptosis, pro-chemoresistance functions, and regulatory mechanisms. In addition, we also utilized patient derived organoids and xenografts, cell line-derived xenografts, and human EAC tissue microarrays. Aberrant expression of PRDX2 was detected in both human EAC tissues and cell lines under ABS exposure. PRDX2 was regulated via an APE1-redox-dependent transcription activation of NF-kB. The knockdown of PRDX2 impaired the recovery of EAC cells from ABS-induced ROS and ROS-dependent lipid peroxidation. Silencing PRDX2 sensitized the chemo-resistant EAC cells to oxaliplatin by enhancing ferroptosis. Mechanistically, we found that PRDX2 inhibits ferroptosis by stabilizing GPX4, a crucial ferroptosis suppressor. PRDX2 enhances GPX4 stability through OTUB1-dependent deubiquitination, preventing its degradation. The APE1-redox inhibitor APX2009 significantly sensitized EAC cells to oxaliplatin by downregulating PRDX2 and inducing ferroptosis. Importantly, the combination of oxaliplatin and APX2009 showed synergistic tumor-suppressive effects in xenograft models. Our findings revealed a novel link between reflux-induced redox rebalance and ferroptosis-related chemoresistance in EAC via APE1-redox/NF-kB/PRDX2/OUTB1/GPX4 signaling cascade. Targeting redox with APE1-redox-specific inhibitors is a potential novel strategy for drug combination in refractory EAC, via inhibition of PRDX2.

Cisplatin is widely employed in the treatment of gastric cancer (GC). However, the resistance mechanisms exhibited by GC cells often result in suboptimal clinical outcomes associated with cisplatin therapy. Autophagy, a self-degradative cellular process, plays a complex dual role in regulating tumor cell death and survival. In recent years, significant attention has been directed toward the relationship between autophagy and cisplatin resistance in GC, fostering the development of various autophagy-related drugs and potential targets aimed at enhancing the sensitivity of GC cells to cisplatin. Nevertheless, a comprehensive analysis of the correlations among relevant studies is still lacking. This review synthesizes recent research examining the impact of autophagy on cisplatin resistance in GC cells, with particular emphasis on existing drugs and potential therapeutic drugs/targets. It briefly explores the fundamental processes of autophagy and clarifies the relationship between autophagy mechanisms and GC. Furthermore, it summarizes the available drugs and potential candidates that can either enhance or inhibit autophagy, thereby improving GC cell sensitivity to cisplatin, alongside their underlying mechanisms. Additionally, it consolidates pertinent research findings to present a more thorough understanding of the intricate relationships between autophagy and cisplatin resistance in GC cells. We hope this review will encourage researchers to investigate novel mechanisms of cisplatin resistance in GC cells, discover new targeted therapies, and propose innovative strategies to tackle this challenge.

Hepatocellular carcinoma (HCC) exhibits diminished capacity for oxidative utilization of long-chain fatty acids (LCFAs). However, the strategic and mechanistic basis by which HCC cells enact metabolic reprogramming to adapt to impaired LCFAs oxidation and sustain viability remains incompletely defined. Here we report that solute carrier family 27 member 5 (SLC27A5), the specific transporter for LCFAs, is broadly downregulated in HCC cells, resulting in reduced LCFAs uptake. In HCC cells with impaired LCFAs oxidation, diminished LCFAs import caused by SLC27A5 loss does not lead to energy deficiency, but instead prevents lipotoxicity derived from unutilized LCFAs, thereby supporting HCC cell growth. Impaired LCFAs oxidation suppresses peroxisome proliferator-activated receptor alpha (PPAR-α) signaling, which in turn represses SLC27A5 transcription, accounting for the widespread downregulation of SLC27A5 in HCC. Owing to reduced LCFAs uptake, HCC cells with low SLC27A5 rely on the glutamine reductive pathway for fatty acid biosynthesis to maintain total fatty acid levels, rendering these cells highly sensitive to glutaminase inhibition. In conclusion, we demonstrate that SLC27A5 downregulation represents a response to defective LCFAs oxidation in HCC, and reduced LCFAs uptake consequent to low SLC27A5 expression constitutes a survival adaptation that enables HCC to tolerate impaired LCFAs oxidation. Glutaminase inhibitors may serve as a precision therapeutic strategy for HCC characterized by low SLC27A5 expression.

Oncolytic virus M1 is a promising anticancer agent; however, its therapeutic efficacy is often limited by insufficient intratumoral viral replication and host antiviral immunity. Sirolimus, an mTOR inhibitor widely used in transplantation immunosuppression, has demonstrated potential to modulate antiviral responses. This study investigates whether sirolimus potentiates the efficacy of M1 virotherapy and elucidates the underlying mechanisms. Sirolimus significantly enhanced the antitumor efficacy of M1 virus in murine prostate cancer and liver cancer models, leading to reduced tumor growth. This synergistic effect remained evident in CD8⁺ T cell-depleted mice, indicating that the therapeutic benefit is independent of adaptive cytotoxic immunity. Mechanistically, sirolimus markedly increased M1 viral replication in tumor tissues, accompanied by enhanced tumor cell-cycle arrest and apoptosis. Notably, sirolimus selectively amplified viral load within tumors but not in normal organs, demonstrating tumor-specific viral enrichment and safety. Further analyses revealed that this increase in intratumoral virus was driven by mTOR pathway inhibition rather than alterations in macrophage or NK cell populations. Transcriptomic profiling and molecular validation indicated that sirolimus-mediated mTOR suppression downregulated key type I interferon-stimulated genes (Ifitm1, Stat1, Ifit3), thereby attenuating intrinsic antiviral defenses and facilitating viral amplification. In summary, sirolimus potentiates M1 oncolytic virotherapy by selectively enhancing viral replication in tumors via mTOR inhibition and suppression of type I interferon signaling, independent of CD8⁺ T cell-mediated immunity. These findings establish a mechanistic rationale for combining mTOR inhibitors with oncolytic viruses to achieve dual benefits of enhanced viral oncolysis and controlled immunosuppression, with translational relevance for cancer patients requiring long-term immunosuppressive therapy.

5-Fluorouracil (5-FU) resistance in colorectal cancer (CRC) involves oxidative stress mechanisms, but the role of epitranscriptomic regulation remains unclear. This study investigates how oxidative post-translational modifications of the N6-methyladenosine (m6A) methyltransferase METTL16 contribute to 5-FU resistance. Parental (HCT8, HCT15) and 5-FU-resistant CRC cells were compared using redox proteomics, m6A-seq, RNA-seq, and functional assays. METTL16 S-glutathionylation was assessed via streptavidin pulldown and mass spectrometry. IGF2BP3's role was validated through knockdown/overexpression, patient-derived organoids (PDOs), and xenograft models. Clinical relevance was evaluated in 112 CRC patient tissues and data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Drug synergy was tested using isoliquiritigenin (ISO) combined with 5-FU. Resistant cells exhibited elevated global protein S-glutathionylation and reduced m6A RNA methylation. METTL16 underwent site-specific S-glutathionylation at Cys548, leading to its degradation and subsequent m6A loss. Integrative omics identified IGF2BP3 as a key METTL16 target: reduced m6A on IGF2BP3 mRNA enhanced its nuclear export, splicing, and overexpression. IGF2BP3 knockdown sensitized resistant cells to 5-FU in vitro and in vivo, while overexpression conferred resistance. Mechanistically, IGF2BP3 destabilized CFTR mRNA (an ABC transporter), reducing 5-FU uptake. Clinically, high IGF2BP3 correlated with poor survival and 5-FU resistance in CRC patients. Targeting IGF2BP3 with ISO synergized with 5-FU, overcoming resistance in cells and xenografts. Oxidative stress-induced METTL16 S-glutathionylation drives 5-FU resistance by reducing m6A modification, enabling IGF2BP3 overexpression and CFTR suppression. IGF2BP3 is a biomarker of clinical resistance, and its targeting with ISO represents a promising combinatorial strategy to restore 5-FU efficacy in CRC.

Although immunotherapy-based regimens have improved overall survival for some patients with advanced hepatocellular carcinoma (HCC), many patients ultimately experience disease progression. Macrophages are abundant in the HCC microenvironment and represent promising therapeutic effectors, yet the potential of combining chimeric antigen receptor macrophages (CAR-M) with macrophage checkpoint targeting remains underexplored. Given that glypican-3 (GPC3), a cell-surface heparan sulfate proteoglycan, is highly expressed in HCC, we developed a GPC3-targeted CAR-M system and evaluated its efficacy alone and in combination with USP22 inhibition. USP22, a deubiquitinating enzyme previously linked to tumor CD24 expression, was selected as a combinatorial target to potentially relieve CD24-associated phagocytic suppression. In vitro assays demonstrated that GPC3 CAR-M cells exhibited specific binding, phagocytosis, and potent cytotoxicity against multiple GPC3-positive HCC cell lines. RNA sequencing and flow cytometry revealed a statistically significant positive correlation between USP22 and the macrophage checkpoint CD24. The addition of the USP22 inhibitor significantly enhanced the tumor-killing capacity of CAR-M in a dose-dependent manner and was associated with reduced CD24 expression on tumor cells. This combinatory strategy robustly suppressed tumor growth in both a murine peritoneal dissemination model and a patient-derived xenograft (PDX) model of HCC. Importantly, the enhanced efficacy was consistently observed across CAR-M cells derived from THP-1, human monocyte-derived macrophages (hMDMs), and human pluripotent stem cells (hPSCs), underscoring its broad applicability. Our findings provide proof-of-concept preclinical evidence supporting further evaluation of combining GPC3 CAR-M therapy with USP22 inhibition as a potential immunotherapeutic strategy for HCC.

Heterojunctions, as semiconductor materials, possess excellent photocatalytic properties and are widely applied in fields such as disease treatment, biosensing, environmental remediation, and energy. However, heterojunctions still encounter issues of environmental sensitivity and low biocompatibility in practical applications, which limit their application in various fields. In recent years, the combination of heterojunctions with hydrogels to form hydrogel composites has been widely used in the medical field, environmental field, and biosensing field. This is because the encapsulation of heterojunctions by hydrogels can provide heterojunctions with good biocompatibility and increase their chemical stability. Heterojunction hydrogel composites can not only achieve efficient carrier separation but also enhance the adhesion and toughness of the interface. This paper focuses on the preparation and classification of heterojunction hydrogels, as well as the electron transfer pathways when different types of heterojunction hydrogels function, and evaluates different heterojunction hydrogels. In addition, the latest progress in the application of heterojunction hydrogels in antibacterial, wound healing, tumor treatment, environmental remediation, and biosensing is summarized. Finally, a deep discussion is conducted on the prospects and challenges of heterojunction hydrogels in practical applications. This review comprehensively introduces heterojunction hydrogels and aims to provide a new perspective for the development and application of heterojunction hydrogels.

Alpha-fetoprotein (AFP) is a key biomarker for hepatocellular carcinoma, but traditional testing methods are associated with radiation risks, high costs, and insufficient sensitivity. In this work, an S-type ZnO-polydopamine (PDA) heterojunction-based photoelectrochemical (PEC) immunoassay was fabricated through a two-step thermal method, combined with cascade enzymatic reactions for the sensitive AFP quantification. The unique S-scheme charge transfer mechanism and Zn-O-C interfacial chemical bonding were rationally designed, and PDA acted as an electron buffer layer with π-π conjugated structure to optimize interfacial charge transport. Through band structure analysis combined with first-principles theoretical calculations, an S-type electron transfer pathway within the heterojunction was identified, which effectively retained the carriers with high redox capacity and accelerated the charge separation. The signal amplification system for the cascading enzymatic reaction further amplified the detectable signal by catalyzing the generation of ascorbic acid, thereby enabling a highly sensitive determination of target AFP at a picogram level (0.497 pg mL-1). Importantly, this strategy integrates S-scheme heterojunctions with enzymatic signal amplification, providing a promising platform for the high-performance PEC detection of low-abundance proteins in biological fluids.

Point-of-care testing (POCT) platforms frequently suffer from a fundamental bottleneck: while advances in molecular amplification improve signal intensity, the reliability of signal readout in complex clinical matrices remains poorly controlled. Here, we present an integrated biosensing framework that treats readout reliability as an explicit engineering objective rather than a post hoc correction problem. The platform integrates three complementary components: (i) a heptameric nanobody probe employed as a multivalent recognition element for target capture, (ii) a DNA-assisted clustering interface that spatially organizes gold nanoparticle reporters for robust signal amplification, and (iii) a few-shot learning module based on Prototypical Networks that enables robust classification with minimal training data while providing interpretable decision-making through metric-based reasoning. Alpha-fetoprotein was selected as the model analyte because it remains a clinically important biomarker for hepatocellular carcinoma screening and follow-up, while also representing a realistic POCT challenge in which clinically meaningful detection must be achieved with low instrumentation burden and reliable readout under matrix variability. In this setting, the system achieves a visual limit of detection of 2 ng/mL and demonstrates quantitative consistency across representative clinical serum samples. Importantly, the AI module functions as an integral system component, identifying diagnostically relevant regions and mitigating readout uncertainty arising from matrix effects and imaging variability. By jointly engineering the sensing interface and the interpretive layer, this work establishes a generalizable strategy for constructing trustworthy POCT systems in which chemical signal generation and digital interpretation are co-designed.

Accurate early screening for hepatocellular carcinoma (HCC) requires multi-target detection methods with high sensitivity and strong anti-interference capability. In this study, we developed an aptamer-mediated SERS sensing method based on bimetallic magnetic nanotubes for the simultaneous detection of HCC markers alpha-fetoprotein (AFP) and Golgi protein 73 (GP73). Tubular Fe3O4 nanostructures were innovatively synthesized as magnetic carriers, onto which a gold-layer, an internal standard molecule (4-mercaptophenylboronic acid), and a silver-layer were sequentially deposited to construct a capture substrate integrating signal calibration and electromagnetic enhancement functions. Raman tags with non-overlapping spectra in the Raman-silent region (1800-2500 cm-1) were screened to establish a sandwich detection system. Using a portable Raman spectrometer and intelligent software, this method achieved rapid quantification of AFP and GP73, with detection limits of 0.433 pg/mL and 0.370 pg/mL, respectively, and recovery rates in spiked serum samples ranging from 95.62% to 106.2%. By simply replacing the aptamers, this method can be readily extended to other targets, demonstrating excellent versatility. This work provides a reliable analytical tool for early HCC diagnosis and establishes a foundation for developing on-site multi-target detection systems for complex samples.

Radiotherapy is essential for cancer treatment, yet radiation-induced skin injury (RISI) remains a major clinical challenge due to reactive oxygen species (ROS) accumulation, metabolic dysregulation, and the limited efficacy of single-modality therapies in modulating the oxidative-inflammatory microenvironment. To overcome this limitation, we have developed a multifunctional injectable hydrogel, HCG@CDs, by cross-linking biocompatible carboxymethyl chitosan (CMCS) with oxidized hyaluronic acid (OHA) conjugated to the Glycyl-L-Histidyl-L-Lysine-Copper(II) complex (GHK-Cu 2 + ) via dynamic Schiff-base linkages. Carbon dots (CDs) possessing superoxide dismutase (SOD)-like activity are uniformly dispersed within this three-dimensional dynamic network, creating an integrated platform for full-cycle therapy. The system exhibits intelligent, pH-responsive release behavior, whereby CDs are rapidly liberated in the acidic wound microenvironment to efficiently scavenge ROS and mitigate early-stage oxidative stress. Subsequently, GHK-Cu 2 + is released in a sustained manner to synergistically promote tissue repair by modulating inflammation, enhancing cell migration and proliferation, and facilitating collagen deposition. In vitro and in vivo evaluations have confirmed that the HCG@CDs hydrogel effectively alleviates radiation-induced oxidative damage and inflammatory responses, significantly accelerating the healing of skin wounds. Overall, this multifunctional hydrogel demonstrates great potential in accelerating the healing of RISI through multi-target synergistic regulation, highlighting its significant promise for clinical wound management and skin regeneration.

The therapeutic gain ratio (TGR) of radiotherapy for hepatocellular carcinoma (HCC) remains limited by two major barriers: insufficient precision in adaptive radiotherapy (ART) on the physical dimension and the lack of effective radiosensitization on the biological dimension. Although advances have been made separately in accurate dose delivery and tumor-sensitizing strategies, no approach has yet integrated both dimensions to achieve a coordinated improvement in TGR, representing a critical gap in current practice. In this study, we propose an integrated physical-biological strategy that combines nanomaterials with artificial intelligence (AI). We first constructed charge-engineered gold-platinum nanoparticles that respond to the acidic tumor microenvironment and enable prolonged, high-contrast computed tomography imaging of HCC. These enhanced images were then used to develop the first Transformer-convolutional neural network hybrid model (3D STS-Net) tailored for this scenario, enabling high-accuracy three-dimensional segmentation of small HCC for image-guided adaptive radiotherapy. In parallel, we systematically evaluated the nanoparticles' radiosensitizing effects in vitro and in vivo. The nanoparticles provided stable imaging enhancement for up to 120 h and markedly improved tumor-liver contrast. The 3D STS-Net achieved high segmentation accuracy, supporting more precise contouring for HCC ART. Moreover, the nanoparticles significantly increased radiation-induced reactive oxygen species and enhanced tumor control in animal models. Together, these findings demonstrate that the proposed strategy simultaneously strengthens radiotherapy performance in both physical and biological dimensions, leading to a coordinated improvement in TGR. This integrated "nanomaterial-AI" framework offers a systematic and generalizable approach for enhancing radiotherapy effectiveness in HCC.

Detecting chromosomal copy-number alterations together with protein-defined cell states in intact tissue is critical for understanding early clonal evolution and microenvironmental interactions in cancer. We developed ORION-FISH, which integrates high-plex tissue imaging with a morphology-preserving DNA fluorescence in situ hybridization (DNA-FISH) workflow and single-cell registration, yielding measurements concordant with clinical FISH. In high-grade serous ovarian carcinoma (HGSOC), ORION-FISH recapitulated known chromosomal changes while revealing subclonal heterogeneity missed by targeted sequencing. Applied to serous tubal intraepithelial carcinomas, precursors of HGSOC, ORION-FISH identified intermixed epithelial cells with MYC or CCNE1 copy-number gains, as well as concurrent alterations associated with distinct immune microenvironments. In addition, epithelial cells with MYC and CCNE1 copy-number gains were detected in morphologically normal fallopian tube epithelium, along with rare MDM4 increases across epithelial lineages. Together, ORION-FISH provides a framework linking chromosomal copy-number states to protein-defined phenotypes within preserved tissue architecture, enabling context-aware interrogation of early copy-number diversification at single-cell resolution. We introduce ORION-FISH, a spatially resolved workflow integrating multiplexed protein imaging with DNA-FISH to map genomic alterations within intact tissues. Applying this approach to ovarian cancer precursors reveals early copy-number diversification and associations with the local immune context, providing a foundation for studying how genomic and microenvironmental states coevolve during tumor initiation.

The efficacy of chimeric antigen receptor T (CAR-T) cell therapy in solid tumors is limited by the scarcity of stable antigen targets. To enhance CAR-T cell immunotherapy for solid tumors, we developed an integrated system that simultaneously introduces a surface antigen target for CAR-T cells and remodels the immunosuppressive tumor immune microenvironment (TIME) using an oncolytic adenovirus (OVs). This system leverages B cell membrane-derived CD19 as an artificial antigen target on tumor cells and enables in situ production of an αCD3e-αEpCAM bispecific T cell engager (BiTEs), which further strengthens the binding and cytotoxicity of CD19 CAR-T cells to cancer cells. The "double strike" therapeutic potential of Epv@CMP, both alone and in combination with CD19 CAR-T cells, was evaluated in colorectal cancer (CRC) and gastric cancer (GC) models. We have designed a lentiviral delivery system (Epv@CMP) that combines the homologous tumor cell membrane with the B cell membrane. The lentivirus has been genetically modified to express the dual-specificity antibody (αCD3e-αEpCAM) in tumor cells (Epv), aiming to enhance the accumulation of the virus in tumors and the efficacy of immunotherapy. Epv@CMP demonstrated efficient tumor-specific delivery and robust viral replication in vivo, leading to tumor regression and immune activation. Successful CD19 target anchor was also confirmed. The combination of Epv@CMP with CD19 CAR-T cells potently eliminated tumors in mouse models. Collectively, Epv@CMP represents a novel synergistic strategy that integrates oncolytic virotherapy with CAR-T cell immunotherapy to achieve potent tumor eradication and immune activation.

Social competition exerts a diverse set of influences on neural development and behavior during adolescence and yet the precise underlying molecular mechanisms remain poorly understood. Here, we demonstrate that individual rodents that rank lower in social hierarchies are more vulnerable to drug abuse. Proteomic analysis revealed a crucial role of the mesocorticolimbic netrin-1/DCC/UNC5 pathway within the nucleus accumbens (NAc) in mediating the impact of social competition. We found that mice with a conditional knockout of the deleted in colorectal cancer (DCC) gene in dopamine neurons were more likely to achieve higher social rank but exhibited increased drug-seeking behaviors. Following dopamine fiber immunostaining, these outcomes were attributed to ectopic mesolimbic dopamine fibers, which enhanced risk-taking behavior in winners that had DCC knockout. Collectively, our work elucidates a molecular mechanism through which social competition influences adolescent brain development and behavior, particularly in relation to drug susceptibility.

Primary liver cancer remains the third leading cause of global cancer-related mortality, with hepatocellular carcinoma (HCC) constituting the predominant histological subtype. The development of malignant ascites in advanced HCC patients signifies metastatic progression and portends poor clinical outcomes, presenting a formidable therapeutic challenge. This study aimed to engineer a natural small-molecule hydrogel for anti-tumor treatment, in order to reduce the production of ascites and achieve the dual therapeutic effect. The EP-GA hydrogel was synthesized by a one-pot method, and its morphology and mechanical properties were characterized by SEM and rheology. UHPLC-Q-Orbitrap HRMS analysis was carried out to identify the active components of EP. The assembly mechanism was analyzed by 1H NMR and molecular dynamics. Small animal imaging techniques were utilized to evaluate the retention of EP-GA in vivo. The H22 ascites tumor model was established, and antitumor mechanism was verified by transcriptomics, metabolomics and flow cytometry. The EP-GA hydrogel formed uniform nanoparticles with an average diameter of approximately 200 nm. Rheological characterization confirmed its excellent injectability. Using marker components euphol (EPH) and glycyrrhizic acid (GA) to analyze the self-assembly process, hydrogen bond interactions occurred between euphol's hydroxyl group and GA's hydrophilic domain. In H22 subcutaneous HCC xenograft models, EP-GA demonstrated significant tumor growth suppression, with a tumor growth inhibition rate (IRG) of 68.63 % at the dosage of 18.75 mg/kg. Combined transcriptomic and metabolomic analysis reveals that it inhibits tumor progression by suppressing choline kinase alpha, regulating the PI3K/AKT/mTOR pathway, and inducing ROS generation and mitochondrial membrane potential decline. This excipient-free nanohydrogel platform effectively circumvents carrier-related toxicity while demonstrating powerful therapeutic efficacy against HCC, and this platform is a promising new strategy for HCC prevention and treatment.

Large-scale multiomics profiling has delineated dynamic molecular landscapes during autophagy, yet translating these complex datasets into mechanistic regulatory insights remains a major challenge. In our recent work, we developed LyMOI, a hybrid artificial intelligence workflow that combines graph-based deep learning and a large language model (LLM) for mechanistic interpretation of autophagy-related omics. The graph model integrates 1.3 TB of autophagy-associated multiomics datasets and prioritizes molecules of interest (MOIs) across 34 autophagy-specific conditions, and then LLM-based chain-of-thought (CoT) reasoning generates mechanistic hypotheses to interpret their potential roles in biological contexts. Using LyMOI, we identified essential regulators, including GIN4, ELM1, RVS167 and STE50, involved in yeast autophagy induced by nutrient deprivation. Furthermore, LyMOI revealed that two cancer-associated proteins, CTSL and FAM98A, are required for maintaining autophagy activity upon disulfiram (DSF) treatment. Silencing either CTSL or FAM98A attenuated DSF-induced autophagy and inhibited cancer cell proliferation. Notably, combination treatment with DSF and Z-FY-CHO, a CTSL-specific inhibitor previously developed against SARS-CoV-2 infection, potently suppressed tumor growth. Collectively, our work presents an LLM-powered platform with biologist-like reasoning for uncovering autophagy regulatory mechanisms.Abbreviations: ARGs: autophagy-regulating genes; ATGs: autophagy-related genes; BECN1: Beclin 1; CoT: chain-of-thought; CQ: chloroquine; CTSL: cathepsin L; DSF: disulfiram; GCNs: graph convolutional networks; GI: graph inference; IHC: immunohistochemistry; LC3-II: lipidated LC3; LGM: large graph model; LLM: large language model; MOIs: molecules of interest; NATs: normal adjacent tissues; SQSTM1/p62: sequestosome 1; 3-MA: 3-methyladenine.

Recent NIH Data Management and Sharing (DMS) policy updates and NIH controlled-access data security requirements have increased attention to facial anonymization and controlled-access handling of shared head imaging data. This is particularly relevant for datasets submitted to or hosted by the Cancer Imaging Archive (TCIA), where NCI Cancer Imaging Program/TCIA implementation practices address imaging data containing potentially reconstructable facial anatomy. While intended to protect patient privacy and strengthen public trust, defacing can distort craniofacial geometry and alter image statistics, potentially compromising the fidelity and reproducibility of artificial intelligence (AI) models trained on such data. Existing studies primarily validate visual anonymization quality, but few have quantified its downstream impact on deep learning-based medical imaging tasks. Understanding this privacy-utility trade-off is crucial for responsible data sharing and compliant AI development. We systematically evaluated three representative defacing algorithms, two invasive (QuickShear and Py-Deface) and one less destructive, facial replacement (mri_reface), across MRI and CT datasets from 600 subjects spanning three institutions. Model performance was assessed on three clinically relevant applications: (1) brain segmentation and Evans ratio biomarker quantification in normal pressure hydrocephalus (NPH) MRI using SLANT and FreeSurfer; (2) representative-slice selection and diagnostic reasoning for brain tumour MRI using vision-language models (VLMs); and (3) automated emergency head CT report generation using a fine-tuned Otter-based vision-language model. Each method's impact was quantified using Dice similarity, correlation metrics, reasoning accuracy, and natural-language generation scores (BLEU, METEOR, ROUGE, CIDEr). Invasive algorithms caused significant degradation across all tasks. QuickShear reduced mean Dice scores by up to 9% and introduced 14-19% failure rates during quality control, while PyDeface induced smaller but measurable performance losses. mri_reface maintained 100% success without any failures and achieved segmentation, diagnostic, and report-generation accuracy within 3-5% of the original data. Evans ratio distributions remained statistically consistent between mri_reface and original images (p > 0.05), whereas invasive methods introduced broader variance. Across all VLM tasks, mri_reface preserved high correlation with radiologist-selected slices (r = 0.979) and stable report-generation quality (BLEU-4 = 0.11 ± 0.06 vs. 0.12 ± 0.07 for original). Facial anonymization introduces a measurable privacy-utility trade-off that must be explicitly considered in the design of AI-ready medical imaging datasets. Invasive defacing compromises geometric and statistical integrity, reducing downstream model accuracy even outside facial regions. Facial replacement anonymization methods, such as mri_reface, effectively reconcile patient privacy with reproducibility, offering a practical path to NIH-compliant open data. Future regulatory and institutional policies should integrate quantitative privacy-utility assessment and mandate transparent reporting of anonymization pipelines to ensure that shared imaging data remain both ethically safe and scientifically valid under emerging digital health frameworks. This work was partially supported by the American Heart Association (Award No. 25IPA1454088), the National Institutes of Health (Award No. 1R03CA286693-01A1 and Award No. 1R01CA291826-01A1), the U.S. Department of Defense (Award No. HT94252510807), and the National Science Foundation (Award No. 2545071).

Non-invasive urine-based cancer biomarker detection holds significant clinical value in cancer screening, treatment efficacy monitoring, recurrence warning, and auxiliary diagnosis. To address the issues of poor sensitivity and selectivity in photoelectrochemical (PEC) biosensing for the detection of cancer biomarkers in urine, oxygen vacancies (OVs)-induced dual-interface modulation strategy is proposed for constructing a TiO2/BiOI-based PEC sensor. The introduction of OVs not only induces the in-situ formation of bismuth nanoparticles at the solid-solid interface but also generates positive surface charge at the solid-liquid interface, which promotes the formation of the Z-scheme heterojunction and the modification with a negatively charged anti-fouling polymer layer (C-PEG), thereby effectively enhancing the photoelectric conversion efficiency and antifouling performance of the PEC sensor. Meanwhile, functionalization with aptamers further improves its selectivity. Leveraging the synergistic effects of the Z-scheme, anti-fouling polymer layers, and aptamers, the developed PEC sensor achieves highly sensitive and selective detection of alpha-fetoprotein (AFP), with a linear relationship in the range of 1.0 pg/mL∼150.0 ng/mL, and a detection limit of 0.3 pg/mL (LOD = 3s/k). In addition, the sensor successfully measures AFP levels in serum and urine samples from healthy individuals and hepatocellular carcinoma patients, and statistical analysis reveals a strong correlation between the serum and urinary AFP concentrations (r = 0.934, P < 0.001, n = 9). These findings indicate the preliminary feasibility of the proposed PEC sensor for non-invasive urine-based AFP detection, which may provide insights for future research.

Aberrant activation of Wnt signaling results in unregulated accumulation of cytosolic β-catenin, which subsequently enters the nucleus and promotes transcription of genes that contribute to cellular proliferation and malignancy. Here, we sought to eliminate pathogenic β-catenin from the cytosol using designer ubiquibodies (uAbs), chimeric proteins composed of an E3 ubiquitin ligase and a target-binding domain that redirect intracellular proteins to the proteasome for degradation. To accelerate uAb development, we leveraged a protein language model-driven algorithm called SaLT&PepPr to computationally design "guide" peptides with affinity for β-catenin, which were subsequently fused to the catalytic domain of a human E3 called carboxyl terminus of Hsp70-interacting protein. Expression of the resulting peptide-guided uAbs in colorectal cancer cells led to the identification of several designs that greatly reduced the abnormally stable pool of free β-catenin in the cytosol and nucleus while preserving the normal membrane-associated subpopulation. This selective knockdown of pathogenic β-catenin suppressed Wnt/β-catenin signaling and impaired tumor cell survival and proliferation. Furthermore, one of the best degraders selectively decreased cytosolic but not membrane-associated β-catenin levels in livers of BALB/c mice following delivery as a lipid nanoparticle-encapsulated mRNA. Collectively, these findings reveal the unique ability of uAbs to selectively deplete abnormal proteins in vitro and in vivo and open the door to peptide-programmable biologic modulators of other disease-causing proteins.

