肿瘤领域文献阅读汇编

First-line (1L) bendamustine plus rituximab (BR) leads to high response rates in follicular lymphoma (FL), but maintaining durable remissions remains challenging. We report the 3-year follow-up from arm 3 of the phase 1b/2 EPCORE NHL-2 trial (NCT04663347) of epcoritamab, a subcutaneously administered CD3×CD20 bispecific antibody, combined with BR in patients with newly diagnosed FL. Twenty-five patients received epcoritamab plus BR, followed by epcoritamab monotherapy for up to 2 years. At a median follow-up of 41.3 months, the best overall response and CR rates were both 96%. The median time to CR was 1.5 (range 1-6) months. At 3 years, 87% of responders maintained CR. High CR rates were observed across subgroups, including 100% of patients with bulky disease (≥7 cm), 93% with Follicular Lymphoma International Prognostic Index score ≥3, and 100% with bone marrow involvement. The three-year progression-free survival and overall survival rates were 83% and 96%, respectively. Three patients progressed within 24 months of initiating treatment. Long-term data were consistent with prior reports and the known safety profiles of the individual agents, with no high-grade cytokine release syndrome or immune effector cell-associated neurotoxicity syndrome events. Infections occurred in 92% of patients; COVID-19 was the most common (84%). Overall, 1L FL treatment with epcoritamab plus BR resulted in deep, durable responses beyond 3 years with a consistent safety profile. These results compare favorably with BR alone, although they require confirmation in further studies, and highlight the versatility of epcoritamab in combination with various standards of care and in improving outcomes in FL.

Across cancer, one of the most frequent examples of histologic transformation is the evolution of follicular lymphoma (FL) to an aggressive large cell lymphoma. Despite recent progress, understanding of the molecular and cellular underpinnings of transformation remains incomplete. Here, we dissect the interplay of tumor and microenvironment cell populations across transformation through a multimodal investigation of 95 FL and transformed FL (tFL) samples, including single-cell and bulk RNA-sequencing alongside spatial transcriptomics and proteomics, and validate findings across independent FL-tFL pairs. Upon transformation, fibroblasts and GPNMB+ macrophages increase while lymph-node organizing follicular dendritic and CCL21+ fibroblastic reticular cells were lost, resulting in an altered spatial distribution of cytokines that impacts T cell infiltration and macrophage differentiation and function. Secreted stromal and macrophage signals were further evident by non-invasive plasma proteomics. Taken together, our data reveal expansion of macrophages and fibroblasts as key features of transformation with potential diagnostic and therapeutic implications.

A streamlined, sensitive, and universal molecular method for MRD detection in pediatric AML remains an unmet clinical need. Here, we propose a novel approach based on tracking somatic non-coding passenger variants. Using whole-genome sequencing (WGS) of diagnostic bone marrow DNA, we identify patient-specific somatic passenger variants, which we term "leukemia-specific passenger variants" (LSPVs). Single-cell DNA proteogenomic analyses demonstrate that LSPVs are specific and robust markers of leukemic cells, present across the entire leukemic cell population. Moreover, we show that LSPVs are accurate markers of disease burden, stable from diagnosis to relapse. Importantly, LSPVs can be detected in all patients, suggesting the universal applicability of this approach. Leveraging these findings, we developed DAISY-MRD, a streamlined WGS-based MRD method that does not rely on the presence of specific trackable genetic aberrations and does not require tailored assay design, making it simple, efficient, and feasible in clinical settings.

The rarer p190 (e1a2) transcript in chronic myeloid leukemia (CML) is associated with atypical presentations; yet, its biological basis remains poorly understood. Using a cohort of 60 patients including 42 chronic phase patients age-matched 1:1 with 42 e13a2/e14a2 patients in the chronic phase, we investigated the clinical, genomic, and clonal features of e1a2 BCR::ABL1 CML. We identified 60 e1a2 BCR::ABL1 CML patients showing distinctive hematologic features including lower leukocyte and platelet counts and higher monocytosis (12.3% vs. 2.0%, P < 0.001). Additional somatic mutations were detected in 37/42 (88%) e1a2 BCR::ABL1 cases compared with 7/42 (17%) e13a2/e14a2 BCR::ABL1 cases. The mutational spectrum was dominated by ASXL1 and TET2, and closely resembled CMML-like profiles. Genomic breakpoint sequences of 34 e1a2 BCR::ABL1 cases showed that BCR and ABL1 coordinates were similar to those observed in 394 B-ALL. Longitudinal mutational tracking revealed two distinct clonal architectures. In 71% of patients, mutations disappeared with molecular response, consistent with BCR::ABL1 as the founding event. In contrast, 29% of patients had mutations with stable VAFs, while BCR::ABL1 transcript levels decreased after treatment, indicating that the fusion had been acquired within a pre-existing mutated clone. Single-cell genotyping experiments confirmed these clonal architectures. These patients frequently developed cytopenias under tyrosine kinase inhibitor therapy and half required red blood cell transfusions, reflecting persistence of the ancestral clone rather than BCR::ABL1-driven disease. These findings show that e1a2 BCR::ABL1 CML frequently arises within complex, premutated clonal backgrounds, providing a biological basis for its atypical presentation and heterogeneous treatment response.

We explored the epigenomic effects of vegan diet (VD) versus meat-rich diet (MR) and identified mechanisms that help to explain how VD affects epigenetic gene regulation. Genome-wide DNA methylation profiles in 48 healthy individuals were investigated after a 1-month randomized isocaloric dietary intervention comparing effects of a VD versus a MR. Genome-wide DNA methylation analysis revealed changes in differentially methylated positions following dietary intervention, with the VD group showing a higher degree of gene-promoter silencing in cancer-related pathways and cell growth-associated pathways (mTOR and Hippo). Cell type deconvolution indicated an anti-inflammatory shift in the VD group, characterized by decreased neutrophils and increased CD4+ T-cell proportions, aligning with whole blood count data. Pathway analysis reflected changes in gene promoter methylation consistent with macronutrient changes according to the diets, in that VD silenced lipid metabolism and increased insulin pathway genes. Interestingly, the health-outcome-trained epigenetic clocks PhenoAge and GrimAge indicated a deceleration of biological aging in the VD group, while the chronologically optimized Blood&Skin clock showed an opposing acceleration. Convergent evidence from pathway analysis and health-outcome epigenetic clocks thus suggests that a 1-month VD drives epigenomic changes associated with reduced cancer risk and decelerated biological aging.

Accurate T-staging is crucial for guiding personalized treatment strategies for laryngopharyngeal cancer. However, current clinical practice relies on invasive biopsy procedures, whereas CT-based staging remains challenging due to the complex patterns of tumor invasion. Recent computer-aided approaches face two key challenges: 1) Structural relationship modeling: existing methods underrepresent anatomically structured patterns of tumor invasion, as they either process whole CT volumes without tumor-specific anatomical constraints or rely on labor-intensive tumor segmentation. 2) Fine-grained cross-modal alignment: while radiology reports contain organ-specific invasion details, current methods that apply global feature fusion struggle to accurately align individual anatomical structures with their corresponding textual descriptions. To address these issues, we propose an anatomy-aware multimodal framework that integrates organ-level CT context and radiology reports into a unified representation for laryngopharyngeal T-staging. The framework first constructs an Anatomy-Structured Organ Graph (AOG) that captures invasion patterns between primary sites and surrounding organs, then performs Organ-Anchored Cross-Modal Alignment (OCA) so that each organ node aggregates textual evidence from the radiology report, and finally refines this graph representation by injecting organ-specific invasion cues extracted from the report via Report-Enhanced Graph-Refinement (REG), yielding a multimodal organ graph that combines spatial and textual evidence. Extensive experiments demonstrate that the proposed framework achieves superior performance in T-staging of laryngopharyngeal cancer.

According to two phase I/II trials, ASCERTAIN-V and SAVE, the all-oral regimen of decitabine-cedazuridine and venetoclax, with revumenib added in the latter study, could be an effective option for patients with acute myeloid leukemia who would otherwise receive intravenous or subcutaneous chemotherapy.

In the phase I inMMyCAR study of the in vivo chimeric antigen receptor (CAR) T-cell therapy KLN-1010, all 18 patients with multiple myeloma responded. None of the evaluable patients had minimal residual disease 1 month after treatment, suggesting that the therapy could prove clinically useful.

Childhood cancer survivors have a heightened risk of developing subsequent neoplasms (SN) related to therapy. We analyzed whole-genome, exome, and RNA sequencing of 200 breast, meningioma, and thyroid SNs, which developed a median of 26.4 years after childhood cancer, among 160 survivors. Meningioma and thyroid SNs were enriched for driver gene rearrangements compared with de novo tumors, including NF2-disrupting alterations and kinase fusions potentially induced by radiation. Radiation correlated with increased insertion-deletion signature ID5. Nitrogen mustard treatment correlated with elevated "flat" signature SBS5 in breast and meningioma SNs; in vitro, these agents caused an unresolved flat signature associated with multiple flat signatures from the Catalogue of Somatic Mutations in Cancer. In meningioma, platinum therapy correlated with NF2 splice-site variants. Analysis of 19 multisample survivors revealed intrapatient heterogeneity in meningioma, including clonally independent tumors. These results demonstrate the long-term impact of childhood cancer treatment on the genomes of SNs developing in adulthood, which may guide SN treatment and prevention. This represents the most comprehensive genomic characterization of SNs from childhood cancer survivors to date, revealing the mutagenic impact of multiple therapies on the SN genome, including the potential impact of nitrogen mustards such as cyclophosphamide. These results may guide the optimization of future cancer treatment regimens to prevent SN development. See related commentary by Bertrums and van Boxtel, p. 1483.

Liquid biopsy has emerged as a transformative development in oncology, enabling the minimally invasive detection and monitoring of cancer through the analysis of tumor-derived material in blood. Moving beyond single-variable analysis, multifeature sequencing-based liquid biopsy (MSLB) integrates diverse classes of data from a single blood sample to provide deep multifactorial insight into tumor biology. In this review, MSLB is defined as the extraction of multiple biological signals from a single sequencing dataset and is put in the context of other layers of multimodal diagnostics. We focus on how recent advances in patient-, and potentially microbe-derived, cell-free nucleic acid analysis expand the biological information that can be extracted from a single blood sample. MSLB enables this by allowing the concurrent assessment of, for example, DNA methylation, copy number, fragmentation, and, in exploratory workflows, microbe-associated signals. This provides a broader view of tumor, immune, and microenvironment states. When combined with emerging bioinformatic and machine-learning frameworks, these complementary signals may improve early detection, disease monitoring, and treatment selection. Addressing challenges in standardization, validation, and regulatory alignment will be essential to determine how MSLB can be integrated into routine oncologic practice.

Nurses contribute across the precision cancer care pathway, providing continuous, person-centred care and supporting genomic testing integration. However, the scope of nursing competencies required for precision cancer care and the extent to which current education prepares nurses for these roles remain unclear. This scoping review maps and synthesises evidence on nurses' roles and competencies in precision cancer care, and educational provision in this field. A JBI scoping review was conducted; MEDLINE, CINAHL, PsycINFO, Embase and Scopus were systematically searched on July 18th 2025. International postgraduate and continuing professional education programmes relevant to nurses were identified via a generative artificial intelligence grey literature search undertaken between July 25th and July 30th 2025. Data were synthesised using convergent integrated thematic synthesis. Fifty-two publications were included, addressing nursing roles and/or competencies (n = 44) and educational interventions (n = 23). Sixty-five education programmes were identified. Five themes captured nursing roles: 1) genomic risk assessment and stratification; 2) genomic communication and shared decision-making; 3) precision cancer care pathway coordination and clinical navigation; 4) interprofessional collaboration and genomic service integration; and 5) professional governance, quality assurance, and advanced practice roles. Educational provision predominantly addressed foundational competencies, with less explicit preparation for roles requiring professional autonomy, interpretive expertise, leadership, and governance. Alignment between nurses' competency expectations and educational preparation is uneven. We propose a three-level model of precision cancer nursing practice to differentiate foundational capabilities from advanced practice roles, distinguishing precision-informed, enhanced, and specialist precision cancer nursing practice. Explicit mapping of postgraduate curricula to stratified competency levels may support safe expansion of nursing scope within mainstreamed genomic cancer care. This research was supported with funding from the Dublin City University Undergraduate Summer Research Internship 2025, which supported AC's time on the project. The authors alone are responsible for the content of this presentation.

Advanced pancreatic ductal adenocarcinoma (PDAC) often progresses rapidly during chemotherapy despite initial assessments of stable disease or partial response by Response Evaluation Criteria in Solid Tumors (RECIST 1.1), underscoring the limitations of the current methods for predicting short-term progressive disease (PD). To address this, the study developed a spatiotemporal deep learning framework that integrates convolutional and long short-term memory (LSTM) neural networks to dynamically predict PD at the next follow-up visit using serial computed tomography (CT) scans and baseline clinical variables. The model was trained on a retrospective cohort of 243 patients (415 predicted events, defined as temporal sequences for the next follow-up PD prediction) and evaluated across internal, external, and prospective cohorts. The model achieved area under the curve (AUC) values of 0.77, 0.76, and 0.74, respectively. Performance remained robust across chemotherapy regimens (AG or Gemcitabine-based, FOLFIRINOX, and SOXIRI; AUC 0.68-0.79), PD subtypes (target lesion growth vs. new metastases; AUC 0.72 vs. 0.77), and baseline disease stages (locally advanced vs. metastatic; AUC 0.85 vs. 0.71). This framework enables the noninvasive, real-time prediction of imminent PD in advanced PDAC, facilitating timely treatment modification. Its validated generalizability and reliance on routine clinical data underscore its potential for seamless integration into chemotherapy management.

Environmental pollutant mixtures are potential risk factors for metabolic dysfunction-associated steatotic liver disease (MASLD), yet their joint effects in complex co-exposure scenarios and toxicological mechanisms remain incompletely elucidated. This cross-sectional study quantified 54 urinary environmental pollutants via mass spectrometry among 1036 participants from the Yinchuan elderly cohort. Logistic regression and mixed-exposure models evaluated epidemiological associations and joint effects. Biological knowledge-driven machine learning algorithms extracted mechanistic features. Bayesian weighted quantile sum regression showed that neonicotinoid and metal(loid) mixtures were significantly associated with elevated MASLD odds (OR = 1.73 (1.37, 2.20) and OR = 1.51 (1.22, 1.84), respectively, per quartile increase), with acetamiprid and zinc as the leading contributors. Interaction analysis identified significant synergistic effects between thiamethoxam and lead, and between nitenpyram and selenium. Machine learning models incorporating biological knowledge-graph features retained ITGAM (integrin alpha-M, CD11b) and LIF (leukemia inhibitory factor) as key target proteins and GO:0006558 and GO:0032125 as core pathways related to inflammatory signaling, DNA damage, and apoptosis. An adverse outcome pathway framework was constructed for six pollutants that were significantly positively associated with MASLD in logistic regression and retained by machine learning models (thallium, 3-hydroxycarbofuran, acetamiprid, zinc, dinotefuran, and thiamethoxam), linking estrogen receptor agonism and NR1I3 (constitutive androstane receptor, CAR) suppression to downstream DNA damage, oxidative stress, and apoptosis as key mechanistic routes. This study provides an integrative approach connecting population-level mixture exposures with mechanistic hypotheses, offering a scientific basis for the health risk assessment of environmental chemicals. SYNOPSIS: Epidemiological models combined with biological knowledge-driven machine learning identified key pollutant-protein-pathway associations, and an adverse outcome pathway framework was constructed to link these findings to mechanistic hypotheses.

Residual disease after cytoreductive surgery is a dominant, surgeon-modifiable determinant of outcome in advanced ovarian cancer, yet white-light inspection cannot reliably identify microscopic implants, plaque-like deposits, or therapy-altered fibrotic foci across complex peritoneal surfaces. Intraoperative molecular imaging aims to close this visibility gap by translating tumor-associated biology into real-time signal that can prompt additional resection, direct sampling, and support intraoperative decision-making. Across probe classes, folate receptor-α-targeted agents represent the most clinically mature approach, while activatable tracers and multimodal platforms are expanding capabilities beyond superficial visualization. However, improved detection alone does not establish patient benefit. We therefore frame molecular guidance as a workflow intervention and organize evidence along a clinically oriented hierarchy-lesion detection, decision impact, and patient outcomes-highlighting the need for standardized acquisition/quantification and pathology-linked validation. We propose pragmatic, trial-compatible definitions for pathology-confirmed completeness and prespecified molecular field clearance, and discuss how AI-assisted interpretation, multimodality, and radionuclide-enabled strategies could strengthen reproducibility and extend management toward microscopic residual disease.

Clonal hematopoiesis of indeterminate potential (CHIP) describes the proliferation of blood cell clones that carry driver mutations, such as DNA methyltransferase 3 alpha (DNMT3A), ten-eleven translocation 2 (TET2), additional sex combs like 1 (ASXL1), and Janus kinase 2 (JAK2), without leading to any obvious malignancy. Its occurrence rate is age-related and associated with cardiovascular and immune aging. In prospective cohort studies, CHIP substantially elevated the risk of myocardial infarction, stroke, and heart failure, with a relative risk increase ranging from 1.4 to 2.0-fold. Furthermore, a larger clone size, as indicated by the variant allele fraction (VAF), is correlated with a heightened risk. Both experimental and human evidence support the association of proinflammatory myeloid reprogramming with the loss of ten-eleven translocation 2 (TET2)/DNA methyltransferase 3 alpha (DNMT3A), NOD-like receptor family pyrin domain-containing 3 (NLRP3), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) signaling, as well as endothelial dysfunction and atherothrombosis. However, the effect sizes vary depending on the specific gene, tissue, and context, and residual confounding remains a concern. This paper reviews the epidemiology and pathophysiology of the disease, its clinical utility, including indications for testing, interpretation of variants, VAF levels, and the added value to conventional risk models and treatment methods, such as cytokine- and mutation-based approaches. Priority areas include genotype- and VAF-stratified trials that focus on outcomes relevant to aging and are rigorously monitored for their safety. Overall, CHIP is associated with somatic mosaicism, inflammaging, and aging of the cardiovascular and immune systems, although significant questions regarding causation and actionability remain unresolved.

Current imaging techniques inadequately delineate surgical margins of well-differentiated liposarcoma (WDLS), increasing risks of incomplete resection or overtreatment. MDM2, a p53-inhibiting oncoprotein amplified in WDLS, represents a dual diagnostic and therapeutic target. Therefore, we developed two radionuclide probes utilizing the MDM2-targeting stapled peptide with high binding affinity to evaluate their suitability for tumor-specific diagnostic purposes, which named [68Ga]Ga-DOTA-7041 and [68Ga]Ga-DOTA-P53-8 to conduct a head-to-head comparative study. Among these, [68Ga]Ga-DOTA-7041 showed superior MDM2-binding affinity (6.7 nM). In vivo imagery in ccRCC tumor-bearing PDX models showed significant and rapid uptake for both radiotracers, with specific radioactive accumulation beginning at 10 min and persisting over time. In contrast, the SW872 liposarcoma mouse model demonstrated slower and lower uptake, visible at 4 h in PET/CT scans. The further Clinical PET/CT in a WDLS patient using the superior probe [68Ga]Ga-DOTA-7041 showed heterogeneous tumor uptake, correlating with postoperative MDM2 immunohistochemistry. In this study, [68Ga]Ga-DOTA-7041 demonstrated diagnostic potential in both animal models and patients with liposarcoma as first-in-human PET tracer targeting MDM2. The disparate tumor uptake observed, compared to the high uptake in renal cancer models, may provide valuable insights into the challenges of targeting MDM2/MDMX with therapeutic agents in liposarcoma treatments.

Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) can bioaccumulate in aquatic food webs and pose potential health risks through aquatic product consumption. This study investigated the occurrence, health risks, and dietary management implications of PCBs and PBDEs in 12 commonly consumed freshwater and marine species from Guangdong, China. Concentrations and congener profiles were determined, followed by population-specific carcinogenic and noncarcinogenic risk assessment. A linear programming approach was applied to develop optimized dietary recommendations by integrating nutritional requirements, contaminant exposure, and economic cost. All samples contained detectable levels of PCBs and PBDEs, with higher concentrations in freshwater species than in marine species. PCBs were dominated by tetra- to heptachlorinated congeners (79.2%-99.97% of ΣPCBs), while PBDEs were mainly composed of tri- to penta-brominated congeners and BDE-209 (70.0%-99.9% of ΣPBDEs). Risk assessment showed that regular consumption of several species exceeded the cancer risk (CR) benchmark of 1 × 10-6 (USEPA), primarily driven by dioxin-like polychlorinated biphenyls (dl-PCBs). The optimization framework provides tailored guidance for different groups in Guangdong, balancing protein intake, cost, and contaminant exposure. Low-risk, protein-rich, and affordable options included Scatophagus argus, Turritella bacillum, and Procambarus clarkii. Tailored consumption advice can reduce exposure without compromising nutrition.

Accurate molecular classification of medulloblastoma is critical for prognosis and treatment planning, but current methods rely on surgical tissue sampling and molecular profiling. This study evaluated whether in vivo proton MR spectroscopy (¹H-MRS) can provide noninvasive metabolic markers to support presurgical molecular group stratification. In this single-center retrospective study, pretreatment ¹H-MRS data were analyzed from 95 pediatric patients with medulloblastoma (median age 7.4 years; 56 male). Single-voxel point-resolved spectroscopy (TE = 35 ms, TR = 1.5-2.0 s) were acquired during routine clinical MRI, adding approximately 5 minutes of scan time. Absolute metabolite concentrations and selected ratios were quantified using automated spectral fitting. Metabolic profiles were compared across molecular groups (group 3, n = 22; group 4, n = 35; sonic hedgehog [SHH], n = 26; wingless [WNT], n = 12) and assessed for qualitative concordance with prior ex vivo high-resolution HR-MAS NMR findings. Group differences were tested using Kruskal-Wallis with Dunn post hoc correction. Significant metabolic differences were observed across molecular groups, with strong group effects for taurine, creatine, choline, glutamate, and γ-aminobutyric acid (GABA) (all P < .0015). Taurine was elevated in group 3 and group 4 relative to SHH (log₂FC = 1.77 and 1.40, adjusted P < 4 × 10⁻⁵). SHH tumors exhibited lower creatine compared with group 3, group 4, and WNT (adjusted P < .05). Glutamate was higher in SHH than in WNT, while WNT tumors showed increased choline and GABA relative to other groups (adjusted P < .05). In vivo patterns were qualitatively concordant with ex vivo NMR findings. In vivo ¹H-MRS is a widely available, clinically feasible imaging biomarker that complements existing diagnostics and supports presurgical stratification of medulloblastoma.

Automatic segmentation of gliomas on amino acid PET is essential for quantitative tumor assessment, a pillar in monitoring gliomas under treatment. This study aimed to develop a deep learning model for the automated extraction of PET RANO criteria from [18F]FDOPA PET, with external validation. A total of 635 static [18F]FDOPA PET scans from 3 European centers were retrospectively included for glioma diagnosis, recurrence assessment, or treatment monitoring. The training cohort comprised 530 scans from Nancy Hospital, with external validation and test sets from Pitié-Salpêtrière Hospital (n = 66) and Turin Hospital (n = 39). Ground-truth segmentations followed international guidelines. A 3D U-Net was trained to segment tumor and healthy brain volumes. Performance was evaluated using the Dice coefficient using the whole tumor volume. Quantitative agreement for PET RANO criteria 1.0 parameters, tumor-to-background ratios (TBRmean, TBRmax) and metabolic tumor volume (MTV), was assessed at the lesion level. Tumor segmentation achieved Dice of 0.925 (0.841; 0.970) in training, 0.885 (0.829; 0.925) in validation, and 0.851 (0.733; 0.911) in the test set. At lesion level, agreement with expert quantification was high, with low bias and strong reliability for MTV (2.293 [-4.734; 9.321] mL), TBRmax (0.056 [-0.189; 0.301]), and TBRmean (-0.139 [-0.424; 0.146]) and intraclass correlation coefficients superior to 0.93. Measurable lesions were correctly identified in more than 97% of cases. Our [18F]FDOPA PET deep learning model (available at https://github.com/IADI-Nancy/FDOPA-PET-GliomaSeg) demonstrates robust multicenter performance and enables fully automated, reproducible quantification, supporting broader clinical adoption of amino acid PET in neuro-oncology.

We evaluated the risk of malignancy following herpes zoster ophthalmicus (HZO) and ophthalmic herpes simplex virus (HSV) in a large, diverse United States cohort. Matched, retrospective cohort study. Adults enrolled in the National Institutes of Health All of Us Research Program. Participants with incident diagnoses of HZO or ophthalmic HSV were propensity score-matched (1:3) to controls based on sociodemographic characteristics and preexisting immune dysregulation (i.e., autoimmune disease or immunodeficient states). Stratified Cox proportional hazards models were applied to estimate relative hazards of incident malignancy within 1-, 2-, and 3-year intervals and over the entire follow-up period. Effect modification was investigated based on age, sex, and baseline immune dysregulation. The HZO cohort comprised 327 patients matched to 981 control participants (mean follow-up, 7.48 ± 5.33 years), and the ophthalmic HSV cohort included 292 patients matched to 876 control participants (mean follow-up, 8.66 ± 5.89 years). Incident malignancy risk did not differ between patients with HZO and matched control participants within 1 year or less (P = 1.00), 2 years or less (P = 0.36), 3 years or less (P = 0.27), or across the full follow-up period (hazard ratio [HR], 1.01; 95% confidence interval [CI], 0.76-1.36; P = 0.93). However, significant effect modification by immune dysregulation was observed, with HZO associated with an increased risk of malignancy among participants with autoimmune disease (HR, 2.91; 95% CI, 1.48-5.74; P < 0.01) or immunodeficient status (HR, 5.75; 95% CI, 2.16-15.35; P < 0.01). For ophthalmic HSV, no increased malignancy risk was observed within 1 year or less (P = 0.86), 2 years or less (P = 0.18), 3 years or less (P = 0.10), or across the full follow-up period (HR, 0.97; 95% CI, 0.72-1.32; P = 0.87), with no evidence of effect modification. Ophthalmic herpes infections were not associated with an increased risk of malignancy in the overall cohort. However, HZO was associated with a higher subsequent risk of cancer among individuals with preexisting immune dysregulation, potentially warranting heightened oncologic vigilance in this patient demographic. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Kidney injury molecule 1 (KIM-1) is a promising biomarker in adjuvant clear cell renal cell carcinoma (ccRCC), but its relevance in advanced ccRCC or papillary RCC (pRCC) remains unclear. CALYPSO (NCT02819596) was a prospective, multi-arm trial that evaluated durvalumab alone or in combination with tremelimumab or savolitinib in metastatic ccRCC and pRCC. Circulating KIM-1 levels were measured at baseline and on-treatment. The primary endpoint was to explore if KIM-1 levels were raised in pRCC. Analyses were exploratory and p values were nominal. KIM-1 was measured in 123 patients with ccRCC and 31 patients with pRCC. Higher median concentrations occurred in pRCC compared to ccRCC (7835 vs 5470 pg/ml, p = 0.05). Reductions in KIM-1 levels occurred with systemic therapy in both ccRCC and pRCC (-59.2% and -32% respectively). In pRCC, radiological responders had significantly lower baseline KIM-1 levels (p = 0.025). In ccRCC, high baseline KIM-1 levels were associated with significantly shorter overall survival (OS) (hazard ratio [HR] 1.77; 95% CI, 1.15-2.72; p = 0.01). Also, an increase in KIM-1 during therapy was linked to worse progression-free survival (HR 1.7; 95% CI, 1.13-2.58; p = 0.01) and OS (HR 1.95; 95% CI, 1.23-3.08; p = 0.004) in ccRCC. This exploratory analysis supports the utility of KIM-1 in advanced ccRCC and pRCC.

Endoscopic ultrasound-guided gallbladder drainage (EUS-GBD) is usually contraindicated in perforated cholecystitis, although initial reports suggest feasibility in contained perforation. This prospective single-center cohort study aimed to explore EUS-GBD outcomes in patients with acute cholecystitis with contained perforation (cp-AC; Niemeier classification type 2). A secondary exploratory comparison was performed with a contemporaneous cohort of patients with intact gallbladder walls. Between January 2021 and March 2025, 23 patients with cp-AC (39.7% of the overall EUS-GBD cohort) were enrolled. This subgroup showed high prevalence of underlying malignancy (87.0%) and previous endoscopic retrograde cholangiopancreatography with placement of metal stents (65.2%). Median operative space and interluminal distance were 30.5 mm (interquartile range [IQR] 26.0-37.0) and 5 mm (IQR 2-8), respectively. Technical and clinical success of EUS-GBD were 100% (95%CI 85.7-100) and 87.0% (95%CI 67.9-95.5), respectively. The adverse event rate was 21.7% (95%CI 9.7-41.9) and did not differ significantly from that in the cohort with intact gallbladder walls; however, two fatal events occurred in the cp-AC group. EUS-GBD appeared to be technically feasible in patients with contained perforation, despite the additional complexity. However, this represents a particularly high-risk population, and the procedure may be considered only for carefully selected patients in expert centers.

The distinction between benign and neoplastic bile duct strictures remains challenging. Pathologic assessment of endoscopic retrograde cholangiopancreatography (ERCP)-obtained specimens has limited sensitivity, particularly among patients with primary sclerosing cholangitis (PSC). Next-generation sequencing of bile duct specimens provides a promising diagnostic approach, but a prospective, multi-institutional, and comprehensive DNA/RNA analysis is lacking. A 6-year, prospective, multi-institutional study was conducted using BiliSeq version 2 (28 cancer-associated genes and 167 fusion genes) and BiliSeq version 3 (161 cancer-associated genes and 763 fusion genes) for 2908 ERCP-obtained brushings, biopsies, and bile from 2116 patients at 28 medical institutions. Molecular results were compared with clinical, imaging, and pathologic parameters including diagnostic pathology and/or at least 1-year follow-up. BiliSeqV2/V3 testing was performed for 2865 (99%) specimens from 2080 (98%) patients. Based on follow-up from 1979 (95%) patients, BiliSeq version 2/version 3 demonstrated 82% sensitivity and 98% specificity for a neoplastic stricture. In comparison, pathologic assessment had a sensitivity of 44% and a specificity of 99%. Combining BiliSeq version 2/version 3 testing with pathologic assessment improved the sensitivity to 88% and maintained a high specificity of 97%. High-risk populations, such as Hispanic, germline carrier, and PSC patients, also showed improvement in sensitivity with BiliSeq version 2/version 3 (74% to 86%) compared with pathologic assessment (26% to 50%). Further, actionable molecular alterations were identified in 20% of BiliSeq version 3-positive neoplasms and modified patient management in 30% of these cases. Applying BiliSeq version 2/version V3 testing to ERCP-obtained specimens improved the diagnostic evaluation of bile duct strictures, achieving higher sensitivity, especially for PSC, and maintained high specificity compared with traditional methods. This study highlights the importance of next-generation sequencing for precise diagnosis and therapeutic intervention.

Helicobacter pylori infection is the principal cause of peptic ulcer disease, mucosa-associated lymphoid tissue lymphoma, and noncardia gastric cancer. The major advances in diagnostics and treatment and eradication success are threatened by rising antimicrobial resistance and concerns about disruption of the normal microbiome. This review summarizes current unresolved issues in H pylori treatment, with a focus on resistance, optimal regimens, ecological impact, and emerging therapies. Extensive review of randomized controlled trials, meta-analyses, international consensus guidelines, molecular epidemiology studies, microbiome analyses, and translational research related to H pylori eradication, resistance, microbiome effects, and novel therapies. Global resistance to clarithromycin, metronidazole, and fluoroquinolones significantly undermines the performance of traditionally effective antimicrobial therapies. The corner stone of successful antimicrobial therapy is susceptibility-guided therapy but remains of limited use with H pylori because of lack of infrastructure. Eradication regimens potentially induce substantial alterations in gut and gastric microbiota and expand the antimicrobial resistome. Probiotics and N-acetylcysteine offer at best very modest improvements in eradication and tolerability. Novel and still experimental platforms, including engineered phage therapy, antimicrobial peptides, nanoparticle-delivered urease inhibitors, biofilm-targeting agents, and vaccines show promise. Implementation gaps, persistent use of suboptimal regimens, and global inequities constrain the impact of available therapies. Optimizing H pylori treatment requires evidence-based regimen selection, precision-guided strategies, antimicrobial stewardship, and equitable access to essential medications. Advances in molecular diagnostics, antimicrobial development, and implementation science are all critical to reducing the global burden of H pylori-associated disease and gastric cancer.