Lysine acetylation affects hepatocellular carcinoma (HCC) malignancy through multiple pathways. Evidence from our laboratory and other groups indicates that long chain acyl CoA synthetase 4 (ACSL4) is a crucial oncoprotein in HCC. However, the precise mechanisms underlying the post-translational regulation of ACSL4 acetylation remains unknown. Here, we report a previously unknown mechanism of ACSL4 regulation involving acetylation at the lysine 49 (K49) site by the acetyltransferase MOF. ACSL4 acetylation hindered its degradation via the ubiquitin-proteasome pathway. Using mass spectrometry and subsequent verification, we demonstrated that TRIM21 is an E3 ubiquitin ligase responsible for ACSL4 proteasomal degradation. Further mechanistic studies revealed that MOF-mediated ACSL4-K49 acetylation counteracted the TRIM21-mediated degradation of ACSL4. Functionally, we demonstrated that acetylated ACSL4 promotes lipid accumulation and HCC progression, both in vitro and in vivo. Clinically, ACSL4-K49 acetylation is frequently increased in HCC samples, and elevated ACSL4-K49 acetylation is associated with poor prognosis in patients with HCC. Together, these findings unveil a novel regulatory mechanism of ACSL4, highlighting its pivotal role in modulating HCC progression.

The Warburg effect, a hallmark of metabolic reprogramming, drives tumor progression, but its upstream regulation remains unclear. Using hepatocellular carcinoma (HCC) as a model, we identify TRIM32, an E3 ubiquitin ligase, as a potent activator of glycolysis that promotes malignancy. TRIM32 is upregulated in HCC tissues and cell lines, correlating with aggressive features and poor prognosis. Gain- and loss-of-function studies show that TRIM32 enhances proliferation, invasion, and migration in vitro and accelerates tumor growth and lung metastasis in xenografts. Mechanistically, TRIM32 mediates K48- and K63-linked polyubiquitination of STING, accelerating its degradation and relieving glycolytic suppression. Reduced STING elevates HK2 and promotes its mitochondrial localization, sustaining glycolysis and bioenergetics. TRIM32 knockdown decreases tumor burden, metastasis, and glycolytic activity, while hepatocyte-targeted STING knockdown rescues tumorigenesis in liver-specific TRIM32 knockout mice. These findings define a TRIM32-STING-HK2 axis that links ubiquitin-mediated suppression of innate immune signaling to glycolytic activation. Although validated in HCC, this mechanism likely applies broadly across solid tumors and nominates TRIM32 as a therapeutic target to reprogram tumor metabolism and limit malignancy.

Hepatocellular carcinoma (HCC) is a highly aggressive malignancy with a poor prognosis, highlighting the urgent need for novel diagnostic and therapeutic targets. In this study, through integrated proteomic profiling, we identify AP4M1 as a potential therapeutic vulnerability for HCC and characterize its upstream regulatory and downstream effector mechanisms. We show that AP4M1 is markedly upregulated in HCC, correlates with unfavorable prognosis, and is functionally involved in driving HCC progression. Mechanistically, the deubiquitinase USP15 stabilizes AP4M1 by removing K11- and K48-linked polyubiquitin chains at lysine 163 (K163). Additionally, MINK1-mediated phosphorylation of AP4M1 at threonine 69 (T69) is critical for its interaction with GPX4. Through upregulating GPX4, AP4M1 reduces intracellular lipid peroxidation, thereby suppressing ferroptosis and facilitating HCC tumorigenesis and progression. Importantly, combined treatment with the USP15 inhibitor USP15-IN-1 and the GPX4 inhibitor RSL3 exhibits potent anti-tumor efficacy in both in vitro and in vivo models, underscoring the therapeutic potential of targeting the AP4M1 axis in HCC. Collectively, this study delineates the USP15/AP4M1/MINK1/GPX4 axis as a previously unrecognized ferroptosis-suppressive pathway that promotes HCC progression, providing a critical theoretical basis for AP4M1-targeted therapeutic strategies in HCC.

Clinical-grade RAS inhibitors raise an unresolved question as to whether KRAS alleles impose constraints on adaptive resistance that can be exploited therapeutically. Using daraxonrasib (RMC-6236), a multi-selective RAS(ON) inhibitor, we compared resistance mechanisms between KRASG12D and KRASG12R, mutants with fundamentally different RAS network dynamics. Daraxonrasib inhibited KRASMUT primarily through steric occlusion of effector binding, while engaging RASWT only modestly (~20%). KRASG12R was marked by its inability to transactivate RASWT, and daraxonrasib resistant KRASG12R PDAC cells utilized EGFR/RASWT-GTP signaling as the dominant adaptive route. In contrast, KRASG12D resistance arose through retained KRASG12D-GTP signaling, with a decrease of cyclophilin A (CypA) protein, the binding partner required for daraxonrasib activity. The shift from KRASG12R dependence to EGFR/RASWT dependence conferred sensitivity to trametinib. As clinical confirmation, a KRASG12R PDAC patient who progressed after 10 months on daraxonrasib showed intratumoral EGFR/RASWT activation, and rapid 3D-bioprinted patient-derived tumoroid modeling predicted sensitivity to trametinib-based combination therapy. Despite the aggressive disease trajectory and lack of response to the two immediately preceding lines of therapy, sixth-line trametinib-based combination therapy achieved approximately 5 months of disease control, and this patient ultimately achieved 40 months of overall survival, far exceeding the 8-12 month median for metastatic PDAC. Collectively, these data establish a framework in which allele-specific RAS network topology dictates the adaptive resistance landscape, enabling rational selection of targeted therapies with meaningful clinical benefit in metastatic PDAC.

Dysregulated mitochondrial dynamics in cancer cells perturbs mitochondrial function and metabolism and promotes cancer progression. Its impacts on the electron transport chain, oxidative phosphorylation, redox balance, and glycolysis are well recognized. However, its influence on tricarboxylic acid (TCA) cycle activity is less clear. In this study, we hypothesized that excessive mitochondrial fragmentation suppresses the expression of succinate dehydrogenase (SDH), resulting in the accumulation and secretion of succinate. We tested this hypothesis in human hepatocellular carcinoma (HCC) cell model, murine xenograft tumor model, human HCC tumor tissues, and serum samples from patients with HCC. Genetic suppression and pharmacological inhibition of dynamin-related protein 1 (Drp1) were employed to examine their effects on SDH expression and succinate levels. The effects of Mdivi-1, a pharmacological inhibitor of Drp1-mediated mitochondrial fission, were evaluated in the xenograft tumor model, and the impact of succinate on mitochondrial dynamics was assessed in Huh7 cells. The results reveal imbalance of mitochondrial fission and fusion proteins and increase in mitochondrial fragmentation which was associated with reduced expression of SDH and increased succinate. Succinate dehydrogenase B subunit (SDHB) mRNA levels were reduced in human HCC tumor tissues, and higher SDHB expression was associated with improved overall and relapse-free survival. Serum succinate levels were increased in patients with HCC. Genetic suppression and pharmacological inhibition of Drp1 resulted in restoration of SDH and reduction of succinate. Administration of Mdivi-1 reduced tumor growth and lung metastasis in the xenograft tumor model, which was associated with reduced p-Drp1 and increased SDHB. Addition of succinate to Huh7 cells enhanced Drp1-mediated mitochondrial fragmentation while succinate antibodies abrogated it. Overexpression of SDHB in Huh7 cells suppressed Drp1 activation and mitochondrial fragmentation through reduction of succinate. By contrast, SDHB silencing with SDHB siRNA enhanced Drp1 activation and mitochondrial fragmentation. These results suggest a positive feedback regulation of mitochondrial fragmentation by SDH/succinate. These findings indicate that the mitochondrial fragmentation-SDH-succinate regulatory loop plays an important role in HCC growth and metastasis and may represent a potential target for new drug development.

Chemotherapy (CT) suffers from poor selectivity and systemic toxicity, while photodynamic therapy (PDT) and photothermal therapy (PTT) are inherently limited by tumor hypoxia and thermoresistance. Herein, we develop a trimodal synergistic nanoplatform (PDM-BDP NPs) for ovarian cancer treatment by integrating PDT, mild PTT (mPTT), and CT into a single azo-bond-linked prodrug system. Upon 808 nm laser irradiation, PDM-BDP NPs generate reactive oxygen species and mild hyperthermia, while simultaneously triggering azo-bond cleavage to release the chemotherapeutic agent phenylenediamine mustard (PDM) and transform into NH2-BDP, which retains photothermal activity for sustained thermal output. PDM-BDP NPs exhibit efficient cellular uptake, predominant mitochondrial co-localization, and potent photocytotoxicity against A2780 cells with low dark toxicity. Transcriptomics reveals multi-target reprogramming involving proteotoxic stress, DNA damage, and TNF/MAPK activation. Laser-activated PDM-BDP NPs achieve subtotal tumor ablation with excellent biocompatibility. This trimodal synergistic strategy provides a promising approach for precise ovarian cancer therapy.

There is broad consensus that the malignant epithelial cells of human pancreatic ductal adenocarcinoma (PDA) comprise multiple, molecularly distinct states. Yet precise characterization of how these are regulated-including their mechanistic determinants, dependencies, plasticity and functional properties-remains elusive. Single-cell master regulator (MR) analysis of multiple PDA cohorts identified malignant cells in three co-existing, molecularly distinct developmental lineage states, with distinct histopathological morphologies and spatial architecture. These include a poorly differentiated lineage driven by epithelial-mesenchymal-transition-related MRs and two well-differentiated states driven by gastrointestinal epithelial development and pancreatic development MRs, respectively. Furthermore, each state comprises two epigenetically distinct substates with low versus high MAPK signaling activity. Barcode-based lineage tracing confirmed both spontaneous and treatment-dependent cross-state plasticity. Furthermore, loss-of-function studies confirmed state-specific MR essentiality, while their ectopic expression effectively reprogrammed cell state, in vitro and in vivo, thus providing a mechanism-based foundation for PDA heterogeneity and a roadmap for pharmacological targeting.

Dysregulated transcription and aberrant cell cycle progression represent core cancer hallmarks, yet not many transcriptional regulators are known to simultaneously govern both pathways as potent tumor suppressors. GDOWN1 is a Pol II-interacting factor mainly sequestered in the cytoplasm in somatic cells, and its nuclear biological functions remain largely undefined. We previously generated a nuclear-targeted GDOWN1 mutant (nGDOWN1(10 M)) that strongly represses transcription and cell proliferation. Here, we systematically dissect the underlying molecular mechanisms and validate its broad-spectrum anticancer potential. Combining genetic manipulation, multi-omics profiling, and multiple functional assays, we demonstrate that nGDOWN1(10 M) disturbs Pol II CTD phosphorylation homeostasis and triggers promoter-proximal premature termination, causing global repression of nascent transcription. Terminated Pol II complexes undergo cytoplasmic translocation alongside associated the CTD phosphatases PP2A and PP1, leading to massive cytoplasmic accumulation of these phosphatases. Concurrently, TP53 and CDKN1A are targeted by nGDOWN1(10 M) and transcriptionally upregulated. The above dual regulatory circuit synergistically activates RB family tumor suppressors: p53-p21 signaling inhibits cyclin-dependent kinases, while cytoplasmic PP2A/PP1 enhance cellular phosphatase activity to dephosphorylate and stabilize RB and p130. Activated RB and p130 form functional RB and DREAM complexes and inhibit the transcription of their target genes to induce robust cell cycle arrest. Notably, nGDOWN1(10 M) exerts profound antiproliferative effects across cancer cell lines, mouse xenograft models, and patient-derived tumor organoids, independent of p53 status. Collectively, this work uncovers a novel interconnected regulatory cascade triggered by nuclear translocation of GDOWN1: it remodels Pol II phosphorylation landscape and induces global transcriptional repression, which further drives cytoplasmic kinase-phosphatase coordination to activate master cell cycle suppressors and ultimately arrest cell cycle progression. Our results illuminate a coordinated molecular circuit governing two central cancer hallmarks and establish nuclear GDOWN1 as a promising therapeutic candidate for both p53-proficient and -deficient malignancies.

Triple-negative breast cancer (TNBC) is an aggressive subtype with an activated tumor immune microenvironment. The multicenter, multinational, double-blinded NSABP B-59/GeparDouze trial evaluated the addition of atezolizumab (atezo) (773 patients randomized) or placebo (777 patients) to sequential taxane-carboplatin-anthracycline-based neoadjuvant chemotherapy in stage II-III TNBC. The addition of atezo did not significantly improve the primary endpoint of event-free survival (EFS) (HR, 0.80 (95% CI, 0.062-1.03); stratified log-rank P = 0.083, 4-year EFS rates difference 3.3%). The HR for overall survival was 0.86 (95% CI, 0.62-1.19), with a 4-year benefit of 0.7%. Prespecified subgroup analyses suggested heterogeneity in EFS, with benefit of atezo in patients presenting with clinical lymph node involvement (Pinteraction = 0.039). Treatment-emergent adverse events with grades ≥3 were reported in 75.3% (atezo) versus 73.4% (placebo), and immune-related adverse events were reported in 27.6% (atezo) versus 11.4% (placebo). A total of 196 (25.5%) patients discontinued atezo and 143 (18.8%) patients discontinued placebo in the neoadjuvant phase. In an exploratory mRNA-based subset analysis, patients with basal-like immune-activated tumors may have benefited from atezo. TNBC subtyping to identify basal-like immune-activated tumors and quantification of tumor-infiltrating lymphocytes to identify tumors with high tumor-infiltrating lymphocyte counts could be a promising strategy to identify patients who benefit from the addition of immune checkpoint inhibitors to neoadjuvant chemotherapy. ClinicalTrials.gov registration: NCT03281954 .

Accurate breast tumor segmentation in dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) is vital for diagnosis and treatment planning. Despite advances in deep learning, its performance remains constrained by the need for extensive voxel-wise annotations. To mitigate this burden, we propose an annotation-efficient framework that jointly optimizes data selection, unlabeled data utilization, and data augmentation under limited annotation budgets. A diversity-aware uncertainty query (DUQ) strategy guides the annotation by jointly modeling data representativeness and informativeness through a representative candidate selector (RCS) and an uncertainty-based decision maker (UDM), ensuring efficient and targeted labeling. To leverage unlabeled data, a cross-decoder consistency regularization (CDCR) mechanism enforces prediction consistency between two decoders with distinct attention mechanisms, enhancing robustness and confidence. Furthermore, a lesion transplant augmentation (LTA) technique synthesizes anatomically valid pseudo samples by transplanting lesion regions from labeled to unlabeled images, effectively expanding training diversity. Experiments were conducted on two DCE-MRI datasets with biopsy-proven breast cancers, one as internal dataset containing 676 subjects and the other as external dataset with 344 subjects. Comparative and ablation results demonstrate that our framework consistently outperforms state-of-the-art semi-supervised and active learning methods, providing a simple yet effective annotation-efficient solution for breast cancer segmentation in DCE-MRI. The code is publicly available at https: //github.com/zouquanling/DUQ_and_CDCR.

Background Photon-counting CT (PCCT) offers improved spatial resolution, contrast to noise ratio, and dose efficiency, but its clinical utility remains incompletely defined for breast cancer. Purpose To evaluate the feasibility of PCCT for pretreatment breast cancer assessment through comparisons with MRI, full-field digital mammography (FFDM), and fluorine 18 (18F) fluorodeoxyglucose (FDG) PET/CT. Materials and Methods In this prospective study (March-May 2025), female participants with breast lesions categorized as Breast Imaging Reporting and Data System 4C or higher at US or FFDM underwent breast MRI and multiphasic contrast-enhanced PCCT. 18F-FDG PET/CT was performed in a subset with locally advanced disease. Four radiologists independently evaluated lesion morphologic characteristics, additional findings, and clinical TNM stage. Agreement was analyzed using intraclass correlation coefficients (ICCs) and κ statistics. The diagnostic performance for additional lesions and nodal metastasis was compared with the reference standard (pathologic examination). Results Among 126 participants (mean age, 58.1 years ± 12.3 [SD]), interreader agreement across PCCT, MRI, and FFDM was good to excellent. PCCT agreed with MRI for lesion characterization (κ = 0.57-0.96) and clinical T categorization (κ = 0.86-0.88), with highest agreement with pathologic size (ICC, 0.70-0.81). For 46 pathologically confirmed additional lesions, PCCT was more sensitive than FFDM (difference, 44% [95% CI: 19, 66]) and similar to MRI (difference, 7% [95% CI: -5, 21]). Additionally, 44% (95% CI: 27, 52) of microcalcifications were missed at PCCT versus FFDM. For pathologically confirmed nodal metastasis, PCCT was more sensitive (difference, 10% [95% CI: 1, 20]) and accurate (difference, 6% [95% CI: 1, 11]) than MRI. For clinical N category, PCCT agreed with PET/CT (κ = 0.82 [95% CI: 0.62, 0.96]; n = 19). Two distant metastases identified at PCCT were consistent with 18F-FDG PET/CT and pathologic findings. Conclusion PCCT demonstrated similar performance to MRI for lesion characterization and detection of additional lesions, with better performance for nodal metastasis evaluation; however, detection of microcalcifications was limited. © RSNA, 2026 Supplemental material is available for this article.

The impact of immune checkpoint inhibitors combined with neoadjuvant chemotherapy (NAC) on surgical outcomes and immune remodeling remains underexplored. To investigate the efficacy and tumor immune microenvironment (TIME) changes of camrelizumab plus NAC for triple-negative breast cancer (TNBC), we collected TNBC patients receiving neoadjuvant immunochemotherapy (NIC) or NAC from clinical trials and routine practice, and set a series of endpoints, including total pathological complete response (pCR), breast pCR and TIME characteristics. After propensity score matching, the NIC cohort showed significantly higher rates of total pCR, breast pCR, and axillary pCR, and a lower rate of axillary lymph node dissection than the NAC cohort (all p < 0.05). Within the NIC group, patients achieving total pCR (p = 0.012) or breast pCR (p = 0.0078) had superior disease-free survival. Analysis of TIME characteristics indicated that the 7 TIME signatures, including tumor inflammation signature, antigen processing machinery and T cell-related features, could predict pCR response to NIC. The finding suggested that in TNBC, NIC enhances pCR achievement and reduces axillary dissection risk. Elevated TIME profiles of the 7 signatures may predict favorable pCR response to NIC.

Hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2-) breast cancer accounts for approximately 70% of breast cancer cases. Despite recent advances with cyclin-dependent kinase 4/6 inhibitors (CDK4/6i), resistance inevitably develops, often driven by activation of the phosphatidylinositol 3-kinase (PI3K)-AKT-mammalian target of rapamycin (mTOR) pathway. Genetic alterations such as PIK3CA mutations (present in ~ 45% of HR+/HER2- tumors), AKT1 mutations, and PTEN loss contribute to endocrine resistance and poor outcomes. This review summarizes emerging strategies targeting this pathway to overcome resistance in advanced disease. Isoform-specific PI3K inhibitors, including alpelisib and inavolisib, have demonstrated clinically meaningful progression-free survival benefits in PIK3CA-mutated populations, with inavolisib showing improved tolerability and efficacy. In contrast, pan-PI3K inhibitors such as buparlisib have been constrained by toxicity. Targeting downstream signaling, AKT inhibitors have also shown benefit: capivasertib has demonstrated clinical efficacy leading to US Food and Drug Administration approval, while ipatasertib has yielded encouraging results, particularly in tumors harboring PIK3CA, AKT1, or PTEN alterations. Mammalian target of rapamycin inhibitors, notably everolimus, have shown efficacy irrespective of mutation status. The dual PI3K-mTOR inhibitor (gedatolisib) has also shown promising progression-free survival benefit in a PIK3CA wild-type population. Next-generation agents, including mutant-selective PI3Kα inhibitors and bi-steric mTOR complex 1 inhibitors, are under active investigation. Optimal sequencing of these agents alongside endocrine therapy and CDK4/6i options remain a critical question, as does integration of genomic testing to guide therapy. Future directions include rational combination strategies, improved biomarker-driven selection, and novel modalities such as proteolysis-targeting chimeras (PROTACs). Collectively, these advances aim to enhance durability of response, minimize toxicity, and improve survival in HR+/HER2- metastatic breast cancer.

Incomplete postmenopausal breast involution leaves persistent epithelial-rich lobules and elevated breast density in about 40% of women and is associated with higher breast cancer risk, but why remodelling stalls remains unclear. We studied a longitudinal cohort of 81 women with paired benign breast biopsies (baseline age 45-55 years; follow-up 2-10 years), all with baseline NanoString transcriptomics and two-timepoint digital morphometry, and with multiplex immunofluorescence in spatial-imaging subsets (baseline n = 14-16 depending on panel; follow-up n = 14). A separate postmenopausal endpoint cohort (12 women: eight noninvoluted, four completely involuted), profiled by genome-wide expression array and multiplex immunofluorescence, defined the persistent-lobule phenotype. Noninvoluted postmenopausal tissue retained a proliferation-competent, tumour-associated epithelial state and showed immune accumulation at lobular boundaries with reduced access to p16+ (senescence-associated) epithelial foci. The same SASP and innate immune programmes that predicted slower involution across the menopausal transition predicted faster involution after menopause. Follow-up boundary CD45→p16 engagement was directionally consistent with this reversal in Pre→Post and Post→Post women. Spatial imaging resolved this reversal into a perimenopausal stall architecture and a postmenopausal clearance-associated architecture marked by direct CD16+ innate-effector engagement of p16+ epithelium; macrophage targeting provided convergent support (two-sided exact permutation interaction p = 0.0077). Menopausal timing conditions whether senescent-immune programmes couple to productive clearance or to spatially uncoupled surveillance and persistent risk-associated tissue. Biomarker interpretation should therefore be anchored to menopausal timing. Casey DeSantis Cancer Fund and US National Cancer Institute.

MicroRNAs (miRNAs) in blood are promising liquid biopsy biomarkers, yet their short length, low abundance, and high intra-family homology hinder sensitive and specific detection. Combining rolling circle amplification (RCA) with CRISPR-Cas12a enables isothermal detection, but existing methods typically depend on auxiliary enzymes or in-assay ligation and rarely encode sequence discrimination within the template itself. Here, we report a one-pot dual-toehold RCA (dtRCA)-Cas12a biosensor driven by a preassembled three-arm toehold-gated (3TG) DNA template for ultrasensitive and selective miRNA detection. The 3TG template adopts a three-arm dumbbell conformation, eliminating the need for a ligase during the assay, and presents two target-complementary toehold domains with a Cas12a-recognition sequence. Target binding triggers strand displacement, initiating dtRCA via a single polymerase. The resulting amplicons activate Cas12a trans-cleavage for fluorescence or lateral flow assay (LFA) readouts. Crucially, a single-base mismatch within the toehold suppressed amplification, whereas a topology-matched circular template lacking the toehold gate failed to distinguish the target, demonstrating that selectivity arises from the template structure. The one-pot dtRCA-Cas12a system achieved attomolar sensitivity, detecting miR-21, miR-375, and let-7a at 2.5, 114.9, and 8.0 aM, respectively. The paper-based LFA maintained femtomolar sensitivity and enabled an instrument-light readout. In plasma, this platform discriminated breast cancer patients (n = 17) from healthy donors (n = 10) with AUC values of 0.97-0.98. Three-marker classification demonstrated robust performance in leave-one-out cross-validation and correctly classified 30 samples in an independent validation cohort, showing performance comparable to RT-qPCR. By embedding selectivity into a preassembled template, this 3TG-driven dtRCA-Cas12a platform provides a highly sensitive and specific strategy for multi-marker miRNA analysis with simplified readout.

Heart failure therapy (HFT) is routinely continued in breast cancer survivors after recovery from anti-HER2 cancer therapy-related cardiac dysfunction (CTRCD), despite limited evidence. HER-SAFE is the first randomised trial to assess HFT withdrawal in this population. In this open-label, multicentre, randomised controlled trial, breast cancer survivors with recovered anti-HER2 CTRCD (asymptomatic, left ventricular ejection fraction [LVEF] ≥50%, normalised biomarkers) were randomised to HFT withdrawal or continuation. Antecedent CTRCD was mild (relative global longitudinal strain [GLS] decline >15%, LVEF ≥50%), moderate (LVEF fall >10% to <50%), or severe (LVEF <40%). The primary endpoint was a moderate or severe CTRCD event over 12 months analysed by intention to treat; secondary endpoints included serial cardiac function, biomarkers, and quality of life. Between July 2023 and June 2025, 90 patients were randomised (all female; median age 50 years [IQR 43-59]; median 17.3 months [IQR 7.7-33.2] since CTRCD recovery; HFT predominantly renin-angiotensin system inhibitors [98%] and beta-blockers [88%]) - 46 randomised to HFT withdrawal (1 lost to follow-up) and 44 to continuation. The primary endpoint occurred in 1 of 45 with withdrawal (moderate asymptomatic CTRCD) versus 0 of 44 with continuation (between-group difference 2.2pp; 90% CI upper limit 9.4pp). No cardiovascular deaths, heart failure hospitalisations or symptomatic CTRCD occurred. Cardiac magnetic resonance-derived LVEF remained stable in both arms at 12 months (between-group difference -0.8% [-2.0 to +0.4], p=0.21). Among breast cancer survivors with recovered anti-HER2 CTRCD, HFT withdrawal was not associated with symptomatic events or significant between-group difference in LVEF at 12-months.

Vepdegestrant (VEPPANU; formerly ARV 471) is a novel PROteolysis TArgeting Chimera (PROTAC) that is able to simultaneously bind the estrogen receptor (ER) and an E3 ubiquitin ligase complex, resulting in polyubiquitination of the ER and its subsequent degradation via a proteasome. It is being developed by Arvinas and Pfizer primarily as a treatment for ER+, HER2- breast cancer. In May 2026, vepdegestrant received its first global approval in the USA for adults with ER+, HER2-, ESR1-mutated advanced or metastatic breast cancer, as detected by an FDA-authorized test, with disease progression following at least one line of endocrine therapy. This article summarizes the milestones in the development of vepdegestrant leading to its first approval in this indication.

The treatment of older adults with chemotherapy for early-stage breast cancer involves a complex decision-making process that weighs the competing risks of disease recurrence against the potential harms of treatment. While older adults continue to be underrepresented in therapeutic clinical trials, there is a growing body of evidence to support optimal management of each subtype of early-stage breast cancer, including novel strategies like adjuvant cyclin-dependent kinase 4/6 inhibitors in hormone receptor-positive disease, and the neoadjuvant use of pembrolizumab in triple-negative breast cancer. The available evidence to support the systemic management of early-stage breast cancer in older adults with consideration of the data gaps for cytotoxic chemotherapy, de-escalated regimens, and targeted therapies is reviewed, and tools to optimize decision-making and supportive care for older adults with early breast cancer are presented. Clinicians should aim to avoid overtreatment when available evidence suggests a less intensive approach may suffice for a lower-risk cancer, and/or the patient is at high risk of serious toxic effects from chemotherapy treatment. On the other hand, undertreatment without consideration of the potential downsides may increase breast cancer-related mortality in older patients with higher-risk cancers. This review will help clinicians best use the available data and decision-making tools to support best practice for treating older adults with early-stage breast cancer and will highlight ongoing clinical trials aimed at optimizing medical care for this patient population.

Triple-negative breast cancer remains an aggressive and biologically heterogeneous breast cancer subtype. Although the therapeutic landscape has expanded, durable disease control remains clinically challenging in many settings. Existing reviews often organize TNBC therapy by drug class or molecular subtype, which can obscure how treatment response is shaped by interacting biological layers. Here, we review current and emerging therapeutic strategies through a three-layer framework: tumor-cell-intrinsic vulnerabilities, the local immune and stromal microenvironment, and host-level systemic modifiers. We summarize established approaches, including chemotherapy, immune checkpoint blockade, antibody-drug conjugates and PARP inhibition in biomarker-defined settings, and distinguish them from maturing or exploratory strategies such as pathway-directed therapy, epigenetic modulation, anti-vascular combinations, regulated cell-death induction, cellular therapy, vaccines, microbiome-related interventions, liquid biopsy, AI-supported multiomics and adaptive trial designs. This framework integrates evidence level, disease stage, biomarker reliability and patient tolerance into treatment selection for TNBC precision therapy.

The management of metastatic breast cancer (mBC) relies on tissue-based immunohistochemical subtypes. However, biopsies are invasive and may not capture metastatic heterogeneity, and subtypes can change over time under treatment pressure. In this study, we developed cell-free DNA (cfDNA) methylation signatures for minimally invasive breast cancer detection, distinction, and estrogen receptor (ER) status classification. Peripheral blood plasma methylomes were analyzed from 79 patients with mBC spanning ER+/human epidermal growth factor receptor 2 (HER2)- (n = 45), HER2+ (n = 13), and triple-negative breast cancer (n = 21). To derive tissue-informed breast cancer and ER-specific features, public 450K methylation array data (n = 9,730) were leveraged, and features were selected using generalized linear models via elastic net regularization with cross-validation. The tissue-informed features were translated to cell-free methylated DNA immunoprecipitation and sequencing (cfMeDIP-seq), and the final signatures were validated across a compendium of cfMeDIP-seq profiles (n = 713) spanning more than 10 cancer types. Across training, validation, and external test cohorts, the signatures demonstrated high accuracy for breast cancer detection versus controls, distinction from multiple other malignancies, and ER status classification. Performance generalized across independent cfMeDIP-seq cohorts and reflected tumor fraction. The sensitivity was reduced in samples with low tumor fractions and bone-only disease while remaining informative for typical tumor fractions observed in the metastatic setting. Promoter-proximal signature regions provided biological insight into tumor phenotypes. This tissue-anchored, platform-translatable framework demonstrates the feasibility of accurate, reproducible cfDNA methylation-based molecular classification in mBC. Tissue-anchored cell-free DNA methylation signatures enable minimally invasive detection, cancer-type distinction, and estrogen receptor classification in metastatic breast cancer, offering an interpretable, cross-platform framework to complement tissue biopsies for guiding subtype-directed therapy.

Decidual protein induced by progesterone 1 (DEPP1), also known as DEPP or C10ORF10, was originally identified as a progesterone-induced protein in endometrial stromal cells. Over the past two decades, research has progressively elucidated its involvement in various biological processes such as energy metabolism, redox regulation, and cellular autophagy. Additionally, DEPP1 has been implicated in the pathogenesis of several diseases, including diabetes, atherosclerosis, ischemic cardiomyopathy, breast cancer, and colon cancer. In this review, we systematically summarise the research progress on DEPP1, with particular emphasis on its molecular mechanisms in the crosstalk between oxidative stress and autophagy. Its cellular localization and functional uniqueness are discussed within the context of the classical redox-autophagy regulatory network. Furthermore, key issues in DEPP1 research and its potential translational applications are discussed to provide insights and perspectives for future studies centred on DEPP1.

The immune-excluded tumor immune microenvironment (TIME) limits responses to ICIs. Cancer-associated fibroblasts are the most abundant stromal population and key regulators of immune suppression; however, the upstream cues that program pathogenic CAF states and the mechanisms of the immune-excluded TIME remain poorly defined. Here, by combining single-cell RNA sequencing and functional validation, we report that tumor cell-released autophagosome (TRAP) programs inflammatory CAFs (iCAFs) and triggers cathepsin L-dependent intracellular cleavage of C3 into C3a via the HSP70-TLR4-MyD88-ERK/p38 pathway. iCAF-derived C3a affects C3a on TAMs, promotes TAM accumulation in the iCAF-rich stroma, limits TIL trafficking into tumor nests, and reinforces an immune-excluded TIME. Disrupting the TRAP-iCAF-C3a/C3aR axis remodels the immune-excluded TIME and sensitizes tumors to anti-PD-L1 therapy. In clinical cohorts, plasma TRAP and C3a levels increased with disease stage, and their combination improved the discrimination of patients with breast cancer from controls (AUC = 0.96). These data define a TRAP-driven stromal-immune circuit that promotes immune exclusion and suggest that the C3a-C3aR axis is a potential target for enhancing ICI efficacy.

Soybean isoflavones (SIs) are pivotal bioactive targets in the growing plant-based functional food market due to their selective estrogen receptor modulator (SERM)-like properties. However, a significant disparity remains between agricultural SI accumulation and ultimate physiological efficacy, constrained by the yield-metabolism genetic antagonism "linkage drag" and the "dual-cage" matrix entrapment barrier. This review establishes an operationalized seed-to-table framework to bridge these gaps through precise profile tailoring. We critically evaluate upstream genetic interventions (e.g., CRISPR/Cas9-mediated flux redirection) and downstream processing (e.g., biotransformation, physical-field intensification, and green solvent engineering). A novel, evidence-based key performance indicator (KPI) matrix is introduced to benchmark these technologies against standardized technology readiness levels (TRLs), economic burden (CapEx/OpEx), and systemic trade-offs. Crucially, we reframe SI bioavailability by accounting for Phase II metabolism, emphasizing that aglycone enrichment optimizes pharmacokinetic consistency and absorption kinetics rather than absolute systemic exposure. By integrating human clinical evidence and safety considerations for vulnerable populations (e.g., ER+ breast cancer survivors), this work bridges the persistent validation gap between laboratory innovation and clinical outcomes. Finally, we outline a paradigm shift toward AI-driven digital twins and host metabotyping to resolve the sensory-bioactivity paradox, providing a scientific roadmap for the rational design of next-generation isoflavone-fortified soy foods.

Apurinic/apyrimidinic endonuclease 1 (APE1) is a multifunctional protein that regulates host responses to oxidative stress. We investigated its role in controlling DNA damage following infection with a genotoxic intestinal microbe. Mice rendered hypomorphic for APE1 were crossed with tamoxifen (TMX)-inducible or constitutive Vil1ACre mice to yield intestinal epithelial cell-targeted knockout mice (Apex1iΔIEC or Apex1cΔIEC). Mice were infected with a genotoxic strain of E. coli (pks + E. coli NC101) to determine the APE1-sensitive effects on the accumulation of reactive oxygen species (ROS) and oxidative DNA damage. Murine and human primary colonic epithelial cell lines genetically altered to reduce APE1 expression were infected with a pks-deficient, the genotoxic E. coli strain or its synthetic colibactin genotoxin. APE1 expression was markedly diminished in enterocytes of Apex1iΔIEC mice, which was accompanied by increased constitutive epithelial cell DNA damage comparable to that observed in APE1 hypomorphic mice. The DNA damage, as indicated by γH2AX levels, was attenuated in germ-free Apex1iΔIEC mice and in APE1-hypomorphic mice treated with broad-spectrum antibiotics, indicating that commensal bacteria exert genotoxic effects that are unmasked by the loss of APE1 function. Levels of ROS and oxidative DNA damage in APE1-deficient primary colonic epithelial cells were increased significantly by pks + E. coli NC101 or its synthetic colibactin 742 genotoxin and this damage was prevented by prior treatment with an antioxidant. APE1 protects epithelial cells by inhibiting the accumulation of ROS and oxidative DNA damage induced by intestinal bacteria, including the colibactin produced by E. coli NC101.

PET-CT lesion segmentation remains challenging due to heterogeneous lesion appearance, small and dispersed lesions, physiological FDG uptake, and limited annotations. Existing self-supervised methods are mostly designed for unimodal imaging and therefore fail to fully exploit the complementary anatomical and metabolic information in PET-CT. Meanwhile, conventional multi-cancer segmentation strategies often treat different cancer types as a unified task, which weakens cancer-specific features, and existing prompt-based methods still have limited task adaptation and sensitivity to small lesions. To address these limitations, a unified two-stage framework for multi-cancer PET-CT segmentation is presented. First, a modality-guided probabilistic masked autoencoder is introduced to enhance cross-modal PET-CT representation learning through modality-specific masking. Second, a dual-prompt downstream segmentation network is designed to model both cancer-specific characteristics and cross-cancer shared knowledge, with prompt-aware heads further improving task adaptation and small-lesion delineation. Experiments on a multi-cancer PET-CT dataset show consistent improvements over the best-performing non-prompt and prompt-based baselines, with average Dice gains of 2.51% and 2.18%, respectively. The framework is further applied to an unannotated breast cancer cohort for survival analysis, demonstrating promising generalizability and improved risk stratification. The code is available at: https://github.com/XinglongLiang08/DpDNet.

Breast cancer remains the second leading cause of cancer-related mortality, with its etiology and pathogenesis involving dysregulated non-coding RNA expression, and the limited efficacy of current targeted therapies underscores an urgent need for novel drug targets and therapeutic modalities. Circular RNAs (circRNAs) are a type of noncoding RNA with a closed loop structure. Emerging evidence suggests that circRNAs function as ceRNAs or miRNA sponges, interact with proteins, and regulate gene transcription and translation. Current research primarily focuses on the ceRNA paradigm, often neglecting issues such as the effective concentration and activity of circRNAs under physiological conditions, as well as the impact of perturbations on individual nodes within complex ceRNA networks. We present a "functional integration" framework. In this model, a pathogenetically validated circRNA molecule serves as an environmentally regulated "molecular toolkit." Under specific pathological conditions, functional modules-such as protein interactions and the encoding of short peptides-can be dynamically activated by multi-level environmental signaling networks to govern phenotypic outputs. When present in a disease-driving circRNA, these modules have the potential to serve as pharmacologically actionable targets, enabling precision interventions-such as small-molecule inhibitors of circRNA-protein interactions, PROTACs targeting oncogenic peptides, and circRNA-based immunotherapies-tailored to the context-dependent primary functional module in each cancer context. This review establishes a foundation for circRNA-based liquid biopsy and therapies targeting specific functional modules.

Developing targeted cancer therapy with minimal side effects remains a significant challenge. Oxidative stress-based cancer therapies have gained traction in recent years. However, challenges such as limited tumor selectivity and therapeutic durability often hinder their clinical application. Here, we report a novel strategy of combining ROS-responsive prodrugs with prooxidants to achieve potent, durable, and selective tumor killing effects. This approach leverages pro-oxidants (i.e. ascorbate) to amplify oxidative stress within tumors, sensitizing cancer cells to ROS-responsive prodrugs. Both in vitro and in vivo studies confirm the anticancer synergism and selectivity of this combination therapy, which achieved complete tumor regression without recurrence, significantly outperforming single-agent treatments. This combination therapy is effective against hard-to-treat cancers like triple-negative breast cancers. Our findings highlight the potential of targeting tumor redox mechanisms through a combination of ROS-responsive prodrugs and pro-oxidants, offering a promising avenue for repurposing these agents in cancer therapy.

Palbociclib, ribociclib, and abemaciclib, which are FDA-approved CDK4/6 inhibitors, constitute the standard first-line treatment for advanced hormone receptor-positive (HR+) breast cancers. Although these drugs share nominal targets and demonstrate comparable first-line efficacy, significant divergences exist in their cross-line therapeutic responses upon first-line progression, indicating distinct mechanisms of action. To systematically elucidate their similarities and differences, we employed quantitative proteomics and phosphoproteomics to investigate and compare the molecular expression characteristics induced by these drugs. As a result, the three CDK4/6 inhibitors exhibited distinct anti-proliferative potencies in HR+ breast cancer cells. Dynamic proteomic and phosphoproteomic profiling demonstrated cell cycle arrest and mTORC1 pathway downregulation by all inhibitors, with abemaciclib exerting the most potent suppression. Subsequent kinase activity analysis indicated an upregulation of AKT1 kinase activity following CDK4/6 inhibitor stimulation. Furthermore, acridine orange staining and flow cytometry indicated that abemaciclib elevated lysosomal proteins and acidification while increasing reactive oxygen species (ROS). Collectively, this study deepens the understanding of shared and drug-specific molecular characteristics and mechanisms of CDK4/6 inhibitors at multi-omic levels. Moreover, it provides an experimental basis and potential directions for customizing combination therapies based on pathway vulnerabilities, as well as exploration of novel therapeutic modes and drugs.

Deregulation of coagulation proteases can lead to a spectrum of hemostatic abnormalities, ranging from subtle subclinical to life-threatening coagulopathies. Additionally, coagulation factors play critical roles in many abnormalities beyond thrombosis, extending to hyperactive inflammatory responses and promoting tumorigenesis. Triple-negative breast cancers (TNBCs) are marked by a pro-thrombotic and highly inflammatory tumor microenvironment that fuels progression and therapeutic resistance, yet the molecular crosstalk linking coagulation signaling to inflammatory transcription remains incompletely understood. Here, we identify DNA topoisomerase I (Top1) as a critical downstream effector of thrombin-PAR1 driven inflammatory reprogramming in breast cancer cells. Thrombin enhances Top1 activity without altering its expression, amplifying NF-κB dependent cytokine production. Mechanistically, this response is governed by suppression of the tumor suppressor Par-4, which resides at the epicentre of this signaling axis and negatively regulates both Top1 and NF-κB. PAR1 activation enforces Par-4 loss through coordinated epigenetic silencing and phosphorylation-dependent cytoplasmic inactivation, thereby relieving transcriptional constraints and sustaining pro-tumorigenic inflammation. Importantly, targeting PAR1 markedly sensitizes TNBC tumors to the Top1 inhibitor irinotecan, highlighting a therapeutically actionable vulnerability. These findings reveal a previously unappreciated convergence of coagulation, epigenetic control, transcriptional activation and DNA topology control in breast cancer-suggesting the possibility of repurposing FDA-approved oral anticoagulants as an effective combinatorial therapy along with Top1 poisons in treating TNBCs.

Tamoxifen has been commonly used in endocrinotherapy for estrogen receptor (ER)-positive breast cancer. In recent years, the immunomodulatory effects of high-dose tamoxifen have been discovered. The immunosuppressive tumor microenvironment (TME), however, remains a major obstacle to the efficacy of radiotherapy. Whether high-dose tamoxifen can reprogram TME to synergistically enhance radiation efficacy is still ambiguous. Here, we have found high-dose tamoxifen could dramatically enhance radiation-induced antitumor effects without significant side effects in immunocompetent mice. Flow cytometry and multiplex immunofluorescence experiments revealed that radiation combined with tamoxifen resulted in significant enrichment of effector CD8+ T cells and M1 tumor associated macrophages (TAMs). Depletion of TAMs and CD8+ T cells impaired the synergistic antitumor effects mediated by combination treatment of radiation and tamoxifen. Furthermore, spatial proximity analysis demonstrated a dramatically reduced nearest-neighbor distance between CD8+ T cells and M1-like TAMs in the combination group, indicating enhanced cellular interaction within the TME. In vitro experiments demonstrated that tamoxifen directly acted on TAMs, rather than CD8+ T cells, to promote M1 polarization and subsequently enhanced the activation and effector function of CD8+ T cells, regardless of ER expression in tumor cells and macrophages. Mechanistically, RNA-sequencing and experimental validation uncovered that tamoxifen and tumor cell-derived TNF-α and IL-1β after irradiation synergistically activated the JNK/c-JUN pathway and promoted M1 polarization of TAMs. In conclusion, our study revealed the immunomodulatory effects of high-dose tamoxifen in the context of radiation and provided preclinical evidence for combination therapy of high-dose tamoxifen and radiotherapy in both ER-positive and ER-negative cancers.

Schizantherin B (SNB), a key bioactive ingredient of the Chinese traditional medicine Schisandra chinensis, possesses anti-inflammatory and antioxidant properties. Cisplatin (CDDP) is typically used to treat various malignant tumors, however, its clinical utility is often limited by significant off-target toxicities, most notably irreversible hearing loss. Various strategies have been explored to mitigate this side effect. In this study, we investigated the protective effects of SNB against cisplatin-induced hearing loss(CIHL) as a potential preclinical therapeutic strategy.Firstly, in the ex-vivo basilar membrane, we found that SNB alleviated CDDP-induced loss of hair cells, cochlear spiral ganglion cells, and ribbon synapses. Secondly, in vivo experiments showed that SNB protected animals against CHL, returning their response close to normal level. Thirdly, in multiple tumor cell lines, we found SNB did not interfere with CDDP's anti-tumor effects. Consistently, in a mouse model for breast cancer, we found that SNB did not obtrude CDDP's effects in reducing tumor mass. Finally, in examining the molecular mechanisms, we found that SNB reduced both oxidative stress and cell apoptosis in the auditory cell line of HEI-OC1 during CDDP treatment, likely through the Bcl-2/Bax/cleaved-Caspase-3 signal pathways. Collectively, our study demonstrated that SNB mitigates CIHL without interfering with its therapeutic effects in treating cancer, suggesting that SNB is a potential drug candidate for preventing CIHL in cancer patients.

In this research, a surface ligand engineering strategy is employed to fabricate UiO-66 confined Au nanoclusters (UiO-66@Au) with controllable multienzyme performances. Especially, ligand PSS and PVP can trigger optimized peroxidase (POD) and glucose oxidase (GOx) mimic activity respectively. Mechanistic studies revealed that electron transferring between Au and ligands can be an effective tactic to regulate the multienzyme activity. Theoretical calculations revealed that electron-withdrawing polystyrene sulfonate (PSS) can decrease the key energy barriers of ·OH desorption in POD process and electron-donating polyvinylpyrrolidone (PVP) decreased the key energy barriers of O2 to OOH∗ in GOx catalysis, which confirmed the enhanced POD and GOx activity correspondingly. For practical application, PSS-UiO-66@Au coupled lateral flow assay (LFA) can visually detect HER2-positive breast cancer exosomes as low as 428 exosomes/μL, about 16355-fold higher sensitivity than that of common LFA. PVP-UiO-66@Au with superior GOx-mimic activity can detect salivary glucose as low as 19 μM, meanwhile, due to the effective gluconic acid adsorption repulsion by PVP, PVP-UiO-66@Au displays superior stability even after 10 reuse cycles This work provides a simple route to regulate the multi-enzyme properties of UiO-66@Au and broadens the application in multiple target biosensing.

Triple-negative breast cancer (TNBC) lacks common receptors and exhibits aggressive behavior, limiting treatment options due to drug resistance and systemic toxicity. TNBC chemotherapy is hindered by poor tumor targeting, drug resistance, and systemic toxicity. Herein, this study presented a cascade targeting exosomal-cisplatin synergistic microneedle nanoplatform (CDDP@RKTExo-MN) as an intelligent wearable therapeutic device for TNBC treatment. Medicinal plant Taxus chinensis derived exosomes (TExo), carrying therapeutic miRNA, was synergized with cisplatin that induced ER stress to trigger a multimodal anti-tumor effects. The cisplatin-loaded TExo was further modified with αvβ3 integrin peptides and an ER-targeting motif for tumor homing and precise subcellular delivery. Leveraging the superficial localization of TNBC, the engineered TExo was integrated into a 3D-printed microneedle patch to construct a closed-loop transdermal delivery system (CDDP@RKTExo-MN). This bioactive architecture ensures precise drug delivery at the tumor site, effectively maximizing therapeutic efficacy while circumventing the systemic off-target toxicity inherent to conventional delivery strategies. CDDP@RKTExo-MN was shown for the cascade targeting capabilities with both cancer cells and their endoplasmic reticulums. By coordinated regulation of MAPK and TNF pathways, the system generated synergistic effects in both significantly amplifying apoptotic signaling and activating immunological protection. In vivo studies conclusively validated its superior tumor suppression efficacy alongside a favorable biosafety.

Breast and prostate cancers are both hormone-driven adenocarcinomas that undergo analogous invasion programs. Using lightsheet microscopy on intact tumors, we identified transitional junctions between precancerous and invasive regions. We then developed a multimodal serial-section workflow integrating volumetric reconstruction with spatial transcriptomics. Analysis of 319 spatial assays from 51 cases revealed gene expression features and novel structural insights defining the shift from precancer to invasive disease. In breast cancer, loss of MGP and PLAT was associated with invasive transition and promoted tumorigenesis in functional assays. In prostate cancer, GDF15, ALDH1A3, ANPEP, and FASN were upregulated along invasive progression, and their knockdown in PC-3 cells suppressed proliferation and migration. Enrichment of tumor-associated macrophages (SPP1+ and MS4A6A+) along non-triple-negative breast cancer breast cancer transitions highlights immune involvement as a potential driver of invasiveness. Our method of defining precise spatial locations of invasive transition allows for the direct interrogation of transition drivers, presenting new therapeutic targets for the two most prevalent cancers and providing a framework for studying spatially defined mechanisms of tumor progression. See related commentary by Jing and Li, p. 1720.

Triple negative breast cancer (require) new treatment strategies due to poor responses to current therapies. While myeloid SIRPα mediates immunosuppression, its cancer intrinsic role remains poorly understood. Human breast cancer scRNAseq profiles were used to examine SIRPα expression across different cell populations and subtypes. TNBC brain-tropic cells were injected into the mouse mammary fat pad for the orthotopic tumor model, and intracardiac-injected for brain metastasis models. Bulk RNA sequencing was used to determine SIRPα-regulated pathway. Stably SIRPα overexpressed and knockout TNBC cell lines were established to determine SIRPα intracellular regulation. Digital spatial profiling was utilized to investigate the orthotopic and brain metastasis tumor immune microenvironment. Human single-cell data showed that SIRPα levels increased in malignant TNBC epithelial cells. We observed that SIRPα is upregulated in patient breast-to-brain metastatic lesions. SIRPα is overexpressed in TNBC brain-tropic cells compared to parental cells. Bulk RNA-Seq showed that targeting SIRPα affects genes involved in mitochondrial dynamics, and that SIRPα upregulates mitochondrial fission and induces metastasis through the SHP2/Erk/Drp1 signaling pathway. In vivo, overexpression of SIRPα in cancer cells significantly increases TNBC systemic metastasis. Next, spatial proteomics revealed changes in the immune microenvironment associated with the SIRPα-regulated ECM protein fibronectin. Fibronectin induces microglial tolerance by impairing inflammatory signaling and metabolic reprogramming, allowing cancer to escape microglial immunosurveillance. Most importantly, SIRPα inhibition reduced TNBC brain metastatic lesions in mouse metastasis models. : Cancer-intrinsic SIRPα promotes TNBC brain metastasis through increased mitochondria fission and triggering microglia tolerance, and targeting SIRPα reduces brain metastasis.

The degradation of cell membrane and extracellular proteins with lysosome-targeting chimeras (LYTACs) is limited by nonrecyclable, receptor-dependent mechanisms that shuttle proteins to lysosomes, restricting the broad use of this emerging technology. Here we developed a recyclable chimera composed of a polyzwitterion and protein of interest (POI) ligand for protein degradation. This chimera could interact with the cell membrane to trigger macropinocytosis together with the POI in a receptor-independent manner. Furthermore, it can dissociate from the POI in the acidic endocytic compartments and subsequently be exocytosed through the endoplasmic reticulum-Golgi transcytosis pathway. Ultimately, the exocytotic chimera initiates the next round of targeted protein degradation. These macropinocytosis-mediated recyclable LYTACs (McR-TACs) durably degrade the cell membrane protein (programmed cell death ligand 1) or the extracellular protein (macrophage migration inhibitory factor) in a triple-negative breast cancer mouse model, thereby inhibiting the tumor growth. Collectively, McR-TACs show the potential of leveraging natural transport pathways to create recyclable protein degraders with wide-ranging applications.

The approvals of four CDK4/6 (cyclin-dependent kinases 4 and 6) inhibitors by the US Food and Drug Administration-palbociclib, abemaciclib, ribociclib and trilaciclib-as anticancer or supportive therapies establish the critical role of CDKs in regulating the cell cycle and cancer progression. However, the emergence of drug-resistant cancers underscores the adaptability and plasticity of this cellular pathway, highlighting the need for additional therapeutic strategies. CDK2, the key kinase activated downstream of CDK4/6, has emerged as a prominent pharmaceutical target, with numerous selective inhibitors reported in recent years. This Perspective discusses the adaptive plasticity of cell-cycle control in cancer cells and then highlights current strategies to target newly emerging CDK2 druggable pockets-including orthosteric, substrate-binding and allosteric sites-with an emphasis on chemical biology approaches, aiming to inspire the next generation of CDK2 inhibitors and kinase-targeted therapies more broadly.

Cardiovascular disease (CVD) is the leading cause of death worldwide. Chemotherapy-induced CVD is increasingly recognized as a contributor to long-term morbidity in cancer survivors. Doxorubicin (DOX) is a widely used chemotherapeutic to treat breast cancer, one of the most common cancers in the United States. However, over 10% of treated women experience acute cardiotoxicity immediately following treatment, and approximately 2% develop severe cardiotoxicity up to 10 years after treatment, yet the mechanisms driving this delayed onset remain unclear. Here, we show that DOX treatment of cardiac cells changes their function and paracrine signaling profile. Subsequent exposure of healthy cells to altered extracellular vesicles (EV) recapitulates the effects of direct DOX exposure in 2D/3D in vitro models, suggesting a mechanism for propagating initial injury. Plasma-EV miRNA profiling of blinded patient samples revealed distinct clustering by DOX-cardiotoxicity risk, with high-risk patients exhibiting miRNA signatures similar to those from DOX-treated models. Pathway analysis of miRNAs linked them to cardiac homeostasis and cardiotoxicity-related mechanisms, supporting the potential of plasma-EV miRNAs as noninvasive biomarkers for early risk stratification and personalized cardioprotective interventions in oncological care, and targeting of key miRNA clusters to enhance understanding of and intervention strategies for preventing the onset of DOX cardiotoxicity. STATEMENT OF SIGNIFICANCE: Doxorubicin is an effective chemotherapy drug, but its use is limited by cardiotoxicity that can appear during treatment or years later. The mechanisms driving this delayed injury remain poorly understood. In this study, we show for the first time that small extracellular vesicles released from doxorubicin-treated cardiac cells can propagate doxorubicin-like dysfunction to healthy cardiac cells even in the absence of the drug itself. We further identify broad miRNA cargo changes in these vesicles and show that related vesicle-associated miRNA shifts are also detectable in patient plasma. These findings provide new mechanistic insight into anthracycline cardiotoxicity and highlight extracellular vesicle-associated miRNAs as promising candidates for minimally invasive diagnosis and future therapeutic intervention.

Triple-negative breast cancer (TNBC) is an aggressive subtype with a high propensity for bone metastasis and limited treatment options. Although many proteins drive metastasis, most remain inaccessible to conventional small-molecule drugs due to lack of suitable binding sites, prompting interest in upstream RNA regulators. Using integrative bioinformatics and machine learning, we identified hsa-microRNA-301a-3p as a central regulator of TNBC bone metastasis. However, current nucleic acid-based therapies are limited by instability and inefficient delivery. To overcome these challenges, we designed a ribonuclease-targeting chimera (RIBOTAC) that recruits endogenous RNase L to degrade the primary transcript pri-microRNA-301a. The resulting RIBOTAC efficiently reduced hsa-microRNA-301a-3p levels, restored NEDD4L expression, and promoted degradation of the oncogenic protein YAP, thereby suppressing TNBC bone metastasis. These findings identify hsa-microRNA-301a-3p as a key driver and demonstrate RNA-targeted degradation as a therapeutic strategy in TNBC bone metastasis.

Despite the major advances in breast cancer management over the past few decades, metastatic breast cancer continues to be a major concern and the main cause of mortality associated with this disease. The Rho GTPase RAC1 has been implicated in breast cancer aggressiveness, yet the mechanisms underlying its prometastatic activity remain poorly understood. Although RAC1 overexpression is correlated with poor prognosis, our data indicate that increased abundance alone is insufficient to drive metastatic dissemination in preclinical models. While RAC1 overexpression in MMTV-ErbB2 mice accelerates primary tumor growth, it does not increase metastasis. Using complementary genetic models, we show that disrupting RAC1 SUMOylation reduces lung metastases; this uncoupling from primary tumor development is clearest in the RAC1K4R knock-in model, whereas the RAC1ΔSUMO1 transgenic mice also show partial attenuation of RAC1-driven proliferative signaling. Mechanistically, loss of SUMOylation impaired the ability of RAC1 to maintain its GTP-bound state in proinvasive contexts. To exploit this dependency, we developed a cell-permeable peptide, TAT-PRASI, that blocks RAC1 SUMOylation. TAT-PRASI decreases RAC1 activity and limits migration/invasion in vitro and metastasis in vivo while reducing the invasion of patient-derived organoids. Moreover, TAT-PRASI diminishes the RAC1-POTEE interaction, which is consistent with impaired invadopodia formation. Collectively, these findings identify SUMOylation as a specific, druggable regulator of RAC1-driven metastasis, providing a preclinical proof-of-concept for selectively targeting this posttranslational modification in metastatic breast cancer.

Short-read sequencing cannot determine whether co-occurring variants within a cancer gene lie on the same allele (cis) or opposing alleles (trans), a distinction with direct therapeutic consequences: trans configurations confirm biallelic tumor suppressor inactivation, whereas cis configurations generate compound oncogenic alleles with enhanced activity. Among 768 patients with prostate, breast, or ovarian cancers, we used mutational signatures to nominate cryptic genomic instability cases lacking a causative biallelic event on short-read sequencing. Long-read nanopore sequencing resolved 32 of 46 cryptic cases (69.6%) through methylation detection, long insertion resolution, and structural variant characterization, confirming trans inactivation in every resolved tumor suppressor case. Analysis of 4,496 MiOncoSeq samples identified 17,519 multi-hit gene pairs, 78.7% of which exceeded the 500 bp short-read phasing limit, and long-read phasing revealed recurrent compound cis alleles in NOTCH1, PIK3CA, PDGFRB, and KIT. Haplotype phasing addresses an overlooked gap in cancer variant interpretation and warrants integration into precision oncology.

Ferroptosis is a cell death mode caused by excessive accumulation of lipid peroxides (LPOs) due to intracellular metabolic pathway disorder, which is closely related to intracellular iron metabolism and lipid homeostasis. In cancer therapy, this metabolic imbalance promotes the clearance of tumor cells, suggesting that ferroptosis exerts a tumor suppressor function. However, the clinical translation of ferroptosis-based strategies in oncology is currently hindered by challenges such as inadequate therapeutic efficacy. In the past five years, the rational design of nano-formulations to induce cancer ferroptosis combined with other emerging therapeutic modalities (chemotherapy (CT), chemodynamic therapy (CDT), photodynamic therapy (PDT), sonodynamic therapy (SDT), and immunotherapy) has achieved promising therapeutic effects in many rodent tumor models. This review summarizes the recent progress of novel nano-formulations for cancer ferroptosis therapy. The multiple mechanisms and detection indicators of cancer cells during ferroptosis, the rational design of novel nano-formulations, and their strategies for inducing cancer ferroptosis will be introduced. Finally, the challenges and prospects of utilizing novel nano-formulations for cancer ferroptosis therapy will be discussed.

Growth hormone-secreting pituitary adenoma (GHPA) can cause excessive growth hormone production, which can lead to clinical symptoms such as acromegaly. Tumor stem cells (TSCs) are postulated to play a significant role in the development of GHPA. We performed RNA sequencing on human pituitary adenoma stem-like cells (hPASCs) and matched bulk tumors to map a differentiation protein-protein interaction (PPI) network. The function of the C-X-C motif chemokine receptor-4 (CXCR4) gene was tested in hPASCs and the GH3 cell line. The CXCR4-based 68Ga-pentixafor tracer was assessed by the positron emission tomography and computed tomography (PET/CT) scanning of GHPA patients. A total of 685 differentially expressed genes were identified between hPASCs and differentiated tumor cells. Four clusters of genes were predicted, each carrying distinct biological functions. CXCR4 was pinpointed as a potential hub gene, which significantly affected the self-renewal and differentiation of hPASCs, as well as the proliferation, invasion, and migration of GH3 cells. CXCR4 suppression decreased intra-tumoral angiogenesis and epithelial-to-mesenchymal transition (EMT). CXCR4-based 68Ga-pentixafor was a more adaptive tracer than 18F-FDG in diagnosing GHPA using PET/CT. This study provides a comprehensive genetic profiling of hPASCs and substantiates the critical role of CXCR4 in tumorigenesis, highlighting its potential in translational medicine.

Cyclins and cyclin-dependent kinases (CDKs) are frequently dysregulated in human cancers and represent compelling therapeutic targets. Beyond their well-recognized roles in cell cycle control, CDK/cyclin complexes orchestrate diverse oncogenic processes, including transcription, genome maintenance, epigenetics, metabolism, and immune regulation. Deciphering the multifaceted biology of CDK/cyclin will provide valuable insights and rationales for the development of CDK/cyclin-targeting strategies and modalities. While the clinical success of CDK4/6 inhibitors has validated CDKs as druggable targets, further efforts are urgently needed to target other CDKs and cyclins. This review critically evaluates recent mechanistic advances in CDK/cyclin biology and their pathological dysregulation across malignancies. We analyze the paradigm shift from conventional enzymatic inhibition toward proximity-induced modulation. Specifically, we highlight emerging approaches including proteolysis-targeting chimeras, HSP90-mediated targeting chimeras, hydrophobic tagging, molecular glues, and autophagy-tethering compounds that achieve selective elimination of CDKs or their cyclin partners. In parallel, we summarize strategies designed to redistribute CDK complexes and rewire transcription without enzymatic ablation, referred to as chemical inducers of proximity and transcriptional/epigenetic modulators. By integrating fundamental CDK/cyclin biology with pharmacological innovation in targeted protein degradation and kinase reprogramming, this review provides a timely roadmap for the CDK/cyclin research field and expands the frontiers of CDK/cyclin-targeted cancer therapy.