IDH-mutant astrocytomas are classified as WHO grade 4 in the presence of conventional high-grade histologic features and/or homozygous CDKN2A/B deletion in the 5th edition of the WHO Classification of Central Nervous System Tumour guidelines. However, work over the past decade has indicated a number of other molecular alterations that warrant consideration as potential prognostic markers. We used univariate Kaplan-Meier and multivariate Cox proportional hazards regression analysis to evaluate the prognostic effects of homozygous CDKN2A/B deletion, CDK4 amplification, CCND2 amplification, PDGFRA amplification/mutation, PIK3R1 mutation, PIK3CA mutation, MYCN amplification, EGFR amplification/mutation, TERT promoter mutation, and grade 4 histologic features in two independent cohorts of WHO grade 2-4 IDH-mutant astrocytoma (n = 840 and n = 367). The presence of CDK4 amplification, CCND2 amplification, PDGFRA alteration, PIK3R1 mutation, MYCN amplification, and EGFR alteration were each associated with reduced overall survival compared to WHO grade 2/3 astrocytomas without these molecular features. 17.7% (148/837) of otherwise grade 2/3 astrocytomas had one or more of these molecular criteria, with resulting intermediate clinical outcome in terms of overall survival (median survival of 67.3-82.0 months) compared to grade 2/3 astrocytomas without these molecular features (median survival of 135.0-140.7 months) and grade 4 astrocytomas (median survival of 35.3-45.0 months). The presence of CDK4, CCND2, PDGFRA, PIK3R1, MYCN, and EGFR alterations result in an intermediate patient survival in IDH-mutant astrocytoma. Adding these molecular alterations should be considered in future diagnostic classification systems to improve stratification of high-risk patients.

Novel immunotherapies and targeted therapies have transformed melanoma treatment, but the geographic distribution of benefits remains unclear. To quantify county-level variation in melanoma mortality declines following the introduction of novel therapies and identify associated patient and health system factors. We conducted a repeated cross-sectional ecological study using county-level mortality data from the Surveillance, Epidemiology, and End Results Program. The primary outcome was percent change in age-adjusted melanoma mortality between 2008-2013 and 2019-2023. County characteristics were obtained from the Area Health Resource File, American Hospital Association Survey, Medicare claims, and National Provider Identifier database. Associations were assessed using ordinary least squares regressions. National melanoma mortality declined 25.6% between 2008-2013 and 2019-2023. Across 751 counties, declines varied widely (median, 23.8%; interquartile range, 7.6% to 35.6%). County population size was the strongest predictor of mortality decline (β = 4.5; 95% confidence interval, 1.6-7.4; P = .002). Had all counties achieved the mortality reductions of the largest counties, 2818 additional deaths could have been prevented. Ecological design and reliance on aggregates limit causal inference and individual-level interpretation. Population size strongly predicts mortality benefits from novel melanoma therapies, underscoring the importance of strengthening health care infrastructure to reduce geographic disparities.

Mycosis fungoides is the most common cutaneous T-cell lymphoma in the pediatric population. We aimed to characterize the long-term outcomes associated with juvenile-onset mycosis fungoides (jMF). Patients were identified from a retrospective database. Clinical and histopathologic data were obtained from medical records, and survival was assessed through contact by telemedicine. Stage progression was defined as progression to a more advanced tumor, node, metastasis, and blood stage or death from disease. From 1987 to 2023, 118 patients were diagnosed with jMF: 97% of patients (115/118) had early-stage (IA-IIA) disease at diagnosis and 3% (3/118) had advanced-stage (IIB-IVB) disease. Thirteen percent of patients (15/118 patients) developed disease progression with a median time to progression of 2 years (range: 2 months to 13 years). Median follow-up time for survival assessment was 7.2 years (range: 2 months to 32 years). Overall, 5-year and 10-year survival rates were 99% and 98%, respectively, and 5-year and 10-year disease-specific survival rates were 100%. No patients died of disease. This is a single-center retrospective study. jMF often presents with early-stage disease and has an excellent prognosis with a 5-year and 10-year overall survival of 99% and of 98%, respectively. Disease progression is uncommon and did not lead to disease-specific death in our cohort.

There are barriers to the clinical use of cisplatin-based neoadjuvant regimens in muscle-invasive bladder cancer (MIBC), especially for patients with renal dysfunction, hearing loss, heart diseases, or other severe comorbidities. Favorable efficacy and safety of disitamab vedotin (DV), an antibody-drug conjugate targeting HER2, have recently been demonstrated in urothelial carcinoma. We explored neoadjuvant gemcitabine combined with RC48 in the MIBC setting. Initially, 26 patients with clinical stage T2-4a Nx M0 MIBC and a HER2 immunohistochemical expression score of 2+/3+ were enrolled. The neoadjuvant regime comprised DV (2 mg/kg intravenous infusion, day 1) and gemcitabine (1000 mg/m2 intravenous infusion, day 2) in a 14-d cycle. Thirteen patients received three cycles, and the other 13 patients received four cycles, in a nonrandomized manner. Subsequently, 22 patients underwent radical cystectomy (RC). Postoperative pathology revealed a pathological complete response (pCR) rate of 59% (13/22) among RC patients, 40% (4/10) of whom had received three cycles and 75% (9/12), four cycles. Intention-to-treat analysis revealed pCR rates of 50% (13/26) overall, 31% (4/13) for the three-cycle group, and 69% (9/13) for the four-cycle group. At median follow-up of 16.9 mo, 25 patients remained event-free. Treatment-related adverse events primarily included aspartate aminotransferase elevation (35%), alanine aminotransferase elevation (38%), a decrease in appetite (23%), alopecia (23%), and sensory neuropathy (23%). Validation of the long-term efficacy of this regimen is required.

FLT3 internal tandem duplications (FLT3-ITD) are major genetic events in acute myeloid leukemia (AML). Although the clinical impact of FLT3-ITD "macroclones" (allelic ratio [AR] ≥0.05) is well established, the significance of low-level FLT3-ITD subclones ("microclones") remains uncertain. We conducted a post hoc analysis of 1733 patients with newly diagnosed AML enrolled in the Backbone Intergroup 1 trial (ClinicalTrials.gov identifier: NCT02416388). Using next-generation sequencing (NGS), we detected FLT3-ITD microclones (AR between 0.0004 and 0.05) in 17.4% of patients without FLT3-ITD macroclones. Microclones and macroclones (low and high AR) were independently associated with increased relapse risk (cause-specific hazard ratio, 1.50 [95% confidence interval (CI), 1.18-1.91]; 1.98 [1.50-2.62]; and 2.33 [1.69-3.22], respectively) after adjustment for age, white blood cell count, other gene mutations, midostaurin treatment, and allogeneic hematopoietic stem cell transplantation. At 2 years, the cumulative incidence of relapse reached 42.5% (95% CI, 37.0-47.9) in patients with macroclones, 45.1% (38.3-51.6) in patients with microclones, and 29.4% (26.6-32.3) in patients without FLT3-ITD. In NPM1-mutated AML, both microclones and macroclones were associated with higher levels of measurable residual disease (MRD) and increased relapse risk, without independent impact on overall survival after adjustment for MRD. An analysis of paired samples further revealed that 41.8% of relapses in patients with FLT3-ITD microclones at diagnosis were associated with a macroclone at relapse. These findings challenge current risk stratification models and support the integration of NGS-based FLT3-ITD detection into the diagnostic and prognostic workflow for AML. Prospective trials addressing the management of patients with FLT3-ITD microclones are warranted, as is their consideration in future European LeukemiaNet guidelines.

Relapse after remission remains the primary cause of treatment failure in acute myeloid leukemia (AML), underscoring the need for strategies to eliminate residual leukemic cells. The bone marrow (BM) microenvironment, largely orchestrated by the CXC chemokine receptor 4 (CXCR4)-CXC motif chemokine 12 axis (CXCL12), enables leukemia cell survival and chemoresistance by anchoring blasts in their protective BM niche. Motixafortide, a selective CXCR4 antagonist, mobilizes leukemic cells and disrupts tumor microenvironment interactions in preclinical models. In this randomized, double-blind, placebo-controlled phase 2 trial, 128 patients in first remission received high-dose cytarabine plus motixafortide or placebo. Median relapse-free survival did not substantially differ between groups: 10.3 months (95% confidence interval [CI], 8.0-12.0) for motixafortide and 11.5 months (95% CI, 8.6-24.1) for placebo (log-rank P = .98). But single-cell measurable residual disease (scMRD) analysis, performed before consolidation, demonstrated heterogeneity of CXCR4 inhibition benefit; in the placebo group, higher CXCR4 expression was associated with increased relapse risk (P = .02), whereas in the motixafortide group, higher CXCR4 expression was linked to a reduced relapse rate (P = .047). Exploratory analyses identified scMRD levels at which higher MRD burden was associated with inferior overall survival. Taken together, combining functional MRD profiling with biomarker-driven patient selection, such as CXCR4 expression, may enable more precise and effective postremission interventions in AML. This trial was registered at www.clinicaltrials.gov as NCT02502968 and at EudraCT as 2014-002702-21.

Conventional receptor-targeted fluorescent probes have shown promise in tumor imaging, yet achieving a high tumor-to-normal (T/N) tissue ratio in vivo remains challenging due to limited biomarker density on tumor cell membranes. Here, we present an in situ assembly strategy of bioorthogonal-functionalized chimeric artificial receptors (BCARs) that locally constructs BCARs on tumor surfaces, which amplify fluorescence signals and enable high-contrast imaging. Rapid, selective membrane engineering under physiological conditions increases effective receptor density, enhancing fluorophore binding and tumor visualization. Mechanistic studies reveal that BCARs exhibit exceptional membrane retention and spatial precision, sustaining signal amplification in heterogeneous tumor microenvironments. In air-pouch and orthotopic bladder cancer models, BCARs notably improve the T/N imaging ratio and tumor boundary delineation. Translational validation with surgical specimens from 14 patients with bladder cancer confirms clinical feasibility. This work establishes a versatile platform for on-site receptor reprogramming and signal amplification, offering a powerful tool for high-contrast tumor margin detection.

Chronic stress is increasingly recognized as a driver of cancer metabolism, highlighting the need to elucidate the mechanism linking chronic stress to metabolic reprogramming. Here, we performed untargeted metabolomics on serum samples from esophageal squamous cell carcinoma (ESCC) patients and integrated the results with clinical and stress-related assessments, revealing pentose phosphate pathway (PPP) enrichment accompanied by elevated epinephrine levels in patients with stress-associated features. β2-adrenergic receptor (ADRB2) activated by chronic stress stabilized MYCBP by competitively displacing VHL, thereby enhancing MYC transcriptional activity and upregulating key PPP enzymes, including G6PD and TKT. Unexpectedly, stress-associated stimulation not only activated ADRB2 signaling but also increased ADRB2 protein abundance. Mechanistically, stress exposure enhanced RNMT-dependent N7-methylguanosine (m7G) cap modification of ADRB2 mRNA, thereby increasing its translational efficiency. Together, these findings define an RNMT-ADRB2-MYCBP-PPP axis that integrates epitranscriptomic regulation with adrenergic signaling to promote metabolic reprogramming and malignant progression in ESCC. The stress-associated RNA modification-metabolism circuitry comprises potential therapeutic targets to overcome stress-associated ESCC progression.

Ubiquitin-specific protease 7 (USP7) is a deubiquitinase that plays critical regulatory roles in multiple signaling pathways by preventing the ubiquitin-mediated degradation of its substrates. Dysregulated expression of USP7 is implicated in tumor progression; however, its role in renal fibrosis remains unclear. In the present study, USP7 was observed to be significantly upregulated in the kidneys of patients with chronic kidney disease (CKD), which correlated with fibrotic lesions and renal dysfunction. Both genetic depletion and pharmacological blockade of USP7 significantly attenuated fibroblast activation and extracellular matrix deposition in two mouse models of kidney fibrosis-unilateral ureteral obstruction and unilateral renal ischemia-reperfusion injury models, indicating a pro-fibrotic function of USP7. Mechanistically, intergrated proteomic sequencing and phosphoproteomic sequencing revealed that USP7 modulated the tuberous sclerosis complex 1 (TSC1)-mTOR pathway. USP7 knockdown restored TSC1 expression and inhibited mTOR activation. However, USP7 did not directly interact with TSC1; instead, it deubiquitinated and stabilized lysine-specific demethylase 5B (KDM5B), which subsequently reduced histone H3K4me3 modification at the Tsc1 promoter to repress its transcription. Conversely, the inhibition of USP7 promoted KDM5B degradation, thereby restoring TSC1 expression and suppressing mTOR-driven fibrogenesis. Thus, these findings identify USP7 as a critical promoter of renal fibrosis, acting at least in part through the KDM5B-TSC1-mTOR axis. This highlights USP7 as a potential therapeutic target for CKD.

Metastasis remains a leading cause of mortality in esophageal squamous cell carcinoma (ESCC) patients, underscoring the urgent need to elucidate the molecular mechanisms driving disease progression. In this study, we delineated that ZBED2, a zinc finger protein, correlates with inferior survival outcomes and metastasis in ESCC patients. ZBED2 formed phase-separated nuclear condensates that functionally promote tumor metastasis, and integrin-linked kinase (ILK) was pinpointed as a critical downstream effector in mediating the pro-metastatic function of ZBED2. Mechanistically, ZBED2 enhanced the transcription of HSP90AA1, promoted the physical interaction between HSP90AA1 and ILK, and consequently stabilized ILK by suppressing its ubiquitination. Subsequently, ILK promoted PD-L1 transcription and CD8+ T cell exhaustion, thereby creating an immunosuppressive microenvironment that facilitates cancer metastasis. Collectively, these findings establish the pivotal role of ZBED2 in driving ESCC metastasis and immune evasion, thereby validating it as a promising therapeutic target for this aggressive malignancy.

Manganese-ion-based nanoparticles have emerged as highly versatile platforms that seamlessly integrate diagnostic and therapeutic functions, enabling precision cancer theranostics. In this review, we discuss manganese-based nanomaterials through a design-oriented framework that connects synthesis, structure, function, application, and limitation. Mainstream preparation strategies, including thermal decomposition, permanganate reduction, hydrothermal/solvothermal synthesis, template-assisted construction, coprecipitation, and mild biomimetic routes, are compared with respect to physicochemical control, Mn activation behavior, scalability, and biomedical suitability. The diagnostic applications of manganese-based nanoplatforms are discussed, including activatable T1-weighted magnetic resonance imaging (MRI) and multimodal imaging systems, as well as Mn-activated DNAzyme platforms for gene regulation. We further review therapeutic modalities enabled by Mn2 +, encompassing chemodynamic therapy (CDT), nanozyme-mediated redox regulation within the tumor microenvironment, stimulus-responsive drug and prodrug delivery, magnetic hyperthermia, and their synergistic combinations. Finally, we critically analyze key biosafety considerations along with current translational challenges and future perspectives for developing clinically viable, image-guided combination cancer therapies based on manganese-ion nanoplatforms.

Cancer and inflammatory diseases are critically influenced by dynamic interactions between pathological tissues and the host immune system. The precise migration of immune cells into the local microenvironments of tumors or inflammation is a fundamental prerequisite for them to exert their functions. Immune reservoirs, including tertiary lymphoid structures, secondary lymphoid structures, bone marrow and the intestinal tract, serve as critical mobilization hubs for diverse lymphoid and myeloid populations to infiltrate tumors or inflamed sites. The directional migration of immune cells is orchestrated through complex regulatory networks involving chemokine or cytokine-receptor pairs, adhesion molecule interactions, extracellular vesicle signaling, metabolic reprogramming and microbiota modulation. In both tumors and inflammation, immune cell trafficking shapes the local immune landscape, contributing to either immune protection or pathological progression. Contemporary therapeutic strategies targeting immune cell migration encompass the following axes: precision modulation of chemokine or cytokine networks, architectural reprogramming of lymphatic structures or extracellular matrix, dietary intervention and strategic manipulation of microbiome. Nevertheless, clinical translation remains hindered by microenvironmental heterogeneity, suboptimal migratory efficiency, and technical limitations in longitudinal tracking of cellular dynamics. This review integrates recent findings from oncology and inflammatory diseases to explore the origins, phenotypes and trafficking mechanisms of migratory immune cells, highlighting how advances in understanding immune migration across cancer and inflammation can inform therapeutic innovation and precision immunomodulation.

Cancers containing histologically distinct cell populations could arise from multiple transforming events, from transdifferentiation late in tumor evolution, or from a single progenitor cell. Poulain and colleagues show that glandular and squamous cells in adenosquamous carcinomas caused by human papillomavirus infection are descendants of a common ancestor.

Liver metastasis is the leading cause of death in pancreatic ductal adenocarcinoma (PDAC), yet its molecular drivers remain poorly defined. Through integrated transcriptomic and proteomic screening of clinical specimens, we identify the membrane protein Cadherin-Related Family Member 2 (CDHR2) as a key orchestrator of hepatic colonization in PDAC. CDHR2 was upregulated in primary tumors versus adjacent tissues, further elevated in liver metastasis versus matched primary tumors, and markedly enriched in patient-derived plasma extracellular vesicles (EVs), peaking in patients with liver metastasis. Knockdown of CDHR2 in human PDAC cells suppressed proliferation, migration, and invasion, and in orthotopic models attenuated primary tumor growth, remodeled the tumor microenvironment, and reduced liver metastatic burden. Mechanistically, CDHR2 associated with PDK1 and promoted activation of the PDK1-AKT (Thr308) axis to drive proliferation. In turn, AKT activity correlated with the packaging of CDHR2 into EVs, coupling intrinsic signaling to EV cargo loading. Tumor-derived, CDHR2-enriched EVs formed a complex with Aminopeptidase N (ANPEP) on liver sinusoidal endothelial cells (LSECs), disrupting endothelial junctions and enhancing tumor-cell adhesion to enable transendothelial migration. Consistently, ANPEP inhibition or liver-specific knockdown curtailed hepatic colonization, whereas priming with CDHR2+ EVs accelerated it. Finally, systemic delivery of EV-encapsulated siCDHR2 suppressed established primary tumors and markedly reduced liver metastasis. Collectively, our findings define a tumor-intrinsic CDHR2-PDK1-AKT (Thr308) axis coupled to an exosomal CDHR2-ANPEP endothelial-priming program and provide proof-of-concept for exosomal siCDHR2 as a candidate strategy against PDAC liver metastasis.

Acute myeloid leukemia (AML) is an aggressive blood cancer with a 5-year overall survival rate of ~30%. Although immunotherapies engaging T cells demonstrate remarkable success in treating many solid tumors and blood cancers, they show little to no efficacy in treating AML. Therefore, immunotherapies are traditionally underappreciated and underdeveloped in AML. Through a drug re-purpose screen, we identified and validated that combined MEK and HDAC inhibitions via trametinib and quisinostat (TQ) potently inhibited the growth of mouse and human NRAS;ASXL1-AML (NA-AML), MLLr, and NPM1 mutated AML cells in vitro. In NA-AML mice, TQ drastically slowed down AML progression and prolonged their survival. The survival benefits of TQ largely relied on T cell functions. We show that TQ synergized to downregulate immune checkpoint ligands and upregulate STAT1- and CIITA-mediated expression of MHC-I and MHC-II in NA-AML cells. In addition, TQ treatment significantly reprogrammed transcriptome and epigenetic landscape of T cells, activated STAT1 signaling, and upregulated genes and pathways promoting activation, survival, and cytotoxicity of CD4 and CD8 T cells. A cytotoxic cluster was thus expanded in central memory and effector memory T cells in TQ-treated NA-AML mice. More importantly, TQ directly acted on AML-associated mouse and human T cells, reverting them from a dysfunctional state to an active state. In leukemia:T cell co-cultures, TQ-treated T cells demonstrated greatly improved MHC-dependent leukemia killing. Our findings suggest that the dual actions of TQ on NA-AML and T cells enhance leukemia recognition and anti-leukemia killing of endogenous T cells, leading to effective AML clearance.

Germline gain-of-function (GOF) mutations in the signal transducer and activator of transcription 1 (STAT1) gene cause a dominantly inherited inborn error of immunity (IEI) characterized by chronic mucocutaneous candidiasis, autoimmunity, severe opportunistic infections and an increased risk of malignancy. Allogeneic hematopoietic stem cell (HSC) transplantation (HSCT) is curative but is associated with increased risk of morbidity and mortality in STAT1 GOF patients compared to other IEI. To develop a curative, autologous alternative to HSCT, we evaluated gene editing strategies in STAT1 GOF model cell lines, primary T cells, and patient-derived HSCs. Universal and mutation-specific strategies using CRISPR/Cas-mediated homology-directed repair (HDR) were limited by low efficacy (<25%), poor viability, and a lack of allele-specificity. In contrast, adenine base editing corrected the recurrent and highly pathogenic p.T385M mutation with upwards of 90% efficiency in patient T cells and HSCs without significant unintended on- or off-target genomic aberrations. Gene editing functionally restored total STAT1 expression (p<0.0217), STAT1 phosphorylation (p<0.0056), interferon-stimulated gene expression (OAS1; p=0.0005) and improved IL-17 production (p<0.0001). Edited HSCs retained multilineage differentiation capacity and sustained engraftment with persistence of the corrected allele at 16 weeks in humanized immunodeficient mice. These data demonstrate efficient and precise correction of STAT1 GOF mutations by base editing, with maintenance of the correction through long-term engraftment in vivo. This represents the first application of gene editing to correct a dominant gain-of-function mutation causing immunodeficiency, with potential applicability to other genetic disorders associated with heterozygous and gain-of-function mutations.

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.

This work demonstrates that the integrity of actin filaments can selectively regulate both lateral motion and in-plane orientation of transmembrane proteins on cell membranes. Using anisotropic gold nanostars functionalized with HER2 (human epidermal growth factor receptor)-targeting aptamers as optical probes, we tracked HER2 receptor dynamics on cancer cells using differential interference contrast microscopy. We found that for cells with an intact actin structure, the lateral mobility of receptors was restricted, consistent with actin-supported membrane compartmentalization. When actin filaments were disrupted after treatment with latrunculin A, HER2 receptors exhibited Brownian motion. In parallel, the receptors exhibited longer rotational dwell times corresponding to reduced orientational freedom. Analysis of receptor dynamics revealed that the coordinated translation-rotation regulation by actin filaments only occurred under directed diffusion modes when the cytoskeleton was intact. Two-dimensional single-nanoconstruct tracking provides a real-time approach to uncover how cytoskeletal architecture can organize receptor dynamics in living cells.

Boron neutron capture therapy (BNCT) is a radiotherapeutic modality in which biological selectivity is dictated not by targeting alone but by the spatial control of nuclear reactions. Following tumor-selective accumulation of 10B, neutron irradiation induces the 10B(n,α)7Li reaction, generating high-linear energy transfer particles with micrometer path lengths that confine cytotoxicity to boron-enriched cells. Despite its intrinsic precision, the clinical translation of BNCT has long been hindered by a fundamental chemical challenge: the inability to achieve sufficient, homogeneous, and durable intratumoral boron distribution. In this Account, we argue that BNCT must be reframed as a problem of chemical architecture across length scales, rather than a simple paradigm of boron delivery followed by irradiation. Building on foundational contributions from Jin-Ho Choy (layered double hydroxide-based confinement and ion-exchange systems), Hiroyuki Nakamura (molecular boron design enabling targeting and functional integration), and Koji Ono (clinical dosimetry and translational constraints), we outline how nanohybrid platforms transform BNCT into an integrated systems therapy in which materials design, biological transport, and radiation physics are co-optimized. We identify three governing design principles for next-generation BNCT. First, structural confinement and high-density boron loading enabled by layered inorganic hosts, carborane clusters, and hybrid nanocarriers, ensure sufficient payload delivery at the cellular and subcellular levels. Second, spatiotemporal control of boron distribution, achieved through surface engineering, targeting ligands, and microenvironment-responsive release, addresses the critical challenge of intratumoral heterogeneity. Third, integration with imaging and dosimetry establishes a "measure-what-you-treat" paradigm, linking molecular design directly to therapeutic outcome. We further highlight that BNCT efficacy is governed not by bulk tumor boron concentration alone but by microdistribution and subcellular localization, reflecting the short path length of high-linear energy transfer(LET) particles. This insight shifts design strategies from maximizing systemic delivery toward controlling intracellular fate and spatial precision. In parallel, the transition to accelerator-based neutron sources imposes additional constraints on boron chemistry, necessitating higher delivery efficiency and tighter coordination between pharmacokinetics and irradiation conditions. Together, these advances position BNCT as a chemically programmable precision radiotherapy, in which therapeutic outcome is dictated by the hierarchical organization of boron from molecular to nanoscale to clinical dimensions. Future progress will depend on scalable chemical design, quantitative imaging of boron distribution, and integration with emerging therapeutic modalities. We propose that the next phase of BNCT will be defined by the convergence of nanochemistry, systems-level transport control, and clinically informed design, enabling broader translation into refractory and heterogeneous cancers.

Human T-cell leukemia virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia/lymphoma (ATLL) and the neuroinflammatory disease, HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-1 Tax regulatory protein plays a critical role in HTLV-1 persistence and pathogenesis; however, the underlying mechanisms are poorly understood. Here we show that Tax dynamically regulates mitochondrial reactive oxygen species (ROS) and membrane potential to trigger mitochondrial dysfunction. Tax is recruited to damaged mitochondria through its interaction with the IKK regulatory subunit IKBKG/NEMO and directly engages the ubiquitin-dependent PINK1-PRKN/parkin pathway to induce mitophagy. Tax also recruits autophagy receptors CALCOCO2/NDP52 and SQSTM1/p62 to damaged mitochondria to induce mitophagy. Furthermore, Tax requires PRKN to limit the extent of CGAS-STING1 activation and suppress type I interferon (IFN) induction. HTLV-1-transformed T-cell lines and PBMCs from HAM/TSP patients exhibit hallmarks of chronic mitophagy, and inhibition of PRKN in HTLV-1-transformed cell lines downregulates p19 Gag expression and induces cell death. Collectively, our findings suggest that Tax manipulation of the PINK1-PRKN mitophagy pathway represents a new HTLV-1 immune evasion strategy important for maintaining viral gene expression and cell survival.Abbreviations: 3-MA: 3-methyladenine; ACTB: actin beta; ATLL: adult T-cell leukemia/lymphoma; BafA1: bafilomycin A1; BECN1: beclin 1; CALCOCO2: calcium binding and coiled-coil domain 2; CCCP: carbonyl cyanide m-chlorophenylhydrazone; CGAS: cyclic GMP-AMP synthase; co-IP: co-immunoprecipitation; DOX: doxycycline; GFP: green fluorescent protein; DNM1L/DRP1: dynamin 1 like; HAM/TSP: HTLV-1-associated myelopathy/tropical spastic paraparesis; HSPD1/HSP60: heat shock protein family D (Hsp60) member 1; HTLV-1: Human T-cell leukemia virus type 1; IFN: interferon; IkB: inhibitor of nuclear factor kappa B; IKBKG/NEMO: inhibitor of nuclear factor kappa B kinase regulatory subunit gamma; IKK: IkB kinase; IRF3: interferon regulatory factor 3; KO: knockout; LAMP2: lysosome associated membrane protein 2; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MT-CO2: mitochondrially encoded cytochrome c oxidase II; mtDNA: mitochondrial DNA; mtROS: mitochondrial reactive oxygen species; NAC: N-acetylcysteine; NBR1: NBR1 autophagy cargo receptor; NFKB: nuclear factor kappa B; OPTN: optineurin; PBMCs: peripheral blood mononuclear cells; PINK1: PTEN induced kinase 1; PRKN: parkin RBR E3 ubiquitin protein ligase; qRT-PCR: quantitative reverse transcription polymerase chain reaction; RFP: red fluorescence protein; ROS: reactive oxygen species; SAR: selective autophagy receptor; SQSTM1: sequestosome 1; STING1: stimulator of interferon response cGAMP interactor 1; TAX1BP1: Tax1 binding protein 1; TEM: transmission electron microscopy; TMRM: tetramethylrhodamine methyl ester; TOMM20: translocase of outer mitochondrial membrane 20; Ub: ubiquitin; VCL: vinculin; WT: wild-type.

Despite a few clinical successes, the efficacy of cancer nanomedicines remains limited by rapid clearance by the mononuclear phagocytic system and poor permeation across the abnormal tumor vasculature. We previously showed that methyl palmitate nanoparticles (MPN) can safely and reversibly inhibit the phagocytic activity of immune cells for several hours, thereby improving tumor accumulation and the efficacy of systemically administered nanomedicines. Here, we demonstrate that, on a shorter time scale, MPN can induce vasodilation, introducing an additional mechanism to enhance the accumulation of therapeutic agents within malignant tissue. Upon internalization by macrophages and endothelial cells, MPN could potentially trigger the release of endogenous nitric oxide (NO), a key mediator of vasodilation, in a concentration-, and time-dependent manner. Following MPN administration, raster-scanning optoacoustic mesoscopy (RSOM) revealed vasodilation across multiple tissues, with a strong effect observed in tumors. To assess enhanced tumor accumulation, we injected 70 kDa fluorescent dextran and demonstrated via histology a markedly increased fluorescence signal exclusively in MPN-treated tumors compared to controls 24 h later. In addition, positron emission tomography (PET) imaging of 89Zr-labeled Feraheme nanoparticles showed significantly greater tumor accumulation after a 15 min MPN pretreatment. Finally, general serum biochemistry panels and histological analyses of major organs in healthy mice revealed that MPN did not induce observable short-term toxicity under the tested conditions (single or repeated MPN dosing). Overall, this study demonstrates that MPN-induced vasodilation occurring within minutes enhances intratumoral deposition of macromolecules and small nanoparticles. Together with their longer-term effects on phagocytosis inhibition, these findings indicate that MPN can improve therapeutic delivery through complementary, time-dependent mechanisms that increase tumor perfusion and vascular permeability.

Melanocytes reside in diverse microenvironments that influence their susceptibility to oncogenic transformation; however, investigation of rare melanoma subsets has been limited by the lack of suitable preclinical animal models. In this study, we developed a primary, immunocompetent zebrafish model to study uveal melanoma using choroidal melanocyte-targeted injection and electroporation of plasmids encoding human GNAQQ209L together with CRISPR/Cas9 cassettes for somatic tumor-suppressor gene deletion. Single-cell transcriptional profiling of primary melanocytes and melanoma derived from the eye and skin revealed distinct transcriptional programs, with epithelial-to-mesenchymal transition pathways enriched in ocular tumors. In addition, choroidal fibroblasts from tumor-bearing eyes exhibited marked transcriptional changes, including increased fibronectin and collagen expression, consistent with stromal remodeling. Given prior associations between mitfa loss and accelerated GNAQQ209L tumor onset, the model was applied to determine whether melanocyte differentiation state contributes to the emergence of GNAQ-driven tumors. The increased susceptibility resulted from expansion of Mitfa-independent melanocyte progenitor populations in germline mitfa-mutant zebrafish, rather than somatic mitfa loss in differentiated melanocytes, as conditional, melanocyte-specific mitfa deletion in adult zebrafish did not accelerate tumor growth. Furthermore, pax3a-positive melanocyte progenitor cells in mitfa-deficient zebrafish embryos and adult eyes and skin were highly susceptible to transformation induced by GNAQQ209L but not BRAFV600E. Analogous PAX3 positive populations were also identified in mouse and human single-cell transcriptomic datasets. Collectively, these findings establish a critical role for Mitfa-independent melanocyte progenitors in uveal melanoma pathogenesis. Choroid-targeted GNAQQ209L expression induces anatomically correct uveal melanoma in adult zebrafish, with germline mitfa deletion expanding mitfa-independent melanocyte progenitors with enhanced susceptibility that are transcriptionally distinct from the subpopulation transformed by BRAFV600E.

High-grade serous ovarian cancer (HGSOC) is a lethal malignancy characterized by profound intratumoral heterogeneity and immune evasion. Although previous research has identified 4 molecular subtypes of HGSOC, defining their spatial distribution across anatomic sites could help identify functional mediators of progression and immune evasion. In this study, we performed single-cell and spatial transcriptomic sequencing on 66 anatomically paired samples across 5 sites from 8 patients and conducted an integrated analysis on the generated datasets. Five distinct gene programs (GP) with specific spatial distributions and functional roles were identified: GP1 (differentiated-invasive), GP2 (differentiated-proliferative), GP3 (immunoreactive), GP4 (mesenchymal), and GP5 (proliferative). Interestingly, molecular subtypes exhibited dynamic spatial transitions that promote HGSOC progression. The differentiated-proliferative subtype dominated tumor cores, whereas the differentiated-invasive subtype localized to the invasive front driven by SDC4-expressing epithelial cells (c40) that were functionally validated to promote migration/invasion. Concurrently, immune evasion operated through distinct mechanisms. Differentiated-invasive and immunoreactive subtype co-localization established immunosuppression via the TNFα-SAA1/2-APOE signaling axis that recruits immunosuppressive myeloid cells across primary and metastatic sites, whereas fibroblast-derived collagen barriers were constructed at the interfaces between mesenchymal and immunoreactive subtypes in omental metastases along with dysregulated chemokines to exclude immune infiltration. This spatially resolved atlas directly links subtype spatial transitions to tumor progression and subtype co-localization to immune evasion, providing a mechanistic framework for targeting spatially organized tumor-immune interactions in HGSOC. Spatially resolved transcriptomic analysis reveals tumor subtype transitions and niche-specific immune evasion mechanisms in ovarian cancer.

Glioblastoma (GBM) infiltration poses a formidable therapeutic challenge, but the mechanisms enabling long-distance tumor invasion remain poorly defined. In a recent study, Nebeling and colleagues have leveraged longitudinal three-photon intravital microscopy to visualize the migratory behavior of invading GBM cells and their interactions with microglia. Using an immunocompetent autochthonous murine GBM model for live imaging of the contralateral cortex and corpus callosum (CC), the team demonstrated that the migration velocity of GBM cells varied by anatomic location, with tumor cells moving faster in the CC than in the cortex. Furthermore, GBM cells with less tumor microtubes (TM) exhibited higher motility and traveled longer distances than TM-rich cells. Interestingly, the authors also uncovered a stage-dependent, biphasic microglial response to invading GBM cells: enhanced surveillance during sparse infiltration, followed by suppressed surveillance as tumor burden increases. Functional analyses identified chemokine receptor CX3CR1 as a key regulator of microglial reactivity and a requirement of microglia to drive GBM invasiveness. These findings align with earlier evidence that spatial constraints and microglial organization influence GBM invasion. Together, these works highlight a critical role of microglia and the tumor microenvironment in shaping invasive GBM phenotypes and offer new avenues for therapeutic strategies to limit GBM invasion.

SETD2 is frequently mutated or deleted in clear cell renal cell carcinoma (ccRCC). Loss of SETD2 could create synthetic lethal dependencies that confer therapeutic vulnerabilities. In this study, we demonstrated that SETD2 deficiency promotes cytoplasmic mitochondrial DNA (mtDNA) leakage, leading to basal activation of cGAS-STING inflammatory signaling and increased apoptotic priming. This inflammatory state upregulated the BH3-only protein NOXA, constrained myeloid cell leukemia 1 (MCL-1) function, and enforced a synthetic lethal dependency on the antiapoptotic protein BCL-xL. Pharmacologic inhibition of BCL-xL further amplified cGAS-STING signaling in SETD2-deficient cells through sublethal mitochondrial outer membrane permeabilization, resulting in increased mtDNA release and robust NOXA induction. Elevated NOXA neutralized the compensatory MCL-1-mediated survival signaling, triggering apoptosis. In contrast, SETD2-proficient ccRCC cells exhibited minimal cGAS-STING activation and failed to induce NOXA following BCL-xL inhibition, rendering them resistant. Genetic ablation of cGAS, STING, IRF3, or NOXA rescued sensitivity to BCL-xL inhibition, confirming that mtDNA-driven innate immune signaling is required for this dependency. In vivo, BCL-xL inhibition suppressed tumor growth and prolonged survival in SETD2-deficient xenograft models. Collectively, these findings establish a mechanistic link between SETD2 loss, mtDNA-driven innate immune activation, and enforced BCL-xL dependence in ccRCC, revealing a therapeutically targetable vulnerability in SETD2-deficient tumors. SETD2 deficiency, which is associated with worse prognosis in kidney cancer, promotes mitochondrial DNA-driven cGAS-STING signaling and NOXA-mediated apoptotic priming, creating a therapeutically targetable dependency on BCL-xL in kidney cancer.

Mitochondrial dynamics is a key regulator of cellular homeostasis, orchestrating metabolic reprogramming that fuels tumor progression and treatment resistance. In multiple myeloma (MM), however, the functional relevance of mitochondrial remodeling has not been fully defined. Using ultrastructural analyses, we reveal that MM cells display a highly fragmented mitochondrial network, a phenotype further exacerbated in both cell lines and primary MM cells resistant to proteasome inhibitors. Transcriptomic profiling across multiple patient-derived datasets consistently demonstrated upregulation of DNM1L gene, which encodes the mitochondrial fission GTPase DRP1, particularly in relapsed and refractory MM, and revealed a significant association with inferior overall survival. Disrupting mitochondrial fission, either through genetic targeting of DNM1L or pharmacologic inhibition of DRP1 with the selective small molecule inhibitor Drpitor1a, resulted in pronounced mitochondrial dysfunction, impaired oxidative phosphorylation, and potent anti-myeloma activity in vitro, culminating in a hybrid cell death program with a predominant apoptotic component accompanied by ferroptotic features. These effects were recapitulated in vivo in a bortezomib-resistant xenograft model, where either DNM1L depletion or DRP1 inhibition produced similar outcomes. Mechanistically, the transcription factor c-MYC upregulated DNM1L expression, and DRP1-dependent mitochondrial fragmentation sustained MYC-driven oxidative metabolism and lipid synthesis. Altogether, these findings establish aberrant mitochondrial fission as a pathogenic hallmark of MM and highlight DRP1 inhibition as a promising therapeutic approach, especially for relapsed or refractory disease.

Photodynamic therapy (PDT) is a minimally invasive cancer treatment strategy. Despite numerous studies confirming its potential advantages, the clinical translation of this technique remains limited owing to the impracticality of using conventional light sources and the instability of photosensitizers. This study presents a novel PDT approach that utilizes organic light-emitting diodes (OLEDs) as compact, low-thermal, wearable, and flexible light sources to activate chlorin e6 (Ce6)-loaded Pluronic nanocapsules (Ce6/Plu NCs) for anticancer therapy. Here, different types of Pluronic polymers are used and optimized to synthesize ∼120 nm-diameter Ce6/Plu NCs. Among the formulated systems, the Ce6/PP123 NCs show optimal spectral characteristics and the highest therapeutic efficacy in vitro, and efficient tumor accumulation after intravenous injection and significant antitumor efficacy upon OLED irradiation with minimal off-target toxicity in vivo. Tissue-attachable OLEDs for PDT can be placed in close proximity to tumor tissue for efficient light delivery. The combination of the Ce6/PP123 NCs with a novel wearable OLED is a promising platform for effective and safe PDT in cancer treatment.

Photodynamic therapy (PDT) is a promising minimally invasive antitumor approach, but its clinical efficacy is severely limited by the single Type‑II reaction of conventional photosensitizers (PSs), insufficient subcellular targeting and poor performance in hypoxic tumors. Herein, a lysosome‑targeted D‑-π-‑A near‑infrared PS Lyso‑IsTp was designed and synthesized, with isophorone as acceptor, methoxy‑modified triphenylamine as donor, benzene as π‑bridge and 4‑(2‑chloroethyl)morpholine as targeting moiety. It emitted fluorescence at 750 nm with good photostability and solvent‑dependent properties. Under 660 nm irradiation, Lyso‑IsTp simultaneously produced Type‑I (•OH, O2 -•) and Type-II (1O2) ROS. Confocal imaging confirmed its excellent lysosome targeting in HeLa cells with a Pearson coefficient of 0.97. It showed negligible dark toxicity and remarkable phototoxicity, inducing apoptosis via ROS‑mediated lysosomal damage. In vivo studies on tumor‑bearing nude mice revealed that Lyso‑IsTp‑mediated PDT significantly inhibited tumor growth with obvious necrosis. Biosafety tests including body weight, H&E staining, and blood analysis demonstrated no obvious systemic toxicity or organ damage. This work provides a high‑performance lysosome‑targeted NIR PS with dual ROS generation, offering a feasible strategy for hypoxia‑tolerant and organelle‑specific antitumor PDT.

The ATP synthase-associated macromolecular complex plays a crucial role in cellular energy homeostasis. However, the regulation and potential function of this complex remain poorly understood in cancer. Here, we identify a 170-amino acid protein encoded by the lncRNA LINC00528, which we term ATP synthase-associated macromolecular complex enhancing microprotein (ATHENA). Mechanistically, ATHENA localizes to the mitochondrial inner membrane, in which ATHENA directly interacts with ATP synthase subunits (β and γ) and solute carrier proteins, including ANT and PiC, thereby promoting the assembly of these macromolecular complexes, enhancing ATP synthesis, and preserving mitochondrial and cristae architecture. Functionally, ATHENA promotes the proliferation, migration, and invasion of renal cell carcinoma (RCC) cells in vitro and drives RCC tumor growth and metastasis in vivo. Clinically, ATHENA is upregulated in RCC tissues and associated with poorer prognosis in RCC patients. Collectively, our findings identify ATHENA as a previously unrecognized mitochondrial microprotein that facilitates the assembly of ATP synthase-associated macromolecular complex and drives RCC progression.

Cuproptosis is a type of recently reported cell death characterized by aberrant accumulation of copper ions within cells, leading to mitochondrial stress and protein aggregation. Recent studies suggest that certain cancer cells are particularly susceptible to cuproptosis-inducing agents. However, the genetic determinants of cellular sensitivity to cuproptosis and the therapeutic potential of cuproptosis inducers in cancer treatment remain unclear. Here, we report the discovery of a small molecule, N1,N1-dimethyl-N4-(4-(pyridin-2-yl)thiazol-2-yl)benzene-1,4-diamine (dPTBD), that targets KRAS-driven cancer via a tetracycline-inducible cell-based drug screening. dPTBD exhibited significant antitumor efficacy in KRAS-driven cancers both in vitro and in vivo. Mechanistic studies revealed that dPTBD acted as a copper ionophore, promoting intracellular copper accumulation particularly in the mitochondria, leading to metabolic disruption and cuproptotic cell death. Adding trace amount of copper massively enhanced the cytotoxic effect of dPTBD, resulting in an immediate mitochondrial dysfunction and cuproptosis. In preclinical models, dPTBD, either alone or combined with physiologically tolerable amount of copper, significantly suppressed tumor growth in KRAS-mutant pancreatic and colon cancer xenografts. Taken together, our study reveals that induction of cuproptosis is a new therapeutic strategy for KRAS-driven cancer and identifies dPTBD as a lead compound for future development.

Hydrogels are highly hydrated three-dimensional polymeric network materials that have attracted considerable attention in medical and biomedical fields owing to their favorable chemical modifiability, physical tunability, biocompatibility, and capacity to mimic key features of native extracellular matrices. With advances in polymer chemistry, crosslinking strategies, stimuli-responsive design, and emerging fabrication technologies, hydrogels have evolved from simple soft materials into multifunctional biomedical systems capable of integrating controlled delivery, tissue support, microenvironmental regulation, biosensing, and disease modeling. These properties make hydrogels particularly relevant for addressing a broad spectrum of human diseases in which conventional therapeutic strategies are often limited by insufficient targeting efficiency, limited therapeutic windows, pronounced systemic side effects, and inadequate restoration of damaged tissue structure and function. In this review, we systematically summarize the design principles and material engineering strategies of hydrogels, including structural construction approaches and functional regulation concepts. We then provide a comprehensive overview of their current biomedical applications, encompassing drug delivery systems, tissue engineering and regenerative medicine, biosensing and diagnostic platforms, as well as cell culture and organoid systems. Building on this foundation, we place particular emphasis on recent progress in hydrogel-based therapeutic strategies across diverse human disease contexts, including wound healing, musculoskeletal system repair, cancer, neurological diseases, cardiovascular diseases, autoimmune disorders, and reproductive system diseases. Finally, we discuss the developmental potential of hydrogels in biomedicine and present an integrated perspective on their clinical translation prospects, with the aim of offering references and insights for the further application of hydrogel materials in future human disease treatment.

Casein kinase 1⍺ (CK1⍺) is a multifunctional serine/threonine kinase, serving as an upstream regulator of the p53 pathway and a potential therapeutic target for acute myeloid leukemia (AML). Small-molecule degraders show preclinical promise in AML by selectively degrading target proteins. Here, we developed and evaluated PinA1, a molecular glue degrader targeting CK1⍺, in preclinical AML models. PinA1 preferentially degraded CK1⍺ at nanomolar concentrations, resulting in enhanced p53 expression, cell-cycle arrest, and apoptosis in TP53 wild-type AML cell lines but not in TP53 mutated cells. PinA1 also induced CK1⍺ degradation and p53-dependent apoptosis in primary AML cells with wild-type TP53. Notably, PinA1 had minimal effects on the viability of human peripheral blood and bone marrow mononuclear cells, including CD34+ cells, with limited activation of the p53 pathway. In both cell lines and primary AML cells, PinA1 enhanced apoptosis when combined with targeted agents, including FLT3, BCL-2, or MDM2 inhibitors. PinA1 demonstrated robust anti-leukemic efficacy as a monotherapy and exhibited synergistic effects when combined with targeted agents in xenografts derived from human AML cell lines and primary AML cells. In conclusion, PinA1 degrades CK1⍺, activates the p53 pathway, and induces cell-cycle arrest and apoptosis in TP53 wild-type AML cells. The robust anti-leukemic efficacy of PinA1, both as monotherapy and further in combination with targeted agents, along with its minimal toxicity to normal hematopoietic cells, underscores its potential for future clinical applications.

Cuproptosis, heavily dependent on copper and mitochondrial tricarboxylic acid (TCA) cycle, can be exploited as a potential oncotherapy. However, the primary metabolic pathway of tumor cells is dependent on glycolysis owing to the overexpression of lactate dehydrogenase A (LDHA), which suppresses high-efficiency cuproptosis oncotherapy. Therefore, inhibiting LDHA to remodel the metabolic pathway of tumor cells could result in sensitizing cuproptosis and, hence, provide a novel strategy to achieve efficient oncotherapy. Accordingly, we herein report a pH-responsive, copper-engineered nanoplatform (ZCIF@GSK) encapsulating the LDHA inhibitor (GSK). ZCIF@GSK initiates the reprogramming of cellular metabolism, thereby achieving efficient cuproptosis. Under the low pH tumor environment, ZCIF@GSK specifically releases GSK to inhibit LDHA. This, in turn, inhibits glycolysis but promotes mitochondrial TCA cycle, to sensitize cuproptosis. Meanwhile, the release kinetics of copper further causes the oligomerization of lipoylated dihydrolipoamide S-acetyltransferase (DLAT) and the downregulation of lipoic acid synthase (LIAS), contributing to efficient cuproptosis. Overall, we propose a novel strategy for sensitizing cuproptosis through an LDHA inhibitor. Additionally, we demonstrate that the inhibition of LDHA upregulates the expression of ferredoxin (FDX1), a key regulator of cuproptosis, further promoting the sensitization of cuproptosis and, hence, a promising cuproptosis-based oncotherapy.

Cancer cells maintain chronically elevated levels of reactive oxygen species (ROS) while relying on robust antioxidant programs to preserve redox homeostasis and viability. Although therapeutic strategies that disrupt this balance to induce lethal oxidative stress and ferroptosis have emerged as promising anticancer approaches, the upstream signaling mechanisms that constrain ROS accumulation under physiologically relevant stress conditions remain incompletely understood. Here, we identify the stress-responsive kinases SMG1 and DNA-dependent protein kinase (DNA-PK) as functionally redundant regulators of redox homeostasis and ferroptosis resistance. Genetic or pharmacological inhibition of either kinase triggers ferroptotic cell death, accompanied by marked accumulation of total ROS, ferrous iron, and lipid hydroperoxides. Mechanistically, under mild oxidative stress, SMG1 and DNA-PK cooperatively phosphorylate the central antioxidant transcription factor NRF2 at serine 13 and serine 40, weakening its interaction with the negative regulator KEAP1 and promoting NRF2 accumulation and transcriptional activation. Transcriptomic profiling of de novo mRNAs revealed that inhibition of either kinase is sufficient to suppress NRF2-driven antioxidant gene expression. In contrast, excessive oxidative stress overrides this pro-survival pathway and redirects signaling toward anti-survival responses mediated by ATF4, ATM-CHK2, and JNK/p38 pathways. Collectively, these findings uncover a previously unrecognized SMG1/DNA-PK-NRF2 signaling axis that functions as a redox stress-intensity-dependent switch governing cell fate decisions between antioxidant adaptation and ferroptotic death. Targeting this axis may represent a promising therapeutic strategy for cancer treatment.

Idiopathic pulmonary fibrosis (IPF) is a progressive and fatal interstitial lung disease with an urgent need for novel therapeutic strategies. M2 macrophage-derived TGF-β1 promotes fibroblast myogenesis, contributing to IPF pathogenesis. Targeting macrophage polarization and fibroblast function thus represents an effective therapeutic approach for treating IPF. Here, we identify B-cell lymphoma 9 (BCL9) as a key upstream regulator implicated in IPF pathogenesis. We demonstrate that BCL9 drives the macrophage M2 program through the MerTK-ERK-SPP1 axis. Notably, pharmacological inhibition of BCL9 with our novel peptide, hsBCL9Z96, effectively attenuates pulmonary fibrosis by reprogramming macrophage-fibroblast crosstalk. Specifically, BCL9 inhibition promotes fibroblast lipogenesis via TGF-β1 signaling, which in turn supports alveolar type 2 (AT2) cell expansion. This macrophage-orchestrated fibroblast phenotypic switch from myogenic to lipogenic is visually corroborated by spatial transcriptomic analyses and immunofluorescence staining of human lung tissues. Functionally, the pathological role of BCL9 and efficacy of hsBCL9Z96 are validated in human cellular models, including IPF patient-derived cells, confirming its translational significance. Collectively, our findings not only elucidate a novel BCL9-driven macrophage-fibroblast-AT2 cell axis in IPF but also establish hsBCL9Z96 as a promising first-in-class therapeutic candidate, providing a strong rationale for targeting BCL9-mediated Wnt signaling in clinical IPF treatment.

Diffuse large B-cell lymphomas (DLBCL) are genetically and phenotypically heterogeneous, making diagnosis and treatment challenging. Current models suggest DLBCLs derive from follicular B cells engaged in adaptive immune responses. By studying cooccurring truncating mutations in SPEN and NOTCH2 in the BN2-DLBCL subtype, our data suggest a previously unrecognized extrafollicular trajectory. Using animal models and human specimens, we find that this cooperative mutational axis supports expansion of putative clonal precursors with features of marginal zone, memory, and a distinct, autoimmune B cell-like state. This trajectory is associated with sex-biased outcomes: Female patients and mice exhibit reduced survival compared with males in our cohorts. Further analysis links this disparity to enhanced X-chromosome-linked expression and functionality of Toll-like receptor signaling. We show that IRAK inhibition represents a potential sex-specific therapeutic strategy in preclinical models. These findings support a distinct developmental origin for BN2-DLBCL and identify a high-risk female population with actionable targets for precision therapy. The findings in this article support a distinct developmental origin for BN2-DLBCL and identify a high-risk female population with actionable targets for precision therapy.

During cancer evolution, derepressed repetitive elements (REs) can trigger innate immune responses via "viral mimicry," imposing selective immune pressure on emerging tumor cells. We propose that RE mimics mediate transitions from initial antitumorigenic immune selection to a tolerized equilibrium shaped by trade-offs that can ultimately be predicted and exploited therapeutically. See related article by Chiappinelli and Burns, p. 1494.

Spatial transcriptomics and proteomics have enabled profound insights into tissue organization, yet these technologies remain largely disparate, and emerging same-slide multiomics approaches are limited in plex, spatial resolution, signal retention, and integrative analytics. We introduce IN-situ DEtailed Phenotyping To High-resolution transcriptomics (IN-DEPTH), a streamlined, resource-efficient, commercially compatible workflow using single-cell spatial proteomics-derived imaging to guide transcriptomic capture on the same slide without RNA signal loss. To integrate modalities beyond niche-level mapping, we developed Spectral Graph Cross-Correlation (SGCC), a proteomic-transcriptomic framework resolving spatially coordinated functional state changes across interacting cell populations. Applied to diffuse large B-cell lymphoma (DLBCL), IN-DEPTH and SGCC enabled stepwise discovery from Epstein-Barr virus (EBV)-positive and EBV-negative tumor comparisons with single-cell resolution, revealing coordinated tumor-macrophage-CD4 T-cell remodeling, immunosuppressive C1Q macrophage enrichment, CD4 T-cell dysfunction, and a candidate IL27-STAT3 signaling axis. Collectively, IN-DEPTH enables scalable spatial multiomics to uncover clinically relevant microenvironmental mechanisms and toward robust spatial multimodal AI models. IN-DEPTH enables same-slide spatial multiomics across commercial platforms via a protein-first strategy preserving protein epitopes, RNA quality, and tissue integrity. Coupled with SGCC, it resolves coordinated spatial immune remodeling, revealing EBV/LMP1-driven C1Q macrophage polarization and CD4 T-cell dysfunction in DLBCL, with broad applicability to other diseases.

Meta-analysis is fundamental to evidence-based medicine, yet traditional workflows remain labor-intensive and susceptible to bias. Although LLM-based research agents offer opportunities for workflow automation, they often lack the data fidelity and methodological traceability required for rigorous quantitative evidence synthesis, particularly when parsing multimodal scientific charts. To address this challenge, we introduce MacAma, a semi-automated multi-agent framework for protocol-constrained and human-verifiable meta-analysis. MacAma operationalizes selected PRISMA 2020 reporting items, PICOS-based eligibility logic, and SYRCLE risk-of-bias domains as structured prompts, decision rules, output fields, and audit records. Critically, MacAma adopts a risk-aware automation strategy: Lower risk, repetitive, and protocol-driven tasks, such as literature screening and drafting, are delegated to AI agents, whereas high-impact steps that directly affect effect-size estimation and statistical conclusions, such as quantitative chart-data extraction, remain subject to expert verification. In a preclinical radiotherapy case study evaluating tumor-related immune outcomes and metastatic potential mediated by circulating tumor cells, MacAma achieved competitive screening performance in the evaluated benchmark and reduced the manual screening burden by over 80% within the current workflow. The case study further demonstrates how structured agent outputs, predefined criteria, and audit records can support transparent screening, data extraction, statistical synthesis, and manuscript drafting. These results suggest that MacAma may provide a scalable and auditable framework for AI-assisted meta-analysis, although important limitations remain in full-text access, quantitative chart data extraction, and expert interpretation of heterogeneity. MacAma is open-source and available at https://github.com/YilinYuan/MacAma.

Tumour necrosis factor (TNF) is a pleiotropic cytokine originally identified for its ability to kill cancer cells. However, a paradoxical tumour-promoting role for TNF emerged when early attempts to exploit its anti-tumour activity in cancer therapy produced conflicting outcomes, raising the question of whether TNF should be viewed as a therapeutic agent or a treatment target in cancer. Here, we demonstrate that expression of cFLIP, a catalytically inactive paralogue of caspase-8 (CASP8), determines the susceptibility of melanoma cells to TNF and thereby controls melanoma growth in a syngeneic, immune-competent mouse model of B16F10 cutaneous melanoma. B16F10 melanoma cells lacking cFLIP (cFlipKO/KO cells) failed to grow in wild-type mice, whereas in TNF-deficient mice, cFlipKO/KO melanoma cells formed palpable tumours and exhibited robust subcutaneous growth. These findings indicate that TNF alone is sufficient to control melanoma growth in the absence of cFLIP. Importantly, the anti-tumour activity of TNF has predominantly been investigated through targeting cellular inhibitors of apoptosis proteins (cIAPs), which promotes RIPK1 activation and TNF-induced cytotoxicity. We show that genomic ablation of cIAPs or RIPK1, in contrast to cFLIP, neither triggered TNF-induced toxicity nor affected melanoma growth in vivo. Collectively, our data underscore the central role of cFLIP in regulating melanoma responses to TNF and suggest that endogenous immune surveillance as well as immunotherapies involving TNF could strongly benefit from cFLIP targeting strategies.

Pancreatic ductal adenocarcinoma (PDAC) is refractory to immunotherapy due to its immunologically cold microenvironment and the scarcity of mutation-derived neoantigens. Here, we introduce NeoAPP, a computational tool designed to systematically decode neoantigens arising from tumor-specific transcripts (TSTs) generated by transcriptional dysregulation. By integrating multi-cohort transcriptomic data from 413 PDAC samples, NeoAPP identified a median of 351 neoantigens per sample, originating from 56 neoantigen-encoding TSTs (neoTSTs), exceeding mutation-derived neoantigens in both abundance and patient coverage. Mechanistic analyses show that noncanonical splicing junctions and transposable element activation drive neoantigen generation, while alternative promoter usage regulated by FOXA2 constitutes a potential major source of neoTSTs. In addition, a subset of neoTSTs was detectable in plasma-derived extracellular vesicles and was associated with patient survival, suggesting their potential as minimally invasive biomarkers. Vaccination with neoTSTs induced CD8+ T cell responses in HLA-A*02:01 and HLA-A*11:01 transgenic mice and suppressed tumor growth in syngeneic PDAC models. Collectively, this work establishes TST-derived neoantigens as a dominant and therapeutically actionable antigen reservoir in PDAC, advancing a transcriptome-guided framework for neoantigen discovery with potential to overcome immune resistance in low-mutation cancers.

Lymphatic endothelial cells (LECs) line the lymphatic vasculature and support interstitial fluid drainage, lipid transport, and immune-cell trafficking. Beyond these classical functions, recent studies now recognize LECs as heterogeneous, spatially organized endothelial regulators that maintain lymphatic identity while adopting tissue-, segment-, and disease-associated states. Advances in lineage tracing, multiomics, functional imaging, and perturbation studies now link LEC heterogeneity to drainage control, immune surveillance, antigen handling, metabolic homeostasis, and tissue repair. Despite these advances, a unified framework explaining how developmental origin, tissue niche, and context-dependent state transitions collectively shape LEC function across physiology and disease remains lacking. In this review, we summarize the developmental origins, identity-maintenance mechanisms, and anatomical deployment of LEC states across the lymphatic network. We then discuss how specialized LEC states coordinate lymphatic transport, immune surveillance, and peripheral tolerance under homeostasis. We further examine how context-dependent LEC reprogramming contributes to lymphatic disorders, inflammation and autoimmunity, cancer, cardiometabolic disease, and central nervous system dysfunction. Finally, we highlight challenges and opportunities for translational LEC-targeted therapy. Together, this state-centered framework reframes LECs as actionable regulators of tissue homeostasis and disease progression, providing a conceptual foundation for precision lymphatic medicine.

Macrophages are sentinel innate immune cells that arise from embryonic precursors and bone marrow monocytes, displaying a functional continuum that transcends the classical M1 (pro-inflammatory)/M2 (anti-inflammatory) dichotomy. Under homeostatic conditions, balanced M1/M2 polarization preserves tissue integrity by coordinating immune surveillance, efferocytosis, and tissue repair. When this equilibrium is disrupted, however, M1-skewed responses drive chronic inflammation and autoimmunity, whereas M2-skewed polarization facilitates tumor immune evasion and organ fibrosis. Although diverse therapeutic strategies-including reprogramming, depletion, blockade of monocyte recruitment, CAR-M cells, and nanomedicine-are being explored to restore homeostasis, clinical translation remains constrained by the lack of pathogenic subset-specific markers, insufficient predictive biomarkers for patient stratification, species divergence between mice and humans, and the temporal complexity of context-dependent intervention windows. In this Review, we systematically delineate macrophage plasticity and the molecular mechanisms underlying polarization imbalance, evaluate existing and emerging macrophage-directed interventions, and dissect these translational bottlenecks in depth, highlighting how single-cell multi-omics, humanized models, and dynamic biomarkers can overcome them. By providing a roadmap for precisely calibrating macrophage functional states and restoring M1/M2 balance, this framework will accelerate the development of precision immunotherapies aimed at re-establishing immune homeostasis across a broad spectrum of human pathologies.

Macrophages orchestrate immune responses through remarkable phenotypic plasticity, which is intrinsically linked to their ability to reprogram intracellular metabolic pathways in response to microenvironmental cues. While recent advances have highlighted the role of aberrant macrophage metabolism in diverse diseases, a systematic synthesis integrating both intracellular and extracellular metabolic signals remains lacking. This review provides a comprehensive framework for understanding how core metabolic pathways-glycolysis, the TCA cycle, oxidative phosphorylation (OXPHOS), fatty acid oxidation (FAO), and amino acid metabolism-are rewired during macrophage polarization under the orchestration of upstream signaling cascades, including NF-κB, PI3K/AKT/mTOR, JAK-STAT, and MAPK. We examine how exogenous metabolites such as succinate, itaconate, lactate, and amino acids reciprocally regulate macrophage function and discuss tissue-specific metabolic signatures of macrophage subsets-including alveolar macrophages （AMs), Kupffer cells (KCs), and tumor-associated macrophages (TAMs)-in the context of obesity, Type 2 diabetes (T2D), metabolic dysfunction-associated steatotic liver disease (MASLD), infections, autoimmune disorders, and cancer. We further evaluate emerging therapeutic strategies targeting macrophage metabolism, summarizing preclinical and clinical advances across signaling pathways, metabolic nodes, cytokines, and cell-based therapies with detailed trial data. By integrating cell-intrinsic metabolic circuitry with extracellular signals, this review establishes a theoretical foundation for metabolism-targeted immunotherapies and identifies key knowledge gaps for future investigation.

Coronary microvascular dysfunction (CMD) is a key contributor to myocardial ischemia and cardiovascular diseases. It is characterized by abnormalities of the coronary microvasculature, leading to impaired myocardial perfusion. Although CMD has a high prevalence in patients with nonobstructive coronary artery disease and is associated with adverse cardiovascular events, its pathogenesis has not yet been fully elucidated. Current diagnostic approaches combine noninvasive and invasive methods, but there remains a lack of effective targeted therapies. This review discusses the epidemiology, pathophysiology, diagnostic strategies, and treatment options for CMD, with a particular focus on the protective role of the neuregulin-1 (NRG-1)/v-erb-b2 erythroblastic leukemia viral oncogene homolog B (ErbB) signaling pathway. We initially outline how the NRG-1/ErbB pathway affects endothelial function, ventricular remodeling, oxidative stress, and myocardial angiogenesis, highlighting its potential as a therapeutic target. In addition, we explore emerging evidence that traditional Chinese medicine (TCM) interventions may regulate the NRG-1/ErbB axis to improve microvascular function and cardiac outcomes. Overall, this review deepens our understanding of the mechanisms underlying CMD and provides new avenues for integrated precision therapies, including TCM, with the aim of improving clinical management and prognosis in patients with CMD.

Sirtuins (SIRT1-SIRT7) are nicotinamide adenine dinucleotide (NAD+) dependent deacylases that serves as metabolic sensors, coupling cellular energy status to chromatin structure, mitochondrial function, and stress responses. Dysregulated SIRT activity has been extensively studied in aging, metabolic syndrome, cardiovascular disease, neurodegeneration, cancer, and immune disorders. However, robust human evidence and SIRT-targeted therapies are lacking. Transgenic mouse models serve as key platforms to study gene function and guide therapeutic development. This review synthesizes evidence from Sirt1-7 transgenic mouse models regarding the core cellular processes governed by SIRTs: metabolism, genome integrity, stress resistance, immunity, and autophagy, and illustrates their operation across different organ systems. By comparing global, tissue-specific, and inducible knockout (KO) and overexpression (OE) models of cardiovascular, respiratory, digestive, nervous, endocrine, urogenital, musculoskeletal, malignant, and immune diseases, we identified central regulatory SIRTs (SIRT1, SIRT3, and SIRT6), context-dependent modifiers (SIRT2, SIRT4, SIRT5, and SIRT7), and their organ- and cell type-specific functions. We also summarize representative small-molecule SIRT activators, inhibitors, and degraders, covering both clinical and preclinical studies, and highlight where contradictions and knowledge gaps remain. Together, these analyses help clarify which aspects of SIRT modulation are most promising and under what isoform, tissue, and disease contexts they should be pursued for the development of SIRT‑targeted therapies in human disease.

Inducing lysosomal membrane permeabilization (LMP) represents an alternative therapeutic strategy that bypasses conventional genetic or protein targets and triggers cell death by physically tumor disrupting. Herein, QS-21, an FDA-approved saponin with membrane-disruptive capability was loaded onto an aluminum-based adjuvant (denoted as QS-NanoAlum) to induce LMP effects and trigger tumor immunogenic cell death (ICD) through a "self-lysis" mechanism. Leveraging its positive surface charge, intratumorally administered QS-NanoAlum could be easily adhered to tumor cell membranes, facilitating efficient cellular internalization. Within the acidic lysosomal environment, QS-NanoAlum underwent degradation and subsequently released QS-21, leading to LMP effect and the cytosolic leakage of hydrolytic enzymes. The released hydrolases and QS-21 would further inflict collateral damage to cellular structures and organelles, including the plasma membrane, nucleus, mitochondria, and endoplasmic reticulum, which ultimately culminated in ICD. Notably, excess QS-NanoAlum could efficiently adsorb the released tumor-associated protein antigens (TAAs) during ICD, forming an antigen depot at the injection site that sustained long-term immune stimulation. Our in vivo studies also demonstrated that QS-NanoAlum significantly suppressed tumor growth and enhanced immune cell infiltration into tumor tissues. Finally, this strategy that physically disrupting the cellular "recycling center" to provoke cell disintegration and elicit robust ICD, offering a promising paradigm for cancer immunotherapy. STATEMENT OF SIGNIFICANCE: Conventional cancer therapies target specific mutations but are limited by tumor heterogeneity and resistance. We introduce a new paradigm that physically disrupts tumor cells via lysosomal membrane permeabilization (LMP), bypassing molecular targets. Our system, QS-NanoAlum (QS-21 loaded to an aluminum adjuvant), triggers immunogenic cell death through a "self-lysis" mechanism. Positively charged QS-NanoAlum is internalized and degrades in acidic lysosomes, releasing QS-21 to induce LMP. Released hydrolases cause widespread organelle damage, leading to robust immunogenic cell death. Excess QS-NanoAlum adsorbs tumor-associated antigens, forming a sustained immune depot. In vivo, QS-NanoAlum suppresses tumor growth and enhances immune infiltration, validating its potential for cancer immunotherapy.