Platelet-derived growth factors (PDGFs) and their cognate receptors (PDGFRα/β) play critical roles in breast cancer progression and metastasis. This review summarizes current evidence of PDGF ligand and receptor expression patterns, oncogenic functions, prognostic significance and therapeutic targetability, with a specific focus on small molecule inhibition. PDGF-PDGFR signaling is known to contribute to epithelial to mesenchymal transition, cancer stem cell maintenance, desmoplasia, angiogenesis, and immune modulation. Additionally, the four PDGF ligands have distinct oncogenic functions. PDGFA and PDGFB have been implicated in breast cancer associated brain metastasis, while PDGFC has been shown to play a crucial role in fibroblast activation. PDGFD, while less studied, may activate epithelial to mesenchymal transition in breast cancer. High expression of PDGFA, PDGFB, PDGFC, and stromal PDGFRβ correlate with poor patient survival, highlighting their potential as candidate biomarkers. We specifically focus on evaluating current therapeutic strategies which target the PDGF-PDGFR axis, including neutralizing antibodies, aptamers, and small molecule inhibitors, which show preclinical promise but limited clinical success in breast cancer to date. We discuss future research directions with emphasis on identifying selective inhibitors, utilizing PDGF-PDGFR signaling components for patient stratification, and combination with immunotherapies.

Collective cell migration plays a critically regulatory role in both physiological and pathological processes, e.g., embryonic development, wound healing, cancer progression and metastasis. Nevertheless, spatiotemporal progression of epithelial and mesenchymal cell migration remains poorly understood. Here, we report an epithelial-mesenchymal transition (EMT)-modulated cell phenotype phase separation (CPPS) within spatially confined microenvironments, where the mesenchymal cells display enhanced boundary-directed colonization in a phenotype-dependent manner. Subsequently, we reveal that the CPPS process correlates with the degree of EMT and cytoskeletal inhibition. With the aid of a boundary attraction potential (BAP), we further develop a reaction-diffusion-based model that can quantitatively capture the spatiotemporal evolution of the EMT-related collective migration. Our findings imply that phenotypic heterogeneity among cells is a major determinant of phase separation in collective migration. This work not only uncovers the CPPS phenomenon and its underlying biophysical mechanism in collective cell migration, but also provides a new perspective for dissecting EMT-regulated intratumoral phenotypic heterogeneity. STATEMENT OF SIGNIFICANCE: Collective cell migration drives development, wound healing, and cancer metastasis, yet how phenotypically heterogeneous populations self-organize remains unclear. Using micropatterned substrates and live-cell tracking, we report cell phenotype phase separation (CPPS) during epithelial-mesenchymal transition (EMT)-mediated collective migration. Inspired by the classical Turing's theory, we develop a reaction-diffusion model with a boundary attraction potential that recapitulates the spatiotemporal dynamics of CPPS across micropattern geometries, substrate stiffnesses, and cell compositions. Our findings not only reveal a comprehensive biophysical mechanism linking EMT status to collective migration, which is crucial for dissecting cellular spatial self-organization in tumors and tissue development, but also suggest that CPPS may serve as a potential target for therapeutic intervention and drug screening.

Cancer is increasingly recognized as a disease of the dysregulated epigenome; however, current epi-drugs are blunt, systemically toxic instruments. Catalytically dead CRISPR nucleases (dCas9) linked to chromatin effectors have now made it possible not only to write and erase epigenetic marks at specified loci without double-strand breaks but also to add an element of optogenetics, or reversible and light-encoded control over the timing and localization of the editors. In this review, the technological underpinnings of light-controlled CRISPR-dCas9 epigenome editing, which include architectures of dCas9 scaffold and guide, blue-to-near-infrared photoswitches, and high-gain epigenetic effector designs, are synthesized, and viral, non-viral, and stimuli-responsive delivery platforms, which have to be co-optimized with clinical light interfaces, are discussed. We then outline four functional routes by which opto-epigenome editors may be used therapeutically in cancer: tumor suppressor reactivation; oncogene and super-enhancer repression with metabolic rewiring; control of cancer stem cell differentiation; and immunomodulation of the tumor microenvironment. Lastly, a translational roadmap is defined in terms of preclinical model tiers, biomarker strategies, regulatory and manufacturing factors, and future directions, including NIR and bioluminescent actuation, implantable μLED devices, and AI-guided closed-loop illumination. Together, these aspects constitute design principles for advancing light-addressable epigenome editors toward first-in-human studies and for integrating them into combination regimens as a new class of precision cancer therapeutics.

Prohibitin 1 (PHB1) and prohibitin 2 (PHB2) are highly conserved, ubiquitously expressed scaffold proteins that play central roles in cellular physiology by forming heterodimeric ring-shaped complexes. Their subcellular localization to mitochondria, the nucleus, cytoplasm, and plasma membrane underpins a remarkable functional pleiotropy that is profoundly exploited in cancer. This review provides a comprehensive synthesis of the current understanding of PHBs in tumor biology, spanning structural features, post-translational modifications, and their integration into multiple oncogenic signaling networks. We systematically describe how PHB1 directly activates the RAS-RAF-MEK-ERK cascade through regulated phosphorylation, how both PHB1 and PHB2 fine-tune the PI3K/Akt/mTOR axis through ubiquitination-dependent scaffolding and degradation of negative regulators, and how they exert bidirectional control over Wnt/β-catenin and NF-κB pathways. A major focus is the dual role of the mitochondrial PHB complex: protecting cristae architecture and regulating the OMA1-OPA1 axis, orchestrating respiratory chain supercomplex assembly, metabolic substrate switching, and mitophagy, while simultaneously suppressing or, in specific contexts, promoting reactive oxygen species signaling and ferroptosis. The review further dissects how dynamic nucleocytoplasmic shuttling of PHBs couples metabolic status to cell cycle progression, stemness, and epigenetic remodeling through interactions with transcription factors (E2F1, p53, Sp1) and chromatin modifiers (MLL2, HDAC1). Within the tumor microenvironment, PHBs emerge as critical immunometabolic hubs that influence macrophage polarization, cGAS-STING activation, and sexual dimorphism in immune responses. We summarize the cancer-type-specific expression patterns of PHB1/2 and their prognostic value, and provide an in-depth analysis of the mechanisms by which PHBs confer resistance to platinum drugs, paclitaxel, PARP inhibitors, and radiotherapy through stabilization of anti-apoptotic proteins, mitochondrial protection, and maintenance of cancer stem cell properties. Finally, we catalogue the expanding armamentarium of PHB-targeted interventions, including small-molecule ligands, stapled peptides, DNA aptamers, and siRNA delivery platforms, and discuss the challenges and opportunities for clinical translation. By integrating molecular mechanisms with translational perspectives, this review highlights PHBs as unique regulatory nodes at the intersection of metabolism, signaling, and immunity, and advocates for precision strategies that exploit context-specific PHB functions to overcome therapy resistance and improve cancer treatment.

CUDC-907, referred to as Fimepinostat, functions as a dual inhibitor of PI3K and HDAC, exhibiting significant anti-tumor efficacy in a range of cancer types. However, its specific role in glioblastoma is not well understood. In this study, we investigated the effects of CUDC-907 on glioblastoma using cellular, organoid, and animal models to assess its inhibitory potential and toxicity. In vitro, we assessed glioblastoma cell proliferation, migration, invasion, and apoptosis using standard assays. glioblastoma organoids were treated to examine three-dimensional tumor growth and cellular changes. For in vivo analysis, animal models with glioblastoma received CUDC-907 to study its systemic impact and toxicity, with tumor progression closely monitored. We also tested the synergy between CUDC-907 and temozolomide to evaluate enhanced chemosensitivity. RNA sequencing was conducted to explore the fundamental molecular mechanisms involved, focusing on alterations in the cell cycle and DNA damage repair pathways. Our findings show that CUDC-907 significantly suppresses the proliferation, migration, and invasion of glioblastoma cells and promotes apoptosis, while exhibiting minimal toxicity. Additionally, CUDC-907 acts synergistically with temozolomide, a chemotherapy drug used for glioblastoma treatment, enhancing glioblastoma's chemosensitivity to temozolomide. RNA sequencing suggests that CUDC-907 achieves its effects by influencing the glioblastoma cell cycle and inhibiting DNA damage repair. Overall, the data suggest that CUDC-907 may be a promising anti-cancer agent for glioblastoma treatment.

Ferroptosis, a unique modality of regulated cell death, has become an emerging strategy for tumor therapy. Multiple cellular pathways, including redox homeostasis, iron handling, epigenetic regulation, and metabolic changes, could mediate ferroptosis. Here, we demonstrate that high expression of iron-responsive element binding protein (IRP1)/A + T rich interaction domain protein 3a (ARID3A) inhibits ferroptosis and enhances chemoresistance of pancreatic cancer cells via handling the promoter region of a ferroptosis gene, cytoglobin (CYGB). Mechanistically, the high level of iron leads to nuclear translocation of IRP1 and ARID3A, thereby mediating ARID3A binding to the promoter region of CYGB and down-regulation of chromatin accessibility. The decrease of CYGB expression results in pancreatic cancer cell resistance to ferroptosis, which makes them more resistant to chemotherapy. Clinically, high expression of IRP1 and ARID3A associates with unsatisfactory chemotherapeutic response and poor survival of patients with pancreatic cancer. Our study highlights the role of IRP1/ARID3A complex as a chemotherapy target and its potential in the combined application of ferroptosis drugs.

Nitric oxide synthase 2 (NOS2) and cyclooxygenase 2 (COX2) lie at a critical intersection between inflammation, metabolism, and oncogenic signaling, where they cooperatively promote and establish a Nitric Oxide (NO)-driven Warburg phenotype in advanced cancers. Early work in macrophages established NOS2-derived NO as both a signaling molecule and metabolic stressor that inhibits oxidative phosphorylation (OXPHOS) by targeting iron-sulfur enzymes and respiratory complexes, forcing neighboring cells to rewire metabolism. In human tumors, sustained NOS2 expression in cancer cells and tumor-associated macrophages (TAMs) enforces a Warburg-like state characterized by high glycolytic flux, glutamine dependence, and enhanced NADPH production, supporting proliferation, biosynthesis, and resistance to oxidative stress. At nitrosative-signaling concentrations (≈100-500 nM), NO breaks carbon entry into the TCA cycle at aconitase and pyruvate dehydrogenase, progressively disables dehydrogenase complexes containing dihydrolipoamide dehydrogenase (DLD) and electron-transport complexes (ETCs), and activates hypoxia-inducible factor 1-alpha (HIF-1), phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt), extracellular signal-regulated kinase (ERK)/pyruvate kinase M2 (PKM2)/c-Myc signaling axis, nuclear factor erythroid 2-related factor 2 (Nrf2), and transforming growth factor Beta (TGF-β)/SMAD pathways. These biochemical and signaling effects convert transient glycolytic adaptation into chemically enforced dependency, further stabilized by metabolite-driven inhibition of ten-eleven translocation (TET) and Jumonji demethylases, creating an "epigenetic lock" that maintains oncogenic transcriptional programs. NOS2 and COX2 form a reciprocal feed-forward circuit in which NO, prostaglandin E2 (PGE2), interleukin (IL)-6, and IL-8 reinforce one another, driving tumor-promoting inflammation, immunosuppression, angiogenesis, and metastasis while depleting nutrients and acidifying the tumor interstitial fluid. Spatially, NOS2/COX2 niches at the tumor-stroma interface and within immune deserts generate gradients of NO, PGE2, oxygen, and metabolites that partition tumors into microdomains with distinct metabolic states, immune composition, and therapeutic vulnerabilities. Integrating these insights with Hanahan's updated hallmarks of cancer, we propose that NOS2-derived NO functions as a node synchronizing deregulated energetics, inflammation, immune evasion, plasticity, and therapy resistance within the tumor microenvironment (TME). Targeting the NOS2-COX2 axis and its downstream NO-iron-epigenetic circuitry may therefore disrupt multiple hallmarks and reveal combinatorial strategies to exploit NO-induced metabolic liabilities in cancer.

WNT2B is canonically characterized as a secreted WNT-family ligand, which is transported to the extracellular space via the endoplasmic reticulum (ER)-Golgi pathway and binds to cell surface FZDs (frizzled class receptors) to trigger downstream signaling cascades. Here, we identify a previously unrecognized non-secretory intracellular function of WNT2B in impairing endosomal trafficking to inhibit macroautophagy/autophagy, as well as a non-canonical LC3B-II-dependent autophagic secretion mechanism for WNT2B. Specifically, the non-secretory intracellular pool of WNT2B via its conserved middle domain (MD) binds to the spectrin repeat domain (SRD) of WASHC5, competitively displacing WASHC1 and thereby disrupting WASH complex assembly and inhibiting WASHC1-mediated actin polymerization on early endosomes. This disruption impairs endosomal cargo trafficking, including the core autophagy protein ATG9A, leading to defective autophagy initiation and subsequent accumulation of pro-inflammatory and pro-fibrotic factors in fibroblasts. We validated this mechanism in vivo using a TNBS-induced mouse model of chronic colitis. Fibroblast-specific wnt2b deletion restores autophagy, reduces pro-inflammatory cytokine secretion, and ameliorates intestinal fibrosis. Consistently, in Crohn disease (CD) patient tissues, elevated WNT2B in fibrotic regions negatively correlates with autophagy activity, and positively correlates with pro-fibrotic phenotypes, and clinical disease severity. Moreover, we identify a novel LC3B-II-dependent autophagic secretion pathway for WNT2B, which is distinct from the conventional ER-to-Golgi-dependent protein secretion. Collectively, our study delineates a novel non-canonical WNT2B-WASH complex-ATG9A regulatory axis through which WNT2B impairs endosomal trafficking and disrupts autophagy, ultimately amplifying inflammation and fibrosis. This study suggests that WNT2B may serve as a promising therapeutic target for CD and autophagy-associated fibrotic disorders.Abbreviations: 3-MA: 3-methyladenine; AAV: adeno-associated virus; ACTA2: actin alpha 2, smooth muscle; ARPC2: actin related protein 2/3 complex subunit 2; ATG: autophagy related; CCN3: cellular communication network factor 3; CD: Crohn disease; CK666: 2-fluoro-N-[2-(2-methyl-1H-indol-3-yl)ethyl]benzamide; COL1A1: collagen type I alpha 1 chain; Co-IP: co-immunoprecipitation; CTNNB1: catenin beta 1; DBcAMP: dibutyryl cyclic adenosine monophosphate; DPT: dermatopontin; EEA1: early endosome antigen 1; EGFR: epidermal growth factor receptor; ELISA: enzyme-linked immunosorbent assay; ER: endoplasmic reticulum; ESCRT: endosomal sorting complexes required for transport; EV: extracellular vesicle; FRAP: fluorescence recovery after photobleaching; FL: full length; FZD: frizzled class receptor; GST: glutathione S-transferase; HIF: human intestinal fibroblast; HMGB1: high mobility group box 1; IKBKB: inhibitor of nuclear factor kappa B kinase subunit beta; IL6: interleukin 6; LDELS: LC3-dependent EV loading and secretion; LPS: lipopolysaccharide; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MD: middle domain; MEFs: mouse embryonic fibroblasts; MTOR: mechanistic target of rapamycin kinase; MVB: multivesicular body; NFKB: nuclear factor kappa B; NFKBIA: NFKB inhibitor alpha; PDCD6IP: programmed cell death 6 interacting protein; PLA: proximity ligation assay; RELA/p65: RELA proto-oncogene, NF-kB subunit; SAFB: scaffold attachment factor B; SES-CD: Simple Endoscopic Score for Crohn disease; SIM: super-resolution structured illumination microscopy; SMAD3: SMAD family member 3; SQSTM1/p62: sequestosome 1; SRD: spectrin repeat domain; TEM: transmission electron microscopy; TFRC: transferrin receptor; TGFB1: transforming growth factor beta 1; TGOLN2: trans-golgi network protein 2; TNBS: 2,4,6-trinitrobenzenesulfonic acid; TNF: tumor necrosis factor; VCA: Verprolin homology, Central and Acidic; WASHC: WASH complex subunit; WLS: Wnt ligand secretion mediator; WCL: whole cell lysates; WNT: Wnt family member; WT, wild type.

Chronic inflammation has emerged as a central driver for the initiation, progression, and therapeutic resistance of prostate cancer (CaP). While androgen receptor (AR) signaling serves as the primary axis for prostate tumorigenesis, mounting evidence suggests that continuous activation of inflammatory signaling pathways, especially nuclear factor kappa B (NFκB) and interleukin-6 (IL6)/Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) pathways, profoundly reshape the transcriptional and epigenetic landscape of advanced disease. These signaling pathways largely converge with AR signaling to form a common axis and promote tumor survival, proliferation, angiogenesis, immune evasion, epithelial to mesenchymal transition (EMT), and metastasis. Furthermore, prolonged exposure to cytokines such as IL6 and tumor necrosis factor-α (TNFα) leads to constitutive activation of STAT3 and NFκB signaling pathways. This persistent inflammatory signaling increases AR transcriptional activity even under androgen-deprived conditions, thereby facilitating the development of castration-resistant prostate cancer (CRPC). Apart from transcriptional crosstalk, inflammatory signaling pathways interact with epigenetic remodeling mechanisms, including histone modifications, DNA methylation, and dysregulation of chromatin modifiers such as EZH2, LSD1, and BRD4. These epigenetic variations stabilize aberrant gene expression programs and promote therapeutic resistance in metastatic CRPC. The present review discusses the crosstalk between inflammatory signaling, AR reprogramming, and epigenetic remodeling, and illustrates emerging therapeutic strategies targeting NFκB and JAK/STAT3 signaling pathways. This review highlights the importance of the integrated inflammatory-AR-epigenetic axis in CRPC progression and emphasizes its potential for improved biomarker stratification and the development of effective combinatorial therapeutic strategies to overcome resistance and improve clinical outcomes in advanced CaP.

Estrogen receptor-positive (ER+) breast cancer exhibits a marked propensity for skeletal metastasis; however, the molecular drivers of bone colonization remain incompletely defined. We investigated the tumor suppressor neurofibromin (NF1), a dual repressor for RAS and ER signaling, whose inactivation promotes endocrine therapy (ET) resistance and is associated with inferior relapse-free survival. NF1 copy number loss was detected in 62% of ER+ patients who subsequently developed metastases and was associated with an increased likelihood of bone metastases at initial diagnosis. In mouse xenograft models, NF1-depleted ER+ breast cancer cells demonstrated enhanced dissemination to skeletal sites following surgical resection of primary tumors. Furthermore, after intra-iliac injection, NF1-depleted cells generated significantly greater tumor burden in bone. Transcriptomic profiling revealed enrichment of bone-related gene signatures in NF1-depleted ER+ breast cancer cells, which more potently induced osteoclast differentiation and bone loss in co-culture systems. In parallel, low NF1 expression correlated with repressed T cell functional states in primary breast tumors and bone metastases. Consistent with these clinical observations, NF1-depleted ER+ breast cancer cells more effectively inhibited proliferation, interferon-γ secretion, and cytotoxicity of human primary CD8+ T cells. Collectively, these findings identify NF1 inactivation as a key driver of bone metastasis in a substantial subset of ER+ breast cancers. By amplifying the osteolytic "vicious cycle" and promoting immune evasion, NF1 loss remodels the microenvironment to favor tumor expansion. These results further suggest that NF1 loss functionally links therapy resistance with increased skeletal metastatic potential.

Angiogenesis supports colorectal cancer (CRC) proliferation and metastasis, yet how lipid metabolic reprogramming intersects with angiogenesis signaling remains unclear. Here, we found that LINC01186 is upregulated in CRC and correlates with advanced stage and poor prognosis. Mechanistically, LINC01186 functions as a molecular scaffold that concurrently interacts with the deubiquitinating enzyme USP4 and long-chain fatty acyl-CoA synthetase ACSL4, facilitating USP4-mediated deubiquitination of ACSL4. Elevated ACSL4 facilitates the accumulation of polyunsaturated fatty acids (PUFAs), namely arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid. Notably, elevated PUFA levels protect VEGFA from autophagy-lysosomal degradation by reducing autophagic flux, resulting in VEGFA overexpression and consequent promotion of angiogenesis. The absence of LINC01186 or ACSL4 impairs this signaling cascade, reduces PUFA levels, reactivates the autophagic degradation of VEGFA, and markedly obstructs the migration, invasion, and angiogenesis of CRC cells both in vitro and in vivo. Exogenous PUFA supplementation fully restores these abnormalities, which supports the essential involvement of lipid reprogramming in this mechanism. The LINC01186/ACSL4/VEGFA axis is essential in the formation and progression of CRC, with the overexpression of LINC01186 likely acting as a significant catalyst for excessive angiogenesis and metastatic advancement. Our findings highlight a unique lncRNA-mediated regulatory mechanism that connects deubiquitination, lipid metabolism, autophagy, and angiogenesis. Notably, although ferroptosis was not directly assessed in this study, the LINC01186/USP4/ACSL4 axis may have broader implications for ferroptosis research given that ACSL4 is a critical regulator of ferroptosis. Taken together, these results suggest that the LINC01186/USP4/ACSL4 axis represents a prospective therapeutic target for anti-angiogenic treatment in CRC.

ATG16L1 (autophagy related 16 like 1) is a core macroautophagy/autophagy protein essential for autophagosome formation. It also functions in non-canonical autophagy pathways such as LC3-associated phagocytosis (LAP) and in other processes including immunity, inflammation, and membrane trafficking. This review synthesizes recent advances and proposes that ATG16L1 functions as a central molecular integrator governed by a multi-layered regulatory code. This framework includes genetic polymorphisms, transcriptional control, and diverse post-transcriptional and post-translational mechanisms. We detail how these regulatory layers collectively fine-tune ATG16L1 function in response to cellular stress. Dysregulation of this network contributes broadly to human diseases including inflammatory bowel disease, cancer, and neurodegenerative disorders. Notably, the functional impact of specific regulatory events is highly context dependent, a principle exemplified by the Crohn disease-associated T300A polymorphism. Deciphering this regulatory landscape and its crosstalk with both autophagy-dependent and autophagy-independent functions positions ATG16L1 as a pivotal node in cellular homeostasis and as an emerging therapeutic target.Abbreviations ATG: autophagy related; CASM: conjugation of Atg8-family proteins to single membranes; CCD: coiled-coil domain; CEBPA/CEBPα: CCAAT enhancer binding protein alpha; CHUK/IKKA: component of inhibitor of nuclear factor kappa B kinase complex; circRNA: circular RNA; CPT1A: carnitine palmitoyltransferase 1A; CREB: cAMP responsive element binding protein; CSNK2: casein kinase 2; FTO: FTO alpha-ketoglutarate dependent dioxygenase; GJA8/connexin 50: gap junction protein alpha 8; H/R: hypoxia-reoxygenation; HDAC: histone deacetylase; KAT2B/PCAF: lysine acetyltransferase 2B; KDM1A: lysine demethylase 1A; LAP: LC3-associated phagocytosis; lncRNA: long non-coding RNA; LRRK2: leucine rich repeat kinase 2; m6A: N6-methyladenosine; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; miRNA/MIR: microRNA; Mtb: Mycobacterium tuberculosis; ncRNA: non-coding RNA; PE: phosphatidylethanolamine; PI3K: phosphoinositide 3-kinase; PRKA/PKA: protein kinase cAMP-activated; PPP1: protein phosphatase 1; RAB33B: RAB33B, member RAS oncogene family; RB1CC1/FIP200: RB1 inducible coiled-coil 1; SETD7: SET domain containing 7, histone lysine methyltransferase; SQSTM1/p62: sequestosome 1; TNF/TNF-α: tumor necrosis factor; ULK: unc-51 like autophagy activating kinase; V-ATPase: vacuolar-type H+-translocating ATPase; VDR: vitamin D receptor; WIPI2B: WD repeat domain, phosphoinositide interacting 2B; YTHDF2: YTH N6-methyladenosine RNA binding protein F2; ZDHHC7: zDHHC palmitoyltransferase 7.

Cancer stemness drives malignant progression and drug resistance in hepatocellular carcinoma (HCC). Although mitochondrial dynamics are known to influence HCC development, the precise mechanisms linking mitochondrial function to stemness remain largely elusive. Integrating bulk and single-cell transcriptomics, we identified Butyrophilin Subfamily 3 Member A3 (BTN3A3) as a novel oncogene driving HCC stemness. BTN3A3 depletion markedly reduced sphere formation, stemness-related gene expression, and the percentage of CD90+/EpCAM+ cancer stem cells. Rescue experiments confirmed that BTN3A3 promotes HCC cell proliferation, migration, and invasion. Furthermore, BTN3A3 depletion sensitized HCC cells to sorafenib by inducing ROS accumulation and apoptosis. Mechanistically, mass spectrometry and Co-IP identified TOMM22 as a key mitochondrial interactor of BTN3A3. Crucially, sorafenib stress actively promotes BTN3A3 mitochondrial translocation, where it shields TOMM22 from ubiquitin-proteasome-dependent degradation. BTN3A3 deficiency led to TOMM22 depletion, mitochondrial fragmentation, and impaired oxidative phosphorylation (OXPHOS) and ATP production. Importantly, silencing TOMM22 reversed BTN3A3-mediated stemness and sorafenib resistance. In vivo orthotopic xenograft models and patient-derived organoids (PDOs) further validated that BTN3A3 correlates with stemness and malignant tumor growth. Utilizing 5E08, a pan-BTN3 monoclonal antibody, markedly suppressed tumor growth and concurrently downregulated TOMM22 expression in vivo. In conclusion, our study unveils a previously unrecognized non-immunological role for BTN3A3 in mitochondrial reprogramming. We demonstrate that BTN3A3 drives HCC stemness and drug resistance by preventing TOMM22 ubiquitination to maintain mitochondrial homeostasis. These findings position BTN3A3 as a promising therapeutic target, with the pan-BTN3 monoclonal antibody 5E08 offering a potential strategy to overcome stemness-driven malignancy and resistance in HCC patients.

Radiation therapy for head and neck cancer often causes xerostomia, a chronic salivary gland (SG) dysfunction driven by excessive reactive oxygen species (ROS), severely impacting the quality of life of patients. Existing treatments, such as artificial saliva or SG stimulants, offer only temporary relief. We developed a novel therapeutic system combining glutathione (GSH)-conjugated gelatin (Gel) hybrid protein-based cell carrier (GC) with adipose-derived mesenchymal stem cell (AdMSC) spheroids. This hybrid protein carrier (GC) extended the antioxidant activity of GSH by overcoming its short half-life and enabled efficient encapsulation and delivery of AdMSC spheroids to irradiation (IR)-damaged SGs. The 3D spheroids enhanced vascular endothelial growth factor (VEGF) expression through hypoxic core formation, promoting angiogenesis. In an IR-damaged mouse model, spheroid-encapsulated GC increased SOD2 expression and decreased 8-OHdG and NOX4 levels, effectively mitigating oxidative stress, fibrosis, and apoptosis, while accelerating angiogenesis. SG structural regeneration and functional recovery were confirmed using histological and immunohistochemical analyses following spheroid-encapsulating GC treatment. These results demonstrate the synergistic effects of ROS-scavenging activity from GC and paracrine signaling from AdMSC spheroids. This groundbreaking spheroid-encapsulating GC improves SG regeneration and functional recovery by integrating antioxidant activity with angiogenic effects, offering a paradigm-shifting solution for other oxidative stress-mediated tissue disorders.

Choroidal melanoma, the most prevalent subtype of uveal melanoma with a bleak prognosis, necessitates safer and more efficient treatment methods. Tumor cell membrane-modified nanodelivery systems have exhibited significant enhancement in photodynamic therapy (PDT). Currently, photosensitizers utilized in PDT encompass porphyrin-related organic molecules and other types, requiring complex and intricate systems construction. This study focuses on harnessing the naturally occurring photodynamic active photosynthetic unit-thylakoid which serve as a major source of singlet oxygen. Initially, thylakoids extracted from fresh leaves were identified based on their absorbance value, morphology, and characteristic proteins. The tumor cell membrane-thylakoid membrane (CM-Thy) photodynamic therapy system was prepared through ultrasonic fragmentation and liposomal extrusion technology. Due to its membrane targeting ability, CM-Thy demonstrated excellent uptake performance towards OCM-1 cells within intraocular tissues. Both in vitro and in vivo experiments confirmed CM-Thy's photodynamic activity. Specifically, Thy in the system generates singlet oxygen, after cellular internalization, the elevated intracellular reactive oxygen species (ROS) triggered by this process induce cell lipid peroxidation and increased membrane permeability, while singlet oxygen further mediates DNA oxidative damage, reduced cell proliferation capacity, and NLRP3 inflammasome-mediated pyroptosis, all these effects synergistically contribute to ultimate tumor cell apoptosis. Furthermore, the anti-tumor effect of CM-Thy was validated through various mechanisms involved in tumor formation such as angiogenesis and vasculogenic mimicry. Interestingly, the results from visible light experiments conducted in vitro also substantiated the remarkable therapeutic efficacy of this system for refractive eye disorders while providing innovative ideas for cross-species biological interventions.

Lysosomal membrane permeabilization (LMP) mediated by pH-responsive materials has demonstrated considerable potential in tumor therapy. However, tumor cells exhibit a pronounced adaptive capacity to remodel lysosomal pH, thereby resisting LMP induction by pH-responsive materials and ultimately compromising therapeutic efficacy. To overcome this limitation, we engineered proton-driven π-π stacked copper-phycocyanin nanoparticles (CuPC NPs) designed to selectively promote LMP in tumor cells through their aggregation within acidic lysosomes. Mechanistically, aggregated CuPC NPs directly engage the RIPK3/p-MLKL signaling axis to trigger LMP, thereby orchestrating the synergistic activation of necroptosis, ferroptosis, and cuproptosis. Due to the coordinated induction of multimodal cell death, these pH-responsive CuPC NPs effectively inhibit primary breast tumor growth and suppress pulmonary metastasis. Collectively, our study establishes a strategy for precise modulation of lysosomal function to enhance antitumor efficacy and provides valuable insights into the development of lysosome-targeted nanotherapeutics for tumor treatment.