More than ten years ago, the World Health Organization's (WHO) International Agency for Research on Cancer (IARC) published a monograph concluding there was limited evidence in experimental animals for carcinogenicity of Radio Frequency Electromagnetic Field (RF EMF). The objective of this review was to systematically evaluate the effects of RF EMF exposure on cancer in experimental animals. Eligibility criteria: Based on pre-established Populations, Exposures, Comparators, Outcomes, and Study Type (PECOS) criteria, studies in experimental animals of the following study types were included: chronic cancer bioassays, initiation-(co-)promotion studies, and studies with tumor-prone animals. MEDLINE (PubMed), Science Citation Index Expanded and Emerging Sources Citation Index (Web of Science), and the EMF Portal. Data abstraction and synthesis: Data are publicly available online as interactive visuals with downloadable metadata. We adapted the risk-of-bias (RoB) tool developed by Office of Health Assessment and Translation (OHAT) to include considerations pertinent to the evaluation of RF EMF exposure and cancer bioassays. Study sensitivity was assessed with a tool adopted from the Report on Carcinogens (RoC). We synthesized studies using a narrative approach. Effect size was calculated as the 1% Bayesian Average benchmark dose (BMD) of a respective study when dose response or a trend was identified (see Supplementary Data 3). Evidence Assessment: Certainty of the evidence (CoE) was assessed using the Grading of Recommendations, Assessment, Developing and Evaluations (GRADE) approach, as refined by OHAT. Evidence from chronic cancer bioassays was considered the most directly applicable to evaluation of carcinogenicity. We included 51 studies with 10 chronic bioassays No studies were excluded based on risk of bias concerns. Studies were not considered suitable for meta-analysis due to heterogeneity in study design, species, strain, sex, exposure characteristics, and cancer outcome. No or minimal evidence of RF EMF exposure-related cancer outcomes was found in most systems or organs in any study (these included gastrointestinal/digestive, kidney, mammary gland, urinary, endocrine, musculoskeletal, reproductive, and auditory). For lymphoma (17 studies), with 5 chronic bioassays (2,186 mice, 2,500 rats) inconsistency between two chronic bioassays was not plausibly explainable, and the CoE for lymphoma was rated 'moderate'. For brain tumors (20 studies), including 5 chronic bioassays (2,575 mice, 8,840 rats), an increase in glial cell-derived neoplasms was reported in two chronic bioassays in male rats. The CoE for an increased risk in glioma was judged as high. The BMD analysis was statistically significant for only one study, and the BMD was 4.25 (95% CI 2.70, 10.24). For neoplasms of the heart (3 chronic bioassays), 3 studies were performed in rats (∼4,304 animals), and 1 in mice (∼1,386 animals). Based on 2 bioassays, statistically significant increases in malignant schwannomas were judged as high CoE for an increase in heart schwannomas in male rats. The BMDs from the two positive studies were 1.92 (95%CI 0.71, 4.145) and 0.102 (95%CI 0.056, 0.244), respectively. Ten studies reported neoplasms in the adrenal gland (5 chronic bioassays). The CoE for an increased risk in pheochromocytoma was judged as moderate. None of these findings were dose-dependent when compared to the sham controls. Fourteen studies investigated tumors of the liver with 5 of these being chronic bioassays. The CoE was evaluated as moderate for hepatoblastomas. For neoplasms of the lung (5 chronic bioassays), 4 studies were conducted in rats (∼2,176 animals) and 9 studies in mice (∼4,171 animals). In one chronic bioassay, a statistically significant positive trend was reported for bronchio-alveolar adenoma or carcinoma (combined), which was rated as moderate CoE for an increase in lung neoplasms with some evidence from 2 initiation-(co-)promotion studies. Meta-analysis was considered inappropriate due to the heterogeneity in study methods. The GRADE/OHAT CoE framework has not been frequently applied to animal studies and experience to date suggests refinements are needed. We deferred to standard methods in environmental health where CoE is framed in the context of strength of the evidence providing positive support for carcinogenicity. High CoE can be interpreted as the true effect is highly likely to be reflected in the apparent relationship. Moderate CoE indicates the true effect may be reflected in the apparent relationship. Cancer bioassays conducted in experimental animals are commonly used to identify potential human carcinogens. We note that the two tumor types with high CoE in animals in this systematic review are the same as those identified with limited evidence in humans by the IARC Working Group. However, even in cases where the animal evidence demonstrates high CoE, the extrapolation of risk from cancer bioassays to humans is particularly complex for RF EMF. Without a better understanding of the mechanism of the carcinogenicity of RF-EMF, the choice of exposure metric for risk extrapolation (whole body versus localized), intensity or cumulative exposure whether or not a monotonic dose response holds for carcinogenic effects, and whether SAR is the appropriate dose metric for adverse effects induced by RF-EMF may be critical. This review was partially funded by the WHO radioprotection programme. The protocol for this review was registered in Prospero reg. no. CRD42021265563 and published in Environment International 2022 (Mevissen et al., 2022).

Inorganic arsenic (iAs) is a widespread environmental carcinogen; however, the mechanisms by which chronic low-dose exposure promotes bladder carcinogenesis in a sex-dependent manner remain insufficiently characterized. Analyses of patient bladder tumors revealed arsenic concentrations consistent with chronic environmental exposure, even when urinary arsenic levels were within current guideline limits. To replicate physiologically relevant exposure, male and female mice received iAs in drinking water for 12 months. This exposure induced urothelial hyperplasia, hemorrhage, inflammation, early neoplastic changes in the bladder, and dose-dependent lung injury. Arsenic speciation demonstrated significant sex differences: males accumulated higher levels of monomethylated arsenic (MMA), while females exhibited increased dimethylarsinic acid (DMA) and greater methylation capacity. Arsenic exposure reduced protective commensal populations and enriched stress-tolerant, pro-inflammatory taxa across the gut, urinary, and bladder microbiomes. In males, proliferative and DNA methylation signatures were predominant, whereas in females, oxidative, endocrine, and redox stress signatures were more pronounced. Spatial transcriptomics identified sex-specific activation of carcinogenic and immune-stromal pathways in the bladder epithelium, with increased populations of cancer-epithelial cells and macrophages. Integrated analyses of the microbiome, metabolome, and spatial transcriptome indicated that chronic arsenic exposure reprograms the gut-bladder axis in both dose- and sex-dependent manners. These findings define a mechanistic, sex-dependent gut-bladder axis through which chronic low-dose arsenic toxicity promotes premalignant bladder pathology.

During the progression from epithelial neoplasms to invasive carcinoma, uncontrolled cellular growth within a confined space generates pronounced cell crowding. Despite its prevalence, how cancer cells sense and respond to crowding during the early stages of tumor invasion remains poorly understood. Here, using a spontaneous crowding model that mimics the progressive compressive stress produced by proliferating cells, we show that cell crowding induces an invasive phenotype in cancer cells. This phenotypic switch is accompanied by a nanoscale smooth-to-corrugated topography transition (nSCTT) of the plasma membrane. By combining biophysical measurements with mechanical modeling, we demonstrate that cell crowding elevates Laplace pressure while reducing membrane tension to drive the nSCTT of the plasma membrane. nSCTT promotes tumor invasion by disrupting the aggregation of lipid raft-like domains. Finally, we demonstrate that strengthening membrane-to-cortex attachment (MCA) effectively blocks the nSCTT and suppresses tumor invasion in both cell crowding models and mouse xenograft models. Together, these findings reveal that cell crowding initiates tumor invasion through a nanoscale plasma membrane topography transition, providing a framework for understanding how mechanical forces are converted into malignant behavior at the nanoscale. STATEMENT OF SIGNIFICANCE: This study reveals how physical crowding of cancer cells triggers invasion through a previously unknown nanoscale mechanism. We discovered that crowding forces cause the cell membrane to transform from smooth to corrugated at the nanoscale. This topography change breaks apart critical signaling hubs (lipid raft domains) on the membrane, ultimately activating invasive behavior. This work provides a framework for understanding how tumors become malignant under mechanical stress and identifies reinforcing the cell membrane as a potential strategy to block cancer invasion.

Accumulating preclinical and clinical evidence identifies ketohexokinase (KHK), the initial enzyme of fructose catabolism, as a core mediator of fructose-driven pathogenesis, positioning pharmacological KHK inhibition as a promising therapeutic strategy for fructose-associated disorders. Functionally divergent KHK isoforms arise from alternative splicing. The high-activity KHK-C, predominantly expressed in the liver, kidney, and small intestine, drives fructose-related metabolic pathologies, including metabolic dysfunction-associated steatotic liver disease, diabetic kidney disease, and obesity. In contrast, the ubiquitously expressed low-activity KHK-A mediates tumor metabolic reprogramming, proliferation, and metastasis via both metabolic and non-canonical protein kinase activities. KHK expression and isoform switching are dynamically regulated by multiple upstream factors, including dietary composition, the uric acid/ChREBP transcriptional axis, hypoxia-inducible factors, and medium-chain fatty acids, with pleiotropic KHK functions extending beyond metabolic regulation to cardiac homeostasis, bone development, and neurocognitive processes. Translational development of KHK inhibitors has achieved notable progress, with the first-generation PF-06835919 completing Phase 2 trials for metabolic dysfunction-associated steatotic liver disease, type 2 diabetes, and hereditary fructose intolerance, and the second-generation LY3522348 finishing Phase 1 evaluation in healthy participants, though key gaps including isoform selectivity and tissue-specific delivery remain unaddressed. Despite growing recognition of KHK's multifaceted roles in health and disease, existing literature lacks a consolidated synthesis of its regulation, tissue-specific functions, and translational progress. This review aims to provide a systematic overview of KHK's molecular characteristics, regulatory networks, pathophysiological roles, and therapeutic potential, establishing KHK as a promising actionable target for fructose-related disorders and offering a foundational framework to guide future research and drug development.

Acute myeloid leukemia (AML) is a blood cancer with poor survival outcomes. Acute respiratory failure frequently occurs due to leukemia infiltration of the lungs. Underlying mechanisms remain unexplored and therapeutic interventions remain empiric. Here we map the AML lung microenvironment at spatial and single-cell resolution. We show that extensive remodeling is coupled with inflammation and impaired tissue integrity and function. Steroid treatment significantly reduces AML burden and lung infiltration, improving oxygenation and pulmonary function. As a mechanistic correlate, the S-type lectin Lgals9 is triggered by inflammation and mediates cell-cell interactions within infiltrated lungs. Also, the alarmin IL-33 and its receptor (IL-1RL1) are involved in cell-cell interactions within the leukemic lung microenvironment. Targeting either the Lgals9 or IL-33 axis significantly decreases overall AML burden and lung infiltration through effects on both the immune microenvironment and AML cells. Our studies delineate pulmonary infiltration phenotypes in acute leukemia, enabling new treatment strategies.

Formaldehyde, classified as a Group 1 human carcinogen by the International Agency for Research on Cancer (IARC), is continuously re-emitted from phenolic foam insulation through Post-curing reactions under ambient conditions. The combined influence of board storage period and thickness-direction position on Formaldehyde emission behavior has not been quantitatively elucidated. Specimens from the same production lot with varied board storage periods of 0-544 days (Cut-Point Series) and naturally aged samples spanning 1697-1932 days (Long-Term Series) were evaluated using three independent analytical methods, including the KS M 1998 small chamber method. The core section exhibited a non-monotonic emission pattern, peaking at the 105-day specimen (0.250 mg/m²·h) among tested time points, with a core-to-surface emission ratio reaching 9.6. Attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FT-IR) indices for methylol groups, ether bridges, and methylene bridges simultaneously peaked at the 105-day specimen, converging with the emission trend. This emission maximum diverged from the residual content maximum, which was found at the core section of the 544-day specimen (12.000 mg/L), indicating that low surface emission rates do not reflect internal Formaldehyde depletion. Two distinct aging trajectories were identified: the surface section showed oxidative resin degradation accompanied by spatial homogenization of residual content, whereas the core section retained elevated reactive group indices without signs of oxidative deterioration even after 1932 days. These results suggest that Formaldehyde exposure risk from phenolic foam insulation may persist across the full service life of the material, from installation through occupancy to demolition. All results are based on single-specimen (n = 1) measurements per condition and are intended to provide an experimental foundation for future reaction-diffusion coupled modeling.

Arsenic, commonly found as an environmental pollutant, poses significant health threats and has become a worldwide public health concern. Extended exposure to arsenic is linked to the development of multiple cancers, particularly bladder cancer. In this study, a rat bladder tumor model was established by combined administration of N-methyl-N-nitrosourea (MNU) and sodium arsenite (NaAsO₂), with MNU serving as a carcinogen and NaAsO₂ as a promoter. Additionally, we constructed arsenic-treated cell model using the SV-HUC-1 human uroepithelial cell line, which was subjected to NaAsO₂ treatment for 40 weeks. This study confirmed that arsenic promoted bladder tumorigenesis induced by MNU in rats. The expression of PLCE1 protein was significantly upregulated in arsenic-treated bladder epithelial cells, and the inhibition of PLCE1 effectively ameliorated the arsenic associated malignant phenotype. Mechanistically, the elevated protein levels of PLCE1 were jointly regulated by OTUD7B-mediated deubiquitination and SENP1-mediated deSUMOylation. Notably, SENP1-mediated deSUMOylation further promoted the deubiquitination modification of PLCE1. Functional experiments demonstrated that either the inactivation of OTUD7B enzyme activity (C194S mutation) or the knockdown of SENP1 significantly alleviated the malignant behavior of bladder epithelial cells induced by inorganic arsenic. This "dual-modification" model provides a new paradigm for studying the carcinogenic mechanisms of environmental toxicants.

The sympathetic nervous system (SNS) plays a crucial role in chronic liver diseases, but its function, particularly in terms of the underlying mechanisms of liver metastasis, remains unclear. Here, we reported that hepatic sympathetic nerve activity increases in the early stage of metastasis and was accompanied by a moderate increase in norepinephrine (NE). Unexpectedly, NE had a dose-dependent effect on colorectal liver metastasis. Only daily administration of low-dose NE could maintain NE at a moderate concentration in the liver during this early phase significantly suppressed metastatic tumor growth. Conversely, extremely low NE concentrations (induced by 6-hydroxydopamine, 6-OHDA) or high-dose NE prompted liver metastasis. Single-cell RNA sequencing (scRNA-seq) demonstrated that NE modulated monocyte recruitment and M2 macrophage polarization through the CCL2-CCR2 signaling axis and influenced the immunosuppressive effects of myeloid-derived suppressor cells (MDSCs) on T cells: low-dose NE inhibited monocyte recruitment and M2 macrophage polarization and relieved the immunosuppressive effects of MDSCs on T cells, whereas extremely low- and high-dose NE supplementation had the opposite effects. Further investigations confirmed that these biological effects were closely associated with the NE-mediated expression ratio of α2a-adrenergic receptor to β2-adrenergic receptor (α2a-AR/β2-AR). Specifically, at moderate NE concentrations, a high α2a-AR/β2-AR expression ratio was correlated with antitumor effects. Collectively, these findings reveal that the SNS mediates U-shaped effects on liver metastasis in an α2a-AR/β2-AR ratio-dependent manner, highlighting moderate sympathetic activation as a potential therapeutic strategy to inhibit the progression of liver metastasis.

Current disease-modifying therapies for multiple sclerosis (MS) primarily target peripheral immune responses but exhibit limited efficacy in mitigating the compartmentalized neuroinflammation driven by central nervous system (CNS)-resident microglia. By integrating clinical sample analysis with experimental autoimmune encephalomyelitis (EAE) model studies, we have demonstrated that the proviral integration site for Moloney murine leukemia virus 1 (PIM1) is significantly upregulated, particularly in microglia, in both MS patients and the spinal cords of EAE mice. This upregulation positively correlates with disease severity and levels of proinflammatory cytokines such as IL-1β, TNF-α, and IL-6. Utilizing a multimodal research approach-including pharmacological inhibition (SMI-4a), genetic knockdown, RNA sequencing, and HIS-SIM super-resolution imaging-we confirmed that PIM1 inhibition effectively attenuates neuroinflammatory responses, improves clinical symptoms in EAE mice, and promotes activation of the mitophagy pathway while suppressing inflammation-related molecules. Mechanistically, PIM1 enhances the phosphorylation of dynamin-related protein 1 (Drp1) at Ser616 while suppressing its phosphorylation at Ser637, which disrupts LC3-mitochondria colocalization and autophagosome-lysosome fusion. This leads to mitophagy dysfunction, loss of mitochondrial membrane potential, and accumulation of reactive oxygen species. Notably, the combined administration of PIM1 and Drp1 inhibitors did not yield synergistic therapeutic effects, suggesting that PIM1 likely functions as an upstream master regulator of Drp1. These findings not only elucidate the molecular mechanism by which PIM1 interacts with Drp1 to regulate microglial activation and mitophagy but also establish PIM1 as a promising CNS-intrinsic therapeutic target for restoring mitochondrial homeostasis in MS.

Microplastics (MPs) are ubiquitous across environments including the Qinghai-Tibet Plateau. Most existing MPs studies focus on aquatic animals and rodents, while MPs influences on yaks (Bos grunniens) remain poorly understood. Using yaks as animal models, we combined metagenomics and metabolomics to explore short-term polystyrene-MPs (PS-MPs) impacts on ruminal microbiota, metabolism, antioxidant capacity and inflammation. Seven-day PS-MPs exposure reshaped rumen microbiota and elevated β-diversity. Four KEGG pathways (peptidoglycan synthesis, vitamin B6/riboflavin metabolism, terpenoid backbone biosynthesis) were enriched alongside altered extracellular polysaccharides composition. Serum metabolomics revealed elevated L-glutamine and indole-3-propionic acid, coupled with reduced 2-C-methyl-D-erythritol 2,4-cyclodiphosphate and indole-3-lactic acid post-exposure. Urinary metabolomics revealed decreased D-erythrose 4-phosphate, dimethyl allyl pyrophosphate, and 2-C-methyl-D-erythritol 2,4-cyclodiphosphate, collectively indicating inhibited terpenoid backbone biosynthesis in yaks. Additionally, PS-MPs triggered inflammatory responses, evidenced by elevated levels of pro-inflammatory cytokines (interferon-γ, interleukin-1β, interleukin-6, interleukin-17, interleukin-22, tumor necrosis factor-α, transforming growth factor-α), yet antioxidant function indexes (total antioxidant capacity, superoxide dismutase, glutathione peroxidase, catalase and malondialdehyde) showed no significant changes. Multi-omics suggested Prevotella ruminicola may help resist PS-MPs invasion. In summary, rumen microbes may alleviate PS-MPs adverse effects, explaining yaks' mild responses to short-term PS-MPs exposure. Long-term MPs effects, tissue deposition and related molecular mechanisms warrant further study.

Rapid industrialization in developing countries often conceals complex environmental and health hazards behind measured total VOC concentration compliance. To address this issue, a "whole-process tracking" campaign for volatile organic compounds (VOCs) was conducted across 24 enterprises in 12 sectors in the Yangtze River Delta（YRD）, China, using synchronized sampling at workshop, outlet, and boundary nodes within each facility. Measurements of 115 VOC species explicitly quantified under-characterized risk signals associated with conventional abatement. First, regarding environmental impacts, a pronounced mass-reactivity decoupling phenomenon was observed. Although thermal oxidizers (e.g., RTOs) are designed for high nominal VOC removal, species-resolved concentration fingerprints showed a 1-2 order-of-magnitude increase in the fractional abundance of small highly reactive unsaturated species under specific high-temperature abatement conditions. Consequently, moderate-concentration outlets (e.g., the Synthetic Leather sector) exhibited a Specific Reactivity (SR) index up to six times higher than bulk-emitting sectors. Second, concerning occupational health, the phenomenon of "regrettable substitutions" was explicitly quantified. Compliance-driven source substitutions in small and medium-sized enterprises (SMEs) reduced measured total VOC concentration by 93%, yet were associated with a 20% higher calculated carcinogenic-risk estimate due to the introduction of unlisted, highly toxic alternatives. Using a reverse dosimetry model, we showed that while screening-level PPE scenarios may extend the carcinogenic safe working duration (EDsafe), massive fugitive emissions of substitute solvents consistently drive cumulative non-carcinogenic hazards above the safety threshold (HI>1.0). Under the HI-based non-cancer safety constraint, this exceedance substantially reduced the integrated EDsafe, indicating that nominal PPE scenarios may not fully offset source-level toxicity in these scenarios. These findings provide regional field evidence from key industries in the YRD suggesting that future VOC management may benefit from supplementing concentration-based compliance with species-specific reactivity, toxicity, and whole-process control considerations.

Prostate cancer is among the top causes of cancer-related mortality in men, according to the latest data from GLOBOCAN, driven primarily by androgen receptor signaling and initially treated with androgen deprivation therapy. However, most patients progress to a resistant state characterized by mechanisms such as receptor amplification, mutations, splice variants, and activation of alternative pathways. This review summarizes the key molecular drivers of resistance and evaluates emerging invasive biomarkers with potential clinical utility, though most remain insufficiently validated. It also outlines current and emerging therapeutic strategies to overcome resistance, highlighting ongoing challenges in clinical implementation.

The therapeutic success of biomacromolecular drugs is fundamentally limited by their poor bioavailability, particularly in hard-to-deliver cells and poorly permeable tissues. To address these challenges, we developed virus-inspired lipopeptide vectors that mimic viral envelopes to enable receptor-mediated endocytosis, and these vectors expose arginine-rich coronas in response to tumor-specific stimuli for direct membrane penetration. The virus-inspired enveloped lipopeptide vectors exhibited remarkable versatility for encapsulating diverse biomacromolecules (including nucleic acids, proteins, and supramolecular assemblies) and achieved efficient intracellular delivery. Furthermore, enveloped lipopeptide vectors facilitated transcytosis and subsequent intercellular transfer, thereby enhancing deep tumor penetration. This work presents a robust virus-inspired delivery platform with enhanced cargo encapsulation and delivery performance, providing a promising strategy to surmount physiological and pathological barriers in biomacromolecule delivery. STATEMENT OF SIGNIFICANCE: Biomacromolecular therapeutics such as nucleic acids and proteins hold great promise, yet their clinical translation is limited by rapid degradation, poor tumor penetration, and inefficient cellular uptake. Inspired by natural enveloped viruses, we developed virus-mimetic enveloped lipopeptide nanoparticles that integrate two complementary entry mechanisms: hyaluronic acid-guided, receptor-mediated endocytosis and tumor microenvironment-activated membrane penetration through exposure of an arginine-rich corona. This dual-entry strategy, combined with transcytosis-enabled intercellular transfer, enables deep tumor penetration and efficient intracellular delivery of diverse biomacromolecules. By recapitulating key principles of viral infection in a synthetic, programmable system, this work establishes a broadly adaptable platform to enhance the bioavailability and therapeutic efficacy of large biologics, offering a new conceptual framework for biomacromolecular drug delivery.

Nanoplastics (NPs) are emerging environmental contaminants capable of interacting with biological systems, yet their effects on intercellular communication in the bone marrow (BM) microenvironment remain unclear. This study investigated the impact of polystyrene NPs on extracellular vesicle (EV) biogenesis, cargo, and function using in vitro models of multiple myeloma (MM) and stromal cells, supported by ex vivo murine bone and human BM aspirates. Under the experimental conditions used in this study, NPs were efficiently internalized by both malignant and stromal cells, inducing oxidative stress and reducing cell viability. NP exposure altered EV biogenesis, leading to increased vesicle release, reduced size, and the incorporation of NPs into a subset of EVs. Additionally, EVs from NP-exposed MM cells showed decreased total miRNA content and modulation of miRNA sorting-related proteins. Among the analyzed miRNAs, miR-505, a candidate linked to osteogenic regulation, was reduced in both cells and EVs following NP exposure. Functionally, EVs from control MM cells downregulated osteogenic markers in pre-osteoblasts, whereas EVs from NP-exposed cells showed reduced regulatory effects. These findings demonstrate that NP exposure modifies EV-mediated signaling under experimental conditions, providing mechanistic insight into nanoplastic interference with BM cellular communication.

Whole Slide Imaging (WSI) has become a gold standard in cancer diagnosis, inspecting multi-scale information from cellular to tissue levels. Processing an entire WSI directly is infeasible due to GPU memory constraints; thus, Multiple Instance Learning (MIL) has emerged as the standard solution by partitioning WSIs into tiles. While recent two-stage MIL frameworks partially achieve memory efficiency by decoupling tile-level extraction from slide-level modeling, they still face four limitations: 1) the conflict between training throughput and inference memory efficiency, 2) the high susceptibility to overfitting on small-scale WSI datasets with sparse supervision, 3) the disruption of spatial structural integrity during sampling-based training, and 4) the inadequate modeling of multi-scale feature interactions within long sequences. We therefore introduce PathRWKV, a novel State Space Model designed for efficient and robust WSI analysis. To resolve the computational trade-off, we propose an asymmetric structure utilizing max pooling aggregation, enabling parallelized training for high throughput and recurrent inference with constant ( $\mathcal {O}\text {(}{1}\text {)}$ ) memory complexity. To mitigate overfitting, we employ random sampling to enhance data diversity, with a multi-task learning module to regularize feature learning on limited data. To restore spatial context, we introduce 2D sinusoidal position encoding to perceive the relative locations of tissue tiles. To capture comprehensive representations, we integrate TimeMix and ChannelMix modules, enabling dynamic multi-scale feature modeling across temporal and spatial dimensions. Experiments on 29,073 WSIs across 11 datasets demonstrate that PathRWKV outperforms 11 state-of-the-art methods on 10 datasets, establishing it as a scalable and solution with application potential.

Skeletally remoulded analogues of natural products enrich compound libraries, accelerating hit identification and drug discovery. Here we present a divergent alkaloid remodelling strategy that is centred on isolable platforms generated by Hofmann elimination. Piperidine deconstruction is thus effected through site-selective cleavage of a C-N σ-bond concomitant with the formation of a second alkene moiety. Ensuing alterations involving Stevens and/or Meisenheimer rearrangements and reconstruction by ring-closing metathesis or intramolecular hydroamination deliver expanded and contracted frameworks, respectively. By using a small selection of readily accessible catalysts and reagents, we prepared 26 altered frameworks in a total of 48 steps (~1.8 steps per reshaped structure). The utility of the approach is highlighted by the discovery of a reverse-Meisenheimer rearrangement and observation of a rare case of anionic [1,6]-sigmatropic rearrangement. In cellulo studies involving ten cancer cell lines reveal that an expanded amine-bridged and a deconstructed tricyclic framework exhibit micromolar and nanomolar activity, respectively.

Two-dimensional cell culture models have long been a cornerstone of biomedical research; however, they often fail to accurately replicate the in vivo environment. In recent years, three-dimensional (3D) cell cultures, particularly 3D spheroid models, have gained recognition for their ability to better mimic the complexities of the in vivo environment, making them valuable tools for studying cellular behavior and responses. Tumor spheroids, in particular, have significant applications in anticancer therapy evaluation, providing a more physiologically relevant model by simulating the spatial architecture and microenvironment of tumors. However, due to the limitations imposed by optical diffraction and background noise in 3D imaging, traditional imaging methods are unable to accurately resolve the growth, morphological changes, and drug responses of tumor spheroids. To address this issue, super-resolution imaging technologies have emerged. Structured illumination microscopy (SIM) combined with reconstruction algorithms can effectively enhance resolution, but challenges such as limited light penetration of single-photon imaging and high background noise remain in 3D imaging. In this paper, an advanced SIM technology with large depth and low noise 3D imaging capability is developed. This study introduces a novel frequency-specific denoising method (FSDM) to effectively reduce noise through adjusting the weights of high-frequency signals to preserve image details. The FSDM optimization significantly reduces background interference from deeper tissue layers, improving image details and the overall quality of 3D imaging. For the first time, scanning SIM is integrated with two-photon microscopy (TPEF-SIM) for 3D imaging, leveraging the strengths of both techniques to enhance resolution and overcome light penetration limitations.

Cancer-associated fibroblasts are major architects of the tumour stroma, where their aligned, elongated morphology forms a capsule that mechanically restrains tumour expansion. However, it is unclear how this supracellular organization emerges and persists. Here we show that fibroblasts generate a fibronectin matrix that progressively acquires the same nematic order as the cell layer, and that this matrix in turn feeds back to immobilize both cells and topological defects. Using long-term live imaging, traction force microscopy, matrix microfabrication and hydrodynamic modelling, we find that this reciprocal coupling induces an ageing process in which cellular flows and defect motion slow dramatically and ultimately freeze. Despite this arrest, the monolayer remains active, with defects concentrating contractile forces that may represent mechanical weak points. Disrupting fibronectin production fluidizes the capsule, reactivates defect dynamics and compromises its barrier-like function. These findings reveal a self-organizing mechanism by which fibroblasts and their matrix co-evolve to create a mechanically stable, yet active, stromal architecture with direct implications for tumour dissemination.

The identification of reliable, non-invasive biomarkers that reflect molecular alterations in deep and inaccessible tissues remains a major challenge in translational medicine. In this context, peripheral blood cells have emerged as valuable surrogates of tissue-level pathophysiology. Among molecular systems expressed in circulating cells, adenosine receptors (ARs) represent particularly attractive candidates due to their central role in regulating inflammation, immune responses, vascular function, and tissue homeostasis. This review focuses on the expression and functional modulation of AR subtypes in peripheral blood mononuclear cells and platelets, highlighting their potential as translational biomarkers across a wide spectrum of human diseases. Accumulating evidence demonstrates that disease-associated alterations of A2A and A₃ ARs in circulating cells frequently mirror receptor remodeling occurring in the brain, cardiovascular system, inflamed tissues, and tumors. In neurodegenerative and cardiovascular disorders, peripheral A2AAR dysregulation reflects central or myocardial changes and correlates with disease severity and progression. In contrast, the A₃AR shows a remarkable and consistent parallel overexpression in both pathological tissues and circulating immune cells in cancer and chronic inflammatory diseases, supporting its value as a circulating biomarker of systemic pathologies. Beyond their diagnostic and prognostic relevance, ARs also represent actionable pharmacological targets, bridging biomarker discovery with therapeutic intervention. Collectively, this evidence positions peripheral ARs profiling as a potential tool for non-invasive disease monitoring and for guiding personalized translational pharmacology strategies.

IR-780 is a lipophilic heptamethine cyanine dye that exhibits strong absorption and fluorescence emission in the near-infrared (NIR) region (700-900 nm). Similar to other cyanine dyes, IR-780 is emerging as a promising fluorophore for fluorescence-guided surgery (FGS). In addition, due to its high molar extinction coefficient, IR-780 can generate reactive oxygen species (ROS), enable efficient conversion of light into heat, and impart additional phototherapeutic capabilities. Consequently, IR-780 represents a promising "all-in-one" single molecule for FGS and phototherapy applications. However, its use is limited by poor aqueous solubility, low photostability, and rapid clearance, which restrict its bioavailability and reproducibility in clinical contexts. Recent strategies to address these drawbacks include both structural modification of IR-780 and nanoparticle (NP)-based delivery approaches. NP encapsulation using lipid-based, polymeric, inorganic, or biomimetic carriers has proven effective in enhancing aqueous solubility and photostability, reducing off-target cytotoxicity, and thereby improving imaging and therapeutic precision. This review systematically compares free and NP-encapsulated IR-780 and its derivatives, highlighting key differences in photophysical and physicochemical properties, in vitro and in vivo outcomes, and NP design considerations. Overall, while the free dye allows straightforward administration, NP formulations provide superior photophysical performance, improved biodistribution control and greater tumor retention. However, they remain limited by challenges in large-scale translation and regulatory approval.

Interpenetrating polymer network (IPN) microspheres composed of alginate and thiolated TEMPO-oxidized cellulose nanofibers (Th-TCNF) were developed as reduction-responsive carriers for Mitoxantrone (MIT) delivery. TCNF was thiolated through covalent coupling with cysteamine, as confirmed by H NMR, FT-IR and XPS which successfully verified thiol grafting. Spray-dried microspheres were subsequently cross-linked via Ca2+ coordination and disulfide bond formation, generating a mechanically reinforced IPN structure. Swelling analysis showed significantly reduced equilibrium swelling in the full IPN compared with single-network controls, indicating restricted polymer mobility and enhanced structural stability. SEM revealed progressively denser surfaces with increasing Th-TCNF content, while TGA demonstrated improved thermal resistance. XPS of the microspheres confirmed reduction-induced disulfide cleavage, supporting redox-triggered release. FITC-dextran and MIT release exhibited reduction-dependent, composition-controlled behaviour, and flow cytometry (FACS) showed sustained intracellular MIT accumulation in MDA-MB-231 cells, leading to prolonged cytotoxicity. In vivo intratumoral administration in BALB/c nude mice bearing MDA-MB-231 xenografts produced significant tumor suppression with extensive necrosis and apoptosis, without systemic toxicity, verified by Histological (H&E) and TUNEL staining. Collectively, these findings highlight alginate: Th-TCNF IPN microspheres as a promising localized, reduction-responsive platform for enhancing chemotherapeutic efficacy while minimizing systemic toxicity.

Cancer transcends a local disease by engaging in systemic crosstalk with the host's physiology. This review redefines the vagus nerve as a central bioelectric processor within a dynamic brain-body-tumor homeostatic network, orchestrating a complex biological dialogue between them. We synthesize evidence illustrating its specialized architecture for encoding tumor microenvironment signals across metabolic, immune, and endocrine interfaces. Functioning as a dynamic, context-dependent integrator, it maintains systemic homeostasis while paradoxically being susceptible to hijacking by tumors, underscoring its dual role as both guardian and accomplice in cancer progression. This mechanistic framework paves the way for novel therapeutic reprogramming strategies. We explore the potential of precision bioelectronic neuromodulation, pharmacological interception of neural signaling, and behavioral interventions to "hack" this dialogue, aiming to enhance anti-tumor defenses, alleviate cancer-related symptoms, and synergize with conventional therapies. Future success hinges on deciphering the vagal neural code and developing personalized, ethically sound applications.

Although histologically benign, invasive pituitary adenomas exhibit malignant-like biological behavior, characterized by local invasiveness and high proliferative activity. Clinical management remains challenging due to the absence of effective targeted therapies, difficulties with surgical resection, and high risks of postoperative residual disease and recurrence. Precise therapeutic targets are urgently needed. To investigate molecular mechanisms underlying pituitary adenoma invasion and identify potential therapeutic targets, we collected 216 whole transcriptome sequencing datasets representing all pituitary adenoma lineages, along with several single-cell transcriptome datasets. The Scissor algorithm was utilized to identify invasion-associated cell subpopulations. Single-cell weighted gene co-expression network analysis delineated the underlying gene regulatory network. Differential gene expression and pathway enrichment analyses explored NNAT-MAPK contributions to pituitary adenoma invasion. Findings were validated through in vitro and in vivo experiments (quantitative polymerase chain reaction, CCK-8, colony formation, Transwell assays, western blotting, immunohistochemistry, and multiplex immunofluorescence). Multi-transcriptomics analysis comprehensively characterized the transcriptomic landscape of invasive pituitary adenomas and revealed active RNA interactions within these tumors, particularly between mRNAs and long non-coding RNAs. Specific cell subpopulations associated with invasiveness were confirmed at the single-cell level. Analysis of phenotype-associated gene regulatory networks within these subpopulations identified the NNAT-MAPK-p38 axis, which promotes epithelial-mesenchymal transition in pituitary adenoma cells and drives invasion. These findings enhance understanding of the mechanisms underlying pituitary adenoma invasiveness and identify NNAT as a candidate for targeted therapy development, offering potential precision treatment strategies for patients resistant to conventional therapies or experiencing recurrent disease after surgery.

In cancer immunotherapy, peptide-major histocompatibility complexes are increasingly exploited as precise therapeutic targets. For instance, T-cell receptor (TCR)-engineered T cells, peptide vaccines, and TCR-mimic antibodies leverage the direct recognition of tumor-specific or tumor-associated antigens presented on human leukocyte antigen (HLA) molecules, offering highly specific and effective approaches to eliminate malignant cells. Classical immunopeptidomic studies rely on tumor tissue samples to identify targetable peptides; however, tissue accessibility and tumor heterogeneity remain major limitations. Liquid biopsy, including circulating tumor cells, extracellular vesicles, and cell-free nucleic acids, represents a minimally invasive alternative to capture tumor-derived material. Recent methodological adaptations allow applying immunopeptidomics to these circulating biomarkers to profile HLA-bound peptides in blood. Integrating immunopeptidomics with liquid biopsy could expand the scope of precision oncology, providing dynamic, patient-specific insights into antigen presentation for developing and monitoring personalized immunotherapies in a minimally invasive manner.

Drug-device combinations (DDCs) have evolved from simple drug-coated implants into sophisticated intelligent platforms capable of real-time sensing, adaptive drug delivery, and closed-loop therapeutic control. This transformation is driven by converging advances in nanotechnology, biomaterials, controlled release mechanisms, wireless communication, artificial intelligence, and additive manufacturing. This review first systematically compares the divergent regulatory frameworks governing DDCs in the United States, European Union, and China. It then examines key enabling technologies and their integration into implantable, wearable, oral, and next-generation DDC platforms, emphasizing how functional convergence enables transition from passive delivery to feedback-controlled, patient-adaptive systems. Clinical applications are evaluated across therapeutic areas including ophthalmology, orthopedics, cardiovascular diseases, diabetes, oncology, and neurology, with attention to mechanistic distinctions and translational status. Finally, the review analyzes persistent technical, regulatory, and translational barriers. By connecting regulatory considerations, technological foundations, and clinical applications, this review offers a coherent roadmap for the next generation of intelligent DDCs.

Redox dysregulation is a hallmark of numerous pathological conditions, including cancer, chronic inflammatory diseases, fibrosis, neurodegenerative disorders and cardiovascular diseases, where aberrant production of reactive oxygen, nitrogen, sulfur and carbonyl species disrupts cellular signalling, metabolism, immune responses and tissue homeostasis. Despite its central role in disease progression, clinical assessment and therapeutic modulation of redox imbalance remain largely indirect, non-specific, failing to adequately capture the spatially heterogeneous and highly dynamic nature of redox dysregulation. Redox-responsive nanosystems offers the potential to bridge this gap by incorporating redox-sensitive materials that can spatiotemporally sense and respond to disease-associated oxidative or reductive microenvironments. Through predictable chemical or structural transformations triggered by endogenous redox cues, these nanoplatforms enable site-specific imaging, on-demand drug release, catalytic therapy and signal amplification. These capabilities position redox-responsive nanomedicine as a powerful approach for enhancing therapeutic precision while minimising off-target toxicity. This review provides a comprehensive and mechanism-informed overview of redox-responsive nanosystems for advanced drug delivery and precision medicine. We systematically connect disease-associated redox mechanisms with nanoplatform design strategies and functional outcomes across diagnostic, therapeutic and combined theranostic applications. In addition, we critically discuss key translational challenges, including biosafety, pharmacokinetics, large-scale manufacturing and regulatory considerations. Finally, we outline future directions toward clinically translatable, redox-guided nanomedicine.

Therapy resistance is a major limitation in therapeutic efficacy for glioblastoma (GBM) patients, positioning GBM among the deadliest tumor types. In this work, we have dissected resistance mechanisms in GBM, which resulted in the identification of FGFR1 pathway as a major controller of the signaling and metabolic rewiring associated to temozolomide (TMZ) resistance. Hence, in FGFR1-positive, p53 WT GBM cells, FGFR1 controls a p53-mediated cell cycle arrest to allow DNA repair in response to TMZ. FGFR1 also regulates a complete metabolic rewiring promoting lipid catabolism and preventing lipid peroxidation. Indeed, FGFR1 inhibition completely abolishes this signaling and metabolic reprograming, restoring sensitivity to TMZ. Our results also indicated a correlation of FGFR1 with poor prognosis in GBM patients, and validated the dual treatment with TMZ and FGFR1 inhibitors as an efficient strategy to induce tumor cell death in FGFR1-positive, p53 WT pre-clinical animal GBM models. These data position FGFR1 as a promising candidate for future clinical evaluation to limit therapy resistance to TMZ in FGFR1-positive GBM patients.

Enhanced cancer prevalence findings restricted to high, cytotoxic dose levels in long-term animal cancer studies are generally assumed to be a consequence of-indirect or secondary-mutational effects, e.g. DNA damage mediated by generation of reactive oxygen species (ROS) or replication errors occurring during regenerative cell proliferation. An alternative explanation is provided by recent findings suggesting that cell lysis caused by cytotoxic doses of non-genotoxic agents may give rise to tumor promotion by selective growth stimulation of pre-existing (already mutated) dormant tumor precursor cells. This growth stimulation is assumed to be mediated by damage-associated molecular pattern (DAMP) signaling ("sterile inflammation"). Here, we discuss the differing views on cancer findings observed only at high cytotoxic doses in particular with respect to the implications for the risk assessment of the agents.

Functionalisation of carbon nanotubes (CNTs), to make them more hydrophilic, is an essential step in their preparation for use in a wide range of industrial processes. The toxicological impact of these modifications has been investigated. However, due to the diversity of experimental models used in studies, no consensus has yet been reached. Here, we studied the toxicity of multi-walled CNT (MWCNT) with or without hydroxyl or carboxyl functional groups in lung cell models-chosen due to the risk of inhalation exposure for employees during handling of nanomaterials. In this study, we investigated the induction of the epithelial-mesenchymal transition (EMT) of human bronchial epithelial cells BEAS-2B following 6 weeks of treatment with MWCNTs ranging from 0.125 to 1 µg/cm2. Epithelial and mesenchymal markers were modified following MWCNT treatment, confirming the induction of the EMT. The effect was more pronounced with non-functionalised MWCNTs than with functionalised ones. In the lung carcinoma cell line A549, IL-8 expression was measured following treatment for 24 h to 0.5-10 µg/cm2 MWCNTs. An equal to or lower level was observed after treatment with functionalised MWCNTs than non-functionalised MWCNTs. Finally, the rat alveolar macrophage cell line NR8383 were exposed to 0.125-10 µg/cm2 to MWCNTs for 24 h. Non-functionalised MWCNTs led to a higher proportion of binucleation than exposure to their functionalised counterparts. Functionalised MWCNTs induce toxicity in lung cell lines to a lower extent than non-functionalised MWCNTs. Because of this toxicity, all MWCNTs should be handled with care regardless of their functionalisation.

Androgen receptor (AR) pathway inhibitors (ARPIs) improve outcomes in advanced prostate cancer (PC) in combination with androgen deprivation therapy (ADT). However, PC rapidly develops ARPI resistance, frequently through expression of truncated AR variants (AR-Vs), like AR-V7, highlighting a need for more effective therapies. The sodium-glucose co-transporter 2 inhibitor (SGLT2i) canagliflozin, an approved diabetes drug, also suppresses PC growth and inhibits AR-related gene expression. Therefore, we hypothesized that canagliflozin may directly inhibit AR. Cellular and tumor models of PC were subjected to proliferation, clonogenic, and xenograft studies. RNA-seq and siRNA knockdown approaches defined molecular mechanisms. Molecular docking, thermal shift, and surface plasmon resonance assays assessed drug-target interactions. Stable sh-AR full-length (sh-AR-FL) and sh-AR-V7 cell lines were generated to interrogate the transcriptomic impact of AR and prognostic analysis was performed using clinical datasets. We found that canagliflozin suppresses PC growth through AR. It interacts with the AR ligand binding domain (LBD) with estimated affinity comparable to ARPIs and blocks AR signaling. Canagliflozin reduces the transcript and protein levels of the HSP70 chaperone and suppresses the cytoplasmic and nuclear levels of AR-FL and AR-Vs through proteasomal degradation. It mediates substantial reprogramming of PC transcriptional activity, including inhibition of AR pathway, cell-cycle, E2F and MYC hallmark targets. Its gene expression profile overlaps with silencing AR-FL or AR-V7 is associated with improved prognosis in clinical datasets. The results of this study demonstrate the potential for canagliflozin to function as a clinically useful ARPI and support prospective clinical investigation of this drug in PC.

Advances in organoid technology have transformed how gliomas are modeled and studied. Recent FDA and NIH initiatives further promote human-relevant organoid platforms for preclinical research. Glioma organoids can be broadly categorized into 3 main classes: Engineered organoids, which enable controlled modeling of gliomagenesis driven by specific mutations; patient-derived organoids, which preserve key molecular and histopathological features of the original tumors; and assembloids, which are designed to model tumor-microenvironment interactions. Together, these systems provide a human cell-relevant framework for investigating glioma biology, tumor-microenvironment crosstalk, and therapeutic responses and resistance. In this review, we provide a comprehensive overview of the spectrum of glioma organoid models, recognizing that different systems offer distinct and complementary strengths, and we offer practical guidance for selecting appropriate models and analytical readouts based on specific basic and translational research objectives. To address increasing methodological heterogeneity and fragmented terminology, we propose a foundational nomenclature framework for glioma organoid models to improve clarity and communication within the field. We highlight applications in technically challenging subtypes, including isocitrate dehydrogenase (IDH)-mutant gliomas and diffuse midline gliomas (DMGs). We also discuss key challenges-including scalability, standardization, microenvironment fidelity, and vascularization-and emerging innovations addressing these limitations. Finally, we call for greater collaboration and standardization within the glioma organoid community to accelerate the integration of organoid models into translational pipelines to redefine and refine preclinical modeling in neuro-oncology.

Gene therapy represents a promising strategy for treating a range of diseases. Non-viral gene delivery systems, including lipid nanoparticles (LNPs), polymer micelles, and liposomes, enable the tissue-specific delivery of genetic sequences and promote the expression of functional proteins in target cells. Compared with traditional carriers, LNPs possess distinct, mechanistically supported advantages: high biosafety with less than 5% cytotoxicity in most primary cells and minimal systemic inflammation in preclinical models, good reproducibility with a coefficient of variation less than 10% for particle size and zeta potential, and efficient delivery of nucleic acids such as DNA, mRNA, and siRNA with transfection efficiencies comparable to those of viral vectors. This review summarizes recent advances in LNP-based delivery platforms, with a focus on their structure-activity relationships and the underlying mechanisms of nucleic acid delivery. It further discusses key challenges in clinical translation, including limited targeting specificity, concerns regarding long-term biocompatibility, difficulties in manufacturing scale-up, and regulatory hurdles. Additionally, the article highlights applications in oncology, for instance the delivery of tumor-suppressor genes, mRNA vaccines, and siRNA-mediated oncogene silencing, as well as applications in other therapeutic areas. Notably, we examine pioneering strategies designed to overcome these limitations, such as selective organ targeting (SORT) nanoparticles and biodegradable ionizable lipids including C12-200, which enhance tissue-specific delivery while reducing inflammatory responses. By integrating insights from lipid chemistry, formulation science, and translational data, this review provides a forward-looking perspective on the role of LNPs in cancer immunotherapy and regenerative medicine, while helping establish a framework for the design and optimization of next-generation nucleic acid delivery systems.

The limited electrocatalytic activity of graphitic carbon nitride (C3N4) constrains its application in electrochemical sensing. Herein, Fe-doped C3N4 (Fe-C3N4) featuring atomically dispersed Fe-N5 sites was synthesized via a one-step solid-state thermal polycondensation approach. Comprehensive structural characterization, including transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and aberration-corrected electron microscopy, confirms both the structural integrity of C3N4 and the atomic dispersion of Fe species. Additionally, synchrotron radiation-based X-ray absorption spectroscopy verifies the Fe-N5 coordination environment. Density functional theory calculations demonstrate that the Fe-N5 sites modulate the local electronic structure, enhance sulfide adsorption, and decrease the limiting free-energy step for hydrogen sulfide (H2S) oxidation. As a result, the Fe-C3N4 electrode exhibits a significantly enhanced anodic response toward H2S, along with excellent selectivity against common biological interferents. Furthermore, reliable electrochemical signals are obtained in bone metastasis tissue homogenates, enabling effective discrimination between tumor and normal tissues. This work presents a scalable strategy for the design of Fe-N5 single-atom catalysts for electrochemical sensing in complex biological environments.

Biomimetic and biocompatible metal-organic frameworks (MOFs) have emerged as versatile platforms for medical applications, enabling drug delivery, bioimaging, biosensing, and catalytic therapy across cancer, infection, wound healing, and neurological disorders. Unlike conventional nanocarriers, MOFs combine modular coordination chemistry with tunable porosity, allowing their structure, surface identity, and degradation behavior to be engineered for specific biological environments. In this review, we critically discuss recent advances in biomimetic MOFs that emulate biological functions such as immune evasion, selective recognition, enzymatic activity, and barrier transport, alongside biocompatible MOFs designed for controlled stability and safe in vivo fate. We highlight how framework composition, surface modification, and physicochemical properties shape interactions with proteins, cells, and tissues, ultimately determining therapeutic performance. Finally, we discuss key challenges limiting clinical translation, including incomplete understanding of long-term biosafety, variable biodegradation pathways, targeting efficiency, and manufacturing reproducibility. Together, this review provides an updated and structured perspective on how biomimetic and biocompatible MOF design can be aligned with biological constraints to advance clinically relevant medical technologies.

Neural functional impairment following stroke is strongly linked to the loss of white matter (WM) integrity, a process critically dependent on the successful differentiation of oligodendrocyte precursor cells (OPCs). Given that hyperhomocysteine (HHcy) aggravates stroke prognosis, we hypothesized that it impairs OPCs differentiation during the recovery period. Using in vivo (MCAO) and in vitro (OGD/R) models, we showed that HHcy hinders functional recovery, impairs OPC differentiation, and compromises WM integrity. Mechanistically, HHcy acts by upregulating tumor necrosis factor-α (TNF-α), which subsequently promotes the nuclear translocation of protein arginine deiminase 4 (PAD4). This leads to the upregulation of nucleosomal citrullinated histone 3 (CitH3) and the subsequent downregulation of myelin regulatory factor (MyRF), resulting in the observed inhibition of OPCs differentiation. Crucially, pharmacological inhibition of PAD4 using the pharmacological inhibitor YW3-56 effectively promoted OPCs differentiation and enhanced WM repair after ischemic stroke. Therefore, our findings identify the TNF-α/PAD4 pathway as a novel therapeutic target for reversing HHcy-induced white matter impairment and improving neurological outcomes after stroke.

Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease characterized by cartilage degradation and synovial inflammation with limited therapeutic options. In our study, cell-free fat extract (CEFFE), a bioactive fraction derived from adipose tissue, is evaluated as a therapeutic strategy for TMJOA. In chondrocytes, CEFFE enhances proliferation and viability, attenuates inflammatory apoptosis, and preserves extracellular matrix homeostasis by upregulating Col2a1 and Sox9 while suppressing matrix metalloproteinases. Integrated transcriptomic and epigenomic analyses demonstrate that CEFFE reduces the activating transcription factor 3 (ATF3) binding at the promoter of actin binding LIM protein 1 (ABLIM1) via repressing ATF3-binding chromatin accessibility, thereby inhibiting nuclear factor kappa-B (NF-κB) signaling. In a rat model of TMJOA, intra-articular administration of CEFFE alleviates cartilage degeneration, subchondral bone loss, and synovitis, without systemic toxicity. CEFFE also suppresses pro-inflammatory M1 macrophage polarization and decreases interleukin-1β and tumor necrosis factor-α expression in synovial tissue. These findings identify CEFFE as a dual-target therapeutic agent that modulates both chondrocytes and macrophages through epigenetic regulation of ATF3/ABLIM1/NF-κB signaling. Our study highlights CEFFE as a safe and accessible biomaterial with potential for clinical translation as a regenerative and anti-inflammatory strategy for TMJOA.

Human tumor organoids represent a paradigm shift in cancer modeling, overcoming critical limitations of conventional systems by faithfully recapitulating genetic heterogeneity, three-dimensional architecture, and tumor microenvironment dynamics of patient tumors. Our review explores how human tumor organoids serve as a transformative preclinical platform, bridging the gap between basic research and clinical translations. We highlight recent advances in tumor organoid generation, spanning patient-derived organoids to genetically engineered platforms from normal tissue and human pluripotent stem cells, and their applications in deciphering carcinogenesis, clonal evolution, and metastatic mechanisms. We further examine technological innovations in culture systems that enhance the interpretability and translatability of tumor phenotypes and drug responses. We present an in-depth exploration of how integrated tumor microenvironment co-culture systems-combining immune cells, cancer-associated fibroblasts, and vascular components-enable novel investigations into tumor-stroma-immune crosstalk. Clinically, human tumor organoid biobanks have shown great promise in predicting personalized therapy responses. Emerging technologies like organoids-on-chip platforms, three-dimensionally bioprinting and artificial intelligence-driven analytics are enhancing high-throughput drug screening efficiency and biomarker identification. Despite advances, complete microenvironmental modeling remains challenging, particularly in replicating vascular complexity and systemic immune responses. Future advancements will demand convergence of synthetic biology, functional genomics, and machine learning to transform human tumor organoids from static avatars into dynamic "living biosensors". In summary, this review provides an in-depth exploration of the organoid field and presents a clear and actionable framework for positioning tumor organoids as indispensable tools in functional precision medicine-a strategy that ultimately bridges mechanistic discoveries with clinical translation.

Cervical carcinoma (CC) mortality remains high due to chemoresistance. Targeting glycolytic reprogramming is promising since CC cells depend on enhanced glycolysis for proliferation and chemoresistance. However, clinical translation faces two barriers: the lack of spatially resolved validation of glycolytic vulnerability in CC specimens, and absence of CC-specific delivery systems for lonidamine (LND), a potent glycolytic inhibitor with poor bioavailability and hepatotoxicity. Here, we aim to establish spatially resolved validation of tumor-associated glycolytic reprogramming and identify differentially enriched receptors within malignant regions in clinical CC specimens. Leveraging these findings, we will design a tumor-targeted nanoplatform for the precise delivery of a glycolytic inhibitor to CC cells, to achieve tumor suppression and reverse chemoresistance. Spatial metabolomics and spatial transcriptomics analyses were employed to validate tumor-associated glycolytic reprogramming and identify specifically overexpressed receptors within malignant regions of clinical CC specimens. Through spatial multi-omics analysis, we demonstrated upregulated glycolysis in malignant regions of CC and identified solute carrier family 1 member 5 (ASCT2), a glutamine transporter, as a superior CC-specific surface marker compared to the pan-cancer nanocarrier targets like CD44. These findings were corroborated through multi-platform validation spanning single-cell RNA-seq dataset, TCGA cohorts, paired patient specimens, matched murine samples, and multiple CC cell lines. In this context, we designed a glutamine-functionalized liposomal system that exploits ASCT2 overexpression to enable CC-selective accumulation of LND. Our findings reveal that this nanoagonist significantly impedes CC growth by disrupting ATP supply and inducing ROS-mediated cellular damage. Moreover, this nanoagonist effectively reverses cisplatin (DDP)-induced chemoresistance in CC by inhibiting MRP2-mediated DDP efflux and blocking ribose-5-phosphate-mediated DNA repair. By integrating spatial multi-omics with rational nanocarrier design, we designed a glutamine-functionalized liposomal system that exploits ASCT2 overexpression for tumor-selective accumulation of LND. Our findings revealed that this nanoagonist achieves significant CC suppression and chemoresistance reversal.

Single-cell copy number variation sequencing (scCNV-seq) is essential for genomic analysis. However, existing methods rely on whole-genome preamplification or special devices, suffering from high cost and biased CNV calling. These challenges are significant in clinical scenarios where cells are precious and limited. We aimed to provide a convenient, efficient, and accurate scCNV-seq method with a data analysis workflow for clinical application with confident output while eliminated limitations above. We developed traceable medium-throughput scCNV-seq (msCNVS). This method directly labels cells in a microplate using barcode-containing modified Tn5 transposomes, enabling early pooling to streamline processing and circumvent preamplification. MsCNVS simultaneously barcoded currently 48 but scable up to 96 or 384 cells, followed by dual indexes (i5, i7) for library construction. A new bioinformatic algorithm, two-dimensional fitting, was also outlined for accurate ploidy and copy number determination. MsCNVS reliably distinguished unique CNV patterns among 5 cell lines comprising 292 cells. Individual msCNVS profiles of 70 K562 cells exhibited high correlation (R = 0.90-0.98) with bulk sequence. Triplicates of HeLa bulk cells demonstrated near-perfect correlation (R = 0.99). No evident cross-contamination of reads was detected when mouse and human cells were processed together. The CNV profiles of 47 single cells from primary amniotic fluid cell cultures were validated via karyotyping and microarray. MsCNVS obtained superior coverage uniformity versus MDA and MALBAC, approached the level achieved by eMDA and DOP-PCR, and showed less fluctuation than PTA. Notably, msCNVS detected CNV deletions in two abnormal blastocysts (totaling 33 cells), including one exhibiting CNV mosaicism. It also revealed CNVs in 24 circulating tumor cells, 35 cancerous pleural effusion cells, and 152 patient-derived xenograft nuclei. All these results clearly set apart confident absolute CNVs from ambiguous fuzzy zones. msCNVS promises a robust and highly efficient CNV-seq approach for precious and rare cells typically encountered in reproductive and cancer clinics.

Neddylation is a critical post-translational modification that is frequently hyperactivated in various human cancers. By modifying key substrates such as Cullins and non-Cullin proteins, this pathway plas a central role in regulating protein stability, cellular signaling transduction, and core biological processes. Its dysregulation directly drives tumorigenesis and progression, making it a highly promising therapeutic target. This review aims to systematically elucidate: (1) the processes of neddylation and its regulatory mechanisms on cellular and protein functions; (2) how this pathway promotes malignant progression and tumorigenesis by driving multiple cancer hallmarks; and (3) the development of drugs targeting different neddylation components, along with associated clinical trial progress and toxicity challenges, while also discussing its future prospects in precision oncology. This review constructs a multi-dimensional pathogenic network of neddylation at a systematic level, providing a comprehensive elaboration of its molecular basis in governing key cancer characteristics through modulating protein functions and cellular activities. Simultaneously, we focus on targeted therapeutic strategies against this pathway, analyzing the clinical potential and future directions of novel cancer treatment approaches based on preclinical and clinical evidence from inhibitors such as MLN4924 and TAS4464.

Carbon dots (CDs), a type of carbon-based nanoparticles with sizes less than 10 nm, have been widely employed in nucleus-targeted drug delivery for cancer therapy and imaging, and some are capable of binding to DNA after entering the nucleus. Our study was designed to investigate the DNA binding mechanisms of CDs and their effect on the cell cycle arrest in cancer cells. Two types of CDs, Y15-CDs and Black-CDs (B-CDs) with similar sizes but carry positive and negative charges, respectively, were selected for investigation. The successful synthesis of CDs was confirmed by UV-vis, fluorescence emission and Fourier-transform infrared (FTIR) spectroscopies, transmission electron microscope (TEM), atomic force microscope (AFM) and zeta potential measurement. DNA binding mechanisms of CDs were investigated using fluorometric competitive displacement assays, circular dichroism spectroscopy, molecular docking, all-atom molecular dynamics (MD) simulations and binding free energy calculations. The in vitro studies including intracellular distribution, cytotoxicity, DNA damage detection and cell cycle distribution were evaluated in osteosarcoma (U2OS) cells. Zebrafish embryos were used as an in vivo mode to assess the influence of CDs on embryonic development. DNA binding results show that Y15-CDs bind to DNA via both intercalative and groove binding modes, while B-CDs engage only in groove binding. In addition, Y15-CDs possess a stronger affinity to DNA. According to in vitro studies, Y15-CDs could induce DNA damage, arrest U2OS cells in S phase, and exhibit higher cytotoxicity than B-CDs. The in vivo experiments demonstrate that after distributing throughout the zebrafish embryonic body, Y15-CDs reduce body length, eye size and head size of embryos. Our results elucidate the DNA binding mechanisms of CDs and their potential influence on the cell cycle profile of cancer cells, offering valuable insights into the design of CDs with both drug delivery and intrinsic anticancer properties.

Recent rapid progress in the field of computational pathology has been enabled by foundation models. These models are beginning to move beyond encoding image patches toward whole-slide understanding, but their clinical utility remains limited. Here we present PRISM2, a multimodal slide-level foundation model trained on 2.3 million whole-slide images and 14 million question-answer pairs derived from 700,000 pathology reports. Through clinical dialogue supervision, PRISM2 aligns histomorphology with diagnostic reasoning, yielding representations that support both prompt-based inference and transferable embeddings for downstream tasks. With prompt-based inference, PRISM2 achieves or exceeds (P < 0.05) the balanced accuracy of clinical-grade products calibrated for cancer detection in the prostate, breast and breast lymph node. Additionally, across comprehensive diagnostic, biomarker and survival benchmarks, PRISM2 embeddings never statistically underperform previous foundation models via linear probing (P < 0.05). Furthermore, task-specific fine-tuning on survival prediction outperforms training from scratch on the same large survival dataset. PRISM2 demonstrates how language-supervised pretraining provides a scalable, clinically grounded signal for generalizable pathology representations, bridging human diagnostic reasoning and foundation model performance.

Here, we examined the consequences of biologically relevant vitamin D deficiency, a known risk factor for aggressive prostate cancer, using ex vivo and in vivo models. Phenotypic and single-cell RNA sequencing of mouse prostate organoids showed that vitamin D deficiency stunted luminal cell differentiation more than androgen deficiency, which is a known driver of prostate development. Mice fed a vitamin D-deficient diet showed significantly altered expression of androgen-responsive genes in their prostate luminal cells, as determined by single-cell RNA sequencing. MDA-PCa-2b and 22Rv1 human prostate cancer cells, when maintained for 6 months in 1,25-dihydroxyvitamin D, had very different responses compared to short-term treatments. Notably, the MDA-PCa-2b cells lost the ability to form xenografts, despite normal proliferation in vitro. RNA sequencing showed that these cells also had disruptions in androgen signaling and in multiple cancer-related pathways, which differed by treatment duration. This study offers new insights and validation of vitamin D's role in both benign and malignant prostate biology, underscoring its essential hormonal functions and supporting strategies for vitamin D supplementation to reduce prostate cancer risk in vulnerable populations.

Diverse genetic and epigenetic factors cooperate to specify cellular fates during development. Establishing these fates is especially critical in the nervous system, which comprises diverse neuronal cell types. How genomic architecture interfaces with epigenetic regulators to drive transcriptional programs underlying neuronal fates remains poorly understood. Here, we show that cohesin, a protein complex that shapes genomic architecture, promotes GABAergic fate specification in a subset of neurons in the nematode Caenorhabditis elegans. This process is facilitated by EOR-1, a homolog of the human promyelocytic leukemia zinc finger (PLZF) transcription factor. The nucleosome remodeling and deacetylase (NuRD) complex and TRA-4, another PLZF homolog, promote tyraminergic fate in the normally GABAergic neurons when cohesin or EOR-1 function is lost, revealing an antagonistic mechanism determining alternative neuronal fates. These findings highlight a critical interplay among genome architecture, epigenetic remodeling, and transcriptional regulation in neuronal fate specification and, given the evolutionary conservation of these factors, suggest a mechanism underlying neural development across species.

Because cytokine storm syndromes such as sepsis, acute respiratory distress syndrome (ARDS), and coagulopathy, including those seen in COVID-19, are fatal, development of highly effective therapeutics is urgently needed. Recent evidence suggests that the hydrolysis of phospholipids in extracellular vesicles (EVs) by secreted phospholipase A2 (sPLA2) can augment the ability of EVs to modulate inflammation, allergy, and cancer. We aimed to apply this phenomenon technically using "sPLA2-reacted EVs (SPLEVs)," which showed high therapeutic efficacies against ARDS and other diseases. Mechanistically, SPLEVs bind to type II alveolar epithelial cells, increase membrane fluidity by reorganizing phospholipid building blocks with more polyunsaturated fatty acids through the sterol regulatory element-binding protein 1 pathway, and elicit "lipid counterstorm" by increasing tissue-protective lipid mediators. The potent therapeutic effects of SPLEVs depend partly on sPLA2-driven generation of lysophosphatidylglycerol. Thus, SPLEVs are expected to be an effective therapeutic tool for treatment of broad ranges of inflammatory diseases including COVID-19 and other new pandemic diseases.

Well-differentiated and dedifferentiated liposarcoma (WD/DD LPS) represent a pathological continuum, often coexisting within the same tumor. While the dedifferentiated component is clinically aggressive, marked by rapid growth and metastatic potential, the evolutionary relationship between WD and DD LPS remains unknown. To investigate this, we performed single-nucleus RNA sequencing on matched WD and DD tumor regions. Both compartments shared a predominant population of undifferentiated mesenchymal cells, but only WD regions contained cells expressing adipocytic differentiation markers and PPARG target genes. Given the central role of PPARG in coordinating lipid metabolism during adipogenesis, these findings suggest that loss of this program may underlie the poorly differentiated, proliferative phenotype of DD LPS. Functional studies confirmed that PPARG activation in DD LPS cells induces lipid accumulation, reduces proliferation, and impairs tumor growth in vivo. These support a model in which impaired adipogenic differentiation underlies DD LPS pathology and identify PPARG as a potential therapeutic target to promote differentiation and suppress tumor progression.

Breastfeeding anchors infant immunity and long-term health, but its benefits are threatened by Staphylococcus aureus (S. aureus) mastitis, an increasingly prevalent condition driven by antimicrobial resistance and therapeutic limitations. Beyond compromising maternal wellness, mastitis threatens the safety and continuity of breast milk, highlighting a critical need for innovative intervention strategies. Herein, we demonstrated that α‑ketobutyrate (α-KB), a metabolite of the transsulfuration pathway, mitigated S. aureus‑induced inflammation, oxidative stress, and blood-milk barrier (BMB) disruption both in vivo and in vitro. α‑KB enhanced macroautophagic/autophagic responses, marked by increased ATG5, BECN1 (beclin 1), and LC3-II:LC3-I conversion and reduced SQSTM1/p62, through a RIPK1-lactate-TFEB axis. Specifically, it directly bound and stabilized RIPK1, elevated lactate production, and drove TFEB nuclear translocation to activate macroautophagy/autophagy and promote intracellular bacterial clearance. Molecular docking and molecular dynamics simulations suggested stable α‑KB and RIPK1 binding via hydrophobic and hydrogen bond interactions; RIPK1 knockout abolished α‑KB-induced autophagy and lactate generation, effects rescued by lactate supplementation. This study identifies a novel immunometabolic circuit linking a metabolite to RIPK1-lactate-TFEB-mediated autophagy, offering therapeutic potential against antibiotic‑resistant S. aureus mastitis and presenting a new paradigm for safeguarding breastfeeding quality and infant health.Abbreviations: 3-MA: 3-methyladenine; α-KB: α‑ketobutyrate; ATG5: autophagy related 5; BMB: blood-milk barrier; CETSA: cellular thermal shift assay; DARTS: drug affinity responsive target stability; IL1B: interleukin 1 beta; IL6: interleukin 6; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MST: microscale thermophoresis; NOS2: nitric oxide synthase 2; OCLN: occludin; PBS: phosphate-buffered saline; PTGS2: prostaglandin-endoperoxide synthase 2; Rapa: rapamycin; RIPK1: receptor interacting serine/threonine kinase 1; RT-PCR: real-time polymerasechain reaction;S. aureus:Staphylococcus aureus; SEM: standard error of the mean; SQSTM1/p62: sequestosome 1; TFEB: transcription factor EB; TJP1: tight junction protein 1; TNF: tumor necrosis factor; WT: wild-type.