Regulated cell death (RCD) is essential for organismal development, as it plays key roles in organ formation, pathogen defense, and the maintenance of homeostasis. However, while RCD subroutines-such as apoptosis, autophagy, pyroptosis, necroptosis, and ferroptosis-are fundamental for health, their dysregulation can be detrimental, potentially triggering or contributing to a variety of diseases. As a result, the modulation of RCD has emerged as a promising therapeutic strategy for numerous conditions, including infectious, neurodegenerative, autoimmune, cardiovascular diseases, and cancer. Over the past two decades, innovative experimental approaches for modulating RCD have proliferated in medicinal research. In parallel, a growing array of online resources-such as databases, web servers, machine learning (ML)/artificial intelligence (AI) models, omics technologies, and systems biology networks, collectively referred to as "in silico methods"-has become available to accelerate research progress. Recognizing the potential of these online resources, we have compiled a comprehensive overview of relevant in silico approaches to RCD, aiming to facilitate their effective use in the development of future therapeutic interventions. RCD-associated databases serve as the foundational data infrastructure for in silico methods, encompassing extensive biological information spanning DNA, RNA, proteins, small molecules, and disease-related data. The continuous curation and expansion of these databases are crucial for elucidating various cell death modalities and visually characterizing the functional dynamics of target molecules across different pathological conditions. Through the integration of AI, ML, multi-omics technologies, and systems biology approaches, we can systematically decode the intricate regulatory networks of RCD and disease progression patterns. This interdisciplinary convergence significantly enhances the discovery pipeline for disease-specific biomarkers and therapeutic targets.

Ferroptosis is an iron-dependent form of regulated cell death driven by lipid peroxidation; however, how selective autophagy regulates ferroptotic sensitivity remains incompletely understood. Here, we identify RAB8A as a selective autophagic substrate and negative regulator of ferroptosis. Quantitative proteomic analyses reveal that ferroptotic stress induces ATG5- and ATG7-dependent degradation of RAB8A. Mechanistically, ferroptotic stimuli induce RNF126-dependent polyubiquitination of RAB8A and subsequent SQSTM1-mediated autophagic degradation. Functionally, loss of RAB8A sensitizes cancer cells to ferroptosis, whereas expression of the degradation-resistant active mutant RAB8AQ67L suppresses ferroptotic cell death. RAB8A interacts with TFRC and facilitates stress-induced redistribution of TFRC from the plasma membrane toward endolysosomal compartments. RAB8A deficiency impairs TFRC clearance, enhances transferrin-dependent iron uptake, and increases intracellular Fe2+ accumulation and lipid peroxidation. In fibrosarcoma and pancreatic cancer xenograft models, RAB8A depletion enhances the antitumor efficacy of ferroptosis-inducing therapy. Clinically, RAB8A is upregulated and associated with poor prognosis and ferroptosis resistance in pancreatic cancer. Collectively, these findings establish an autophagy-RAB8A-TFRC axis that regulates ferroptotic sensitivity.

Cancer cells undergo profound metabolic reprogramming to sustain uncontrolled proliferation within a nutrient-limited and often hypoxic tumor microenvironment (TME). Metabolic rewiring is an active driver of oncogenesis, immune evasion, epigenetic remodeling, and therapy resistance. Over the past century, our understanding of tumor metabolism has grown from Warburg's seminal description of aerobic glycolysis to a comprehensive adaptive network. Cancer cells coordinate glucose catabolism, mitochondrial oxidative metabolism, fatty acid synthesis and oxidation, amino acid catabolism, nucleotide biosynthesis, and one‑carbon metabolism into an integrated metabolic framework. These pathways form a deeply interconnected web in which metabolic intermediates serve as biosynthetic building blocks, bioenergetic substrates, redox buffers, signaling molecules, and epigenetic cofactors. Within the TME, metabolic competition between tumor cells and immune cells, together with the accumulation of immunosuppressive metabolites such as lactate, kynurenine, and adenosine, creates a profoundly immune-hostile landscape. Recent work has further revealed that key post-translational modifications, directly driven by metabolic flux, reshape the chromatin and proteome of both cancer cells and tumor-infiltrating immune cells, linking metabolism to gene regulation in previously unanticipated ways. Therapeutically, the FDA approval of IDH1/IDH2 inhibitors for acute myeloid leukemia demonstrated that metabolic enzymes are tractable oncology drug targets. Yet the broader effort to translate metabolic insights into robust clinical benefit has encountered formidable obstacles, including metabolic plasticity, intratumoral heterogeneity, overlap with normal tissue function, and inadequate biomarkers. This review traces the evolution of our understanding of cancer metabolism from its origins to therapeutic targeting. It further examines how anabolic and catabolic pathways, energy production, redox balance, and metabolic crosstalk across intracellular, intercellular, and systemic domains shape tumor biology and therapeutic response. It also critically analyzes approved and investigational metabolic therapies and charts a course for the emerging era of precision metabolic oncology.

Cancer progression is driven by epigenetic reprogramming, where promoter hypermethylation of tumour-suppressor genes and global hypomethylation reshape gene regulation and cellular phenotypes, promoting oncogenesis and disease advancement. We previously introduced the Methylscape, a cancer-specific DNA methylation landscape characterized by clustered promoter hypermethylation and gene body hypomethylation that enhances DNA's physical affinity for gold surfaces. Here, we demonstrate that Methylscape can be leveraged to monitor cancer progression. In a TGF-β-induced breast cancer epithelial-mesenchymal transition (EMT) model, we observe increased Methylscape enrichment of mesenchymal-state DNA, indicating that this method can sensitively detect subtle epigenetic remodelling linked to tumour progression. Using a gold-based DNA desorption enrichment strategy coupled with methylation sequencing and qPCR, we show that hypermethylated regions are preferentially enriched on gold surface. Finally, we developed a low-cost, disposable screen-printed electrode platform for stage-specific breast cancer monitoring. Together, these findings establish Methylscape as a promising biophysical biomarker for non-invasive, real-time monitoring of cancer progression, advancing its potential for clinical translation.

Radiotherapy augments immunotherapy, yet conventional 2 Gy x-ray fractions largely induce apoptosis, whereas robust immunogenic cell death (ICD) is more commonly associated with higher-dose irradiation regimens. Here, we reveal a dose-independent, high-LET-specific paradigm: a physical dose of 2 Gy of carbon ions provokes antitumor immunity. Carbon ions redirect death wiring to MLKL-dependent necroptosis and trigger an NF-κB-driven inflammatory cascade (IL1A/B, CXCL1/2/3), surpassing 8 Gy x-rays in immunogenicity despite lower direct tumor cell-killing capacity. Multi-omics analyses identify clustered DNA damage-driven super-enhancer remodeling as an epigenetic switch that suppresses cIAP1/2-caspase-8-mediated apoptosis while licensing inflammatory necroptosis. Pharmacologic inhibition of MLKL abolishes these responses to carbon ion radiotherapy (CIRT). In vivo, 2 Gy CIRT drives abscopal response with increased CD8+ T cell infiltration and function. Together, these findings establish CIRT as a feasible, immune-stimulatory radiotherapy converting local exposure into systemic control via super-enhancer-mediated reprogramming of death and inflammation, providing a mechanistic rationale to integrate carbon ions into next-generation radio-immunotherapy.

Acquired resistance to poly(ADP-ribose) polymerase (PARP) inhibitors limits the durability of niraparib maintenance therapy in epithelial ovarian cancer. Here, integrated multi-omic and functional analyses of paired parental and niraparib-resistant models identified DNMT3B-dependent promoter hypermethylation and silencing of argininosuccinate synthase 1 (ASS1), establishing an arginine-auxotrophic state in resistant cells. Despite impaired de novo arginine synthesis, resistant cells maintained intracellular arginine through enhanced extracellular acquisition. Arginine deprivation alone induced a largely cytostatic state characterized by mitochondrial dysfunction, oxidative stress, and energetic imbalance, whereas its combination with niraparib produced marked synergy, amplifying DNA damage, suppressing homologous recombination, and driving cell death. Mechanistically, arginine restriction depleted NAD(P)H and acetyl-CoA pools, reduced histone H3 acetylation, and compressed chromatin accessibility at DNA repair loci, thereby intensifying niraparib-induced genotoxic stress. Resistant cells also displayed persistent mTORC1 activation and blunted autophagic flux; mTORC1 inhibition restored autophagy and niraparib sensitivity but attenuated the incremental benefit of arginine deprivation, identifying mTORC1-constrained autophagy as a key determinant of this vulnerability. In vivo, a well-tolerated 50% arginine-restricted diet lowered circulating and intratumoral arginine and potentiated niraparib across resistant xenograft, patient-derived, and maintenance-mimicking models. Clinically, DNMT3B-high/ASS1-low tumors were associated with shorter progression-free survival, and serial plasma arginine declined prior to progression during niraparib maintenance, supporting plasma arginine dynamics as an exploratory candidate monitoring biomarker. Together, these findings define an epigenetically fixed arginine dependency in niraparib-resistant ovarian cancer and support partial dietary arginine restriction as a practical resensitization strategy.

Clear-cell renal cell carcinoma (ccRCC) is defined by cancer cells with lipid-filled cytoplasm that appears "clear" in tissue preparations; however, the origin of these lipids and their role in tumor progression remain unclear. Notably, cultured ccRCC cells rarely exhibit the clear-cell phenotype in vitro, suggesting that lipid accumulation in ccRCC is not cell-autonomous. Here we show that tumor-associated macrophages (TAMs) undergo immunometabolic reprogramming and serve as major suppliers of lipids for ccRCC cells. Upon activation by the tumor suppressor gene VHL-deficient kidney tubule cells, TAMs acquire adipogenic and cholesterol metabolic signatures, accumulate lipids, and differentiate into lipid-laden macrophages (LLMs) via a TGF-β-APOE-dependent pathway. LLMs then transfer lipids directly to tumor cells through tunneling nanotubes (TNTs). Lipidomic analyses revealed that LLMs and recipient tumor cells share nearly identical lipid profiles enriched in cholesterol and phosphatidates, but not triglycerides. In patient cohorts, elevated APOE expression levels in macrophages (MΦs) correlated with advanced disease stages. In vivo, disruption of MΦ-specific APOE expression abrogated the clear-cell phenotype, reduced tumor growth, and suppressed metastasis in autochthonous and orthotopic xenograft ccRCC models. These findings identify a previously unrecognized MΦ-tumor crosstalk in which reprogrammed TAMs supply lipids to tumor cells, driving the clear-cell phenotype and disease progression. Targeting the TGF-β-APOE axis or TNT-mediated lipid transfer represents a potential therapeutic strategy. More broadly, this work supports a "metabolic checkpoint" paradigm, revealing a therapeutically amenable vulnerability in ccRCC.

Ferroptosis represents a promising strategy to overcome apoptosis-associated drug resistance, yet access to diverse and drug-like ferroptosis-inducing chemical space remains limited. Among potential scaffolds for populating this high-value space, cyanamide-bearing molecules are attractive but synthetically challenging due to the long-standing difficulty of constructing the N-CN bond. Here we disclose a thiocyanate-enabled cascade cyclization that leverages inexpensive, low-toxicity inorganic thiocyanate as an unconventional electrophilic CN surrogate. This transformation proceeds through a cascade sequence involving SCN installation, remote S → N cyano migration, and subsequent intramolecular thiazine annulation under mild, metal-free conditions. The resulting cyanamide 3,4-dihydro-2H-1,4-thiazines exhibited nanomolar ferroptosis-inducing potency; lead compounds 3b and 3c achieved IC50 values of 6.8 nM and 14 nM, respectively; and significantly suppressed tumor growth in a lung cancer PDX model.

Retinoblastoma (RB) represents the most common primary intraocular malignancy in childhood and stands as a paradigm for translating molecular oncology into precision clinical management. This review synthesizes the comprehensive evolution in the understanding and treatment of RB. First, we deconstruct the intricate oncogenic circuitry that extends far beyond Knudson's classic "two-hit" RB1 inactivation model, describing non-classical MYCN-driven pathogenesis, multi-layered epigenetic reprogramming (including chromatin, RNA and histone changes), and distinct histological subtypes with defined clinical correlates, such as the favorable-prognosis cavitary RB. Single-cell genomics has elucidated the cellular origin from cone precursor cells and intratumoral heterogeneity. Risk stratification has been refined through well-defined classification systems, from the therapy-guiding International Intraocular Retinoblastoma Classification (IIRC) to the comprehensive American Joint Committee on Cancer Tumor-Node-metastasis (AJCC TNM) staging.Furthermore, the diagnostic paradigm has advanced from conventional anatomical imaging to liquid biopsies, enabling non-invasive molecular staging and monitoring via tumor-derived cell-free DNA analysis. Concurrently, the therapeutic landscape has undergone a radical shift, moving from enucleation and external-beam radiotherapy to an era dominated by local sight-preserving strategies. We provide a critical synthesis of the evidence for intravenous chemotherapy and the transformative role of super-selective intra-arterial chemotherapy (IAC), and describe essential randomized controlled trials, technical innovations, and optimized drug regimens. Finally, we explore emerging targeted molecular therapies and future directions. By integrating cutting-edge molecular insights with robust, high-level clinical evidence, this review offers the framework for achieving patient and eye survival as well as vision preservation in children with Retinoblastoma.

Bevacizumab plus chemotherapy is the standard first-line therapy for metastatic colorectal cancer (mCRC). To date, no phase 3 trial has compared first-line oral multitargeted TKI versus bevacizumab plus chemotherapy in RAS/BRAF wild-type mCRC. The open-label, noninferiority, randomized, phase 3 trial (ANCHOR; NCT04854668; CTR20210940) evaluated first-line anlotinib versus bevacizumab plus oxaliplatin and capecitabine (CapeOX) in this setting. Patients were centrally randomized (1:1) to receive 4-8 cycles of CapeOX in combination with either anlotinib (12 mg once daily on days 1-14) or bevacizumab (7.5 mg/kg on day 1) every 3 weeks, followed by maintenance therapy with anlotinib or bevacizumab plus capecitabine until unacceptable toxicity or disease progression. The primary endpoint was progression-free survival (PFS) assessed by an independent review committee in the intention-to-treat population. The hazard ratio (HR) for the noninferiority margin was 1.09. Between May 25, 2021, and August 30, 2023, 373 patients were assigned to the anlotinib group and 375 to the bevacizumab group. As of February 2, 2025, the median follow-up was 25.1 months (95% confidence interval [CI] 23.8-26.3). The median PFS was 11.0 months (95% CI 9.8-11.2) in the anlotinib group versus 11.0 months (9.7-11.2) in the bevacizumab group (stratified HR, 1.00; 95% CI 0.84-1.18; p = 0.87). The incidences of grade ≥3 treatment-related adverse events were 64.9% and 44.8%, respectively. Compared with bevacizumab plus CapeOX, anlotinib plus CapeOX showed similar antitumor activity but failed to reach the prespecified noninferiority margin for PFS and was associated with increased manageable toxicity.

We previously reported higher failure-free survival (FFS) with induction paclitaxel, cisplatin, and capecitabine (TPC) compared with cisplatin and fluorouracil (PF) in high-risk locoregionally advanced nasopharyngeal carcinoma (LA-NPC). We now report 5-year FFS, with prespecified secondary outcomes. In this multicenter, randomized trial, patients with high-risk LA-NPC (T4N0-2M0 or TanyN3M0) received two 21-day cycles of induction chemotherapy with TPC or PF, followed by concurrent chemoradiotherapy. The primary endpoint was FFS; secondary endpoints included distant metastasis-free, locoregional relapse-free, and overall survival (OS), tumor response, and safety. Among 238 patients (TPC, n=118; PF, n=120), with median follow-up of 89.1 months, 5-year FFS was 77.6% in the TPC group and 62.9% in the PF group (hazard ratio [HR], 0.52; 95% confidence interval [CI], 0.34-0.82). At 5 years, distant metastasis-free survival was 87.8% in the TPC group and 78.8% in the PF group (HR, 0.51; 95% CI, 0.28-0.95); locoregional relapse-free survival was 92.0% and 82.1%, respectively (HR, 0.43; 95% CI, 0.21-0.88); and OS was 89.6% and 82.2%, respectively (HR, 0.51; 95% CI, 0.27-0.95). Among patients with pretreatment Epstein-Barr virus (EBV) DNA <3000 copies/ml, 5-year OS was 92.3% in the TPC group and 84.4% in the PF group. Among those with higher EBV DNA levels, rates were 84.2% in TPC group and 81.1% in PF group. Grade 3 or 4 adverse events occurred in 68 patients (57.6%) receiving TPC and 79 patients (65.8%) receiving PF. Treatment adherence was similar in the two groups. Among patients with high-risk LA-NPC, TPC induction chemotherapy was associated with higher 5-year FFS than PF. (Funded by National Natural Science Foundation of China and State Key Laboratory of Respiratory Disease; ClinicalTrials.gov number, NCT02940925.).

The Lancet Oncology Commission on cancer genomics and precision oncology identified workforce capability, education, and training as major constraints to equitable implementation. As genomic and multi-omic technologies become integrated into cancer care, structured approaches are needed to define competencies across the multidisciplinary workforce. The Core Competency Framework for Precision Oncology was developed using a narrative review and iterative expert-informed framework development process. Competency frameworks, professional standards, policy guidance, organisational reports, and educational literature relevant to genomic medicine, precision oncology, and workforce development were identified through purposive searches of organisational repositories, peer-reviewed literature, and reference list screening between January 2025 and March 2026. Search terms included combinations of "precision oncology", "cancer genomics", "genomic medicine", "competenc∗", "education", "training", and "workforce". Competency domains, role functions, progression structures, and implementation concepts were extracted, compared, and synthesised into shared and role-specific competencies mapped across foundational, intermediate, and advanced levels. The framework was refined through iterative multidisciplinary expert consultation involving contributors to the Lancet Oncology Commission. The framework defines three components; shared core competencies applicable across professional groups; role-specific competencies aligned with functions across the precision oncology pathway; and a progression model reflecting increasing interpretive complexity, autonomy, accountability, and leadership. An associated tiered workforce model links competencies to service contexts and levels of practice, supporting distribution of genomic responsibilities while preserving escalation pathways and governance structures. The framework is designed to be adaptable across professions, health systems, and resource settings. This framework provides an implementation-oriented structure to support workforce development, curriculum design, continuing professional education, role mapping, competency assessment, and service planning in precision oncology. Its application may support scalable, equitable, and sustainable integration of genomics into routine cancer care. No specific funding was received for this work.

Patients with non-muscle invasive bladder cancer (NMIBC) frequently experience recurrence and may progress to muscle-invasive disease. Although several common germline variants associated with bladder cancer risk have been identified, their prognostic value in NMIBC remains unclear. We performed an updated genome-wide association study (GWAS), incorporating additional cohorts and analysing multiple NMIBC recurrences, to identify germline genetic variants associated with recurrence and progression. We analysed eight cohorts (N = 5009) from the Netherlands, UK, Canada, and Spain. Cohort-specific GWAS were conducted using Cox regression for recurrence-free survival (RFS) and progression-free survival (PFS), including recurrent-event analysis and gene-based analyses. Analyses included chromosome X and were stratified by sex and Bacillus Calmette-Guérin (BCG) treatment. Previously reported variants for bladder cancer risk and prognosis were also evaluated. We observed 4237 recurrences, of which 2145 were first recurrences, and 742 cases of progression in stage and/or grade. No genome-wide significant associations were identified in the overall population, chromosome X, or sex-stratified analyses. In BCG-treated patients, two loci reached genome-wide significance for RFS, with the strongest signal for rs72744118, an intron variant in DISP-1 (HR = 0.43; 95% CI (0.32, 0.56), p = 5.8 × 10-10). Gene prioritization identified 101 candidate genes from SNP associations (p < 1 × 10-6) and colocalization analyses, of which 16 genes showed nominally significant association between gene expression and NMIBC outcome in UROMOL. In the largest GWAS of NMIBC prognosis to date, we prioritized a set of 16 genes. Future research should independently validate the prognostic and functional roles of identified genes.

Cutaneous squamous cell carcinoma exhibits a significant disease burden due to local relapses and metastasis. Despite advances in prognosis, little evidence exists on prognostic factors of early and late relapse, which could provide more personalized management and follow-up schedules. We conducted a multicenter retrospective cohort study including 23,166 cases of cutaneous squamous cell carcinoma. Patients were categorized into those with early recurrence (≤4 months), late recurrence (>24 months), and no recurrence. Risk factors and tumor characteristics were analyzed to identify patterns associated with early and late relapses. Some risk factors show a different impact depending on the type of recurrence and its timing. Lymphovascular invasion (subdistribution hazard ratios [SHR] = 3.01) and large caliber and deep perineural invasion (SHR = 2.55) show a statistically significant association with early locorregional metastasis, losing impact with time. Poor differentiation shows impact early (SHR = 3.26) and late (SHR = 4.63), with less relevance from 4 to 24 months (SHR = 1.68). Other features keep a similar impact along the whole period. Retrospective cohort study. Lymphovascular invasion and large caliber and deep perineural invasion are risk factors associated with early locorregional metastasis and should encourage nodal staging at diagnosis. Poor differentiation keeps importance beyond 24 months, and longer follow-up might be recommended in patients with this feature.

Colorectal cancer screening reduces mortality, yet uptake remains suboptimal. Various interventions aim to improve screening rates, but their comparative effectiveness is unclear. We aimed to evaluate the effectiveness of colorectal cancer screening uptake interventions using a systematic review and network meta-analysis. We analyzed data from 76 randomized clinical trials across 8 intervention strategies: patient navigation, mailed fecal immunochemical testing (FIT) outreach, educational multimedia, reminder only, choice-based outreach, colonoscopy outreach, multistep, and usual care. Network meta-analysis compared interventions using risk ratios (RRs) and 95% CIs. P scores and rankograms assessed intervention rankings. Risk of bias was assessed, and certainty of evidence was graded using the Grading of Recommendations Assessment, Development and Evaluation framework. Patient navigation (RR, 1.58; 95% CI, 1.23-2.02; P score .81) and mailed FIT outreach (RR, 1.36; 95% CI, 1.07-1.74; P score = .79) were the most effective strategies, significantly outperforming usual care. Educational multimedia (RR, 1.27; 95% CI, 0.91-1.78) and reminder-only interventions (RR, 1.24; 95% CI, 0.98-1.57) showed modest effects. Choice-based outreach and colonoscopy outreach were not significantly more effective than usual care. Mailed FIT outreach was superior to colonoscopy outreach (RR, 1.35; 95% CI, 1.11-1.63), and patient navigation outperformed reminder-only interventions (RR, 1.48; 95% CI, 1.14-1.94). In low baseline uptake settings (<30%), mailed FIT outreach was most effective (RR, 3.12; 95% CI, 1.70-5.71), and educational multimedia performed best in higher uptake populations (≥30%) and in recent studies (2021-2024). The majority of studies were at low risk of bias and the certainty of evidence mostly ranged from moderate to low. Patient navigation and mailed FIT outreach are the most effective strategies for increasing colorectal cancer screening uptake, particularly in low baseline uptake populations. Educational multimedia has shown promise in recent years and high baseline uptake settings, offering a scalable alternative.

Gastric cancer remains a major global health burden, with high mortality driven by late-stage diagnoses that limit treatment options and reduce survival. Current diagnostic methods such as endoscopy and biopsy are invasive, resource-intensive, and impractical for large-scale early detection. This study aimed to develop and validate an ensemble machine learning model integrating four cell-free DNA (cfDNA) fragmentomic feature classes derived from 5 × whole genome sequencing (WGS) data to non-invasively differentiate malignant gastric cancer from benign gastric lesions in high-risk or symptomatic patients. A total of 681 plasma samples were prospectively collected, comprising 329 from patients with gastric cancer or high-grade intraepithelial neoplasia (HGIN) and 352 from individuals with benign gastric conditions. The dataset was divided into a training cohort (n = 333) and a temporally independent validation cohort (n = 348). An external validation cohort of 305 participants was also included. The ensemble model achieved an AUROC of 0.920 in cross-validation testing on the training cohort, 0.912 in the independent validation cohort, and 0.896 (95% CI 0.860-0.932) in the external cohort. At a pre-specified prediction threshold of 0.402, the model demonstrated 93.3% sensitivity and 71.9% specificity in the validation cohort, yielding a PPV of 71.3% and an NPV of 93.5%. In the external cohort, sensitivity and specificity were 91.7% and 69.1%, respectively (PPV 75.7%, NPV 88.8%). Model scores correlated with clinical stage, tumor grade, and histopathological subtype. Approximately 71% of non-cancer patients could have been spared unnecessary endoscopy. The cfDNA fragmentomics-based ensemble model enables accurate, non-invasive differentiation between gastric cancer and benign gastric lesions in high-risk or symptomatic patients. This approach demonstrates strong potential as a pre-endoscopy triage tool, supporting earlier detection and more efficient use of diagnostic resources.

Recently we observed high numbers of marginal zone B cells (MZBs) within murine head and neck squamous cell carcinoma (HNSCC) with immunosuppressive potential. To date, MZBs have not been linked to tumor development or tumor prevention. Based on our previous findings the present study aimed to validate the presence of MZB in HNSCC and to investigate their possible implications in tumorigenesis and prognosis. Flow cytometry was used to uncover MZB within tumors and blood of HNSCC patients. A single-cell RNA sequencing cohort of 118 HNSCC patients across different disease stages and 6 healthy donors (HDs) was compiled. Comparative transcriptomic profiling of B lymphocytes between HNSCC and HDs were performed. Downstream analysis, such as pathway enrichment, cell-cell communication, pseudotime trajectory inference, survival correlation, and spatial transcriptomics were applied. Two MZB subsets were revealed in tissues and blood of HNSCC patients and HDs. The tumor-associated MZBs were featured with hypoxia stress and viral-related hallmark genes. MZB-2, characterized by elevated expression of activation markers and immune-regulatory genes, displayed strong interactions with CD4+ T cells and antigen-presenting cells. These interactions were supported by costimulatory signals in HDs but were absent in HNSCC patients. Co-localization of MZB-2, germinal center B cell (GCB), and CD4+ follicular helper T cell (Tfh) was detected in HNSCC, suggesting the presence of an intratumoral MZB-Tfh-GCB axis. Clinically, MZB-2 abundance was associated with favorable prognosis in early-stage HNSCC, but not in advanced disease. Immunosuppressive gene signatures were not exclusive to MZBs, indicating that they do not represent a purely regulatory B cell phenotype. Our findings demonstrate an immunomodulatory role of MZBs in tumor immunity, balancing antigen presentation, cytokine signaling, and immune suppression. The association of MZB-2 with improved prognosis in early-stage HNSCC highlights its potential as a beneficial regulator of antitumor immunity during early tumor progression.

Advances in cancer care have improved survival, increasing the importance of comorbidities at diagnosis for longer-term outcomes. Pre-existing cardiovascular disease (CVD) can complicate management and prognosis, yet its long-term trends and future burden at cancer diagnosis are unclear. This study aimed to quantify temporal trends in pre-existing CVD among patients with newly diagnosed cancer and project prevalence through 2050. Linked primary care, cancer registry, and hospitalization records were used to identify 773 590 adults (≥18 years) diagnosed with the five most common cancers between 2001 and 2020 in England. Serial cross-sectional analyses estimated annual crude and age-standardized prevalence of pre-existing CVD and comorbidities. Temporal trends were assessed using logistic regression adjusted for demographics and cancer type. The contributions of changes in risk factor domains were evaluated using sequential modelling, and CVD prevalence was projected to 2050 using demographic-adjusted spline models. The age-standardized prevalence of pre-existing CVD rose from 31.4% [95% confidence interval (CI) 31.2-31.6] in 2001-2005 to 39.2% (95% CI 39.0-39.4) in 2016-2020, with the steepest increases in lung and haematological cancers. Valvular disease, heart failure, atrial fibrillation, and diabetes showed the largest subtype rises. Sequential modelling showed the greatest attenuation of the temporal trend after adjustment for cardiometabolic conditions and chronic comorbidities. Projection analyses suggest that, if current temporal patterns continue, approximately half of patients may present with pre-existing CVD at cancer diagnosis by 2050. Pre-existing CVD at cancer diagnosis has increased substantially over 2 decades and is projected to affect nearly half of patients by 2050. These trends underscore the need for the expansion of cardio-oncology services.

Patients with prostate cancer have a high burden of cardiovascular risk factors, often suboptimally controlled, and adverse cardiovascular outcomes. To determine whether the routine referral of patients with prostate cancer to a cardiovascular specialist to implement a systematic risk factor strategy is more likely to reduce adverse cardiovascular outcomes and improve risk factor control than usual care. This randomized clinical trial included patients with prostate cancer from 55 sites in 8 countries between 2015 and 2025. Eligible patients were diagnosed with prostate cancer during the past 12 months; had received treatment with androgen deprivation therapy (ADT) for the first time within the past 6 months; or planned to start ADT in the next month. Patients taking a statin with a systolic blood pressure of 130 mm Hg or lower were ineligible. Data were analyzed from May 25 to August 7, 2026. Patients were allocated in a 1:1 ratio to receive usual care alone or usual care plus routine referral to an internist or cardiologist. The specialists provided a systematic intervention, including a target of systolic blood pressure of 130 mm Hg or lower and a statin medication, irrespective of the patient's cholesterol levels (even if not usual or guideline-driven practice); encourage smoking cessation; and provide guidance on diet and exercise. Hierarchical composite of cardiovascular death, myocardial infarction, stroke, heart failure, suboptimal cholesterol (total cholesterol, >155 mg/dL [to convert to mmol/L, multiply by 0.0259]) and suboptimal blood pressure (systolic blood pressure, >130 mm Hg) as evaluated by the win ratio. The analysis included 2487 patients with prostate cancer (mean [SD] age, 68 [8] years). During median (IQR) follow-up of 5.8 (2.7-8.2) years, the win ratio in favor of the intervention was 1.60 (95% CI, 1.42-1.81), mostly attributable to lower cholesterol in the intervention group (mean difference, 12 mg/dL; 95% CI, 9-15 mg/dL) as a consequence of greater protocol-mandated statin use. Mean (SD) close-out systolic blood pressure values were 131.1 (16.9) mm Hg in the intervention group and 132.9 (18.3) mm Hg in the control group. There was no difference in time to cardiovascular death, myocardial infarction, stroke, or heart failure between groups (subdistribution hazard ratio, 1.08; 95% CI, 0.79-1.49). In this randomized clinical trial, routine referral of patients with prostate cancer to a cardiovascular specialist lead to improved outcomes, specifically through better cholesterol control. However, it is uncertain whether this reduced clinical cardiovascular events. ClinicalTrials.gov Identifier: NCT03127631.

This systematic review and meta-analysis aimed to comprehensively evaluate the prognostic impacts of seven classes of antidiabetic medications in patients with cancer, predominantly in the setting of concomitant type 2 diabetes mellitus (T2DM), and to elucidate their potential differential effects. A systematic search of PubMed, Embase, Cochrane Library, and the Web of Science was conducted from inception to May 2025, utilizing keywords including "antidiabetic drugs," "cancer," and "prognosis." Based on predefined criteria, eligible English-language randomized controlled trials and cohort studies were included if they compared the prognostic impacts of metformin, insulin, sulfonylureas, dipeptidyl peptidase-4 (DPP-4) inhibitors, sodium-glucose cotransporter 2 (SGLT2) inhibitors, glucagon-like peptide-1 (GLP-1) receptor agonists, and thiazolidinediones in patients with cancer. The primary outcome domain was all-cause survival, represented by all-cause mortality (ACM) or overall survival (OS) according to the terminology of the original studies. Secondary cancer-related outcomes included cancer-specific mortality (CSM), disease-free survival (DFS), progression-free survival (PFS), and recurrence-free survival (RFS). Multiple independent investigators performed data extraction and quality assessment. Statistical analyses were performed using OnlineMeta V1.1. DerSimonian-Laird random-effects models were used for all primary meta-analyses to calculate hazard ratios (HRs) and 95% confidence intervals (CIs), while fixed-effects estimates were examined as complementary sensitivity analyses. Study quality was assessed using the Newcastle-Ottawa Scale and the Cochrane risk-of-bias tool for randomized trials. A total of 61 studies comprising 1,106,966 patients with cancer were included. Metformin use was associated with lower ACM (HR=0.82, 95% CI: 0.74-0.91, P=0.0001) and CSM (HR=0.77, 95% CI: 0.69-0.87, P<0.0001), whereas insulin use was associated with higher ACM (HR=2.03, 95% CI: 1.63-2.51, P<0.0001). Among the drug classes analyzed, SGLT2 inhibitor use showed a nonsignificant inverse trend with ACM (HR=0.44, 95% CI: 0.11-1.71, P=0.24) and was associated with lower CSM (HR=0.21, 95% CI: 0.20-0.22, P<0.0001), although these estimates were based on few observational studies with study-specific comparator groups and do not establish superiority over other drug classes. In adjusted-HR-only sensitivity analyses, the direction of association was generally consistent for the major metformin outcomes and insulin-related ACM, whereas several other comparisons were attenuated or could not be pooled because too few studies reported adjusted estimates. This meta-analysis identified heterogeneous associations between antidiabetic drug use and cancer-related survival outcomes, with variation across drug classes, cancer types, and study-level mean age groups. Because the evidence was derived predominantly from observational studies with study-specific comparator groups, substantial between-study heterogeneity, and limited data for newer agents, the pooled estimates should not be interpreted as evidence of treatment superiority or used to rank drug classes. These findings are hypothesis-generating and warrant confirmation in prospective studies using clinically comparable treatment groups, standardized outcome definitions, and rigorous control of confounding.

The optimal self-expandable metallic stent (SEMS) choice for endoscopic drainage of unresectable extrahepatic malignant biliary obstruction (MBO) with endoscopic retrograde cholangiopancreatography (ERCP) remains unclear due to limited randomized controlled trial (RCT) evidence. Prior meta-analyses have relied on observational data, introducing heterogeneity and bias. To overcome these limitations, an RCT-only network meta-analysis (NMA) was performed to evaluate the efficacy and safety of SEMS types. A literature search was conducted to identify RCTs assessing fully covered SEMS (FCSEMS), partially covered SEMS (PCSEMS), and/or uncovered SEMS (UCSEMS) for ERCP drainage of unresectable extrahepatic MBO. Study outcomes included recurrent biliary obstruction (RBO), specific causes of RBO, overall survival (OS), and adverse events (AEs). A frequentist NMA was performed to estimate hazard ratios (HRs) and incidence rate ratios (IRRs) with their 95% confidence intervals and overall P-scores. Ten RCTs (1,659 patients) were included. There were no significant differences in time to RBO, incidence of any RBO, or OS between stent types. UCSEMS had the lowest rates of stent migration, tumor overgrowth, and sludge occlusion, whereas FCSEMS most effectively prevented tumor ingrowth. AE rates were similar across groups; however, only FCSEMS, but not PCSEMS, had significantly higher rates of acute cholecystitis than UCSEMS in patients with an intact gallbladder. There were no differences in RBO, OS, or AE excluding acute cholecystitis between each SEMS type, although each type has its advantages and disadvantages. FCSEMS was associated with a higher risk of acute cholecystitis, emphasizing careful patient selection, particularly regarding gallbladder status.

Antibody-drug conjugates (ADCs) represent a breakthrough in tumor-targeted therapy, showing promise in both solid tumors and hematological malignancies. By 1 September 2025, more than 370 ADC candidates had entered clinical development, 17 unique products had received at least one regulatory approval worldwide, and more than 150 candidates had been discontinued. This study analyzes 3,077 ADC trials from Citeline Trialtrove, revealing that most trials are concentrated in the United States and China. At the data cutoff, 35% of trials were ongoing and 12% were classified as terminated in Trialtrove. ADC trials primarily focused on solid tumors, with brentuximab vedotin, disitamab vedotin, and gemtuzumab ozogamicin being the most frequently investigated agents. Early-phase trials predominated, whereas Phase III and IV trials were less frequent. EGFR, BRCA1, and TP53 were frequently recorded across trial-design and outcome-related fields. Future research should focus on advancing later-phase trials, refining safety and efficacy, and exploring new therapeutic targets with continued biomarker integration.

In the randomized, double-blind, phase III RATIONALE-306 trial, patients with unresectable, locally advanced, recurrent, or metastatic esophageal squamous-cell carcinoma (ESCC) treated with tislelizumab plus chemotherapy in the intent-to-treat population and in the programmed death-ligand 1 (PD-L1) Tumor Area Positivity (TAP) score ≥5% subgroup experienced clinically meaningful overall survival (OS) benefit compared with placebo plus chemotherapy at the interim analysis and minimum 3-year follow-up. We report post hoc, exploratory, longer-term outcomes at study closeout with minimum 45.2-month follow-up in patients with PD-L1 TAP score ≥5% per European Medicines Agency recommendation. Patients were randomly assigned (1 : 1) to receive tislelizumab 200 mg or placebo every 3 weeks plus investigator-chosen chemotherapy. The primary endpoint was OS. Secondary endpoints included progression-free survival (PFS), objective response rate (ORR), duration of response (DoR), safety, and quality of life (QoL). Of 649 randomly allocated patients, 55.2% had PD-L1 TAP score ≥5% (tislelizumab plus chemotherapy, n = 172; placebo plus chemotherapy, n = 186). At 45.2-month minimum follow-up, tislelizumab plus chemotherapy improved OS [median 19.1 versus 10.0 months; hazard ratio (HR) 0.61] and PFS (median 8.2 versus 5.5 months; HR 0.50) compared with placebo plus chemotherapy. ORR was 71.5% versus 41.4%; median DoR was 7.1 versus 5.4 months, respectively. QoL was similar overall between arms, with tislelizumab plus chemotherapy trending toward better global health and pain reduction. Any-grade treatment-related adverse events (TRAEs) occurred in 97.7% (tislelizumab plus chemotherapy) versus 98.4% (placebo plus chemotherapy) and grade ≥3 TRAEs in 70.2% versus 66.5%, respectively. The most common grade ≥3 TRAEs were decreased neutrophil count (35.1% versus 31.9%), anemia (13.5% versus 11.4%), and decreased white blood cell count (12.3% versus 17.8%). Treatment-emergent adverse events with ≥5% difference in incidence between arms at interim analysis decreased substantially after 12 months. First-line tislelizumab plus chemotherapy provided sustained and clinically meaningful efficacy benefits over placebo plus chemotherapy and was tolerable, with no new safety signals for patients with unresectable, locally advanced, or metastatic ESCC and PD-L1 TAP score ≥5%.

Parity and breastfeeding reduce lifetime breast cancer (BC) risk, but the postpartum period is associated with transiently increased risk and poor outcomes. The prognostic significance of reproductive timing in women with germline pathogenic susceptibility variants (PVs) remains unclear. We evaluated the association between time since last childbirth, breastfeeding, and other reproductive variables and overall survival (OS) in a large Australian hereditary BC cohort. We conducted a retrospective cohort study of 1200 female PV carriers with invasive BC enrolled in the Kathleen Cuningham Foundation Consortium (1980-2022). Eligible participants carried class 4-5 PVs in high- or moderate-penetrance BC susceptibility genes and had no pregnancies after BC diagnosis. Time since last childbirth was categorised as nulliparous, <10 years, or ≥10 years, with further analyses using five postpartum intervals. Cox proportional hazards models, adjusted for age, oestrogen receptor (ER) status, chemotherapy, surgery, and radiotherapy, were used to assess associations with OS. Of 1200 women (median follow-up, 13 years), 52% were diagnosed ≥10 years postpartum, 32% were diagnosed <10 years, and 16% were nulliparous. Women diagnosed <10 years postpartum were younger (median 36 compared with 48 years, P < 0.001), more likely to carry BRCA1 PVs, and had more ER-negative disease. Diagnosis ≥10 years postpartum was associated with improved OS compared with nulliparity [hazard ratio (HR) 0.63, 95% confidence interval (CI) 0.43-0.92, P = 0.02]. In parous women, breastfeeding compared with no breastfeeding (HR 0.49, 95% CI 0.33-0.73, P < 0.001) and diagnosis ≥10 vs <10 years since last childbirth (HR 0.48, 95% CI 0.31-0.74, P < 0.001) independently predicted better OS. In women with PVs, longer time since last childbirth and breastfeeding were independently associated with improved survival following BC diagnosis. These findings identify reproductive timing and lactation as important prognostic modifiers in hereditary BC and may inform risk stratification and survivorship care.

Age is a nonmodifiable risk factor for atherosclerosis and cardiovascular disease (CVD), with their prevalence increasing over time. Dyslipidaemia plays a key role in coronary artery disease (CAD) across all age groups, including older adults. However, limited evidence exists regarding the efficacy of lipid-lowering therapy, particularly statins, for primary prevention in older populations. This review underscores the significance of statins for primary prevention in older adults. The PROSPER study and similar trials demonstrated a 24% reduction in CAD mortality with statins, without notable cognitive or functional impairments. Similarly, the EWTOPIA 75 trial reported a 34% reduction in major cardiac events with ezetimibe, though benefits diminished in those aged >85 years. A meta-analysis revealed a 26% reduction in major vascular events per 1 mmol/L decrease in low-density lipoprotein cholesterol in older adults, with no heightened risks of cancer, haemorrhagic stroke, or cognitive decline. The Danish Contemporary Primary Prevention Cohort identified individuals aged 80-100 years as having the highest myocardial infarction risk, emphasizing the benefits of statin therapy in this group. Ongoing STAREE and PREVENTABLE trials aim to provide further evidence on statins for primary prevention in older adults. Effective CVD management in older populations should begin early, prioritizing a healthy lifestyle and addressing modifiable risk factors. Statin therapy should not be discontinued based on age alone and requires careful consideration for individuals with established CVD.

Adhesions commonly occur after abdominal and pelvic surgery, causing infertility, intestinal obstruction, and pelvic pain. Existing anti-adhesion materials are often difficult to use on uneven organ surfaces, highlighting the need for novel solutions; SI-449, a cross-linked, powder-formed chondroitin sulfate (CS), may address these limitations. To evaluate the anti-adhesion effect of the SI-449 adhesion barrier system (cs barrier) in patients undergoing open rectal resection for rectal cancer. This multicenter, prospective, randomized, open, and blinded-end point (PROBE) method pivotal clinical trial was conducted at 53 hospitals in Japan from September 2020 to June 2023. The study participants and efficacy assessors on the central review board were blinded. Patients aged 20 years or older with clinical stage I through IV rectal cancer who underwent open rectal resection and were scheduled for follow-up ileostomy closure were eligible for inclusion. Data were analyzed from July 2023 through September 2023. The control group received no additional procedure before abdominal closure, while the experimental group received 2 to 3 g of SI-449 applied to the abdominal or pelvic cavity immediately before closure. The primary end point was the incidence of adhesions under the incision. Secondary end points included adhesion number, severity, and extent; number and type of adhered organs; and number of adhesions per organ. Safety and operability were also evaluated. Among 155 screened patients, 131 met the inclusion criteria and were randomized. Of these, 128 underwent the intervention (safety analysis population; control group: n = 64; experimental group: n = 64), and 118 completed a second surgery (modified intention-to-treat population; control group: n = 60; experimental group: n = 58). Overall mean (SD) patient age was 68.2 (10.3) years, and 34 patients of 118 (29%) were female. Adhesion incidence was significantly lower in the experimental group (26 of 58 [45%]) compared to the control group (56 of 60 [93%]) (P < .001). The experimental group also showed significantly reduced adhesion severity and extent. No safety concerns were identified, and operability was good. In this randomized clinical trial, SI-449 in the powdered form reduced the incidence of adhesions, and the adhesion barrier system had good operability and no safety concerns. Thus, this system may provide a new anti-adhesion option for use in open surgery. jrct.mhlw.go.jp Identifier: jRCT1080225247.

The development of therapy resistance compromises the long-term efficacy of lenvatinib in advanced hepatocellular carcinoma (HCC), highlighting the need to characterize the molecular drivers of resistance. Through integrated multiomics analysis combining genome-wide CRISPR screening, transcriptomics, and proteomics, we identified Heat shock protein family A member 6 (HSPA6) as a critical driver of lenvatinib resistance. HSPA6 was consistently upregulated in resistant cell lines and patient tumors, and high expression correlated with poor treatment response and survival. HSPA6 recruited the deubiquitinase USP9X to stabilize the antioxidant enzyme thioredoxin reductase 1 (TXNRD1), thereby suppressing lenvatinib-induced ferroptosis. Furthermore, HSPA6 underwent drug-enhanced liquid-liquid phase separation through its intrinsic disordered region 1 domain, facilitating the assembly of biomolecular condensates that reinforce TXNRD1 stability and ferroptosis resistance. Leveraging these insights, canagliflozin, an FDA-approved sodium-glucose cotransporter 2 inhibitor, was repurposed as a direct HSPA6-targeting compound that disrupted this resistance axis, restored ferroptosis sensitivity, and synergized with lenvatinib in patient-derived models. This work unveils a dynamic, condensate-driven mechanism of drug resistance and offers a readily translatable strategy to overcome lenvatinib resistance in HCC. HSPA6 drives lenvatinib resistance and can be targeted by repurposing canagliflozin to restore ferroptosis sensitivity and overcome therapy resistance in hepatocellular carcinoma.

Metabolic dysfunction-associated steatohepatitis (MASH) is characterized by progressive inflammation and fibrosis. Left untreated, MASH can progress to cirrhosis, hepatocellular carcinoma, and liver failure. MASH has become the leading indication for liver transplantation worldwide. While global prevalence is increasing, effective and mechanism-oriented therapies remain limited. Based on earlier independent studies showing that lysosomes are impaired and autophagy is dysregulated in MASH, our aim was to finely map autophagy dysfunction in an experimental mouse model of MASH and explore the capacity of a modulator of chaperone-mediated autophagy to mitigate the course of the disease. We effectively identified a number of markers whose expression was pathologically increased or decreased in various autophagy pathways. In vivo, pharmacological modulation with the phosphopeptide P140 -currently evaluated in phase III-clinical trials for lupus- corrected the expression of some of these markers and restored lysosomal and mitochondrial autophagy programs. It reduced steatohepatitis and fibrosis, and improved systemic inflammatory features without, however, broadly correcting metabolic parameters. Mechanistically, consistent with its established HSPA8 interaction, P140 restored lysosomal/autophagy markers, supporting modulation of this proteostasis network. Our data indicate that this pharmacological restoration of lysosomal proteostasis engages key transcriptional regulators (Mediator complex), leading to the selective remodeling of pro-fibrotic and inflammatory pathways. Collectively, we identified a coordinated disruption of lysosomal quality control networks across multiple autophagy pathways in a validated mouse model of advanced MASH. We established lysosomal autophagy as a druggable vulnerability in MASH and support therapeutic repositioning of P140 as a safe strategy to counter progressive liver diseases.

Infection with hepatitis B virus (HBV) remains a severe concern to public health, with roughly 292 million people worldwide suffering from the chronic form of the disease, for which there is no cure. Chronic HBV infections frequently lead to hepatocellular carcinoma (HCC), one of the world's leading causes of cancer-related deaths. Although the process of hepatocarcinogenesis is complex and not fully understood, various studies have identified numerous long non-coding RNAs (lncRNAs) as contributing to the formation of HCC. These host-derived lncRNAs are frequently dysregulated as a result of viral infection. Numerous lncRNAs have been linked to HBV carcinogenesis and replication, particularly those that are dysregulated in HBV-associated HCC. HBV X protein regulates the majority of these dysregulated lncRNAs. Certain lncRNAs have been found to exert regulatory functions in HBV replication and carcinogenesis. The prognosis for HCC remains poor, and early detection of novel tumor markers is critical for effective HCC therapy. Understanding the biological activities and regulatory mechanisms of HCC-associated lncRNAs will aid in disease diagnosis and therapy and help elucidate the disease etiology. In HBV-related HCC, certain dysregulated lncRNAs may develop into biomarkers for early detection or potential targets for HCC treatment. This review provides a brief overview of the recent findings on lncRNAs in HBV with a focus on current developments. We also investigated the possible relevance of dysregulated lncRNAs in HCC as biomarkers for diagnosis and treatment and assessed their carcinogenic and tumor-suppressive effects.

Lung metastasis remains a determinant of poor prognosis and survival in breast cancer and is understood to depend on a permissive pulmonary immune niche rather than tumor cell traits alone. Here, we developed a host-directed RNA interference strategy to modulate this niche by reprogramming pulmonary B cells for breast cancer lung metastasis treatment. IF7C peptide-decorated cationic liposomes were constructed, which preferentially accumulated in the lung, and were internalized by pulmonary B cells, enabling selective silencing of annexin A1 (ANXA1). In tumor-conditioned primary B cells, ANXA1 knockdown reshaped the transcriptional landscape and shifted cytokine output away from an immunosuppressive profile characterized by IL-10, TGF-β, and IL-35. Functionally, ANXA1-silenced B cells lost their capacity to drive CD4⁺ T cells toward Foxp3⁺ regulatory differentiation and instead promoted Th1 features, while concurrently relieving suppression of CD8⁺ T-cell proliferation. In two postoperative syngeneic breast cancer models, perioperative administration achieved ANXA1 silencing in pulmonary B cells, reduced lung Treg accumulation, enhanced CD8⁺ T-cell infiltration and effector activity, and suppressed metastatic outgrowth with favorable systemic safety. These findings identify pulmonary B cells as an actionable regulator of the lung metastatic niche and establish perioperative, B-cell-focused ANXA1 silencing as a practical approach to prevent postoperative lung metastatic recurrence.

KRASG12D-selective and pan-RAS inhibitors have shown promise in pancreatic ductal adenocarcinoma (PDAC), yet adaptive resistance is anticipated to limit durability of response. Exportin 1 (XPO1), a nuclear export protein frequently overexpressed in PDAC, represents a potential vulnerability in KRAS-mutant cancers. We evaluated whether pharmacologic inhibition of XPO1 enhances therapeutic efficacy and durability of KRAS pathway inhibition. KRASG12D inhibitor- and pan-RAS inhibitor-resistant PDAC cellular models were generated and assessed for sensitivity to the second-generation XPO1 inhibitor Eltanexor. Antiproliferative synergistic effects of Eltanexor combined with MRTX1133, Zoldonrasib (RMC9805), or Daraxonrasib (RMC6236) were evaluated in PDAC 2D cultures, 3D spheroids, patient-derived organoids, and tumor-fibroblast co-culture models. Eltanexor sensitized KRAS inhibitor-resistant PDAC cells and synergistically enhanced growth suppression across multiple KRASG12D-mutant models. Combination treatment reduced clonogenic survival, disrupted 3D spheroid integrity, and significantly inhibited viability of patient-derived organoids. The in vivo efficacy of the combination was tested in PDAC cell-derived xenograft/allograft and patient-derived xenograft models. Combining sub-therapeutic doses of Eltanexor with allele-specific inhibitors or pan-RASi resulted in significant tumor regression, prevention of metastatic spread and prolonged survival in vivo. Notably, Eltanexor maintenance therapy suppressed tumor regrowth following RAS inhibitor withdrawal and preserved responsiveness upon re-challenge. Mechanistically, molecular and phosphokinome profiling showed that the combination broadened suppression of MAPK- and mTOR-associated signaling and reduced activity of multiple oncogenic kinases. In conclusion, XPO1 inhibitor Eltanexor enhances the efficacy and durability of KRAS and pan-RAS inhibition in PDAC models. These findings provide a preclinical rationale for clinically evaluating Eltanexor in combination with RAS-targeted therapies to delay or overcome adaptive resistance in KRAS-mutant PDAC.

Targeted therapies have improved outcomes in EGFR-, KRAS G12C-, and BRAF V600E-driven non-small cell lung cancer (NSCLC), but acquired resistance is molecularly and spatially heterogeneous. Driver-specific algorithms and validated biomarkers underpin post-progression care. Determining pathway activity, causal dependency, and a tractable therapeutic vulnerability requires evidence beyond detecting an acquired alteration. We propose a lesion- and time-specific pathway-dependency framework centered on the RAS-RAF-MEK-ERK mitogen-activated protein kinase (MAPK) pathway and the PI3K-AKT-mTOR pathway. It distinguishes three provisional biological states: MAPK-dominant resistance, shared-input MAPK-PI3K reactivation, and a candidate PI3K-enriched/MAPK-low state. A clinical management branch encompasses histologic transformation, central nervous system-limited progression, oligoprogression, and diffuse polyclonal progression and may coexist with a biological assignment. Spatially discordant mechanisms support a mixed assignment, whereas insufficient evidence remains indeterminate. Biological assignment integrates contemporaneous lesion-level findings, clonality, histology, and exploratory pathway readouts; progression pattern guides the clinical branch. The framework complements genotype-based classification and may clarify when better-supported systemic, histology-directed, or local treatment should take precedence. Evidence remains uneven: some driver-specific interventions have established clinical evidence or clinically supported activity, whereas most downstream MAPK strategies and all approaches matched to the candidate PI3K-enriched/MAPK-low state remain investigational. Limited tissue availability, spatial heterogeneity, unstandardized assays, and combination toxicity constrain implementation. Prospective studies should determine whether a locked classifier adds predictive value beyond the initiating driver and acquired genomic alterations. Until prospective validation is available, the framework is best suited to mechanistic interpretation and trial design; routine treatment continues to rely on validated biomarkers and established clinical evidence.

Based on the assumption that radiation dose escalation to the primary tumor improves local control and therefore produces survival benefits under the condition of limiting the dose to the organ at risk (OAR), we launched a study to evaluate the effect of simultaneous integrated boost (SIB) radiotherapy implemented in stage III locally advanced non-small cell lung cancer (LA-NSCLC) patients. A prospective, single-center, open-label, randomized phase II trial was performed in patients with stage III LA-NSCLC. Patients were randomized to receive SIB thoracic radiation: 50 Gy in 25 fractions of 2.0 Gy for planning target volume (PTV), 55 Gy in 25 fractions of 2.2 Gy for clinical target volume (CTV), 60-70 Gy in 25 fractions of 2.4-2.8 Gy for gross tumor volume (GTV), or control group: 30 fractions of 2.0 Gy for PTV. A platinum based concurrent chemotherapy was administered to patients in both groups. The primary endpoint was progression-free survival (PFS). The secondary endpoints were overall survival (OS), objective response rate (ORR), failure patterns, and treatment-related toxicities. Exploratory analyses of epidermal growth factor receptor (EGFR) and programmed death-ligand 1 (PD-L1) were performed for PFS and OS. From April 2015 to April 2018, 168 patients were included in the analysis, with 84 in the control group and 84 in the SIB group. The median PFS was 21.0 months in the SIB group and 11.0 months in the control group (P < 0.001, Hazard Ratio (HR) 0.48, 95% CI 0.34-0.68). The median OS was 42.0 months and 26.0 months(P = 0.002, HR 0.55, 95% CI 0.37-0.81) in the SIB group and the control group, respectively. The SIB group had a significantly lower risk of brain and other distant metastasis, with adjusted sub-distribution HR of 0.35 (95% CI, 0.14-0.87, P = 0.025) and 0.65 (95% CI, 0.45-0.94, P = 0.023), respectively. There was lower radiation-related pneumonitis (P = 0.015) in the SIB group because of the lower dose for the irradiated lung (irradiated-lung V5, 55.38±18.11 Gy vs. 61.95±15.45 Gy, P = 0.015). Most other OAR doses and incidences of grade ≥2 disease were similar between the two arms. Exploratory biomarker analysis reveals a trend toward superior survival benefit from SIB in patients with PD-L1 ≥1% or EGFR-mutant. We demonstrated that the routinely adaptive SIB strategy could significantly improve PFS and OS and reduce the dose to the irradiated lung without increasing the risk of tumor recurrence. Exploratory analyses of EGFR mutation status and PD-L1 expression suggested potential differences in treatment outcomes across biomarker-defined subgroups. This strategy represents a potentially feasible regimen to improve survival outcomes for patients with stage III LA-NSCLC eligible for definitive concurrent chemoradiotherapy and warrants further validation in larger prospective trials.

Histopathology is the cornerstone of oncology diagnosis, while whole-slide images (WSIs) enable the transition to digital, quantitative pathology. Leveraging WSIs to accurately predict therapeutic response is increasingly vital for advancing precision oncology and optimizing clinical workflows. However, existing artificial intelligence models for WSI-based immunotherapy response prediction often yield suboptimal performance, as they frequently fail to fully capture the complex multi-scale morphological features and heterogeneous information inherent in clinical pathology data. To address this challenge, we propose ViGNet, a Visual-Global Relation Fusion Network designed for immunotherapy response prediction directly from WSIs. ViGNet is a multimodal framework that effectively integrates histopathological image features with specific clinical data modalities, including gene expression profiles and cancer type text. The proposed architecture features two primary encoders: a multi-scale visual encoder and a gene-driven encoder. The visual encoder utilizes pre-trained CTransPath and an MSCNNPath to extract pyramidal image features, which are integrated with cancer-type diagnostic priors. Meanwhile, the gene-driven encoder converts gene-guided information into a slide-level global relational prior. This global relational prior was then injected into the patch-level multi-scale visual representation learning via a top-down SRFF module, and the resulting representations were subsequently passed to a classification head to predict immunotherapy response. Across one internal and two independent external cohorts, ViGNet achieves 82.55% discrimination ability for immunotherapy response prediction, outperforming the baseline methods. These results underscore ViGNet's potential as a clinical decision-support tool, aiding personalized treatment planning and enabling informed clinical decision-making through accurate and interpretable predictions.

Patients with unresectable stage III non-small cell lung cancer (NSCLC) have an unmet need for new therapies that improve survival. This phase III trial investigated the safety and efficacy of concurrent ociperlimab and tislelizumab plus concurrent chemoradiotherapy (cCRT) in treatment-naïve patients with stage III NSCLC. In this phase III, multicenter, randomized, multiarm, open-label trial, patients with unresectable stage III NSCLC received concurrent ociperlimab plus tislelizumab and cCRT, followed by ociperlimab plus tislelizumab (arm A), tislelizumab and cCRT, followed by tislelizumab (arm B), or cCRT, followed by durvalumab (arm C) (NCT04866017). Objectives were to compare progression-free survival (PFS), overall survival (OS), objective response rate (ORR), duration of response, disease control rate, clinical benefit rate, time to death or distant metastasis (TTDM), and safety and tolerability for arms A versus C, B versus C, and A versus B. 63 patients were randomized to arms A (N=22), B (N=19), and C (N=22) prior to early trial termination. In A, B, and C, respectively, 95.5% (21/22), 84.2% (16/19), and 95.5% (21/22) were current or former smokers, and 68.2% (15/22), 73.7% (14/19), and 68.2% (15/22) had PD-L1 expression in tumor cells of ≥1%. Median PFS (95% CI) was not reached (NR) (6.3-not estimable (NE)) in A, 15.0 months (7.4 to NE) in B, and 10.4 months (5.7 to NE) in C. Median OS and TTDM were NR in any arm. ORR (95% CI) was 68.2% (45.1%-86.1%) in A, 68.4% (43.4%-87.4%) in B, and 59.1% (36.4%-79.3%) in C; all responses were partial responses. Treatment-emergent adverse events (TEAEs) occurred in all patients; in arms A, B, and C, respectively, pneumonitis occurred in 18.2% (4/22), 5.6% (1/18), and 9.1% (2/22) of patients, and interstitial lung disease occurred in 13.6% (3/22), 11.1% (2/18), and 0% of patients, of which the majority of events for each were grade 1/2. Grade ≥3 treatment-related TEAEs occurred in 68.2% (15/22), 66.7% (12/18), and 68.2% (15/22) of patients in arms A, B, and C, respectively. There was a trend toward improved efficacy when adding tislelizumab with or without ociperlimab to cCRT followed by tislelizumab with or without ociperlimab compared with cCRT followed by durvalumab; however, efficacy data were for descriptive purposes only. No unexpected or new safety signals were identified.

The effectiveness of any cancer therapy depends on its dose and its treatment schedule. A potential third dimension of regimen design is whether intentional fluctuations in administered dose might improve treatment outcomes. In this issue, West and colleagues demonstrate that dose fluctuations can alter both tumor drug response and the evolution of drug resistance, in ways related to the shapes of dose-response functions. In mouse xenograft models of ALK-fusion non-small cell lung cancer treated with the ALK inhibitor alectinib, steady dosing produced the best immediate tumor response, but fluctuations in dose were better at delaying the evolution of drug resistance. Treatment schedules that switched between even and uneven dosing navigated this tradeoff, achieving comparable tumor control with steady dosing while better preserving drug sensitivity. By showing how dose-response measurements can guide when doses should remain steady or fluctuate, this work expands how cancer treatment schedules might be optimized. See related article by West et al., p. 4434.

Deep learning can extract predictive and prognostic biomarkers from histopathology whole-slide images. However, explainable artificial intelligence approaches widely used in digital pathology, such as attention heatmaps and class activation mapping, provide limited insight into the image features associated with classifier outputs. In this study, we developed Morphing histoPathology Diffusion (MoPaDi), a framework for generating counterfactual explanations for histopathology images that help identify morphologic or stain-related features linked to model predictions. MoPaDi combined diffusion autoencoders with task-specific multiple instance learning classifiers to manipulate images and induce prediction shifts by modifying classifier-associated features. The framework was evaluated on multiple datasets spanning colorectal, breast, liver, and lung cancers, including tasks for tissue type, cancer subtype, and biomarker [microsatellite instability (MSI)] classification. MoPaDi generated perceptually realistic counterfactual histopathology images, enabling pathologists to identify morphologic features associated with changes in model predictions, complementing the conventional inspection of highly attended regions in digital pathology. In the MSI status prediction task, MoPaDi highlighted morphologic features linked to classifier predictions, including mucinous differentiation, altered glandular architecture, and lymphocytic infiltration, consistent with prior literature. Analyses separating stain-related from morphology-related components suggested that in this setting, prediction changes were predominantly associated with morphology-related rather than stain-related alterations. Overall, MoPaDi is a practical framework for counterfactual explanations in computational pathology that supports the evaluation of model-specific decision cues and hypothesis generation. MoPaDi is a diffusion-based tool for counterfactual image generation in cancer histopathology that reveals features associated with deep learning classifier predictions and supports transparent auditing of computational models in biomedical research.