Sulfur dioxide and its physiological forms bisulfite (HSO3-) and sulfite (SO32-), along with intracellular viscosity, are involved in regulating many physiological processes and are closely related to diseases and cancer development. Therefore, developing probes capable of selectively responding to HSO3-/SO32- and viscosity is of great significance and practical application value for exploring the pathological mechanism of related diseases and realizing early biological detection. An aggregation-induced near-infrared emission-based Ir(III) complex probe named Ir-TH was designed and synthesized, in which the ester-linked thiophene unit serves a dual role: it not only acts as an AIE-active rotor to track viscosity changes through the restriction of intramolecular rotation but also provides the supramolecular binding sites for HSO3-. The probe exhibits a high sensitivity and selectivity toward viscosity, good photostability, and a favorable luminescent response to HSO3- with a fast response time (5 min), low detection limit (0.022 μM, 3σ/k) and wide linear range (0-80 μM). Moreover, Ir-TH can specifically track changes in intracellular HSO3-, as well as monitor dynamic changes in intracellular viscosity during ferroptosis, providing a valuable reference for the construction of multifunctional sensors.

Osteosarcoma is widely concerned because it is a malignant tumor with a high incidence rate among teenagers. Although some chemical, physical, and synergetic technologies have been developed to treat malignant osteosarcoma, molecular mechanism of structural nanomechanics is indistinct, especially for inducement mechanism, malignant evaluation, and distant metastasis of osteosarcoma. We fabricated photo-responsive biomaterials as the biofunctional UV-mediated "cell adhesion" switch to construct the advanced microarrays on cell culture plates. The adhesion-automatic microarrays were modified with fibronectin and osteosarcoma cells were micropatterned on the microarrays by microscopy and AFM observation. The focal adhesion (FA) was matured in microarrayed osteosarcoma cells by adhesion phase separation. The influence of phase-separated FA-induced cytoskeleton bias on spatial mechanical property and force heterogeneity is investigated to disclose its interaction with nuclear activity in microarrayed osteosarcoma cells. Heterogeneous force remodeling could monitor nuclear force-sensing mechanotransduction, based on nuclear ectopia configuration by the evaluation of LaminA/C, Ki67 and YAP analysis. This study will provide theoretical potential for understanding phase-separated adhesion, heterogeneous mechanical remodeling, and nuclear force-sensing mechanotransduction of malignant osteosarcoma.

The nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) fusion constitutively activates ALK tyrosine kinase, driving oncogenesis in anaplastic large cell lymphoma (ALCL). While arsenic trioxide (ATO) exhibits therapeutic potential, its precise mechanism remains elusive, limiting clinical application. Here, we used a combined approach of E3 ligase library screening, ATO chemical proteomics, and NPM-ALK immunoprecipitation-mass spectrometry to reveal that ATO regulates NPM-ALK stability via the E3 ligase STUB1. Clinically, high STUB1 expression correlates with improved survival, identifying it as a candidate favorable prognostic biomarker in NPM-ALK+ ALCL. Mechanistically, ATO promotes a glue‑like interaction that stabilizes the ternary complex of STUB1 and NPM-ALK, promoting ubiquitination at K174 and subsequent proteasomal degradation. Furthermore, structural modeling, domain mapping, and point mutations support a proposed arsenic-dependent interaction model involving STUB1 Cys83, Cys103, and NPM-ALK Cys599, which may modulate the STUB1-NPM-ALK interface and enhance their interaction, thereby promoting ubiquitin-mediated degradation. Crucially, we demonstrate that STUB1 overexpression or pharmacological activation using the FDA-approved cardiac glycoside Deslanoside synergizes profoundly with ATO to inhibit tumor growth both in vitro and in vivo. These findings provide novel mechanistic insight into ATO's action in NPM-ALK+ ALCL and reveal new therapeutic strategies and candidate prognostic biomarkers for patient management.

Cellular senescence is a consequence of many chemotherapeutics that plays context-dependent roles in cancer. Senescent cells secrete an array of factors collectively known as the senescence-associated secretory phenotype (SASP). Here we show that the cisplatin-induced SASP enhances the detachment of high-grade serous ovarian cancer (HGSOC) cells in vitro and dissemination in vivo. We identify fructose as a metabolic component of the SASP that facilitates cell detachment and show that a high-fructose diet increases HGSOC dissemination in vivo. We identified complex I as the driver of SASP-mediated cell detachment and HGSOC dissemination. Mechanistically, this effect was driven by SASP-mediated inhibition of an NAD+-SIRT-SREBP axis, leading to decreased plasma membrane cholesterol that increased cell detachment. These findings reveal that the SASP reprograms the metabolic microenvironment, promoting metastatic dissemination in a paracrine fashion, and highlight a pro-tumorigenic metabolic effect of fructose in the SASP that may contribute to the high recurrence rate of HGSOC.

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.

Mechanical forces in solid tumors strongly affect tumor progression, invasion and treatment. Over the years, several approaches have been developed to investigate tumor mechanobiology, however, existing techniques to quantify mechanical forces remain invasive and rely on indirect estimations of mechanical stress through mathematical modeling and entailing several assumptions. To address these challenges, we introduced a non-invasive method for in vivo measurement of macroscopic mechanical forces in solid tumors and the surrounding normal tissue. The proposed approach relies on a multi-sensor tactile imaging system based on fiber Bragg grating sensors (FBGs) to measure tumor forces and reconstruct 2D spatial force distributions. Unlike existing methods, this technique does not require the use of mathematical modeling, providing a clinically translatable framework for longitudinal monitoring of tumor mechanics. The method was applied to orthotopic 4T1 and E0771 murine breast tumor models at four time points during progression. The results demonstrated the capability of this technique to quantify the magnitude of forces in both tumor and surrounding normal tissue, allowing the reconstruction of spatial distributions that reflect the heterogeneous mechanical responses across the investigated area. Higher force levels were detected at tumor sites, and their temporal evolution was correlated with tumor volume and stiffness. The corresponding stress values were consistent with previous computational predictions, providing in vivo evidence of solid-stress accumulation in growing tumors. Overall, this study offers a complementary and directly translated to humans' strategy, expanding the experimental tools available for investigating tumor mechanics. STATEMENT OF SIGNIFICANCE: Tumor mechanical forces strongly affect tumor progression, invasion, and drug delivery. However, current approaches for quantifying these forces remain invasive and rely on indirect estimations through mathematical modeling. To address these limitations, we propose a method for the direct, non-invasive, in vivo measurement of tumor mechanical forces using fiber Bragg grating sensors. The method was assessed in orthotopic 4T1 and E0771 murine breast tumor models. Two-dimensional force maps revealed heterogeneous mechanical responses across tumor and surrounding normal tissue. Higher forces were detected at tumor sites and correlated with tumor volume and stiffness. This study provides a clinically translatable strategy that expands the experimental tools available for investigating tumor mechanics, with potential implications for cancer diagnosis, treatment, and therapy monitoring.

Amino acid levels fluctuate across diverse pathological conditions. Whether such amino acid modulations directly shape pathophysiology by regulating host gene expression remains unknown. We found that extracellular arginine restriction, observed in cancer and infection, represses specific arginine tRNAs-directly suppressing translation of major histocompatibility complex I (MHC class I) and antigen presentation. Arginine regulation of MHC class I was codon-usage dependent, as synonymous codon mutations prevented MHC class I modulation. Dietary arginine restriction impaired anti-viral immunity against influenza and SARS-CoV-2 and increased colon tumorigenesis. Conversely, increasing arginine availability via dietary supplementation or myeloid-specific arginase 1 deletion enhanced MHC class I protein levels, suppressed colon tumorigenesis, and improved viral infection outcomes. These disease-modulating effects were abolished in β2-microglobulin (B2m)-deficient mice. Thus, dietary modulation of a single amino acid critically influences codon-biased translation and MHC class I-mediated immunity to respiratory viral infections and cancer, revealing an unexpected mechanism and disease hazard for arginine deficiency and highlighting potential for amino acid-based translation modulation therapy.

The liver-secreted protein fibrinogen-like protein 1 (FGL1) is a ligand of the coinhibitory receptor LAG3 on T cells; however, Fgl1-/- mice exhibit autoimmune features distinct from those of Lag3-/- mice. Here, we examined whether FGL1 acts via receptors beyond LAG3 to regulate autoimmunity. Recombinant FGL1 administration reduced autoimmune symptoms in B6/lpr lupus-like mice. This was associated with diminished antigen-specific IgM responses and B cell numbers. Genome-wide surface proteome screening identified the tumor necrosis factor receptor (TNFR) family member transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI), a receptor for B cell-activating receptors BAFF and APRIL, as a receptor for FGL1. TACI bound FGL1 via an N-terminal site not required for BAFF and APRIL interaction. The impact of FGL1 administration on B cell numbers and autoimmune phenotypes was lost in Taci-/- mice. Mechanistically, FGL1 promotes TACI internalization, thereby regulating receptor availability for activating ligands. Thus, FGL1 regulates an innate-like subset of B cells via modulating TACI availability, with implications for autoimmunity and inflammation.

Intratumor transcriptional heterogeneity (ITTH), defined as the coexistence of diverse cell states within a single tumor, complicates cancer treatment and contributes to variable therapeutic responses. Although single-cell RNA sequencing (scRNA-seq) can resolve this complexity, its cost and technical demands limit large-scale use. Bulk RNA sequencing (bulk RNA-seq) provides a scalable alternative but requires computational methods to deconvolve bulk transcriptomes into distinct cell states. Existing supervised approaches rely on accurate reference data, which are lacking for many cancer types, while unsupervised methods are not tailored to capture heterogeneity within the malignant compartment. To address these limitations, we developed CDState, an unsupervised deconvolution method based on nonnegative matrix factorization with a sum-to-one constraint and a cosine-similarity-based optimization, which infers malignant cell states using bulk RNA-seq data. CDState demonstrated robustness using pseudobulk scRNA-seq datasets from five cancer types, outperforming existing unsupervised methods in estimating both state-specific gene expression and cell proportions. Applied to 33 cancer types from The Cancer Genome Atlas, CDState revealed recurrent gene programs, including epithelial-mesenchymal transition, MYC targets, and oxidative phosphorylation, as major contributors to malignant ITTH. The malignant state proportions were linked to clinical features, including patient survival and therapeutic response. Finally, mutations and copy number alterations in genes such as TP53, KRAS, PIK3CA, SOX2, and SATB1 were identified as potential genetic drivers of malignant cell ITTH across cancer types. This study demonstrates the utility of CDState for characterization of malignant cell states from bulk RNA-seq data, establishing a framework for investigating malignant cell ITTH in large-scale cancer atlases.

Targeting angiogenesis represents a cornerstone in the development of antitumoral therapy, yet the suboptimal responses and inevitable resistance significantly limit its clinical efficacy. To better understand the mechanisms underlying anti-angiogenic resistance, this study systematically characterized the pancancer angiogenic landscape based on 1.24 million individual cells across 13 cancer types. Our analyses revealed that pericytes (PCs) and PC-derived non-classical angiogenic factors PGF/ANGPT2 correlate with angiogenesis more prominently than the classical VEGFA-centric model. Tumor-educated PCs, namely MCAM+ immature PCs (imPCs), were spatio-transcriptionally identified as the primary source of PGF/ANGPT2, driving alternative angiogenesis and serving as a major contributor to αVEGFR resistance. MCAM+ imPCs were phenotypically shaped under the dysregulated Notch signaling and hypoxia stress. To address the limitations associated with single-targeted endothelial cells (ECs) inhibition via αVEGFR, we adopted MCAM-directed antibody-drug conjugate (MCAM ADC) to specifically eliminate pro-angiogenic PCs, whose combination with αVEGFR showed enhanced anti-angiogenic effects. This dual-targeting approach demonstrated superior tumor control, offering MCAM ADC as a promising translational solution to circumvent anti-angiogenic resistance. Overall, our findings redefine the mechanisms of angiogenic resistance and suggest a dual EC/PC inhibition strategy for more effective anticancer therapy.

Hematopoietic stem cells (HSCs) and multipotent progenitors (MPPs) are sustained by networks of transcription factors and epigenetic regulators that prime lineage-specific programs yet maintain multipotency. Two epigenetic regulators, MLL3 and MLL4, play important but distinct roles in maintaining this balance. MLL3 promotes HSC differentiation, whereas MLL4 opposes differentiation. These activities are essential for both normal homeostasis and leukemia suppression, yet it is not clear how MLL3 and MLL4 regulate HSC and MPP gene expression to control HSC/MPP fate decisions. To resolve these mechanisms, we performed an extensive series of single-cell genomic studies after conditionally deleting Mll3, Mll4 or both genes together. Mll3 deletion had only limited effects on HSC/MPP enhancer networks at steady state, whereas Mll4 deletion led to precocious activation of myeloid enhancers. Surprisingly, compound Mll3/4 deletion eliminated all myeloid, erythroid, and megakaryocytic potential within the hematopoietic hierarchy and caused all progenitors to rapidly default to a B-cell-like identity. These changes were accompanied by widespread inactivation of HSC/MPP enhancers and superenhancers and ectopic activation of B-cell superenhancers. Disabling MLL3/4 histone methyltransferase activity did not recapitulate the pervasive changes in cell identity that were observed when MLL3 and MLL4 were fully inactivated, indicating that MLL3 and MLL4 activate HSC/MPP enhancers independently from their enzymatic activities. Our findings show that HSC/MPP multipotency requires sustained tension between MLL3/4-dependent enhancers that maintain myeloid, erythroid, and megakaryocyte potential and MLL3/4-independent enhancers that prime B-cell identity. MLL3 and MLL4 therefore serve as critical linchpins of multilineage hematopoiesis.

Intercellular communication between multiple myeloma (MM) cells and osteoclast precursor cells (pre-OCs) contributes extensively to the occurrence and development of myeloma-related bone destruction. However, key interacting modes and the exchanged substances involved in this communication remain unclear. In this study, we discover that tunneling nanotubes (TNTs) directly connect MM and pre-OCs. Using the stable isotope labeling with amino acids in cell culture assay and a positive-negative double selection strategy, we identify filamin A (FLNA) as a major protein transported from MM to pre-OCs. FLNA acts as a molecular clutch linking extracellular matrix-bound MAC1 to the cytoskeleton, activating Rho and MAPK signaling pathways and promoting F-actin polymerization, which subsequently enhances osteoclast differentiation by modulating cellular stiffness, traction force, and deformability. In addition, FLNA directly binds vinculin and promotes its recruitment to podosomes, thereby enhancing the functions of podosomes and the bone resorption capacity of osteoclasts. The conditional depletion of Flna in mice suppresses podosome activity and reduces stiffness, traction force, and deformability in pre-OCs, leading to significantly impaired osteoclast differentiation and increased bone mass. In the Vk∗MYC mouse model of myeloma, the administration of the TNT inhibitor latrunculin B disrupts FLNA transport to osteoclasts and alleviates osteolytic bone disease. These findings highlight the critical role of MM-transferred FLNA in osteoclastogenesis and suggest that targeting TNTs may represent a therapeutic strategy to limit pathological bone resorption associated with MM.

Cachexia is a complex, multifactorial syndrome driven by anorexia and systemic inflammation, ultimately resulting in a catabolic state characterized by weight loss, decreased muscle mass, and impaired physical function. Many chronic comorbidities are implicated in the pathophysiology of cachexia, with malignancy being one of the most common. Although approved cachexia-directed therapies are limited to date, the growth differentiation factor 15 (GDF-15)-glial cell-derived neurotrophic factor family receptor alpha-like (GFRAL) signaling axis has been identified as a primary mediator of cachexia and is an interesting pathway for cachexia-directed therapies and antibodies. Ponsegromab, a monoclonal antibody that is a highly selective and potent inhibitor of GDF-15, has resulted in improved body weight and other cachexia-related symptoms in patients with cancer. Other anti-GDF-15 and anti-GFRAL therapies are currently under investigation. Through the development of these treatments, many unexpected potential therapeutic applications have been identified, highlighting the broad, systemic impacts of cachexia. As research continues, it is imperative to focus on clear trial design and endpoints.

Glucocorticoid receptor (GR) signaling is critical to our physiology but is dysregulated in cancer by the pain, psychologic distress, iatrogenic morbidity, and pathology associated with an advanced, invasive disease. Many tumors exploit glucocorticoid signaling to provide antiapoptotic survival signals, stimulate growth and metastasis, suppress immune surveillance, and drive treatment resistance. Endogenous cortisol and exogenous glucocorticoids are typically associated with poor responses to cytotoxic chemotherapy, targeted therapy, and immunotherapy in patients with solid tumors. Translational and nonclinical data show that selective GR antagonists (SGRA) synergize with anticancer agents, including taxanes, androgen receptor inhibitors, PARP inhibitors, and anti-programmed cell death ligand 1 (PDL-1) immune checkpoint inhibitors, driving improved anticancer activity. In a tumor-intrinsic manner, SGRAs downregulate the antiapoptotic proteins SGK1 and DUSP1, which induce resistance to cytotoxic chemotherapy. This synergy has been confirmed in several randomized controlled trials, which showed improved efficacy when SGRAs were added to standard-of-care taxane therapy in platinum-resistant ovarian cancer. In a distinct cellular context, SGRAs inhibit resistance to antiandrogen therapy mediated through the GR and have the potential for additive anticancer activity in prostate cancer. Herein, we summarize the role of glucocorticoid signaling in solid tumor biology, providing mechanistic insights into recent clinical advances and emphasizing key outstanding translational and clinical questions.

The tumor microenvironment, particularly the tumor stroma, plays a critical role in tumor progression, immune evasion, and therapeutic resistance. However, its interaction with the immune landscape in rectal cancer (RC) remains incompletely understood. This study aimed to comprehensively characterize the stromal-immune ecosystem associated with the tumor stroma ratio (TSR) in RC and to evaluate its clinical and therapeutic relevance. We analyzed a multicenter cohort of 498 patients with treatment-naïve RC in whom TSR was assessed on H&E-stained sections. Integrative multi-omics analyses were performed, including bulk RNA sequencing (n=118) and single-cell RNA/T-cell receptor (TCR) sequencing (n=10). Key findings were validated by immunohistochemistry (n=114) and multiplex immunofluorescence (n=20). Survival analyses and statistical comparisons were conducted to evaluate clinical associations and treatment responses. High TSR was an independent predictor of unfavorable disease-free survival and cancer-specific survival and was associated with aggressive clinicopathological features. Single-cell analyses revealed that TSR-high tumors exhibited a profoundly immunosuppressive microenvironment, characterized by clonally expanded terminally exhausted CD8+ T cells (CD8+ Tex-CXCL13) and activated CD4+ regulatory T cells (CD4+ Treg-TNFRSF4). Two LRRC15+ cancer-associated fibroblast (CAF) subsets (mCAF-CTHRC1 and mCAF-FAP) were enriched in TSR-high tumors. Among them, mCAF-CTHRC1 was associated with increased Treg abundance and activation features, with predicted interactions with CD4+ Treg-TNFRSF4 cells through the LGALS9-CD44 signaling axis. In addition, SPP1-expressing monocytes (Mon-SPP1) and malignant epithelial cells were prominent in TSR-high tumors and showed a predicted SPP1-CD44 interaction with T-cell subsets, suggesting potential involvement in immunosuppressive stromal-immune interactions. In patients receiving neoadjuvant therapy, pretreatment TSR-low tumors showed improved pathological response and survival outcomes compared with TSR-high tumors. In the neoadjuvant chemoradiotherapy plus immunotherapy cohort, TSR-low tumors were associated with a significantly higher major pathological response rate, whereas pathological complete response showed a non-significant trend in the same direction. High TSR identifies a clinically aggressive subtype of RC characterized by a profoundly immunosuppressive stromal-immune ecosystem enriched for exhausted T cells, immunosuppressive CAF programs, and SPP1-associated stromal-myeloid interactions. These findings highlight LGALS9-, LRRC15-, and SPP1-related stromal-immune pathways as candidate stromal-immune therapeutic vulnerabilities that warrant further mechanistic and preclinical validation.

Brady and colleagues investigated the mutational consequences of cancer treatment on the genomes of 160 childhood cancer survivors who developed a subsequent neoplasm (SN). Their research aids in directing the next steps toward the prevention of SNs. See related article by Brady et al., p. 1590.

The added value of comprehensive genomic and transcriptomic profiling (CGTP) with whole-exome sequencing (WES) and whole-transcriptome sequencing (WTS) as compared with conventional targeted panel-based sequencing is not well-characterized for patients with cancer. We thus sought to determine the potential clinical utility of CGTP in a prospective clinical study in patients with advanced or metastatic solid tumors. We performed WES and WTS for patients who had prior targeted panel sequencing and had no DNA alterations with approved biomarker-matched therapies. We analyzed CGTP data of 99 patients with advanced cancers across 19 solid tumor types and assessed the presence of actionable DNA alterations and RNA expressions linked to approved or investigational agents. CGTP identified actionable genomic and transcriptomic alterations in 69.7% and 100% of cases, respectively. In this pilot study in which CGTP was incorporated into routine care, 19.2% of patients received biomarker-matched therapy. This study demonstrates the feasibility and utility of extensive genomic and transcriptomic profiling to match patients with advanced cancer to molecularly informed treatments. Transcriptomic actionability analysis identifies actionable RNA expressions for patients beyond those with actionable DNA alterations, highlighting the potential of transcriptional profiling to enhance therapeutic opportunities in precision oncology.

Immune checkpoint inhibitors and adoptive cell therapies have revolutionized cancer treatment, yet their success is accompanied by immune-related hepatotoxicity that can range from asymptomatic enzyme elevation to life-threatening liver failure. Unlike conventional drug-induced liver injury, immune-mediated hepatotoxicity arises from complex, therapy-specific mechanisms that remain incompletely understood, creating critical knowledge gaps in risk prediction and prevention. This review incorporates current evidence on the clinical presentation, mechanistic pathways, and risk factors underlying hepatotoxicity across major immune and cell therapy platforms, with emphasis on translating mechanistic insights into actionable management strategies. We systematically examine hepatotoxicity patterns for immune checkpoint inhibitors, CAR-T cell therapies, bispecific T-cell engagers, and tumor-infiltrating lymphocyte therapy, integrating clinical trial data, real-world evidence, and mechanistic studies. Our analysis shows distinct injury mechanisms: T-cell-mediated hepatocyte destruction following checkpoint blockade, cytokine-driven bystander injury during cytokine release syndrome, and emerging on-target/off-tumor toxicity from engineered lymphocytes. Critical risk modifiers include pre-existing liver disease, concomitant hepatotoxic medications, gut microbiome dysbiosis from antibiotic exposure, and host pharmacogenomic variation. We propose three priority research directions: development of predictive biomarkers enabling pretreatment risk stratification, microbiome-directed interventions to preserve hepatic immune tolerance, and implementation of Safety-by-Design engineering strategies that integrate hepatotoxicity prevention into therapeutic design. This review provides a mechanistic framework for transitioning from reactive toxicity management to predictive, personalized prevention, essential for maximizing the therapeutic potential of immune and cell therapies while protecting patient safety in this rapidly expanding treatment landscape.

Rheumatoid arthritis (RA) remains a challenging autoimmune disease with variable treatment responses to tumor necrosis factor-α (TNF-α) inhibitors. This study investigates the clinical significance of PD-1hiCXCR5-CD4+T peripheral helper (Tph) cells in RA and their potential utility as biomarkers for predicting etanercept (ETN) therapy response. We enrolled 58 RA patients, 12 age- and sex-matched osteoarthritis patients, and 15 healthy controls, with Tph cells quantified by flow cytometry. Among 25 RA patients with inadequate response to conventional synthetic disease-modifying antirheumatic drugs receiving ETN therapy, treatment outcomes were stratified by ACR20 response criteria. Tph cell frequency was significantly elevated in RA patients and correlated positively with multiple disease activity indicators. At baseline, ETN nonresponders exhibited higher Tph proportions than responders (13.23 ± 2.60% vs. 10.82 ± 3.08%, p = 0.0467). Following ETN treatment, responders demonstrated significant Tph reduction (10.82% decreased to 7.97%, p = 0.0105), paralleling serum IL‑21 dynamics. In an exploratory analysis, baseline Tph levels showed an association with ETN response. These findings suggest that circulating Tph cells may serve as a candidate biomarker for RA disease activity and therapeutic response monitoring, warranting further validation in larger prospective cohorts.

Although immune checkpoint inhibitors (ICIs) have transformed cancer treatment, their potential to disrupt maternal-foetal immune tolerance raises important safety concerns during pregnancy. This study used global pharmacovigilance data to compare pregnancy-related adverse events (PRAEs) following ICI monotherapy vs combination therapy. A cohort based case-noncase disproportionality analysis was conducted, using VigiBase®, the WHO's global Individual Case Safety Report database through January 21, 2025. ICI exposure was classified as monotherapy or combination therapy. Cases were defined as pregnancy reports including at least one PRAE from the nine predefined pregnancy-related adverse event categories (premature birth, abortion/stillbirth, gestational age or birth weight abnormalities, congenital anomalies, neurodevelopmental disorders, neonatal respiratory events, eclampsia, delivery complications, and other pregnancy-related adverse events), while noncases were defined as pregnancy reports with ICI exposure that did not include the specific PRAE category under analysis, although they may have included other reported adverse events. Reporting odds ratios (RORs) and 95% CIs were estimated to compare PRAE reporting between combination and monotherapy exposures. Multivariable Firth logistic regression was used to calculate adjusted RORs, controlling for reporter qualification and report completeness score. Generalised estimating equations with a binomial distribution were applied to account for clustering of multiple adverse events within the same pregnancy. Levels of significance are unadjusted for multiple testing. 337 case reports were initially retrieved from VigiBase, of which 164 met the inclusion criteria. Of these, 22 (13%) involved combination ICI therapy and 142 (87%) monotherapy exposure. Overall, 72 (43.9%) of 164 reported at least one PRAE, with a significantly higher frequency among combination therapy cases compared with monotherapy (17 [77.3%] of 22 vs 55 [38.7%] of 142; p < 0.001). Premature delivery was the most common pregnancy-related adverse event across all subjects, occurring in (14 [63.6%] of 22) of the combination therapy group and (23 [16.2%] of 142) of the monotherapy group (p < 0.001). Gestational or weight abnormalities followed, occurring in (4 [18.2%] of 22) and (7 [4.9%] of 142) of the combination therapy and Monotherapy groups, respectively (p = 0.04). In multivariable analyses adjusted for reporter qualification and completeness score, combination therapy remained strongly associated with foetal and neonatal complications, particularly premature birth (adjusted ROR 9.10, 95% CI 3.38-24.55). Reports with higher completeness scores were significantly more likely to include adverse pregnancy outcomes across several categories. Findings were consistent in generalised estimating equations models, where combination therapy showed a twofold higher population-averaged risk of any PRAE (ROR 2.26, 95% CI 1.39-3.68; p < 0.001). ICI combination therapy was associated with a higher reporting of PRAEs compared with monotherapy, particularly premature delivery and gestational or foetal growth abnormalities. This study received no external funding.

Thymoma represents the predominant malignant tumor arising from the anterior mediastinal region, constituting roughly 30% of primary tumors in this anatomical area with an estimated annual occurrence of 1.2-2.6 cases per million population. Research consistently highlights the prognostic significance of achieving complete surgical removal (R0 resection) for optimizing survival outcomes in affected individuals. Current clinical practice prioritizes surgical intervention as the frontline therapeutic approach for most thymoma cases. Notably, approximately one-third of patients present with advanced-stage disease (stage III/IV) at diagnosis due to its insidious progression, with the majority of these cases being deemed unresectable during initial evaluation. This clinical reality underscores the critical importance of implementing neoadjuvant therapeutic strategies to facilitate subsequent complete tumor excision and improve long-term prognosis. The current analysis comprehensively examines contemporary management approaches for advanced-stage thymoma cases where primary surgical resection is not feasible, encompassing chemotherapy, targeted therapy, immunotherapy, and systemic glucocorticoids. This study focuses on clarifying the beneficial effects of glucocorticoids in thymoma treatment while examining the underlying mechanisms and identifying potential challenges for future research.

Dermatofibrosarcoma protuberans (DFSP) is a rare subcutaneous soft tissue malignancy associated with high local recurrence rates but low propensity for distant metastases, thus mandating aggressive loco-regional control. Diagnosis depends on histopathological evaluation, with CD34 staining needed for confirmation. There is no known sex predilection, however Black patients are disproportionately affected. When subclinical extension is suspected, magnetic resonance imaging or computed tomography may be performed, but are not routinely indicated. Patients with extensive tumor invasion or fibrosarcomatous DFSP, an aggressive variant associated with worse prognosis compared to DFSP, necessitate management by a multidisciplinary team including soft tissue sarcoma experts. We recommend Mohs micrographic surgery as the preferred treatment for DFSP given its comprehensive margin assessment, resulting in significantly lower local recurrence rates. Radiation therapy and imatinib may be considered for unresectable tumors, and we advise that COL1A1-PDGFB fusion is confirmed before starting imatinib. Possible therapeutic targets, including EGFR, PD-L1, and PRAME, have been identified by molecular analyses. Advances in organoid models and single-cell RNA sequencing have improved understanding of the tumor microenvironment. However, racial disparities in diagnosis and treatment emphasize the need for a high index of suspicion in skin of color patients to reduce misdiagnosis and delays in care.

Cancer vaccines have demonstrated potential as a key immunotherapy strategy based on stimulating immune responses targeting tumor-specific antigens and tumor-associated antigens. Therapeutic cancer vaccines have been proven effective in the treatments of genitourinary cancers, such as Sipuleucel-T (Provenge) and Bacillus Calmette-Guérin (BCG). However, the full fulfillment of these potentials remains an ongoing pursuit. In this review, we provide a comprehensive overview on vaccines for genitourinary cancers, encompassing bladder, renal, and prostate cancer, addressing their mechanisms and the current status of preclinical/clinical outcomes. The combination strategies based on those cancer vaccines are also summarized, which may provide longer-lasting and more effective treatment. Moreover, we discuss the emerging technologies and future directions for genitourinary cancer vaccines. Through different platforms (peptides, DNA, RNA, dendritic cells, etc.), therapeutic cancer vaccines can provoke or strengthen anti-tumor immunity with the assistance of cytokines, chemokines as well as other adjuvants to therefore prevent the deterioration of cancer and eradicate tumor cells. Only a limited number of therapeutic genitourinary cancer vaccines have received regulatory approval, indicating that there are still many critical challenges for the clinical translation of cancer vaccines, which mainly focus on finding effective antigens, immunogenic platforms and suitable combinational clinical strategies.

Aggressive pathologic features are closely associated with early recurrence of hepatocellular carcinoma (HCC), and the association between vessels encapsulating tumor clusters (VETC) and poor prognosis has received increasing attention. This study aimed to investigate the role of adjuvant Sintilimab (AS) with or without Lenvatinib (ASL) compared to active monitoring (AM) in high-risk HCC, with emphasis on VETC-positive subtypes. Patients identified as being at high risk for recurrence and who had undergone either AM, AS, or ASL were retrospectively enrolled from four medical centers. Propensity score matching (PSM) was adopted to minimize bias. Digital spatial profiling (DSP) and multiplex immunofluorescence (mIF) were employed to elucidate the molecular underpinnings of VETC-positive HCC. This study included 620 patients with a median follow-up of 46.9 months. After PSM, 135 patients were allocated to each cohort. Both the AS and ASL regimens significantly prolonged median disease-free survival compared to AM (26.0 and 59.2 months vs. 12.2 months, respectively; p < 0.001). Critically, subgroup analysis revealed that the benefit of AS was absent in VETC-positive HCC (p = 0.681), whereas ASL demonstrated a substantial reduction in recurrence (p = 0.001). This specific efficacy of Lenvatinib against VETC-positive HCC was further corroborated by data from first-line treatment for recurrent disease and an independent neoadjuvant therapy cohort. Mechanistically, DSP revealed that VETC-positive tumors possess an immunosuppressive microenvironment, characterized by downregulated antigen presentation and impoverished CD8+ T cell infiltration. This immune contexture was validated by mIF, which confirmed sparse CD8+ T cells alongside enriched CD4+ and regulatory T cells in VETC-positive HCC. This study validates VETC-positive HCC as a distinct entity with a high risk of postoperative recurrence and validates the combination of PD-1 inhibitors with Lenvatinib as its preferred therapeutic strategy.

d-amino acids (D-AAs), the enantiomers of proteinogenic l-amino acids, are detectable in mammals, yet their biological roles in cancer immunity remain largely unexplored. Whether specific D-AAs modulate tumour progression or influence responsiveness to immunotherapy in gastrointestinal cancer is unknown. We aimed to determine how D-AAs, particularly d-serine (D-ser), shape the tumour immune microenvironment and affect clinical outcomes in gastrointestinal cancers. Mechanistic studies were conducted using murine MC38 tumours and orthotopic gastric cancer (GC) organoid allografts with or without D-AAs supplementation. Immune landscape alterations were assessed using single-cell RNA sequencing of tumour-infiltrating immune cells, flow cytometry, ex vivo macrophage-T cell co-culture assays, and microbiome manipulation experiments. D-AAs concentrations in plasma, urine, and stool were quantified in healthy controls (HCs; n = 87) and patients with GC across three cohorts (Cohort 1, n = 14; Cohort 2, n = 108; Cohort 3, n = 28). Associations between plasma D-ser levels, disease stage, immune cell infiltration, and clinical outcomes following anti-PD-1 antibody therapy were analysed. D-ser promoted tumour progression by suppressing CD8+ T cell immunity and enhancing SPP1-associated immunosuppressive macrophage signalling in murine model. Across all clinical cohorts, the plasma, urine, and stool levels of several D-AAs, most prominently D-ser, were significantly elevated in patients with GC compared to HCs. Plasma concentration of D-ser strongly correlated with disease stage (I-IV). Elevated plasma D-ser is associated with an immunosuppressive tumour microenvironment and poor response to anti-PD-1 monotherapy in patients with advanced gastric cancer. This study identifies D-ser as a previously unrecognised immunosuppressive metabolite that promotes tumour immune evasion by increased macrophages and reduced CD8+ T cell effector function, thereby shifting the tumour microenvironment toward an immunosuppressive phenotype. Clinically, D-ser is a potential metabolite to predict cancer progression and immunotherapy resistance. This work was supported by The Japan Science and Technology Agency (JST) Fusion Oriented Research for Disruptive Science and Technology (FOREST)[JPMJFR210P], Grants-in-Aid from the Japanese Society for the Promotion of Science (JSPS) (25K10430, 21K18272, 23H02899, 23K27590, 25K22627), KGRI challenge grant, Sakaguchi Memorial Foundation, Japan Agency for Medical Research and Development (CREST 21gm1510002h0001), and Miyarisan Pharmaceutical Grant.

This study aimed to determine the prognostic significance of circulating proteins and the effects of nintedanib on these proteins in patients with systemic sclerosis-associated interstitial lung disease (SSc-ILD) in the safety and efficacy of nintedanib in systemic sclerosis study (SENSCIS) trial. Patients had SSc with ≤7 years since their first non-Raynaud symptom. Candidate biomarkers of inflammation, epithelial dysfunction, extracellular matrix (ECM) synthesis, and ECM turnover were measured in serum/plasma. We assessed associations between baseline biomarker levels and decline in forced vital capacity (FVC) over 52 weeks and change in modified Rodnan Skin Score (mRSS) at week 52 in the placebo group, and changes in biomarker levels in nintedanib and placebo groups. Baseline Krebs von den Lungen-6 (KL-6) and citrullinated vimentin (VICM) degraded by MMP-2/8 levels were significantly associated with the rate of decline in FVC over 52 weeks in uncorrected analyses. A baseline KL-6 concentration of >1000 vs ≤1000 U/mL was associated with a greater rate of decline in FVC (mL) over 52 weeks (estimates: -132.5 [95% CI: -174.1, -91.0] vs -56.4 [-95.2, -17.6]; P = .009). Higher baseline N-terminal propeptide of type III collagen (Pro-C3), chemokine (C-C motif) ligand 2, and C-reactive protein levels were significantly associated with less improvement in mRSS at week 52 in analyses corrected for multiple comparisons. Decreases in cancer antigen (CA)-125 and N-terminal propeptide of type VI collagen (pro-C6) over 52 weeks were observed in patients who received nintedanib vs placebo. In mediation analysis, 48.0% of the effect of nintedanib on change in FVC at week 52 was attributed to the treatment-related decrease in CA-125 at week 24. Applying a dichotomised threshold for KL-6 can aid in identifying patients with SSc-ILD with more progressive ILD. Nintedanib reduced levels of the epithelial dysfunction marker, CA-125, and collagen synthesis epitope, pro-C6.

Brain metastases (BM) in renal cell carcinoma (RCC) are associated with poor prognosis and limited clinical guidance. We aimed to identify prognostic factors for overall survival (OS) in RCC BM and develop an interpretable machine-learning (ML) model for individualized risk prediction. We retrospectively analyzed 929 patients with histologically confirmed clear cell RCC BM treated with stereotactic radiosurgery or surgical resection and stratified them by treatment era [interferons (IFN), tyrosine kinase inhibitors (TKI), and immune checkpoint inhibitors (IO)]. Univariate Cox regression analyses were carried out, and a prognostic classification tree was constructed for the IO era. ML analyses used a comprehensive survival modeling framework integrating multiple algorithms and ensemble strategies, with final model selection based on nested cross-validation performance and interpretation using SHapley Additive exPlanations. Model performance was evaluated using the concordance index (C-index), time-dependent area under the receiver operating characteristic curve (AUC), Brier score, and calibration metrics, and an interactive web-based calculator was developed. Median OS increased from 0.8 years in the IFN era to 1.1 years in the TKI era and 2.0 years in the IO era. The CoxNet (elastic net, α = 0.2) survival model achieved a C-index of 0.64, a 6-month AUC of 0.75, and a 95% confidence interval of 0.47-0.92 in the test cohort. Model interpretation identified extracranial disease status and age at BM intervention as the dominant predictors, with additional contributions from functional status and intracranial disease burden. Limitations include a lack of patient-level systemic therapy details, tumor volume measurements, and laboratory variables used in prior prognostic models. This study reports the largest cohort of patients with RCC BM and presents an interpretable ML model for individualized survival prediction to support clinical decision making.

Although adding immune checkpoint inhibitors to neoadjuvant chemotherapy improves outcomes in high-risk early-stage breast cancer, opportunities remain to further enhance response. Dual checkpoint blockade offers a potential strategy to further enhance efficacy. To evaluate the combination of anti-programmed cell death 1 protein (PD-1) cemiplimab and anti-lymphocyte activation gene 3 (LAG-3) added to neoadjuvant therapy in ERBB2-negative early-stage, high-risk breast cancer. The I-SPY2 (Investigation of Serial Studies to Predict Your Therapeutic Response With Imaging and Molecular Analysis 2) is an ongoing randomized clinical platform trial being conducted at multiple US clinical sites including patients with early-stage (II or III) ERBB2-negative, high-risk breast cancer. Participants, continuously enrolled since 2010, were adaptively randomized from February 2, 2020, to December 9, 2021, to one of several experimental neoadjuvant therapies or control groups based on receptor subtypes defined by hormone receptor (HR), ERBB2 status, and MammaPrint (Agendia Inc) molecular risk, categorized as high (MP1) or ultrahigh (MP2). Data were analyzed from January 1, 2022, to August 5, 2025. Both groups received weekly paclitaxel for 12 weeks, then doxorubicin and cyclophosphamide followed by surgery; concomitant with paclitaxel, the intervention group also received 4 doses of cemiplimab and fianlimab (PCF) every 3 weeks. Pathologic complete response (pCR). Treatments graduated when they achieved 85% bayesian probability of success in a subtype-specific phase 3 trial. Pathway-specific biomarkers were assessed for response prediction. A total of 78 participants (mean [SD] age, 47 [39-54] years) were randomized to the intervention group, with 350 participants (mean [SD] age, 48 [39-57] years) randomized to the historical control population. PCF graduated in all clinical signatures, with pCR rates vs control of 44% (95% CI, 34%-53%) vs 21% (95% CI, 17%-25%) in all ERBB2, 53% (95% CI, 39%-67%) vs 29% (95% CI, 22%-36%) in triple-negative, and 36% (95% CI, 23%-49%) vs 14% (95% CI, 9%-19%) in HR-positive and ERBB2-negative disease. Among the total participants, 16 (21%) experienced adrenal insufficiency, including hypophysitis (11% grade 3 or 4), mostly occurring after immunotherapy completion. PCF was found to be highly effective in the subset of patients with immune signature positive status (ImPrint positive). In this randomized clinical trial, the combination of PD-1 and anti-LAG-3 inhibition with standard NAC was effective in early-stage ERBB2-negative breast cancer, particularly in patients displaying a positive ImPrint immune signature. These results warrant further definitive trials. ClinicalTrials.gov Identifier: NCT01042379.

Neutrophils are increasingly recognized as key orchestrators of the immunosuppressive tumor microenvironment, yet their intrinsic plasticity, short lifespan and resistance to genetic manipulation have impeded therapeutic targeting. Here, we report a non-pharmacological, biophysical immunomodulatory strategy based on cold atmospheric plasma (CAP) to reprogram tumor-associated neutrophils and restore antitumor immunity. We show that CAP simultaneously delivers reactive oxygen species and redox cues that inhibit mitophagy, thereby restoring mitochondrial membrane potential and oxidative metabolism in neutrophils. This metabolic reinstatement drives a shift from immunosuppressive to immunostimulatory phenotypes. In both syngeneic and humanized bladder cancer models, intravesical CAP reshapes the myeloid landscape, enhances T cell infiltration, suppresses tumor progression, and sensitizes tumors to PD-1 blockade. These findings establish CAP as a locoregional, drug-free biophysical modality capable of overcoming neutrophil-mediated immune suppression and provide a materials-based framework for modulating innate immunity in solid tumors.

Metabolic reprogramming and immune regulation are tightly interconnected processes that critically influence cancer progression. The efficacy of immunotherapy is limited in triple-negative breast cancer (TNBC) by metabolic abnormality and immunosuppressive microenvironment. However, the molecular mechanisms through which these alterations cooperate to drive immune evasion and tumor progression in TNBC remain poorly defined. Through transcriptomic profiling, we identified glycolysis and chemokine signaling as the dominant intersecting metabolic and immune pathways that distinguish TNBC from non-TNBC subtypes. Phosphoglycerate kinase 1 (PGK1) is associated with poor prognosis and with myeloid-derived suppressor cells (MDSCs), acting as a key metabolic node linking metabolic regulation to immune modulation. Functionally, PGK1 knockdown inhibited tumor growth in vitro and in vivo and reduced MDSC recruitment. Notably, PGK1 knockdown exerted a more pronounced antitumor effect under immune surveillance, accompanied by decreased infiltration of both monocytic and polymorphonuclear MDSCs and recovered CD8+ T cell function. Mechanistically, PGK1 increased lactate production and global lysine lactylation. Notably, histone H3 lysine 18 lactylation (H3K18la) at the CCL5 promoter served as a dominant and required epigenetic modification for PGK1-driven CCL5 transcription, thereby driving CCL5-dependent MDSC recruitment. P300 and class I HDACs were identified as candidate "writer" and "eraser" enzymes for PGK1-dependent H3K18la modification. Notably, combining the PGK1 inhibitor ABT-E79 with anti-PD-1 therapy synergistically decreased MDSC infiltration, recovered CD8+ T cell function, and elicited superior antitumor responses compared to monotherapy. Collectively, this study shows a mechanistic link between metabolic reprogramming and immune evasion, offering new therapeutic insights for TNBC. Starting from analyzing metabolic and immunological signaling pathways in cancers lacking clear therapeutic targets and treatment options, we aimed to identify metabolism-associated regulators of immune responses as potential therapeutic targets. Focusing on triple-negative breast cancer (TNBC), through transcriptomic profiling, we identified glycolysis and chemokine signaling as the dominant intersecting metabolic and immune pathways that distinguish TNBC from non-TNBC subtypes. Functional assays demonstrated that PGK1-driven glycolysis in TNBC cells promotes lactate accumulation and H3K18la, which subsequently induce transcriptional activation of CCL5 and recruitment of MDSCs, thereby impairing CD8+ T cell function and fostering an immunosuppressive tumor microenvironment that facilitates immune evasion. Furthermore, treatment with ABT-E79, a PGK1 inhibitor, enhances the antitumor immune efficacy of anti-PD-1 therapy.

Gastric cancer (GC) remains a major clinical challenge, with most patients exhibiting primary resistance to anti-programmed cell death protein-1 (anti-PD-1) immunotherapy and a lack of effective predictive biomarkers. Most advanced GC presents as immune-excluded "cold" tumors that respond poorly to immune checkpoint blockade. Traditional Chinese medicine (TCM) "Yong (abscess)" syndrome and the "treating GC as Yong" theory are widely applied in clinical practice, yet lack clear molecular and immunological mechanisms. Here, by translating these clinical observations into modern biological terms, we present an original, testable hypothesis proposing the "Yong" syndrome-cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) axis as the central molecular bridge connecting TCM syndrome subtypes, GC histological subtypes, and TME reprogramming. We hypothesize that heat-clearing and blood-activating TCM monomers activate cGAS-STING in a subtype-selective manner to convert "cold" tumors to "hot" immunogenic phenotypes, directly addressing the critical clinical dilemmas of primary anti-PD-1 resistance and insufficient biomarkers in GC immunotherapy. This hypothesis translates universal cGAS-STING mechanisms into a clinically actionable, syndrome-based precision strategy for GC.

Focused ultrasound thermal ablation (T-FUS) is a clinically accessible, non-invasive modality capable of inducing rapid tumor cytoreduction while mobilizing early immunologic danger signals. However, its capacity to synergize with potent co-stimulatory immunotherapies in breast cancer (BC) remains undefined. Here, we investigated whether subtotal T-FUS cooperates with CD40 agonism to elicit durable, T cell-dependent tumor control across immunologically and molecularly distinct murine BC models. Subtotal ultrasound-guided T-FUS was applied to E0771, BRPKP110, EMT6, and 4T1 tumors in mice treated with systemic agonistic αCD40. Histology, ATP bioluminescence imaging, and multispectral flow cytometry were used to define acute tumor injury and immune remodeling. Therapeutic efficacy was evaluated by tumor growth, survival, T-cell depletion, and contralateral tumor rechallenge. Partial thermal ablation triggered canonical immunogenic cell-death signatures and acute remodeling of intratumoral myeloid populations, while expanding circulating CD4+ and CD8+ T cells. When layered onto this immunogenic milieu, αCD40 markedly constrained tumor outgrowth, yielding significant reductions in tumor burden across all models and complete tumor eradication in 30% of E0771 tumors, with additional complete responses in BRPKP110 and EMT6. Combination treatment also extended survival across three of four models. Efficacy required both CD4+ and CD8+ T cells, as depletion of either compartment abrogated tumor control. Complete responders mounted robust systemic immunity, rejecting contralateral tumor rechallenge with 100% protection and displaying persistent effector-memory T-cell activation. Subtotal T-FUS cooperates with CD40 agonism to drive durable, T cell-dependent BC regression and immunologic memory. These findings establish T-FUS as an immune-potentiating partner for CD40 agonism and position this combination as a clinically scalable, in situ vaccination-like strategy with potential to broaden immunotherapy benefit across breast cancer subtypes, including luminal tumors that remain largely refractory to immune checkpoint blockade.

Resistance to anti-programmed cell death protein-1 (PD-1) treatment in gastric cancer (GC) is closely associated with an immunosuppressive tumor microenvironment. However, the role of neutrophils in resistance to anti-PD-1 therapy remains unclear. Single-cell RNA sequencing was performed on tumor samples from patients with advanced GC receiving anti-PD-1 therapy to identify neutrophil subsets associated with neutrophil extracellular traps (NETs). Multilevel experimental validation was conducted using multiomics analysis, flow cytometry, multiplex immunofluorescence, and in vitro co-culture. Therapeutic strategies targeting NETs and CD8+ T-cell exhaustion were evaluated in a mouse model of YTN16 tumors. We identified a NETs-associated neutrophil subset enriched in patients with GC resistant to anti-PD-1 treatment. This subset was marked by CD177, and it exhibited a high potential for NETs release. Peripheral blood NETs levels and CD177+ neutrophil ratios in patients with GC act as markers for evaluating the efficacy of PD-1 inhibitors. Furthermore, transforming growth factor-β1 (TGF-β1), which was highly expressed in GC and spatially colocalized with CD177+ neutrophils, might induce neutrophils to release NETs via the Smad3-NFE2 axis. NETs promoted CD8+ T cell exhaustion by activating the MEK/ERK-c-Fos/JunB axis, as evidenced by increased PD-1/TIM3 expression and reduced interferon-gamma/tumor necrosis factor-alpha secretion. In vivo experiments confirmed that targeted inhibition of NETs formation using DNase I or TGF-β1 inhibitors significantly suppressed tumor growth and CD8+ T cell exhaustion. Notably, the MEK inhibitor trametinib reversed the immunosuppressive microenvironment associated with CD8+ T cell exhaustion and synergistically enhanced the antitumor efficacy with anti-PD-1 therapy. TGF-β1 drives CD177+ neutrophils to release NETs, which induce CD8+ T cell exhaustion via the ERK-c-Fos-JunB pathway, thereby mediating resistance to anti-PD-1 treatment in GC. Furthermore, targeting NETs formation and combining trametinib with PD-1 inhibitors can significantly reverse CD8+ T cell exhaustion, exert synergistic antitumor effects, and offer a potential therapeutic strategy for overcoming resistance to anti-PD-1 therapy in GC.

Although immunotherapy has revolutionized cancer treatment, hepatocellular carcinoma (HCC) continues to demonstrate limited clinical responses, highlighting the urgent need for novel immunomodulatory strategies. Trained immunity, an emerging paradigm wherein innate immune cells develop a memory-like phenotype through epigenetic and metabolic reprogramming, offers a promising avenue to remodel the immunosuppressive tumor microenvironment. This study investigated whether β-glucan-induced trained immunity could potentiate antitumor immunity against HCC. We established orthotopic HCC mouse models to investigate the role of trained immunity induced by whole β-glucan particle (WGP) in the HCC microenvironment, particularly in modulating hepatic apolipoprotein E (APOE)-positive monocytes/macrophages. Transcriptional changes in trained monocytes/macrophages were identified by analyzing single-cell RNA sequencing and bulk RNA-sequencing data from the livers of WGP-treated and control mice. Mechanistic studies were performed using Apoe -/- mice and in situ monocyte/macrophage engineering. Flow cytometry was performed to assess immune cell phenotypes and phagocytosis, while luminescence-based assays were used to evaluate cytotoxic activity. The translational potential was assessed using human monocyte training assays. This study demonstrated that preconditioning with WGP, a trained immunity inducer, increased the accumulation of trained monocytes/macrophages in the liver and suppressed tumor progression in HCC mouse models. Mechanistically, WGP-trained APOE+ monocytes/macrophages exhibited a decrease in lipid accumulation and endoplasmic reticulum stress, thereby enhancing their antitumor function. Genetic deletion of Apoe in monocytes/macrophages abrogated the antitumor effects of WGP, demonstrating that APOE+ monocytes/macrophages are essential mediators of WGP-induced trained immunity. Adoptive transfer of WGP-trained bone marrow-derived macrophages suppressed the growth of HCC in recipient mice. Furthermore, WGP induced trained immunity in human monocytes, leading to enhanced killing of HCC cells. Notably, combination therapy with WGP and anti-programmed death-ligand 1 antibody achieved superior tumor control compared with either monotherapy. These findings identify a critical role for trained APOE+ monocytes/macrophages in WGP-mediated antitumor immunity in the liver. Harnessing WGP-induced peripheral trained immunity represents a novel therapeutic strategy for HCC.

Single-cell transcriptomics identifies a convergent activation state of conventional dendritic cells (cDCs) shared by type 1 and type 2 cDCs (cDC1s and cDC2s). These activated DCs (actDCs) are characterized by co-expression of T cell-stimulating and inhibitory molecules. Here, we examined the functional contribution of actDCs to anti-tumor immunity by developing mouse models that leverage CCR7 expression to conditionally label or ablate actDCs. The capacity of cDCs to stimulate tumor-specific cytotoxic T lymphocytes (CTLs) was restricted to the actDC state. cDC1- and cDC2-derived actDCs supported CTL priming through cross-presentation and cross-dressing, respectively, with the latter occurring in a cancer type-dependent manner. actDCs were required for the activation of naive CTLs in tumor-draining lymph nodes and for sustaining effector CTL function within tumors. Consequently, ablation of actDCs impaired spontaneous tumor control and responses to immune checkpoint blockade or adoptive T cell therapy. Thus, the actDC state emerges as a critical determinant of cDC-mediated anti-tumor immunity.

Cancer remains a leading cause of morbidity and mortality worldwide. While classical psychedelics have been used clinically to treat cancer-associated psychiatric disorders, their impact on tumor progression is unclear. Here, we show that by targeting the serotonin receptor 5-HT2AR, lysergic acid diethylamide (LSD) enhances CD8+ T cell-mediated antitumor immunity and suppresses colorectal cancer (CRC) growth. To harness this activity while avoiding psychedelic effects, we developed IHCH-8110, a non-brain-penetrant 5-HT2AR agonist that selectively targets peripheral 5-HT2AR. We show that IHCH-8110 inhibits CRC progression by activating 5-HT2AR on enteric glial cells, thereby inducing CXCL10 and interleukin (IL)-18 expression to promote CD8+ T cell recruitment and effector polarization within the tumor microenvironment. By converting immune-cold CRC into a more immunologically responsive state, IHCH-8110 enhances the efficacy of PD-1 blockade. Together, our findings identify enteric 5-HT2AR signaling as a regulator of antitumor immunity and support peripheral 5-HT2AR agonists as a therapeutic strategy for CRC immunotherapy.

The emerging advance of targeted protein degradation (TPD) technology offers a novel option for protein modulation, ensuring a more durable and precise therapeutic impact for cancer management. While promising, complete degradation of the proteins of interest (POIs) in both pathological and normal tissues may cause severe side effects. Furthermore, insufficient accumulation of the protein degraders at the target tissues also limits the clinical translation of TPD. It remains an unmet need to achieve spatiotemporally tunable degradation of the POI at the tumor lesion. In recent years, our group has extensively exploited the potential of the stimuli-activatable TPD technology for precise cancer therapy. The stimuli-activatable protein degraders were rationally designed for achieving tumor-specific enrichment in vivo to maximize their therapeutic effects while minimizing the side effects. Several kinds of stimuli-labile prodrugs of the proteolysis targeting chimeras (PROTACs) were rationally designed for restoring their protein degradation functions with the endogenous or exogenous stimulus of tumor while remaining "silent" elsewhere, resulting in precise therapies and reduced side effects. Leveraging the advantages of nanomedicine delivery systems, several kinds of tumor acidity and enzymatic-activatable nanodegraders were developed to achieve tumor-targeted protein degrader distribution and POI degradation. In particular, photothermally activatable protein degraders were developed to perform spatiotemporally controllable degradation of various POIs. In this Account, we systematically summarize recent advances from our group regarding the rational design of stimuli-activatable protein degraders, and strategically outline the "when and how" of integrating these degraders with nanomedicine platforms to tailor precise cancer therapy. We discuss the crucial role of the tumor microenvironment-responsive moieties for stimuli-triggered degradation of both intracellular and membrane POIs, highlighting the distinct design rationale for their respective prodrugs. Furthermore, we summarize our advances of strategic integration of the TPD technology with nanomedicine to augment the therapeutic outcomes of phototherapy, radiotherapy, chemotherapy, and immunotherapy of solid tumors. It is envisaged that the tumor microenvironment-activatable protein degradation approaches will achieve tumor-specific protein degradation and precision therapy, thereby facilitating the clinical application of TPD. By outlining optimized design strategies and future challenges, this Account aims to serve as a roadmap for researchers seeking to develop next-generation activatable TPD technologies that are modular, functionally versatile, and translatable.

Glioblastoma (GBM) is a highly lethal brain tumor, with therapeutic efforts hampered by the restrictive blood-brain barrier (BBB) and a profoundly immunosuppressive tumor microenvironment (TME). Driven by bioinformatics analysis identifying epidermal growth factor receptor (EGFR) and caspase-3 as key regulators of an immune-evasive pyroptosis pathway, we screened natural compounds and identified quercetin (Q) and chlorogenic acid (C) as dual-targeting agents, thereby laying a therapeutic foundation for amplifying pyroptosis in GBM treatment. The compounds were conjugated into a glutathione-responsive prodrug (QSSC) and encapsulated in a tumor-derived exosome-liposome nanoplatform (QSSC@Exo-LNP), enabling enhanced BBB penetration and intracranial targeting. Mechanistic studies revealed a dual-pathway amplification of pyroptosis, in which C directly activates caspase-8 to initiate gasdermin E (GSDME)-mediated pyroptosis, while Q/C-mediated EGFR inhibition activates mitochondrial pro-apoptotic protein, thereby augmenting caspase-3 and intensifying pyroptotic cell death. Upon intravenous injection, QSSC@Exo-LNP triggers robust pyroptosis, releasing DAMPs and tumor antigens for immune activation and macrophage reprogramming, converting the TME from "cold" to "hot" state. Moreover, this treatment strategy can significantly inhibit the distant tumors in the primary-distal orthotopic GBM model. This study proposes a strategy for the precise immunotherapy of GBM by exploiting natural products to target overexpressed GSDME and induce the pyroptotic cascade.

Glycolysis and protein lactylation both drive hepatocellular carcinoma (HCC) progression, yet their mechanistic interplay remains unclear. Through integrated single-cell and spatial transcriptomic analyses stratified by glycolytic activity, we identified alanyl-tRNA synthetase 1 (AARS1), a recently characterized protein lactyltransferase, as a key metabolic-immune regulator in HCC. Clinically, AARS1 is upregulated in tumors, correlates with elevated glycolytic flux measured by 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT), poor prognosis, and immunotherapy resistance. In murine models, hepatocyte-specific knockout of AARS1 suppressed tumor growth and reduced the abundance of regulatory T cells (Tregs). Mechanistically, AARS1 catalyzes the lactylation of activating transcription factor 6 (ATF6) at lysine 424, preventing its degradation and leading to transcriptional activation of TDO2. This process promotes L-kynurenine production and supports Treg differentiation and function. Furthermore, L-kynurenine-AHR signaling drives eNAMPT secretion from Tregs, which augments tumor cell glycolysis and lactate production, thereby reinforcing a feedback loop that sustains AARS1-catalyzed ATF6 lactylation. Pharmacological inhibition of AARS1 with β-alanine sensitized tumors to PD-1/PD-L1 blockade.

Chimeric antigen receptor (CAR) T-cell therapy has shown early promise against glioblastoma, which lacks effective treatment options. However, two key challenges curtail efficacy: tumor-antigen heterogeneity and an immunosuppressive tumor microenvironment. CAR T cells engineered to secrete combinations of immunomodulatory proteins can reverse immune suppression and engage endogenous immunity. Through head-to-head in vivo comparisons of potentially synergistic armor combinations, we demonstrated that T cells expressing a CAR plus IL12 and the decoy-resistant form of IL18 (CAR-12.DR18 T cells) show strong efficacy against antigen-heterogeneous glioma in immunocompetent mice. Robust antitumor efficacy with effective toxicity mitigation was achieved via combined administration of CAR-12.DR18 T cells with CAR T cells that secrete an anti-vascular endothelial growth factor (anti-VEGF) single-chain variable fragment (scFv). This combination therapy presents a clinically applicable strategy to overcome key barriers to the effective treatment of glioblastoma. CAR-T cells armored with cytokines and anti-VEGF single-chain variable fragments can control orthotopic, antigen-heterogeneous glioma with minimal toxicity, providing a therapeutic strategy for glioblastoma patients in urgent need of efficacious treatments.

The N-terminal fragments of gasdermin (GSDM-NT) form pores on the plasma membrane that initiate pyroptosis. However, the presence of GSDM-NT pores does not necessarily result in cell death, allowing some cells to survive GSDM-mediated pyroptosis. Understanding the regulators of response to the formation of GSDM-NT pores is crucial for revealing the strategies to harness the potential of pyroptosis for treating cancer. In this study, we found that myosin 1G (MYO1G) enabled cells to resist death driven by membrane rupture. Mechanistically, MYO1G tethered GSDME-NT pores to caveolin 1 (CAV1), thus facilitating CAV1-mediated endocytosis of pyroptotic pores. Pyroptosis triggered the transcriptional upregulation of MYO1G through stabilization of hypoxia-inducible factor 1α (HIF1α), which resulted from the release of intracellular α-ketoglutarate. Moreover, pharmacologic inhibition of MYO1G or cholesterol synthesis promoted pyroptosis, boosted antitumor immunity, and synergized with chemotherapy to eradicate tumors. Clinically, MYO1G was validated as an indicator of poor short-term response to cisplatin-based chemotherapy and unfavorable long-term survival in patients with nasopharyngeal carcinoma. Overall, these findings provide further understanding of a protective mechanism against pyroptosis and identify potential therapeutic targets for cancer treatment. MYO1G-facilitated endocytosis of gasdermin E pores protects cancer cells against pyroptotic death and contributes to cancer chemotherapy resistance, which provides potential biomarker and therapeutic strategies for cancer.

Pancreatic ductal adenocarcinoma (PDAC) is notorious for its aggressive, therapy-resistant nature that is in part driven by the desmoplastic, hypoperfused, and immunosuppressive tumor microenvironment (TME). In this study, we demonstrated that the αv integrin and neuropilin-1 (NRP-1) dual targeting iRGD peptide reverses some of these TME features by inhibiting transforming growth factor β (TGFβ) activation in the tumor, a process mediated by the αvβ5 integrin. In addition to PDAC epithelial cells and fibroblasts, regulatory T cells (Treg) in PDAC tumors also expressed the αvβ5 integrin and NRP-1. The αvβ5+ Tregs potently inhibited T-cell proliferation, and systemic iRGD therapy not only depleted αvβ5+ Tregs from PDAC tumors but also reduced their αvβ5- counterparts. Mechanistically, iRGD inhibited the activation of TGFβ mediated by the αvβ5-rich TME, thereby depriving Tregs of the cytokine essential for their development and maintenance. NRP-1-dependent tumor penetration was required for this effect because a traditional RGD peptide without an NRP-1-binding motif failed to inhibit TGFβ signaling or deplete Tregs in vivo. Treatment with iRGD induced a series of additional TME changes, such as improved vascular patency and perfusion, reduced stromal fibers, and increased CD8+ T-cell entry into the core of the tumors. Combining iRGD with immune checkpoint blockade led to an enhanced antitumor effect. Together, these findings support targeting the αvβ5 integrin with affinity ligands such as iRGD as a potential approach to enhance immunotherapy efficacy against PDAC and other desmoplastic tumors with high TGFβ and αvβ5 expression. The iRGD tumor penetrating peptide inhibits αvβ5 integrin-dependent TGF-β activation in pancreatic cancer to normalize the desmoplastic and immunosuppressive tumor microenvironment.

The gut microbiota is increasingly recognized as a contributor to breast cancer progression. Here, we report that the gut bacterium Enterocloster bolteae, a member of the Lachnospiraceae family, is progressively enriched during tumor development and is associated with increased levels of the microbiota-derived metabolite deoxycholic acid. Deoxycholic acid accumulates in tumors and activates the farnesoid X receptor in tumor cells, inducing interleukin-6 production through nuclear factor κB signaling. Interleukin-6 promotes the recruitment of granulocytic myeloid-derived suppressor cells and T helper 17 cells, establishing an immunosuppressive microenvironment. Inhibition or knockdown of the farnesoid X receptor, as well as blockade of interleukin-6 signaling, attenuates these effects. These findings identify a microbiota-metabolite-immune axis driving breast cancer progression and uncover microbial metabolites as potential therapeutic targets.

Microsatellite stable (MSS) rectal cancer exhibits intrinsic resistance to immunotherapy. Although radiotherapy is frequently combined with immune checkpoint inhibitors (ICI) to augment immunotherapy responses, numerous immunologically cold tumors remain unresponsive. In this study, we observed a significant increase in electron transport chain activity, acetyl-CoA levels, and global lysine acetylation levels in patients achieving a pathologic complete response following immunotherapy administered after radiotherapy. Transcriptomic screening and in vivo experiments revealed that SIRT1, a key regulator of protein acetylation, restricted the immunostimulatory effects of radiotherapy. Mechanistically, SIRT1 deacetylated DDX5, promoting the unwinding of irradiation-induced R-loops and inhibiting the accumulation of cytoplasmic RNA:DNA hybrids to suppress cGAS/STING pathway activation and T-cell infiltration. Moreover, radiotherapy induced a tryptophan-SIRT1-SLC36A4 positive feedback loop that enhanced SIRT1 activity and promoted competitive tryptophan uptake from the microenvironment, thereby inhibiting tertiary lymphoid structure (TLS) formation and radioimmunotherapy efficacy. Finally, combining both an SIRT1 inhibitor and aspirin with radiotherapy converted ICI-unresponsive rectal cancer into immunogenic tumors that were sensitive to ICI. Together, this study identifies SIRT1 as a potential biomarker and therapeutic target to overcome radioimmunotherapy resistance in MSS rectal cancer. Radiotherapy activates a tryptophan-SIRT1 metabolic feedback loop in microsatellite stable rectal cancer that suppresses T cell infiltration and tertiary lymphoid structures formation, which can be overcome with SIRT1 inhibition and aspirin.