Cancer cachexia is a devastating wasting syndrome with no approved therapies. In this study, we identify the tumor-derived glycoprotein ADAMTSL4 as a circulating factor associated with body weight loss in preclinical cachexia models and patients with colorectal and lung cancers. In mice, Adamtsl4 overexpression converted non-cachexia-inducing tumors into cachexia-inducing tumors, whereas its deletion in cachexia-inducing tumors spared fat and muscle, blunted muscle atrophy signatures, and reduced cachexia severity. ADAMTSL4 engages the latency-associated peptide (LAP) of TGFβ1, promoting local activation of TGFβ1 at muscle cell membranes. Genetic blockade of proTGFβ1 or pharmacologic inhibition of TGFβ signaling reduced ADAMTSL4-dependent wasting in adipocytes and muscle cells. Suppression of tumor-derived ADAMTSL4 attenuated skeletal muscle fibrosis in mice. Together, the association between increased circulating ADAMTSL4 levels and TGFβ-driven muscle atrophy and fibrosis gene signatures in patients with cachectic cancer identifies ADAMTSL4 as an upstream regulator of TGFβ1 and a potential therapeutic target in cancer cachexia. Cancer cachexia lacks effective therapies and remains a major cause of cancer-related morbidity and mortality. We identify tumor-derived ADAMTSL4 as an upstream regulator of latent TGFβ activation via LAP engagement that promotes multiorgan wasting and fibrosis-related remodeling. Targeting ADAMTSL4 may provide a selective therapeutic strategy without systemic TGFβ pathway blockade.

The intratumoral microbiota is increasingly recognized as an active component of the tumor microenvironment, yet whether it directly drives tumor mutagenesis remains unclear. Here, integrated multi-omics analysis of human non-small cell lung cancer (NSCLC) identifies Mycobacterium abscessus as a microbial determinant of APOBEC3A-associated mutagenesis. Mechanistically, the bacterial effector nucleoside diphosphate kinase (NDK) directly targets the host transcription factor IRF3 and installs a non-canonical 1-phosphohistidine modification at H263, thereby amplifying type I interferon signaling and sustaining APOBEC3A expression. This inter-kingdom phosphotransfer event links intratumoral microbial colonization to an endogenous mutational process that promotes genomic diversification. Genetic inactivation of NDK, or pharmacologic elimination using an engineered NDK-PROTAC, suppresses APOBEC3A activation and attenuates microbe driven mutagenesis. Together, these findings establish a direct microbial effector mechanism that promotes APOBEC3A-associated mutagenesis and provide a therapeutic framework to intercept microbiome driven mutagenesis in NSCLC.

Across many solid tumor types, cancer-associated fibroblasts (CAFs) are abundant and heterogeneous, with distinct subpopulations exerting immunomodulatory functions. Here we identify a novel population of immunomodulatory CAFs (imCAFs) in primary lung adenocarcinoma and pulmonary metastases, characterized by cell adhesion molecule L1-like (CHL1) expression and enriched in immune regulation and chemokine signaling programs. Through single-cell and spatial transcriptomics, we demonstrate that imCAFs are spatially colocalized with CXCR3+ regulatory T (Treg) cells, a hyper-suppressive subset accumulating at the tumor border. imCAFs produce CXCL9, driving CXCR3+ Treg cell recruitment and promoting an immunosuppressive microenvironment. CXCR3+ Treg cells display enhanced proliferative and suppressive capacity and are transcriptionally distinct from CXCR3- counterparts. Genetic ablation of Cxcr3 in Treg cells or Cxcl9 in stromal cells reduces Treg cell accumulation, enhances CD8+ T cell activation and decreases tumor burden. Analogous CHL1+ imCAF-like fibroblasts in human non-small cell lung cancer colocalize with Treg cells, and elevated CHL1 expression is associated with reduced cytotoxicity and decreased progression-free survival, highlighting the imCAF-CXCL9-CXCR3+ Treg axis as a promising therapeutic target.

Arsenic exposure is closely associated with lung carcinogenesis, but how cells transition from early stress buffering to later adaptive remodeling remains unclear. Stress granules (SGs) are key cytoplasmic condensates formed during acute stress, yet whether arsenic-responsive circular RNAs (circRNAs) participate in this process in non-small cell lung cancer (NSCLC) is poorly understood. Here, we identified hsa_circ_0000118 as an arsenic-responsive circRNA that was upregulated in both arsenic-exposed human subjects and arsenite-treated NSCLC cells. Silencing hsa_circ_0000118 impaired cell viability, promoted apoptosis, suppressed proliferation, and increased vulnerability to sustained arsenic stress. Mechanistically, acute arsenite exposure induced mitochondrial dysfunction and SG assembly before overt cytotoxicity. Under these conditions, loss of hsa_circ_0000118 increased SG susceptibility and delayed SG clearance during recovery. The RNA-binding protein quaking (QKI) acted upstream in this process, as QKI knockdown reduced hsa_circ_0000118 expression, weakened its association with G3BP1-related complexes, and phenocopied the SG-sensitive state. In contrast, under sustained arsenic stress, loss of hsa_circ_0000118 disrupted nuclear factor kappa B (NF-κB)-related adaptive output, characterized by accumulation of p105 and p65 but reduced downstream survival-related signaling. P65 also showed partial colocalization with G3BP1-positive granules after acute arsenite exposure, suggesting a possible spatial link between early SG remodeling and later NF-κB-related signaling dysregulation. Overall, hsa_circ_0000118 is associated with both acute SG dynamics and sustained NF-κB-related survival output in arsenite-treated NSCLC cells. These findings identify hsa_circ_0000118 as a candidate component of arsenic-associated stress adaptation.

Small cell lung cancer (SCLC) is an aggressive malignancy with limited effective therapeutic options. DLL3-targeted CAR-T therapy shows promising anti-tumor potential but is often restricted by T-cell exhaustion, which impairs its long-term efficacy. In the present study, we found that golidocitinib, a highly selective JAK1 inhibitor, induces apoptosis in SCLC cells in vitro via inhibiting STAT3 phosphorylation and regulating apoptosis‑associated genes. It also reduced the expression of exhaustion markers in anti-DLL3 CAR-T cells, promoted the formation of memory T cell phenotypes, and enhanced CAR-T cell persistence both in vitro and in vivo. When combined with anti-DLL3 CAR-T therapy, golidocitinib significantly augmented anti-tumor efficacy in both in vitro cytotoxicity assays and in vivo models, without obvious organ toxicity. These findings collectively demonstrate the dual anti-tumor effects of golidocitinib, thus providing a novel and promising strategy for SCLC treatment.

Trametinib, a selective MEK1/2 inhibitor, is approved for melanoma, BRAF-mutant non-small cell lung cancer, and thyroid cancer. Its favorable pharmacologic profile has prompted broader evaluation across cancers driven by MAPK/ERK signaling. However, its efficacy as monotherapy in breast cancer remains limited due to intrinsic resistance. Here, we investigated the molecular basis of intrinsic trametinib resistance and sought strategies to enhance therapeutic response. Trametinib responsiveness was associated with ETV4 expression. Notably, trametinib reduced ETV4 expression in three cell lines with high basal ETV4 expression (MDA-MB-453, SKBR3, and T47D). Consistent with ERK/MAPK-dependent regulation of ETV4, trametinib-mediated MEK inhibition was associated with stabilization of Capicua (CIC), a transcriptional repressor of ETV4, thereby suppressing ETV4 expression. In ETV4-high cells, trametinib-induced ETV4 downregulation promoted autophagic flux. Mechanistically, trametinib treatment and ETV4 silencing induced AMPK Thr172 phosphorylation, leading to ULK1 Ser555 phosphorylation and mTOR inhibition, thereby activating protective autophagy. RNA-seq analysis revealed that trametinib treatment and ETV4 knockdown produced highly overlapping transcriptomic profiles. Notably, trametinib reduced the expression of PPM1E, a phosphatase that negatively regulates AMPK, along with canonical MAPK effector genes. ChIP-PCR analysis and public ChIP-seq data demonstrated that ETV4 directly occupies the PPM1E promoter region and enhances PPM1E transcription, whereas trametinib-induced ETV4 suppression reduced PPM1E expression, limiting AMPK dephosphorylation and thereby promoting AMPK activation. Pharmacological inhibition of autophagy using chloroquine (CQ) or 3-methyladenine (3-MA) enhanced trametinib-induced apoptosis in vitro and suppressed T47D xenograft tumor growth in vivo. Collectively, our findings define a CIC-ETV4-PPM1E-AMPK signaling cascade through which trametinib-induced ETV4 downregulation drives AMPK-ULK1-dependent protective autophagy, thereby conferring a survival advantage in ETV4-high breast cancer. Autophagy blockade restores trametinib sensitivity and induces apoptosis, supporting a combinatorial strategy to improve MEK1/2-targeted therapy.

Multidrug resistance (MDR) is one of the main causes of chemotherapy failure and cancer recurrence. Among the ATP-binding cassette (ABC) transporter family, ATP binding cassette subfamily C member 1 (ABCC1/MRP1) is a key member that significantly reduces intracellular drug accumulation by effluxing various chemotherapeutic drugs, leading to treatment failure. Recent studies have found that microRNA (miRNA) can inhibit ABCC1 expression at the post-transcriptional level by targeting its 3' untranslated region (3'-UTR) region, thereby affecting chemotherapy sensitivity. However, miRNA itself is regulated by upstream molecules, particularly circular RNA (circRNA), which acts as competing endogenous RNA (ceRNA) and can bind miRNA through the sponge mechanism, relieving their inhibition of ABCC1 and promoting the MDR phenotype. Multiple studies have identified specific circRNA/miRNA/ABCC1 regulatory axes in various cancers such as non-small cell lung cancer, colorectal cancer, breast cancer, and ovarian cancer. This axis is not only involved in chemotherapy resistance but also affects tumor proliferation, migration, and invasion. Due to their high stability and tissue specificity, circRNAs and miRNAs have the potential as liquid biopsy biomarkers and therapeutic targets. Despite challenges posed by the complexity of the ceRNA network, intervention strategies targeting this axis, such as circRNA knockdown or miRNA mimics, are still considered important approaches to reverse cancer MDR. In addition, emerging computational approaches, such as graph learning, hypergraph networks, and pre-trained models, now enable large-scale prediction of circRNA-miRNA interactions, offering powerful tools to further dissect the regulatory axes underlying MDR.

The emerging focus on epigenetic regulation in cancer biology has unveiled the significant role of CXorf67, a protein encoded by a gene on the X chromosome. CXorf67 interacts with core components of PRC2, namely EZH2 and SUZ12, thereby influencing histone modifications like H3K27me3. Research indicates that CXorf67 is overexpressed in specific malignancies, including posterior fossa ependymomas, diffuse midline glioma, endometrial stromal sarcoma, non-small cell lung cancer, and Merkel cell carcinoma. In posterior fossa ependymomas and diffuse midline glioma, CXorf67 mimics the oncogenic histone H3K27M, inhibiting PRC2 function and altering chromatin states. In endometrial stromal sarcoma, CXorf67 forms fusion genes with MBTD1, potentially disrupting polycomb group (PcG) functions. Additionally, CXorf67's interaction with PALB2 affects the BRCA1-PALB2-BRCA2 complex, influencing DNA repair mechanisms. These findings highlight CXorf67's dual role in epigenetic regulation and DNA damage response, suggesting its potential as a therapeutic target. However, further research is needed to explore its functions in other cancers and clarify its molecular mechanisms. Our review synthesizes current knowledge on CXorf67's biological significance, particularly in epigenetics and DNA damage, and its implications in oncogenesis.

Tuft cells are rare, chemosensory epithelial cells present in various tissues, including the respiratory and gastrointestinal tracts. Recent studies have revealed their significant role in cancer biology, particularly through the expression of the transcription factor POU2F3, which serves as a master regulator of tuft cell lineage. In several cancer types, including small cell lung cancer, gastric cancer, and breast cancer, POU2F3 expression defines a distinct molecular subtype termed "tuft cell-like" tumors. These tumors exhibit unique transcriptional programs and altered tumor-immune interactions, contributing to their distinct therapeutic sensitivities. In this review, we first analyze the expression patterns of POU2F3 across cancer types using the TCGA datasets, revealing differential expression profiles and supporting the classification of tuft cell-like subtypes. We further explore cancer-type-specific signaling pathways regulating tuft cell differentiation and function, such as IL-25, acetylcholine, and taste receptor-related pathways. Finally, we propose that tuft cell-like signatures may serve as promising biomarkers for diagnosis, prognosis, and treatment stratification. Understanding the tuft cell-like-POU2F3 axis could open new avenues for targeted therapies in lineage-defined cancers.

Alectinib serves as an indispensable treatment for ALK (ALK receptor tyrosine kinase)-positive non-small-cell lung cancer (NSCLC), yet its hepatotoxicity and dermatotoxicity pose significant clinical concerns due to poorly understood mechanisms. This study demonstrated that alectinib-induced dermatotoxicity was secondary to hepatotoxicity. Integrated multi-omics analysis revealed that alectinib triggered excessive macroautophagic/autophagic degradation of hepatic BTD (biotinidase), causing systemic biotin deficiency that drove both hepatocyte apoptosis and skin barrier dysfunction. Mechanistically, we discovered increased phosphorylation of NBR1 at Ser656, a previously uncharacterized site, which conferred protein stability and contributed to selective BTD degradation. Importantly, exogenous biotin supplementation concurrently mitigated alectinib-induced hepatotoxicity and dermatotoxicity, providing a strategy for safer clinical application. These results uncovered a novel paradigm in drug-induced multi-organ toxicity, in which dysregulated inter-organ crosstalk served as a central mechanistic element.Abbreviations: AAV: adeno-associated virus; ALK: ALK receptor tyrosine kinase; BTD: biotinidase; c-CASP3: cleaved CASP3; c-PARP: cleaved poly(ADP-ribose) polymerase; CQ: chloroquine; DAPI: 4',6-diamidino-2-phenylindole; DHE: dihydroethidium; ELISA: enzyme-linked immunosorbent assay; GOT1/AST: glutamic-oxaloacetic transaminase 1; GPT/ALT: glutamic--pyruvic transaminase; GSEA: gene set enrichment analysis; H&E: hematoxylin and eosin; HPH: human primary hepatocyte; HRP: horseradish peroxidase; KEGG: Kyoto Encyclopedia of Genes and Genomes; KRT1: keratin 1; KRT5: keratin 5; KRT10: keratin 10; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MPHs: mouse primary hepatocytes; MSEA: metabolite set enrichment analysis; OCR: oxygen consumption rates; PBS: phosphate-buffered saline; PCA: principal component analysis; PLS-DA: partial least squares discriminant analysis; PTMs: post-translational modifications; ROS: reactive oxygen species; RSD: relative standard deviation; SD: standard deviation; SQSTM1/p62: sequestosome 1; SRB: sulforhodamine B; TBIL: total bilirubin; TUNEL: terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling.

Targeting redox homeostasis represents a promising strategy to selectively eliminate cancer cells through oxidative stress-mediated apoptosis. However, developing specific molecular entities capable of effectively disrupting this balance remains a key challenge. This study aimed to construct a novel library of cytisine derivatives through rational drug design to discover lead compounds with potent redox-disrupting activity and validate their potential as precision therapy candidates for lung cancer. A focused library of 77 derivatives was designed and synthesized via rational modification of key pharmacophores in cytisine. The antitumor activity and mechanism of action of candidate compounds were evaluated using in vitro cellular models and in vivo animal models. Among these, YU-C-ThioU-9 emerged as a lead compound with potent antitumor activity. Mechanistic investigations revealed that YU-C-ThioU-9 disrupts intracellular redox balance, leading to a marked accumulation of reactive oxygen species (ROS) and consequent oxidative damage. This redox perturbation, marked by oxidative stress accumulation, ultimately induces apoptotic cell death in lung cancer cells. Importantly, YU-C-ThioU-9 exhibited robust anticancer efficacy in both in vitro and in vivo models. These findings establish redox disruption as a clinically actionable avenue for precision oncology and position YU-C-ThioU-9 as a lead cytisine-derived candidate for lung cancer therapy.

Cardiac muscle wasting is a significant complication observed in lung cancer patients receiving radiotherapy. Radiotherapy, a commonly used anticancer treatment, is known to cause cardiovascular complications; however, the mechanisms linking tumor irradiation to cardiac wasting remain poorly understood. Lewis lung carcinoma (LLC) and CT26 tumor-bearing mice received localized tumor irradiation. Conditioned medium or EVs from irradiated tumor cells were collected and used to treat cardiomyocytes. Autophagy, protein synthesis, and atrophy were assessed. The roles of tumor Thbs1 and cardiac PERK signaling were determined via shRNA-mediated knockdown and PERK mutation in vitro and in vivo. We demonstrated that localized tumor irradiation induces cardiac muscle wasting in mice, which is associated with PERK-eIF2α-Atf4 pathway activation and increased Thbs1 protein-but not mRNA-levels in cardiomyocytes. Mechanistically, Thbs1 is delivered via extracellular vesicles (EVs) derived from irradiated tumors. Tumor-derived Thbs1+ EVs are necessary and sufficient to trigger autophagy, suppress protein synthesis, and cause atrophy in cardiomyocytes, which is dependent on the Thbs1-PERK interaction and downstream signaling. These results indicate that radiotherapy promotes the release of Thbs1+ EVs, which drive cardiac muscle wasting via PERK-eIF2α-Atf4 signaling, revealing a novel mechanism underlying cancer-associated cardiac damage.

Non-small cell lung cancer (NSCLC) remains the leading cause of morbidity and mortality from malignant tumors in China with therapeutic limitations. Developing novel therapeutic agents and innovative treatment strategies is critical for advancing NSCLC management. Gymconopin C, a compound isolated from Bletilla striata, has demonstrated potential anti-NSCLC activity; However, the underlying mechanism remains elusive. This study aimed to decipher the anti-NSCLC mechanism of Gymconopin C by demonstrating its induction of PINK1/Parkin-mediated mitophagy via the miR-6777-5p/ADRB2 pathway, and to establish the fundamental regulatory role of miRNA/mRNA axis in NSCLC suppression. This study establishes a novel therapeutically targetable pathway and offers a mechanistically grounded candidate for NSCLC therapy. Integrated in vitro and in vivo strategies were used to elucidate the anti-NSCLC mechanism of Gymconopin C. Inhibitory effects on two NSCLC cell lines were assessed using CCK-8 proliferation, transwell migration and invasion assays. Subsequently, transmission electron microscopy and mitochondrial functional analyses were conducted to assess mitophagy. Transcriptomic profiling revealed dysregulation of the miR-6777-5p/ADRB2 axis, and gain/loss-of-function experiments were performed to investigate its functional role in mitophagy. Finally, the therapeutic efficacy and pathway modulation were evaluated in vivo using zebrafish xenograft and murine ectopic tumor models. Gymconopin C significantly inhibited the proliferation, migration and invasion of NSCLC cells; induced cell cycle arrest and enhanced apoptosis of A549 cells. Mechanistically, it modulated the miR-6777/ADRB2 axis to promote PINK/Parkin-mediated mitophagy, ultimately leading to NSCLC growth arrest in both in vivo and in vitro experiments. Gymconopin C exerts tumor-suppressive effects by activating PINK1/Parkin-mediated mitophagy via the miR-6777-5p/ADRB2 axis, highlighting its potential as a therapeutic agent for NSCLC.

Osimertinib resistance remains a major obstacle to durable disease control of epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC), yet the regulatory circuitry that sustains a stem-like drug-tolerant state is incompletely understood. Here, the deubiquitinase (DUB) USP48 was identified as a critical determinant of acquired resistance through stabilization of Y-box binding protein 1 (YBX1) and reinforcement of Wnt/β-catenin signaling. CRISPR-Cas9 dropout screening of patient-derived organoids under osimertinib selection, integrated with multi-omics profiling and clinical correlation analyses, identified USP48 as a functionally selected vulnerability factor in resistant NSCLC. Mechanistically, USP48 associated with YBX1 and removed K48-linked polyubiquitin chains to prevent proteasomal degradation, thereby enabling YBX1-dependent transcriptional activation of PTK7 and subsequent amplification of Wnt-driven stemness programs. Notably, the canonical Wnt effector TCF7L2 directly upregulated USP48 transcription, establishing a positive feedback loop that locked resistant cells in a stable stem-like state. Therapeutically, genetic or pharmacological disruption of this axis, including inhibition of USP48 activity, blockade of downstream Wnt transcriptional output, disruption of the USP48-YBX1 interaction with glycyrrhizic acid, and lipid nanoparticle-mediated delivery of siUSP48-restored osimertinib sensitivity, achieved robust antitumor efficacy with survival benefits in orthotopic and patient-derived xenograft models. Collectively, our findings defined a DUB-centered positive feedback circuit that sustained osimertinib resistance, and highlighted USP48 as a therapeutically useful node to overcome EGFR-TKI resistance of EGFR-mutant NSCLC.

Lung neuroendocrine tumours (NETs, also known as carcinoids) are rapidly rising in incidence worldwide but have unknown aetiology and limited therapeutic options beyond surgery. The current WHO classification, based on mitotic count and presence or absence of necrosis, divides lung NETs into grade-1 typical, and grade-2 atypical tumours. This dichotomous classification however does not account for recently described molecular entities nor is it sufficient for clinical management. Here we conducted integrative multi-omic analyses on over 300 lung NETs including whole-genome sequencing, transcriptome profiling, and DNA methylation arrays, followed by archetype analysis, to identify and characterise molecular groups. We further investigated molecular groups using spatial RNA sequencing and proteomics, and deep learning analysis of whole slide images. The integration of multi-omic data provided definitive proof of the existence of four strikingly different molecular groups that vary in patient characteristics, genomic and transcriptomic profiles, microenvironment, and morphology. Among these, we identified a new molecular group, enriched for highly aggressive supra-carcinoids that displayed an immune-rich microenvironment linked to tumour-macrophage crosstalk. We uncovered an undifferentiated cell population within supra-carcinoids and show the transcriptomic similarities between supra-carcinoids and the recently identified atypical small cell lung cancer tumours, further demonstrating their molecular link to high-grade lung neuroendocrine carcinomas. Multi-regional genomic analyses identified distinct evolutionary trajectories, suggesting that molecular groups are determined early in tumourigenesis by genomic events, and that transitions between groups, though infrequent, are possible for supra-carcinoids. Deep learning models accurately identified these groups based on morphology alone, outperforming current histological criteria. Together with the validation of a panel of immunohistochemistry markers, we demonstrated that these molecular groups can be accurately identified based on morphological features, facilitating their future implementation in the clinical setting. Our proposed morpho-molecular classification highlights potential group-specific therapeutic opportunities, with differences in expression to DLL3, EGFR, FGFR and TERT inhibitor targets. Overall, our findings unify previously proposed molecular classifications and refine the lung cancer map by revealing novel tumour phenotypes with potential implications for prognosis and therapeutic management.

The prognosis of non-small cell lung cancer (NSCLC) with leptomeningeal metastasis (LM) is dismal. The regulators of LM progression remain elusive, thus impeding effective clinical intervention. Here, we performed in vivo genome-wide CRISPR-based screens and found that fused in sarcoma (FUS) ablation promoted LM in both PC9 and A549 cells. FUS repressed CD36 expression by directly interacting with and destabilizing PPARA (peroxisome proliferator-activated receptor α) messenger RNA. CD36 augmented fatty acid uptake and oxidative phosphorylation in NSCLC cells. Matrix metallopeptidase 2 (MMP2) was up-regulated through CD36-mediated fatty acid metabolism, which enabled NSCLC cells to disrupt the endothelial barrier. FUS-deficient NSCLC cells increased the expression of neuroendocrine differentiation (NED)-related markers, including SRY-box transcription factor 2 (SOX2), microtubule-associated protein 2 (MAP2), enolase 2 (NSE), and synaptophysin (SYP). They also exhibited neurite-like extensions and expanded in cerebrospinal fluid-supplemented medium in a CD36-dependent manner. Mechanistically, fatty acid uptake increased acetyl-coenzyme A and H3K27ac modifications to promote the expression of NED signature genes and MMP2 in NSCLC cells. Genetic or pharmacological inhibition of CD36 or inhibition of NED-related markers in NSCLC cells hindered LM and prolonged survival in mice. In patients with NSCLC, down-regulation of FUS or up-regulation of CD36 or SYP was associated with progression to LM. Thus, CD36-mediated fatty acid metabolism and NED signature are crucial for progression to LM in NSCLC, highlighting CD36 as a promising therapeutic target to limit the progression of LM.

While opioids remain the primary pharmacological intervention for cancer pain management, their clinical utility is frequently compromised by dose-limiting toxicities. This study aimed to determine the comparative efficacy, opioid-sparing potential, and clinical hierarchy of non-opioid adjuvant drug classes. The study was structured around the PICO framework to evaluate the pharmacological strategies currently utilized in multimodal clinical oncology. A systematic search of electronic databases (PubMed, Embase, Cochrane) was conducted for randomized controlled trials (RCTs) published between 2000 and 2025. The primary outcome was global analgesic efficacy (standardized mean difference [SMD]), while secondary outcomes included the opioid-sparing effect, defined as the percentage reduction in morphine equivalent daily dose (MEDD) and the incidence of treatment-emergent adverse events (Harms). A Bayesian network meta-analysis (NMA) was performed to rank treatments using SUCRA values. The methodological quality was assessed using the Cochrane Risk of Bias (RoB 2.0) tool. Twenty-three RCTs (n = 1845) met the inclusion criteria. Nonsteroidal anti-inflammatory drugs (NSAIDs) (-1.10) and anticonvulsants (-1.06) demonstrated the most robust analgesic effects. The SUCRA ranking confirmed a clear hierarchy, with the combination of anticonvulsants and antidepressants showing the highest probability of efficacy. A significant opioid-sparing effect was observed for gabapentinoids and ketamine, facilitating MEDD reduction. While serious adverse events were rare, minor harms (somnolence, dizziness) were more frequent in the most effective classes. Our NMA provides a robust evidence base for a "Clinical Tier" system, ranking adjuvants by their balance of efficacy and safety. These findings support the early integration of Tier I agents (anticonvulsants and NSAIDs) to optimize pain control and reduce opioid-related toxicities in chronic cancer pain management.

The purpose of this study was to determine whether liver magnetic resonance imaging (MRI)-derived tumor burden and growth rate at 3 months better predict overall survival (OS) than baseline imaging, circulating tumor DNA (ctDNA), and RECIST objective response in patients with hepatic metastases from metastatic uveal melanoma (mUM) treated with tebentafusp. A total of 88 patients with mUM treated with tebentafusp between 2018 and 2022 who underwent MRI examination at baseline and at three months following initiation of tebentafusp were retrospectively included. There were 52 women and 36 men, with a median age of 59.0 years (Q1, 52.0; Q3, 66.5; range: 30-77 years). Hepatic metastases were segmented to compute total tumor volumes at baseline (TTV0) and at 3 months (TTVm3) and the exponential tumor growth rate between scans (TGRm3, %/month). When available, ctDNA was quantified at the same time points. Associations with OS were assessed using Cox models and Harrell's concordance index (C-index) with 95 % confidence intervals obtained by 1,000-iteration bootstrap resampling. A three-tier MRI risk score combined median-based TTVm3 and TGRm3 cutoffs. In multivariable analysis, greater TTVm3 and TGRm3 were independently associated with shorter OS (hazard ratio [HR] per doubling of TTVm3, 1.25; 95 % confidence interval [CI]: 1.15-1.35; HR per +10 %/month TGRm3, 1.18; 95 % CI; 1.08-1.29; both P < 0.001), whereas baseline tumor volume and RECIST objective response were not. The MRI risk score yielded three well-separated OS curves (log-rank P < 0.001) and a C-index of 0.79 (bootstrap 95 % CI: 0.71-0.86), outperforming ctDNA-based models (C-index, 0.73; P = 0.01) and RECIST objective response (C-index, 0.65; P < 0.001). Dynamic liver MRI biomarkers in patients with mUM under tebentafusp, particularly month-3 total tumor volume and growth rate, provide independent prognostic information beyond baseline imaging, ctDNA, and RECIST and support MRI-based risk stratification as a central decision tool in mUM.

Animals coexist with complex microbial communities that influence their development, immunity, metabolism, and behavior. Evidence shows these effects arise not just from metabolic and immune signaling but also from epigenetic mechanisms that alter host gene expression. Microbial signals can modulate DNA methylation, histone modification, chromatin accessibility, and RNA pathways, reshaping transcription across tissues. This review synthesizes evidence from diverse animal systems to demonstrate how microbial communities influence epigenetic landscapes and contribute to immunity, development, metabolism, and neurobiology. We explore data suggesting that microbial epigenetic interactions extend into the tumor microenvironment, where intratumoral microbes may shape disease progression by remodeling epigenetic states. Comparative studies indicate that microbial regulation of host epigenetics is an evolutionarily conserved mechanism linking environmental signals to phenotype. Despite recent advances, questions remain about causality, cell-type specificity, persistence, and inheritability of these effects. We propose microbial epigenetic regulation as a key interface integrating microbial cues with host physiology and pathology, providing a framework for understanding host-microbe interactions across species.

Transplant-associated cutaneous squamous cell carcinoma (TSCC) is one of the most common malignancies in solid organ transplant recipients and is strongly associated with long-term immunosuppression. However, the transcriptional programs and molecular mechanisms through which chronic immunosuppression reshapes intratumoral CD8⁺ T-cell differentiation remain poorly understood. We integrated single-cell transcriptomic and T-cell receptor repertoire analyses of TSCC and immunocompetent cutaneous squamous cell carcinoma (SCC). An independent SCC cohort containing paired tumors and adjacent tissues, together with matched spatial transcriptomic data, was used to evaluate the association between EOMES and antitumor CD8⁺ T-cell differentiation. Single-cell transcriptomic analysis revealed marked accumulation of precursor effector-memory CD8⁺ T cells (Pre-Tem) in TSCC, accompanied by reduced cytotoxic programs and clonal expansion. Trajectory and regulatory-network analyses revealed retention of precursor-like differentiation states associated with reduced EOMES expression and regulon activity. In silico perturbation modeling predicted EOMES as a candidate transcriptional regulator linked to cytotoxic gene expression and effector differentiation. An independent SCC cohort further supported the biological relevance of EOMES-associated programs in antitumor CD8⁺ T-cell states. Cell-cell communication analysis identified CXCL10-CXCR3 signaling as a dominant pathway targeting Pre-Tem cells and related with diminished EOMES-associated differentiation programs. Spatial transcriptomics further identified spatial co-localization among CXCL10 signaling, EOMES-associated states, and Pre-Tem cells within a tumor-margin niche. Pre-Tem accumulation and altered differentiation states represent a previously unrecognized feature of immune dysfunction in TSCC. EOMES-associated programs were closely linked to precursor-to-effector CD8⁺ T-cell differentiation, while CXCL10-CXCR3 signaling may contribute to impaired precursor-to-effector differentiation. These findings provide a conceptual framework for understanding immune dysfunction in transplant-associated malignancies and establish a rationale for future mechanistic and translational studies.

Microwave ablation is a crucial option for liver tumors, with success hinging on generating a suitably sized ablation zone for complete tumor eradication. Mathematical modeling supports ablation zone prediction and clinical decision-making, but its utility is limited by high computational cost and modeling complexity. We propose a physics-informed ablation modeling framework based on a homogenization-perturbation strategy that separates device-driven physics from patient-specific variability. A unified bioheat solver efficiently computes a semi-analytical temperature baseline induced solely by device-configurable parameters. A dual-branch network learns first-order corrections from pre-ablation MRI to align this baseline with individual anatomy and physiology. Ground-truth ablation zones delineated on post-ablation MRI provide high-fidelity image-based supervision. The framework combines physical interpretability with data-driven adaptability, enabling real-time (∼20 ms), patient-specific prediction and offering broad potential for clinical deployment and extension to other thermal therapies.

Accurate segmentation of multiple organs is essential for the diagnosis and treatment of head and neck cancer. However, the intricate anatomical structure and dense organ distribution in the head and neck region pose significant challenges for existing automated segmentation models, which predominantly target single organs and rely on single-modality imaging. Achieving comprehensive, one-step segmentation of organs-at-risk (OARs) remains challenging. To this end, we propose a Point-cloud Matrix Fusion-based Segmentation Model (PMFM) that leverages an improved multi-modal data fusion strategy for the automated full segmentation of OARs in head and neck cancer. The proposed PMFM involves three core modules: 1) a camera model-based 3D feature mapping and point-cloud extraction module (PEM) that enables vertical decoupling of modalities and objects; (2) a Point Cloud Matrix Module (PMM) utilizing PointNet and a virtual point cloud-based attention mechanism to facilitate horizontal association and global feature learning across modalities; and (3) a Cross Fusion Module (CFM) based on virtual point clouds to achieve deep intermodal object fusion and enhance inter-organ correlation. PMFM effectively integrates multi-modal image information, transforming them into a unified virtual point cloud matrix, and enables precise, comprehensive segmentation of OARs in head and neck cancer. Extensive validation and comparative experiments on the HaNSeg dataset demonstrate that PMFM significantly outperforms state-of-the-art methods, achieving an average Dice coefficient of 79.8% and an average Hausdorff distance of 2.47 mm. The source code for this study will be publicly available on GitHub at https://github.com/zhouxinyu1028/PMFM.

Nucleic acid modification constitutes a pivotal regulatory mechanism in cancer, influencing the entire process of tumor development, diagnosis, treatment, and prognosis. This review delineates the role of diverse chemical modifications-including methylation, demethylation, N6-methyladenosine, and 5-methylcytosine-in governing genomic stability and cellular dysfunction across DNA, coding RNA, and non-coding RNA levels. The research paradigm of nucleic acid modification in cancer is transitioning from static modification maps to dynamic, interconnected modification networks. Moreover, the crosstalk between nucleic acid modifications and nucleic acid processing further intensifies epigenetic remodeling and oncogenic risk. Elucidating the fundamental mechanisms underlying these modifications will provide critical insights into their overarching significance in cancer initiation and maintenance, while offering promising diagnostic and therapeutic targets for precise targeted treatment of cancer at the nucleic acid level.

Multiple instance learning (MIL) has enabled substantial progress in computational histopathology, where a large amount of patches from gigapixel whole slide images are aggregated into slide-level predictions. Heatmaps are widely used to validate MIL models and to discover tissue biomarkers. Yet, the validity of these heatmaps has barely been investigated. In this work, we introduce a general framework for evaluating the quality of MIL heatmaps without requiring additional labels. We conduct a large-scale benchmark experiment to assess six explanation methods across histopathology task types (classification, regression, survival), MIL model architectures (Attention-, Transformer-, Mamba-based), and patch encoder backbones (UNI2, Virchow2). Our results show that explanation quality mostly depends on MIL model architecture and task type, with perturbation ("Single"), layer-wise relevance propagation (LRP), and integrated gradients (IG) consistently outperforming attention-based and gradient-based saliency heatmaps, which often fail to reflect model decision mechanisms. We further demonstrate the advanced capabilities of the best-performing explanation methods: (i) We provide a proof-of-concept that MIL heatmaps of a bulk gene expression prediction model can be correlated with spatial transcriptomics for biological validation, and (ii) showcase the discovery of distinct model strategies for predicting human papillomavirus (HPV) infection from head and neck cancer slides. Our work highlights the importance of validating MIL heatmaps and establishes that improved explainability can enable more reliable model validation and yield biological insights, making a case for a broader adoption of explainable AI in digital pathology. Our code is provided in a public GitHub repository: https://github.com/bifold-pathomics/xMIL.

Osteosarcoma is a highly malignant bone tumor with a complex immune evasion mechanism involving various molecules and signaling pathways. This study aims to clarify how APE1 regulates the expression of miR-513a and miR-149 and to explore its role in osteosarcoma immune evasion. The results indicate that APE1 levels are inversely related to those of miR-513a and miR-149. Exosomal miR-513a and miR-149 significantly impair T-cell function by decreasing TNFα secretion. APE1 contributes to osteosarcoma's immune evasion by modulating exosomal miR-513a and miR-149, influencing the PD-L1 and TCR pathways, thus presenting a potential molecular target for therapy.

To enable simple and rapid detection of circulating tumor cells (CTCs), we developed a targeted colorimetric biosensor based on MiRGD-functionalized Fe3O4-PEI-Au nanoparticles conjugated with TGA and Ni+2-NTA (MiRGD@NTA@Au-Fe3O4 nanoparticles). The biosensor enables magnetic separation of target cells and visual signal generation through the peroxidase-mimicking activity of Au-Fe3O4 nanoparticles, using TMB as a substrate. Colorimetric signals were monitored at 650 nm and discerned by the naked eye. Spectral data and fluorescence microscopy confirmed the specific binding of MiRGD peptides to cancer cells, ensuring discrimination from non-cancerous cells. Under optimized conditions, the assay exhibited a linear response for MDA-MB-231 cells suspended in PBS in the range of 5-5000 cells/mL, with a limit of detection (LOD) 6 cell/mL. The biosensor also demonstrated robust performance in human blood samples, with LOD 8 cell/mL, confirming its applicability in complex biological environments. These results highlight MiRGD@NTA@Au-Fe3O4 nanoparticles as a promising platform for simple, rapid, and high-sensitivity detection of CTCs, offering potential for clinical diagnostics and personalized cancer monitoring.

Single-molecule localization microscopy (SMLM) enables visualization of chromatin architecture at nanoscale resolution. However, high-performance DNA probes suitable for SMLM in both live cells and tissues remain limited. We developed Hoechst-6-Carboxytetramethylrhodamine (6-TAMRA) derivative (HoT) probes-rhodamine-based derivatives conjugated to a Hoechst moiety-through structural fine-tuning of rhodamine spirocyclization. HoTs are self-assembling, auto-blinking probes with excellent photostability and high temporal resolution. They permeate live cells, enabling long-term, real-time nanoscopic chromatin imaging in live and fixed cells and in tissue sections. In live cells, we identified nanoscale features in the 3D organization of chromatin and quantified DNA fiber kinetics at high resolution. We quantified DNA compaction in single cells within retinal and colon cancer sections. OligoSTORM (stochastic optical reconstruction microscopy)-labeled gene loci can be visualized and measured within their HoT-labeled chromatin footprints. Our work provides powerful tools for investigating chromatin structure and functions in living cells and tissues, with applications ranging from cancer diagnosis to retinal regeneration.

Glioblastoma radioresistance is linked to glutathione, a key antioxidant protecting against oxidative stress. Sulfasalazine inhibits the xCT antiporter mediating cystine uptake required for glutathione synthesis in glioma, potentiating radiotherapy in preclinical studies. This trial evaluated safety and therapeutic potential of combining sulfasalazine with stereotactic radiosurgery (SRS) for recurrent glioblastoma. Adults with recurrences were enrolled in a 3 + 3 dose-escalation trial receiving 1.5, 3.0, 4.5, or 6.0 g sulfasalazine orally for 3 days before SRS. Primary endpoint was safety. Secondary endpoints included changes in quality of life (QOL), tumor glutathione-levels, metabolism, and volumes assessed via Functional-Assessment-of-Cancer-Therapy-Brain-questionnaire, glutathione-edited-MR-spectroscopy,11C-Methionine-PET, and MRI. Eleven eligible patients treated with GKRS only due to closed research facilities served as concurrent controls for time to local tumor progression, progression-free survival (PFS), and overall survival (OS). Twelve patients were enrolled and followed until death or consent withdrawal (n = 1). Of 20 adverse events, two were grade 3 (transient lymphocytopenia), four grade 2, and fourteen grade 1. QOL remained stable for 6 months (p = 0.292). Tumor glutathione-levels and metabolic activity were reduced on day 3 (p = 0.010) and 1 month (p = 0.052), respectively. Best RANO-responses were objective (5/11), stable (5/11) or progressive (1/11) for participants vs. stable (2/11) or progressive (9/11) for controls, p = 0.001. Participants had longer local tumor control (median difference: 5.3 months, 95% CI: 1.0-9.6, p < 0.001) and PFS (1.7 months, 95% CI: 0.3-3.0, p = 0.009), with equivalent OS (p = 0.915). In conclusion sulfasalazine and SRS were well tolerated, reducing intratumoral glutathione-levels and associated with more durable local control than SRS alone supporting phase II investigation. TRIAL REGISTRATION: NCT04205357.

Glioblastoma (GB) is the most aggressive brain cancer, with poor prognosis due to infiltrative invasion of glioma stem cells (GSCs) and the immunosuppressive tumor microenvironment (TME). We have previously demonstrated that pericytes (PCs), specialized cells in the blood microvessels surrounding GB, are conditioned by infiltrating tumor cells to aberrantly upregulate their chaperone-mediated autophagy (CMA). Elevated CMA in PCs promotes stable cell-cell interactions with tumor cells and a pro-tumoral immune phenotype that supports tumor progression. In this work, to test if inhibition of CMA in PCs might be an effective strategy to reduce tumor survival, we have used the phosphopeptide P140, known to restore aberrant CMA upregulation in specific immune cells. We found that administration of P140 peptide in an immunocompetent GB mouse model with both patient-derived GSCs and GB cell lines, neutralizes GB-induced PC CMA upregulation resulting in ablation of PC-tumor cell interactions and triggering of a secretome toxic to tumor cells. We identified MAPT/tau, a known CMA substrate, as one of the main components of this secretome, and discovered that CMA-dependent PC MAPT/tau secretion within the GB TME plays a key role in cancer progression and recurrence. Furthermore, we found that perivascular accumulation of MAPT/tau is an effective way to monitor peptide P140 treatment efficacy in GB and for prognosis of the patient evolution. Our findings validate P140 peptide treatment as a safe and specific strategy to halt GB progression and establish the clinical relevance of extracellular MAPT/tau as a biomarker for therapeutic success in GB.Abbreviations: ACTA2/ASMA: actin alpha 2, smooth muscle; BBB: blood-brain barrier; CMA: chaperone-mediated autophagy; CTLA4: cytotoxic T-lymphocyte associated protein 4; DAB: 3-3'diaminobenzidine; GB: glioblastoma; GBCPC: GB-conditioned pericyte; GSCs: glioblastoma stem cells; HSPA8/HSC70: heat shock protein family A (Hsp70) member 8; LAMP2A: lysosome associated membrane protein 2A; LI: lysosomal inhibitors; MAPT/tau: microtubule associated protein tau; NAc: N-acetylcysteine; PC: pericyte; PCGB: pericyte-glioblastoma coculture; PDGFRB: platelet derived growth factor receptor beta; ROS: Reactive oxygen species; SPARC: secreted protein acidic and cysteine rich; TME: tumor microenvironment; TNT: tunneling nanotubes.

Gliomas are aggressive and treatment-resistant tumors of the central nervous system, characterized by molecular heterogeneity, diffuse infiltration, rapid progression, and persistent poor prognosis despite multimodal therapy. Emerging evidence highlights the role of complex genetic changes (such as IDH1 and H3F3A mutations) and epigenetic interactions in reshaping chromatin structure and activity in gliomas, increasing their reliance on epigenetic regulators for their growth and resistance. The nuclear scaffolding protein WD repeat domain 5 (WDR5), is a core component of the MLL/SET1 (WRAD) methyltransferase complex, which has been recently validated as a molecular target for cancer. WDR5 implication in H3K4 trimethylation (H3K4me3)-mediated gene regulation sustains transcriptional programs linked to proliferation, ribosome biogenesis, stemness, and MYC-driven oncogenic activity. In gliomas, WDR5 expression is increased, promoting proliferation and migration, as well as maintaining the glioma stem cell population, contributing to tumor progression. Targeting of WDR5 through specific WIN-site and WBM-site inhibitors or PROTAC degraders has been shown to impair WRAD assembly, reduce H3K4me3 levels, weaken MYC-associated transcription, and suppress tumor growth. In this review, we highlight the significant role of WDR5 in gliomas as part of a tumor-specific epigenetic vulnerability network, providing a critical update on the major WDR5-targeted inhibitors and degraders for future therapeutic applications.

Despite successful immuno-oncology therapies in other cancers, they have largely failed in glioblastoma (GBM). In this study, natural killer (NK) cells from patients with glioma show impaired oxidative phosphorylation and mitochondrial complex I activity. Multiomics profiling identified complex I subunit NDUFA9 as a critical mediator of NK cell metabolic fitness. The abundance of NDUFA9+ NK cells informed patient outcomes. Ndufa9 knockout in NK cells compromised mitochondrial function, antitumor efficacy, and the memory-like phenotype of NK cells by triggering a metabolic reprogramming toward glutamine dependence. The decreased α-ketoglutarate/succinate ratio in Ndufa9-deficient NK cells mediated widespread epigenetic reprogramming by inducing the transcriptionally repressive histone mark H3K27me3 on key immune function genes. Resveratrol-mediated NDUFA9 activation or its overexpression enhanced NK cell anti-GBM function by restoring complex I activity. Together, these findings reveal the critical role of mitochondrial complex I activity in NK cells and highlight its potential as an actionable target to enhance NK cell-based immunotherapy for patients with GBM. This study reveals that NDUFA9 deficiency in mitochondrial complex I compromises metabolic fitness and impairs the antitumor activity of NK cells in GBM. We identify the mitochondrial complex I subunit NDUFA9 as a key pharmacologically targetable node for NK cell-based immunotherapy in GBM, leveraging metabolic reprogramming to enhance antitumor efficacy. See related commentary by Tiberti et al., p. 1727.

Glioblastoma (GBM) is a highly aggressive brain tumor for which cell-free DNA (cfDNA) has shown promise as a minimally invasive biomarker, yet the biological processes governing cfDNA release and composition in GBM remain incompletely understood. We investigated cfDNA release dynamics, fragmentation patterns, variant allele frequencies (VAF), and copy number profiles under controlled experimental conditions using patient-derived GBM cultures. Longitudinal sampling of conditioned media from monocultures revealed progressive increases in cfDNA yield that correlated more strongly with viable cell numbers than with cell death, suggesting that cfDNA production in these models is not solely driven by apoptosis. In co-culture experiments combining GBM cells with normal human astrocytes (NHA), distinct tumor- and astrocyte-specific variants enabled deconvolution of mixed-cell populations, and cfDNA composition shifted over time, consistent with increasing astrocyte death under competitive co-culture conditions. Temozolomide (TMZ) treatment altered cfDNA release dynamics, shifting the dominant source from viable cells to cell-death-associated pathways, accompanied by increased cfDNA yield, nucleosomal fragmentation, and evidence of reduced variant diversity under therapeutic pressure. Tumor-derived cfDNA was also detected and deconvolved from plasma in GBM orthotopic xenograft models, with copy number profiles recapitulating those of the parental tumor cells. These findings suggest that cfDNA composition is shaped by tumor proliferation, microenvironmental context, and therapeutic stress, establishing a preclinical foundation for interpreting cfDNA-based liquid biopsy signals in GBM.

Diffuse middle glioma (DMG or DIPG) is a fatal pediatric brain tumor. Although chimeric antigen receptor (CAR) T-cell therapy shows promise, clinical outcomes remain inconsistent due to premature exhaustion, underscoring a critical need to improve CAR-T persistence. A major barrier to CAR-T efficacy is antigen-independent tonic signaling, yet the extent to which tonic signaling shapes CAR-T durability and clinical outcomes, particularly in DMG, remains incompletely defined. Using a clinically investigated B7-H3 MGA271-based CAR as a reference platform, we generated alternative B7-H3 CARs incorporating either a human codon-optimized 376.96 (B7H3.BC) or Hu8H9 scFv antigen binding domain to systematically assess scFv-dependent effects on tonic signaling and therapeutic efficacy. CAR-T cells were evaluated using integrated in vitro and in vivo functional assays, alongside multi-omics profiling and computational modeling. We further derived a tonic signaling-associated gene signature and evaluated its predictive performance across independent clinical datasets. B7H3.BC CAR-T cells exhibit markedly restrained tonic signaling compared with MGA271- and Hu8H9-based counterparts, accompanied by superior antitumor activity and enhanced persistence across patient-derived DMG cells. Integrated multi-omics and single-cell profiling further identified a tonic signaling-associated gene signature that outperforms conventional T-cell exhaustion signatures in predicting therapeutic efficacy across multiple clinical trials, including DMG and other tumors. Our findings establish that scFv-dependent modulation of tonic signaling critically governs CAR-T persistence and antitumor efficacy in DMG. By linking CAR design to transcriptional and epigenetic programs, our study provides a principle-based and predictive framework to inform rational CAR engineering and improve therapeutic outcomes.

Isocitrate dehydrogenase-mutant (IDH-MUT) gliomas exhibit distinct metabolic profile marked by 2-hydroxyglutarate (2-HG) accumulation at the expense of α-ketoglutarate. How these tumors maintain the tricarboxylic acid (TCA) cycle, however, remained unclear. We conducted comprehensive metabolomic profiling using clinical cohorts, cell lines, and patient-derived organoids (PDOs). The metabolic dynamics of TCA and urea cycles were interrogated using stable isotope tracing with 13C-aspartate, U5-13C-15N-aspartate, 15NH4Cl and 15N-glutamate. The functional role of carbamoyl-phosphate synthase 1 (CPS1), the key enzyme of the urea cycle, was validated through inhibition experiments in vitro, in vivo and in PDO model, followed by seahorse respirometry and electron microscopy. Metabolomic profiling of two glioma cohorts consistently showed elevated urea cycle metabolites in IDH-MUT tumors. We identified CPS1 as a metabolic checkpoint sustaining TCA cycle through half urea cycle (from ammonia to arginine). Of note, CPS1 upregulation drove fumarate anaplerosis to sustain TCA flux in IDH-MUT gliomas. Thus, CPS1 inhibition not only reduced fumarate levels but also decreased oncometabolite 2-HG both in vitro and in vivo. Consistently, CPS1 inhibition impaired mitochondrial respiration and suppressed tumor growth in vitro, in vivo and in PDOs. Taken together, metabolic checkpoint CPS1 orchestrates half urea cycle to replenish the TCA cycle in IDH-MUT gliomas. Targeting CPS1 represents a promising metabolic therapeutic target for this glioma subtype.

Diffuse midline glioma, H3K27-altered (DMG), is a lethal midline brain tumor. Most DMG, unlike glioma arising in other regions, harbor histone H3.3 K27M (K27M) mutations. The basis for this anatomical selectivity remains unclear. Stem-like DMG cell transcriptomes most resemble oligodendrocyte precursor cells (OPCs). Using conditional K27M knock-in mice, we show that K27M reduces oligodendrocyte differentiation, altering the proportions of oligodendrocytic cell states in a region-specific manner. In vivo EdU labeling in tissue-cleared whole brains revealed greater K27M-driven increases in proliferation within pons, midline, and hindbrain regions than in telencephalon. In vitro, wild-type brainstem OPCs proliferate more slowly and differentiate later than telencephalic OPCs. K27M enhances brainstem OPC proliferation and disrupts regional transcriptional programs, selectively restraining full maturation of brainstem OPCs while inducing brainstem-selective upregulation of bivalent Bmp, Wnt, and Notch pathway genes. These findings suggest that K27M exploits intrinsic regional differences in oligodendrocyte development, creating a brainstem-selective window for gliomagenesis.

Cerebrospinal fluid contributes to homeostasis in the central nervous system, but how its dynamics are altered in glioblastoma is unclear. We find that glioblastoma drives leptomeningeal perivascular fibrosis that is associated with impaired fluid transport and clearance. Lineage tracing and single-cell analyses in male tumor-bearing mice identify leptomeningeal fibroblasts as the principal source of this fibrotic response, with limited contribution from pericytes. Fibrosis is linked to activation of nuclear factor kappa B signaling in collagen-producing fibroblasts, extracellular matrix deposition around perivascular spaces, reduced intratumoural cytotoxic T cell accumulation and a less permissive immune microenvironment. Here, we show that inhibiting nuclear factor kappa B signaling in leptomeningeal fibroblasts reduces fibrosis, improves fluid clearance, enhances intratumoural cytotoxic T cell accumulation and restores responsiveness to programmed cell death protein 1 blockade, identifying this pathway as a therapeutic target in glioblastoma.

Relacorilant (LIFYORLI™) is a non-steroidal, selective glucocorticoid receptor II (GR II) antagonist being developed by Corcept Therapeutics for the treatment of various solid tumours (including ovarian cancer, fallopian tube cancer, peritoneal cancer, pancreatic cancer and prostate cancer) and Cushing syndrome. Relacorilant received its first approval in the USA on 25 March 2026 for use in combination with nab-paclitaxel for the treatment of adults with platinum-resistant epithelial ovarian, fallopian tube or primary peritoneal cancer who have received 1-3 prior systemic treatment regimens, at least one of which included bevacizumab. This article summarizes the milestones in the development of relacorilant leading to this first approval for platinum-resistant ovarian, fallopian tube or peritoneal cancer.

Breast and prostate cancers share notable similarities in their progression to invasive disease, providing a unique opportunity to uncover common mechanisms of malignant transition. By integrating volumetric reconstruction with multimodal spatial profiling, Storrs and colleagues revealed the precancer-to-cancer transition as a continuous three-dimensional process and identified shared molecular programs associated with invasion. See related article by Storrs et al., p. 1819.

This study investigated how the natural compound celastrol (CEL) impedes bladder cancer (BLCA) progression by irreversibly occupying the ATP-binding pocket of hexokinase 2 (HK2). CEL binds directly to the ATP cleft of HK2, permanently inactivating its enzymatic function and consequently suppressing glycolytic flux and lactate output in tumor cells. The diminished availability of lactate leads to reduced histone lactylation-a recently recognized epigenetic mark-which in turn downregulates the expression of the RNA methyltransferase METTL3 and decreases the global N⁶-methyladenosine (m⁶A) abundance. Importantly, we revealed a self-reinforcing circuit linking HK2‑driven glycolysis, histone lactylation, and METTL3‑dependent m⁶A deposition that fuels malignant growth. By stably engaging the ATP-binding pocket of HK2, CEL dismantles this metabolic-epigenetic feed‑forward loop, a mechanism that was substantiated through in vitro and in vivo assays and an analysis of clinical samples. Collectively, these results define a pivotal metabolic-epigenetic axis in BLCA and identify CEL as an agent that concurrently blocks energy production and post-transcriptional regulation. The results of the present study underscore the promise of irreversible HK2 blockade as a novel therapeutic avenue for managing BLCA.

Neuroblastoma is a pediatric solid malignancy that primarily arises from poorly differentiated sympathoadrenal cells in the developing autonomic nervous system. The neuroblastoma genetic landscape is characterized by a paucity of somatic mutations and a far more common occurrence of chromosome copy number variations (CNVs). Two of the most common genetic abnormalities in neuroblastoma are the amplification of MYCN and the unbalanced gain of the long arm of chromosome 17 (17q+), both of which correlate with poor prognosis. Although mouse models exist for MYCN-driven neuroblastoma, no mouse models exist for 17q+. Thus, the mechanism by which 17q+ contributes to poor prognosis remains largely unknown. Here, we used a human pluripotent stem cell (PSC) model of neuroblastoma to show that 17q+ cooperates with MYCN amplification to accelerate tumorigenesis compared to MYCN amplification alone. We identified six genes on 17q that are upregulated in both our 17q+ PSC-derived tumors and 17q+ neuroblastoma patient tumors compared to WT 17q, correlate with poor prognosis in neuroblastoma patients, and are overexpressed at the protein level in our MYCN/17q+ tumors compared to MYCN alone. Of these six genes, knockdown of BIRC5, CDC6, IGF2BP1, or TK1 in MYCN/17q+ cells increases tumor latency in vivo and contributes to tumor progression through diverse mechanisms. Furthermore, knockdown of CDC6, IGF2BP1, and TK1 in neuroblastoma patient-derived xenografts with 17q+ slowed growth in vivo. Our findings provide new insights into potential candidate 17q drivers of neuroblastoma tumorigenesis and possible therapeutic vulnerabilities.

Multimodal artificial intelligence, albeit showing great potential in computational pathology, remains limited to isolated patch-level interpretation and often fails to analyze gigapixel-scale whole-slide images (WSIs) essential for clinical utility. Here we present SlideChat, a multimodal generative artificial intelligence assistant for whole-slide computational pathology across cancer types. SlideChat integrates patch-level and slide-level pathology encoders with a pretrained large language model. Using 274,233 multimodal instruction samples, SlideChat is trained to learn the associations between WSIs and diagnostic reports and interpret complex queries in clinical practice. Evaluated on 8,836 closed-ended questions, 129 open-ended questions and 3,149 WSI reports from five cohorts spanning 31 cancer types, SlideChat outperformed leading baselines by 19.1% in closed-ended accuracy and by 7.7% in report-generation Metric for Evaluation of Translation with Explicit Ordering score and received the highest expert ratings across five dimensions in open-ended question answering, showing the potential to enhance diagnostic workflows, medical education and clinical decision-making.

Tumor-associated macrophages (TAMs) play important roles in cancer progression and resistance to therapy. Recent studies have shown that TAMs include both long-lived resident tissue macrophages (RTMs) and short-lived monocyte-derived macrophages (MDMs) with limited proliferative potential. RTMs and MDMs have been suggested to play divergent roles in tumorigenesis; RTMs are aligned with trophic functions, whereas MDMs are enriched for immune-regulatory pathways. Here we established a specific role for the AP-1 factor JUN in the differentiation and maintenance of MDMs and the specification of pro-tumoral trophic functions during tumor development. Alternatively, the immune-regulatory functions of TAMs remained JUN-independent. JUN was required for the specification and maintenance of pro-tumoral TAMs that support blood vessel maturation and tumor growth. Single-cell transcriptomics analysis uncovered the alternative fates for tumor-infiltrating monocytes and the development of distinct TAM states associated with trophic functions and immune-regulation. These studies demonstrate an important role for JUN in the specification of pro-tumoral monocyte-derived TAMs that could offer opportunities for selective TAM-targeted therapies for cancer. JUN promotes the specification of pro-tumoral macrophages but is dispensable for pro-inflammatory macrophages, suggesting targeting the JUN pathway in myeloid cells could suppress pro-tumoral macrophages while sparing antitumor inflammatory myeloid cells.

Lactate, known for its metabolic functions, has recently emerged as a signaling molecule. In a recent issue of Immunity, Guo and colleagues report that lactate is a competitive antagonist of STING and propose an epidermal growth factor receptor (EGFR)-mediated signaling cascade that enhances lactate dehydrogenase A (LDHA) activity, lactate production, and immunosuppression.1.

Quinones are natural compounds widely distributed in animals, plants, and microorganisms, where they play essential roles in cellular energy metabolism. Ascorbate (vitamin C) can exert antioxidant or pro-oxidant effects depending on concentration and cellular context. Studies have shown that quinones can undergo redox cycling, leading to the generation of reactive oxygen species (ROS), a process that is further enhanced in the presence of ascorbate. Due to the elevated production of reactive oxygen species (ROS) and compromised antioxidant defenses in cancer cells, redox imbalance has been proposed as a potential "Achilles' heel" of tumors. In this context, the combination of ascorbate and quinones represents a promising strategy for selectively targeting cancer cells through enhanced ROS generation. This review discusses the mechanisms and biological effects of natural and synthetic quinoid compounds, particularly their interactions with ascorbate in cancer cells.

Simultaneous acquisition of quantitative phase and fluorescence information is essential for comprehensive cellular analysis, as these complementary modalities provide structural and biochemical insights, respectively. However, conventional dual-mode imaging systems require fluorescent labeling, complex optical architectures, and multiple acquisition channels, which limit scalability, increase cost, and hinder deployment in low-resource or high-throughput settings. Here, we present an AI-driven dual-mode phase and label-free fluorescence imaging platform using a single-shot Gabor hologram. The proposed framework enables simultaneous reconstruction of quantitative phase images and virtual fluorescence channels from a single low-cost holographic measurement, eliminating the need for fluorescent staining, multi-shot acquisition, or multimodal optical hardware. To achieve this, we introduce a one-sided unsupervised diffusion model that learns a unidirectional mapping from Gabor holograms to dual-mode outputs without requiring paired training data or cycle-consistency constraints. The model integrates contrastive learning-based hologram synthesis with hologram-conditioned denoising diffusion to ensure high structural fidelity and robust cross-modal reconstruction. Ground-truth phase and fluorescence images acquired from a conventional dual-mode optical system are used only during training, while inference relies exclusively on a single-shot Gabor hologram captured with a minimal optical configuration. Experimental validation across multiple cancer cell lines and organelle-specific fluorescence channels demonstrates that the proposed platform accurately recovers cellular morphology and subcellular distributions, achieving an FID of 57.74, SSIM of 0.76, PSNR of 26.89 dB, and LPIPS of 0.12, and further generalizes to unseen conditions including higher magnification, different cell type, low-illumination, and defocused acquisitions. These results support quantitative analysis, cell-type discrimination, and drug-response assessment, establishing a scalable and cost-effective platform that paves the way toward compact, high-content label-free imaging solutions.

Rare cancers collectively account for approximately a quarter of diagnosed cancers. However, these tumors remain understudied due to research constraints, including underfunding, scarce samples, and reliance on resource-intensive mouse models. We propose a Zebrafish-First Triage Strategy to screen potential therapeutic targets before mammalian validation and accelerate research and drug development.