The prevailing view of cold tumors as a singular immune-insensitive state is not merely imprecise, which is clinically misleading. Currently, we contend that cold tumors comprise a heterogeneous spectrum of resistance ecosystems by distinct, coexisting, dominant barriers. We argue that a universal "heating" strategy is unlikely to succeed; instead, durable clinical benefit will depend on barrier-directed immune reprogramming. We dissect this heterogeneity by linking a set of core mechanistic drivers to canonical hot, immune‑desert, and immune-excluded phenotypes. The drivers include defective antigen processing and presentation, impaired T-cell priming, physical exclusion by aberrant vasculature, stromal components, metabolic exhaustion, and adaptive resistance. Our evaluation of emerging conversion strategies focuses not on their novelty but on capacity to neutralize a specific rate-limiting barrier. We also examine why combination and sequencing are indispensable, highlight translational gaps between preclinical models and human disease and propose a dynamic biomarker framework that extends beyond static PD-L1 assessment to capture real-time shifts in dominant barrier. Our central conclusion is that durable cold-to-hot conversion will require iterative, biomarker-guided and barrier-adaptive interventions that evolve alongside tumor counter-adaptation. The reframing positions immunotherapy resistance not as a terminal obstacle but as a manageable, dynamic challenge, offering a practical strategy for clinical translation.

EGFR-mutant lung adenocarcinoma (LUAD) is typically associated with an immunosuppressive tumor immune microenvironment (TIME) and poor responses to PD-1 blockade. However, the contribution of drug-tolerant persister cells (DTPs) to immunotherapy resistance remains unclear. We hypothesized that DTPs-derived chemokine (C-C motif) ligand 20 (CCL20) promotes immune evasion and impairs PD-1-based immunotherapy in EGFR-mutant. To test this, humanized NSG mice engrafted with EGFR-mutant H1975 cells and human peripheral blood mononuclear cells (n = 6 per group) were administered with anti-PD-1 (200 µg) and/or anti-CCL20 (20 µg) every other day for a total of four doses. The combination reduced mean tumor volume by 65% compared with control treatment (p < 0.01). Single-cell RNA sequencing showed that cotreatment selectively suppressed CCL20 expression in DTP clusters and activated interferon-α and -γ signaling (ISG15, CMPK2). Multiplex immunofluorescence revealed combination treatment increased infiltration of M1-like macrophages, plasmacytoid dendritic cells, and memory B cells, alongside spatial segregation of CD4+ regulatory T cells (p < 0.05). These findings identify DTPs-derived CCL20 as a mediator of immunosuppressive TIME and support combined CCL20/PD-1 blockade as a potential therapeutic strategy for EGFR-mutant lung adenocarcinoma.

Triggering protease-activated cell death is a promising strategy for cancer treatment. Here, we used phage-assisted evolution to reprogram botulinum neurotoxin serotype X proteases to cleave and activate procaspase-1 and gasdermin D, key effectors of inflammatory cell death. We also developed an efficient system to broadly characterize the substrate specificity of wild-type and evolved botulinum neurotoxin serotype X protease variants. Evolved proteases triggered robust cell death across multiple cancer cell lines. The gasdermin D-cleaving protease exclusively induced lytic death, whereas the procaspase-1-cleaving variant initiated both lytic and apoptotic cell death. To enable self-delivery into mammalian cells, we reconstitute evolved proteases with a native BoNT translocation domain, selectively killing cultured cancer cells while sparing non-cancerous cells. Expression of the evolved protease targeting caspase-1 reduced tumor growth in a highly drug-resistant tumor mouse model. These findings establish an evolving protease system to modulate inflammatory cell death and highlight the potential of BoNT proteases as programmable tools for targeted cancer therapy.

Antibody-drug conjugates (ADCs)-composed of a monoclonal antibody linked to a payload-were designed to deliver high concentrations of cytotoxic agents to cancer cells. Since their inception, a deeper understanding of the mechanisms of action and resistance to ADCs in patients has generated a wealth of advances in the chemistry of ADC constructs, together with rational therapeutic combinations. However, the pace of innovation now exceeds clinical trial capacity. Also, any single modification or combination is unlikely to generate clinically meaningful benefit on its own. In this context, there is a need to integrate multiple chemistry advances into individual ADCs and to develop frameworks, infrastructures and tools to accelerate and de-risk the preclinical and early clinical development of these agents. In addition, the development of multidimensional molecular tools to predict ADC sensitivity, together with the optimal use of new ADCs in early-stage cancers, should contribute to improved outcomes for patients. We discuss these opportunities and challenges and predict that in the longer term, the development of diversified ADC libraries incorporating distinct constructs and drug-to-antibody ratios will enable personalized treatment strategies aligned with individual tumor biology.

T/B cell receptors (T/BCR), coordinating antigen-targeting immune response, play crucial roles in anti-tumor immune response. Tracking T and B cell clonal evolution in situ at single-cell resolution is essential for understanding the adaptive immune responses. To address the lack of tools for in situ single-cell T/BCR (XCR) sequencing, we develop Stereo-XCR-seq, an efficient method for retrieving and sequencing TCR and BCR from spatial transcriptome cDNA libraries at subcellular resolution. Stereo-XCR-seq enables high-fidelity, unbiased recovery of XCR sequences alongside spatial transcriptomics in extensive tissues, facilitating the identification of heterogeneous lymphoid aggregates with distinct clonal activities in situ. Using Stereo-XCR-seq, we uncover that IgG+ plasma cell aggregates display features of ectopic germinal center-like (GC-like) niches to select tumor-reactive clones from distal immature tertiary lymphoid structures in 11 lung adenocarcinoma (LUAD) tumors. The presence of these IgG+ plasma cell aggregates in LUAD indicates an improved anti-tumor immune surveillance and sensitivity towards immune checkpoint blockade (ICB) therapy. Collectively, Stereo-XCR-seq enables in situ single-cell profiling of T and B cell clonal activities and their interactions with local microenvironment, links tumor reactivity to intratumoral distribution of lymphocytes, and thus offers a versatile tool for dissecting lymphocyte clonal dynamics across human diseases.

Conventional antibody discovery approaches that do not account for enrichment-driven biases, such as epitope immunogenicity, PCR amplification bias, or protein expression efficiency, may result in under-representation of rare yet functionally relevant clones, necessitating labor-intensive in vitro screening to identify agonistic antibodies among a large number of dominant clones. Thus, efficient screening methods for agonistic antibodies are urgently needed. OX40 is a promising target for cancer immunotherapy due to its role in enhancing T-cell activation and survival. However, effective anti-OX40 agonistic antibodies have not yet been developed. We developed a novel screening strategy that involves the selection of nanobody clone pools enriched by biopanning against gp34-engaged and non-engaged OX40-expressing cells, next-generation sequencing, and computational clustering and subtraction analysis to identify clones recognizing the ligand-receptor interface. Representative nanobody clones underwent in vitro validation, including epitope mapping, binding affinity measurements, and functional assessments. Furthermore, we engineered the selected nanobody to enhance its in vivo efficacy. We also performed structural analysis of the nanobody-OX40 complex. Our epitope-directed approach efficiently identified nanobody clones recognizing functionally relevant epitopes distinct from dominant immunogenic regions. Notably, clone Nb479 demonstrated robust agonistic activity, closely mimicking the natural ligand gp34 with extensive OX40-binding interactions. Trimerization of Nb479 facilitated potent OX40 activation without the need for a cross-linking scaffold. Conjugation of the Nb479 trimer with an anti-serum albumin nanobody exhibited significantly improved pharmacokinetics in vivo and enhanced antitumor activity in a mouse model treated with CD19 chimeric antigen receptor T cells. This study presents an innovative epitope-directed approach that greatly accelerates the discovery of functionally potent agonistic nanobodies by effectively circumventing enrichment-driven epitope bias. Our approach and engineered multivalent anti-OX40 nanobody offer a powerful platform to advance immunotherapeutic strategies for cancer treatment.

While classical tumor suppressors in colorectal cancer (CRC) are predominantly recognized for restraining cell-autonomous proliferation, their extrinsic mandate in orchestrating the tumor immunometabolic niche remains poorly defined. Clinically, we document that APC membrane recruitment protein 1 (AMER1) downregulation correlates with advanced progression and cytotoxic CD8+ T cell spatial exclusion in CRC patients. Using parallel homograft models in diverse host immune backgrounds, we show that tumoral AMER1 confers robust in vivo tumor-suppressive effects that are dependent on a fully functional immune system. Single-cell RNA sequencing reveals that tumoral AMER1 enrichment actively preserves CD8+ T cell effector stemness by expanding the CXCR5+ precursor exhausted subset (Tpex) across regional lymph nodes and primary tumor microenvironments. Integrated multi-omics and biochemical tracking identify dopamine (DA) as the conserved neurometabolic effector driving this niche remodeling. Mechanistically, AMER1 physically binds and rescues dopa decarboxylase (DDC) from post-translational degradation to sustain tumoral DA secretion; conversely, AMER1 loss creates a localized DA void. Cell-autonomously, tumoral DA accumulation triggers Gasdermin D (GSDMD)-dependent tumor pyroptosis. Therapeutically, local DA administration halts multi-lineage carcinoma progression by reversing CD8+ T cell terminal exhaustion and reinforcing central memory differentiation. Collectively, our findings redefine AMER1 as a critical immunometabolic gatekeeper and establish neurotransmitter metabolic bypassing as a promising therapeutic strategy for CRC.

Glioblastoma remains a challenging disease to approach with immunotherapy due to pronounced antigen heterogeneity, immunosuppressive tumor microenvironment, and barriers to effective molecule delivery within the central nervous system. T-cell engagers provide an off-the-shelf approach to redirect endogenous T cells toward tumor cells. However, bispecific formats are constrained by intra- and interpatient antigen heterogeneity, which can limit therapeutic efficacy. In this review, we examine trispecific T-cell engagers (TriTEs) as an emerging strategy to address this limitation by simultaneously targeting multiple tumor-associated antigens. We discuss principles guiding antigen selection in glioblastoma, summarize available preclinical evidence supporting multispecific engagement, and outline key design considerations, including molecular architecture, stability, half-life extension, and safety optimization. We further review delivery strategies, such as gene-encoded expression, cellular carriers, and blood-brain barrier modulation, that may improve tumor access and the durability of TriTEs. Together, these considerations position TriTEs as a modular immunotherapy platform relevant to glioblastoma and other heterogeneous solid tumors.

Immune evasion remains a major barrier to effective immunotherapy in cancer. Coordinated activation of different immune cell types shapes immune responses within the tumor microenvironment (TME). In this study, we uncovered a spatiotemporal synergy between cytotoxic T lymphocytes (CTL) and natural killer (NK) cells in tumors with low T-cell infiltration. An integrative multiomics approach combining spatial transcriptomics, single-cell RNA sequencing, and high-dimensional flow cytometry demonstrated that vaccination, capable of activating both innate and adaptive immunity, rapidly reshapes the TME. Early infiltration of IRF8+KLRG1+ NK cells established a chemokine-rich niche together with myeloid cells that promoted the recruitment of antigen-primed CXCR3+ CTLs. The accompanying inflammatory monocyte recruitment and dendritic cell activation led to the coordinated structural and chemotactic remodeling of the TME. Functionally, NK-CTL clustering at the tumor margin was associated with enhanced cytotoxic activity and sustained immune engagement. These spatially organized immune interactions involved CCR5-CCL5 and CXCR3-CXCL9 signaling pathways that coordinate communication between innate and adaptive immune compartments. Together, these findings reveal a previously unrecognized NK-CTL cooperative program that promotes the transition of poorly infiltrated tumors toward an inflamed, immune-responsive state. More broadly, this study illustrates how spatially resolved multiomics approaches can uncover immune interactions and provides a conceptual framework for designing next-generation immunotherapies that mobilize coordinated innate and adaptive immunity. Early infiltration of NK cells followed by T cell accumulation reshapes the tumor microenvironment to promote antitumor immunity following vaccination, providing a mechanistic rationale for the development of effective vaccine-based immunotherapy strategies.

Ever since immune checkpoint blockade showed activity in the treatment of cancer, the search has been on for combination regimens that make this therapy more effective. In this issue, Blagg and colleagues describe an unorthodox approach to increasing the effectiveness of cancer immunotherapy. See related article by Blagg et al., p. 1649.

Despite remarkable advances in cancer drug treatment, including chemotherapy, targeted therapy, and immunotherapy, therapeutic resistance remains a formidable clinical barrier, limiting durable responses and long-term survival. Drug resistance can be broadly categorized as intrinsic, where tumors fail to respond to initial treatment, or acquired, which emerges during or after therapy due to adaptive or evolutionary processes. A comprehensive understanding of the multifactorial and dynamic nature of resistance is essential for improving treatment efficacy. In this review, we systematically examine the molecular and cellular determinants of drug response and resistance across 22 cancer types, highlighting key resistance mechanisms such as compensatory pathway activation, phenotypic plasticity, immune evasion, enhanced DNA damage repair, and the survival of drug-tolerant persister cells. These mechanisms are further contextualized across major therapeutic modalities, supported by clinical trials. We also present emerging strategies to overcome resistance, including rational drug combinations, novel agents, microbiome modulation, adaptive and intermittent therapies and advanced drug delivery systems, each illustrated with representative clinical studies. Moreover, we discuss cutting-edge tools that are revolutionizing resistance research, including single-cell and spatial multiomic profiling, patient-derived tumor organoid and xenograft (PDO/PDX) models, and artificial intelligence (AI)-powered predictive analytics. By integrating insights across molecular, cellular, and clinical dimensions, this review offers a strategic framework for understanding and tackling cancer drug resistance, with important translational implications for the future of precision oncology.

DNA mismatch repair (MMR) detects and corrects postreplicative DNA alterations; it is deregulated in up to 20% of human cancers. MMR-deficient (MMR-d) cancers display increased tumor mutational burden (TMB) and microsatellite instability (MSI) and are eligible for checkpoint inhibitor (CPI) immunotherapy which commonly elicits durable responses. We reasoned that pharmacologic blockade of MMR could broaden the patient population eligible for immunotherapy. Here, we reveal MMR protein PMS2 as a druggable target and describe the discovery and characterization of first-in-class small-molecule MMR pathway modulator NP1867. In vitro treatment of murine cancer cells abrogates MMR function and elicits an MMR-d genotype, including increased TMB, MMR-d mutational signatures, and MSI-high (MSI-H) status. Inoculation of syngeneic immunocompetent mice with cancer cells pretreated with NP1867 leads to CPI sensitivity, tumor growth delay, and complete responses. For the first time, we demonstrate pharmacologic targeting of MMR to proactively rewire the tumor-host relationship for therapeutic purposes. We present the discovery and characterization of NP1867, a selective covalent small-molecule inhibitor of MMR protein PMS2. Pharmacologic blockade of PMS2 leads to an MMR-deficient genotype characterized by increased TMB, enriched MMR-d mutational signatures, and MSI-H status. Treatment with NP1867 converts immunoresistant, MMR-proficient cancers into CPI-responsive tumors. See related commentary by Bernards, p. 1486.

Lymph node (LN) metastasis (LNM) in early-stage pancreatic ductal adenocarcinoma (PDAC) predicts systemic dissemination and poor survival, yet its underlying mechanisms remain elusive. In this study, we demonstrated that senescent cancer-associated fibroblasts (senCAF) drive lymphatic remodeling and LNM in early-stage PDAC. Mechanistically, senCAFs increased glucose metabolism and lactate production, which activated lactylation-mediated serine metabolism to protect lymphatic endothelial cells from oxidative stress. Moreover, we discovered CCR4+ regulatory T cells from the draining LNs accumulated around lymphatic vessels, which established an immunosuppressive perilymphatic niche. High-throughput drug screening determined selective clearance of senCAFs via chidamide, attenuated tumor progression, and improved chemoimmunotherapeutic efficacy. We subsequently initiated a clinical trial (chidamide and nab-paclitaxel/gemcitabine plus anti-PD-1/CTLA-4) in patients with metastatic PDAC and reported its preliminary promising results. Collectively, these findings reveal a closed link between cellular senescence and PDAC metastasis, offering the potential senolytic means to improve chemoimmunotherapy efficacy. Our findings have revealed a closed link between cellular senescence, metabolic reprograming, and spatial immunosuppressive niche and PDAC metastasis, offering the potential senolytic drugs to improve chemoimmunotherapy efficacy in patients with PDAC.

The role of intratumoural plasma cells in immune checkpoint blockade (ICB) therapy has never been tested although their presence is linked with improved response and survival in people with cancer. Malignant peripheral nerve sheath tumours (MPNSTs) are deadly sarcomas with minimal responsiveness to ICB therapies. Strikingly, drugs inhibiting cyclin-dependent kinases 4/6 (CDK4/6) and MEK sensitise de novo MPNSTs to immunotherapy targeting programmed death-ligand 1 (PD-L1), which correlates with increased intratumoural plasma cells. Here, we tested if plasma cells mediate MPNST response to anti-PD-L1 therapy and how they modulate tumour immune responsiveness to CDK4/6-MEK inhibition. Anti-tumour activity of PD-L1 inhibition, with or without CDK4/6-MEK inhibition, was measured in de novo MPNSTs within wild-type versus plasma cell-deficient mice. Plasma cell-dependent effects of CDK4/6-MEK inhibition on priming the MPNST immune environment were determined by single cell transcriptomics and immune cell analyses. Plasma cell-deficient MPNSTs failed to respond to anti-PD-L1 monotherapy and were no longer sensitised by CDK4/6-MEK inhibition to immunotherapy. Following kinase inhibitor treatment, plasma cells were necessary for a pro-inflammatory response marked by increased tumour infiltration and activation of natural killer (NK) and CD8+ T cells, major histocompatibility class I antigen presentation, and decreased M2 macrophages. By comparison, elevated CD4+ T cell and B cell infiltration dominated the plasma cell knockout phenotype. Plasma cells favourably remodel the tumour immune environment for anti-tumour immunity and are critical for successful ICB therapy in MPNSTs. These findings may inform ICB treatment strategies and patient stratification for therapy of many tumour types. This research was supported by University of Iowa Sarcoma Research Program awards, the Gilbert Family Foundation, and NIH grants T34-GM141143, T32-GM067795, F31-CA281312, P30-CA086862, and R01-NS119322.

Metabolic reprogramming is a defining feature of cancer and a major contributor to immune escape. Beyond the well-defined glycolysis, dysregulated amino acid and lipid metabolism also regulate tumor growth, stress adaptation, and therapeutic resistance. Amino acids such as glutamine, arginine, tryptophan, methionine, serine, and cysteine shape biosynthesis, redox balance, one-carbon metabolism, epigenetic control, and nutrient competition in the tumor microenvironment. Lipid uptake, de novo lipogenesis, fatty acid oxidation, cholesterol remodeling, COX-PGE2 signaling, sphingolipid metabolism, and ferroptosis further influence antigen presentation, immune cell fitness, and checkpoint regulation. However, most studies still consider these metabolic axes separately, leaving the coordinated amino acid-lipid crosstalk across tumor and immune compartments insufficiently defined. This review synthesizes recent advances in amino acid metabolism, including the glutamine axis, arginine-polyamine biology, the tryptophan-kynurenine-AHR pathway, and methionine-dependent methylation programs. It then discusses lipid metabolic programs that regulate dendritic cell cross-presentation, suppressive myeloid polarization, CD8+ T cell exhaustion, PD-L1 palmitoylation, MHC-I stability, and lipid-peroxidation-linked ferroptosis. We further integrate nutrient competition, immunometabolic checkpoints, and dual metabolic targeting strategies with immune checkpoint blockade. This review provides a unified framework for identifying metabolic vulnerabilities, designing rational combination therapies and refining precision cancer immunotherapy.

The MYC oncogene family constitutes a master regulatory hub in tumorigenesis, with functional complexity extending far beyond individual gene activities. Recent advances unveil cooperative yet context-dependent antagonism and dynamic interplay among MYC family members, fundamentally reshaping our understanding of lineage specific oncogenic programs. This review synthesizes emerging insights into MYC orchestrated tumor microenvironment remodeling, reciprocal regulation with noncoding RNAs, and the transcriptional and epigenetic governance of metabolic reprogramming. We delineate mechanisms by which MYC drives therapeutic resistance and critically evaluate current strategies targeting MYC or its downstream networks, encompassing direct MYC-MAX disruptors, upstream pathway inhibitors, synthetic lethality, and combinatorial regimens with immune checkpoint blockade or conventional chemotherapy. We further discuss MYC's prognostic significance across diverse cancer types, the critical gap between preclinical efficacy and clinical outcomes, and emerging combination strategies aimed at overcoming acquired drug resistance. By integrating these rapidly evolving biological dimensions, we posit MYC as a highly multidimensional regulatory node whose context-dependent functions present both formidable challenges and promising new opportunities for effective therapeutic intervention.

Glucose Transporter 1 (GLUT1) is the quintessential facilitator of basal glucose uptake, indispensable for maintaining cellular energy homeostasis, particularly across the blood-brain barrier. Beyond physiological necessity, GLUT1 dysregulation drives a broad pathological spectrum. While genetic haploinsufficiency precipitates severe neurological energy crises like Glut1 deficiency syndrome (Glut1DS), oncogenic networks hyperactivate GLUT1 as the central executor of the Warburg effect to fuel malignant proliferation. Despite its immense therapeutic potential, severe on-target toxicity in normal tissues and adaptive metabolic plasticity remain critical roadblocks to systemic GLUT1 inhibition. This review comprehensively synthesizes GLUT1's multidimensional regulatory networks in health and disease, dissecting how its overexpression fundamentally remodels the tumor microenvironment (TME). We elucidate how GLUT1-driven "metabolic competition" fosters metabolic immune exclusion and drives therapeutic resistance. Furthermore, we map the paradigm shift from traditional systemic blockades to emerging precision interventions. Specifically, we highlight "Trojan horse" glycan-functionalized nanocarriers, targeted protein degradation technologies like PROTACs, and metabolically engineered CAR-T cells. By conceptualizing GLUT1 as the linchpin of the immunosuppressive ecosystem, this work provides a strategic roadmap for precision metabolic immuno-oncology, guiding the development of novel therapies that maximize durable efficacy while minimizing collateral physiological damage.

Metabolic dysregulation is a hallmark of tumorigenesis and profoundly impacts immune surveillance; however, the underlying mechanisms and targeted treatments remain limited in colorectal cancer (CRC). Here, we utilize public databases, mouse models, and multi-omics analyses to identify N-acetyltransferase 1 (NAT1) as a critical prognostic-associated gene that emerges as a significant modulator of tumor immunity in CRC. We demonstrate that NAT1 suppresses glycolysis and lactate production, thereby promoting an immune-activated tumor microenvironment (TME). Mechanistically, NAT1 interacts with enolase 1 (ENO1), a key glycolytic enzyme, acetylating it at lysine 343 (K343) and thereby inhibiting its activity. However, loss of NAT1 in tumor cells leads to enhanced ENO1 activation, which in turn drives glycolysis and lactate production. Lactate then binds to TNF receptor-associated factor 6 (TRAF6), promoting its oligomerization and activation, leading to K63-linked ubiquitination of PD-L1, which enhances PD-L1 stability and facilitates immune evasion. Importantly, low NAT1-expressing tumors from CRC patients show increased sensitivity to anti-PD-1 therapy, and NAT1 deficiency significantly improves the efficacy of anti-PD-L1 antibody treatment in preclinical mouse models. Collectively, our findings highlight the critical role of NAT1 in reshaping the TME and suggest that targeting the NAT1-ENO1-lactate axis represents a promising therapeutic strategy to enhance immunotherapy in CRC.

Metastasis remains the leading cause of cancer-related mortality and is increasingly recognized as a consequence of dynamic interactions between tumor cell plasticity and a heterogeneous tumor microenvironment (TME). Rather than being genetically fixed, cancer cells exhibit phenotypic flexibility, enabling reversible transitions among epithelial, mesenchymal, and stem-like states in response to intrinsic programs and extrinsic microenvironmental cues, central to this adaptability. This review synthesizes emerging evidence that tumor progression is governed by reciprocal feedback loops between plastic tumor cells and distinct microenvironmental niches, including hypoxic cores, invasive margins, and perivascular regions. We highlight how stromal components, immune infiltrates, endothelial cells, and extracellular matrix (ECM) remodeling dynamically shape tumor cell states through biochemical and biophysical signals. Advances in single-cell and spatial transcriptomic technologies have revealed the spatial organization and reversibility of these plastic phenotypes, uncovering rare but clinically significant drug-tolerant persister populations. Importantly, we discuss plasticity-mediated therapy resistance as an adaptive, nongenetic process driven by transcriptional and epigenetic reprogramming, metabolic flexibility, ECM stiffening-induced mechanotransduction, and immune-checkpoint plasticity under therapeutic pressure. Together, these findings establish tumor plasticity and microenvironmental heterogeneity as an integrated, evolving system that fuels metastasis and limits durable treatment responses. Targeting this tumor-TME plasticity axis represents a promising strategy to disrupt metastatic progression and overcome therapeutic resistance.

Clear cell renal cell carcinoma (ccRCC) remains a therapeutic challenge due to tyrosine kinase inhibitor (TKI) resistance and an immunosuppressive tumor microenvironment (TME). To avoid the limitations of systemic nanotherapeutics, which often suffer from inadequate tumor accumulation, we developed a pH-activatable injectable gelatin methacryloyl (GelMA) hydrogel (termed SUN/CuONPs@GelMA) for localized co-delivery of sunitinib and copper oxide nanoparticles. The photocrosslinkable GelMA matrix enables minimally invasive intratumoral administration and sustained drug retention. Unlike ferroptosis, which is frequently migigated by the antioxidant defenses inherent in ccRCC, cuproptosis circumvents this resistance by targeting mitochondrial metabolism. In the weakly acidic TME (pH 5.3-7), CuONP assemblies dissolve to release CuO nanoparticles and Cu2+, triggering dual mechanisms: (1) cuproptosis via mitochondrial proteotoxicity and the depletion of iron-sulfur cluster protein, and (2) Fenton-like reaction-mediated reactive oxygen species (ROS) generation and the release of damage-associated molecular patterns (DAMPs). Concurrently, sunitinib inhibits angiogenesis and synergizes with copper-driven immunomodulation to reprogram macrophages toward tumor-suppressive M1 phenotypes. In vitro and in vivo studies demonstrated potent tumor growth inhibition, migration suppression, and prolonged survival in ccRCC. The SUN/CuONPs@GelMA platform overcomes TKI resistance through the synergy of cuproptosis and immunotherapy. This localized co-delivery strategy provides a multi-mechanistic approach for the treatment of advanced ccRCC. STATEMENT OF SIGNIFICANCE: Systemic treatments for advanced clear cell renal cell carcinoma (ccRCC) are restricted by tyrosine kinase inhibitor (TKI) resistance, systemic toxicity, and an immunosuppressive tumor microenvironment (TME). We herein report a pH-activatable, injectable photocrosslinkable GelMA hydrogel for localized co-delivery of sunitinib and copper nanoparticles, enabling minimally invasive delivery, sustained drug retention, and reduced systemic toxicity. This hydrogel triggers cuproptosis via acidic TME-responsive copper release, and synergizes with sunitinib to reverse TKI resistance, suppress tumor progression and migration, reprogram immunosuppressive macrophages to antitumor phenotypes, and extend preclinical survival. This work establishes a combinatorial ccRCC therapeutic platform, providing a promising translational strategy for advanced ccRCC management.

The immunoregulatory role of androgen receptor (AR) in hepatocellular carcinoma (HCC) is still poorly understood. Herein, we found that inhibition of AR function could induce immunogenic cell death (ICD) and trigger endoplasmic reticulum (ER) stress in vitro. Mechanistically, AR transcriptionally regulated the valosin-containing protein (VCP) expression via directly binding the promoter of VCP. AR inhibitor promoted dendritic cells (DCs) maturation in vitro and increased tumor-infiltrating DCs and CD8+ T cells in vivo. Combination of AR inhibitor with anti-PD-1 therapy remodels tumor immune microenvironment (TIME) and reduces tumor growth in mouse models of HCC. Addition of oncolytic virus further augments the therapeutic activity of the combination regimen. Our study shows the molecular mechanism for regulating VCP expression by AR and suggests that inhibition of AR has the potential to increase the efficacy of immunotherapy in HCC.

T cells are crucial for defending against viral infection and cancer by eliminating infected or transformed cells and establishing immune memory. However, persistent antigenic stimulation in chronic infections or tumors drives T cells into a dysfunctional state known as exhaustion. This state is characterized by reduced proliferation and effector functions, upregulation of inhibitory receptors like PD-1, LAG-3, and CTLA-4, and alterations in transcriptional, epigenetic, and metabolic programs. T cell exhaustion is driven by both intrinsic factors, including changes in transcription factor networks and metabolic dysfunction, and extrinsic factors, such as continuous antigen exposure and an immunosuppressive microenvironment. Key molecules like PD-1, TOX, and TCF-1 are central to this process, though the complex interactions between intrinsic and extrinsic signals in chronic viral infections and cancers remain poorly understood. This review summarized T cell exhaustion in chronic viral infections, such as HIV, HBV, and SARS-CoV-2, as well as in tumors, emphasizing shared mechanisms and context-specific differences. We focused on the roles of transcriptional networks, metabolic changes, immune checkpoints, and exhaustion-related signaling. Additionally, we discussed emerging therapeutic strategies, such as immune checkpoint inhibitors, CAR-T cell therapies, cytokine supplementation, and metabolic interventions, based on recent high-impact studies. By integrating insights from both chronic infection and cancer, this review aims to identify common principles of T cell exhaustion and propose strategies to improve clinical outcomes in chronic viral diseases and cancer immunotherapy.

The tumor microenvironment (TME)-marked by hypoxia, acidosis, and nutrient scarcity-creates a metabolically restrictive niche that undermines sustained antitumor immunity. Within this niche, infiltrating immune cells could become functionally exhausted to limit the efficacy of existing immunotherapies. Recently, metabolic competition between tumor cells and immune cells for nutrients has attracted great attention in immunometabolism to explain the immune dysfunction. Notably, micronutrients, particularly vitamins, have been increasingly revealed to serve as active immunoregulatory agents rather than working solely as metabolic precursors or intermediates. In this review, we highlight the role of vitamins as immune-metabolic modulators that coordinate metabolic reprogramming, epigenetic remodeling, and signal transduction in tumor-infiltrating immune cells. We further discuss how vitamin activity is shaped by tumor-specific metabolic states, molecular forms, microbiome-dependent regulation and microenvironmental conditions, resulting in context-dependent immunological outcomes. By integrating preclinical mechanistic insights with the current landscape of clinical trials, the translational challenges arising from the pleiotropic effects of vitamins have also been evaluated. Finally, we also summarize emerging biomarker-guided and tumor-targeted intervention strategies that may help improve the therapeutic utility of vitamins and overcome immunotherapy resistance.

Chimeric antigen receptor (CAR)-T cell therapy has emerged as a revolutionary treatment for hematologic malignancies and solid tumors. Multiple approved products and ongoing clinical trials have demonstrated its remarkable antitumor efficacy. However, its clinical application is severely limited by prominent cardiotoxicity, which is closely associated with high morbidity and mortality, posing an urgent clinical challenge. This review systematically elucidates the pathophysiological mechanisms underlying CAR-T cell therapy-induced cardiotoxicity, which can be primarily categorized into acute and long-term adverse effects. Acute cardiotoxicity is driven by cytokine release syndrome (CRS) through macrophage and monocyte activation, endothelial dysfunction, and subsequent myocardial injury, leading to hypotension, reduced left ventricular ejection fraction, and cardiac arrhythmias. Long-term cardiotoxicity is mainly caused by B-cell aplasia-induced immune deficiency and secondary hemophagocytic lymphohistiocytosis, which trigger infectious complications, myocardial ischemia, and apoptosis. We have further summarized clinical pharmacological interventions for such cardiotoxicity, including interleukin (IL)-6 receptor antagonists (tocilizumab), interferon-γ inhibitors (emapalumab), and Janus kinase inhibitors (ruxolitinib). However, none of these drugs can completely alleviate CRS or CAR-T cell therapy-induced cardiotoxicity in all patients. Preclinical agents targeting inflammatory cytokines (IL-1β, tumor necrosis factor-α) and signaling pathways have shown efficacy in ameliorating cardiotoxicity in various animal models. These findings provide novel directions and drug candidates for the treatment of CAR-T cell therapy-induced myocardial toxicity. This review provides a comprehensive theoretical basis for optimizing cardiovascular safety management and developing novel targeted interventions for CAR-T cell therapy.

Trained immunity, an emerging treatment strategy, confers long-term anti-tumour memory by modulating epigenetic and metabolic reprogramming of innate immune cells, offering a novel pathway for immunotherapy of haematological malignancies. This article systematically outlines the mechanisms by which inducers such as β-glucan, Bacille Calmette-Guérin (BCG) and cytokines train key effector cells including macrophages and natural killer (NK) cells. It further analyses how ageing-encompassing both organismal ageing and cellular senescence-specifically undermines the efficacy of relevant pathways at multiple levels, from haematopoietic stem cell function and intracellular signalling to the tumour microenvironment, thereby systematically impairing the induction efficiency, durability of effect, and safety of trained immunity. Building on this, the article evaluates the clinical translation prospects and associated risks, including graft-versus-host diseases (GVHDs), of strategies such as cytokine-induced memory-like (CIML) NK cells, and proposes concrete, testable translational ideas aimed at overcoming ageing-related defects. We argue that to realise the full potential of this therapy in elderly patients with haematological malignancies, future work must focus on developing individualised training protocols capable of circumventing or reversing immunosenescence.

While the long-term efficacy of checkpoint inhibition is established, the lack of response in IFN-rich tumors presents a critical challenge. These findings suggest that immune sensitivity depends on variables beyond cytokine levels and should be assessed in a temporal context. We propose the "IFN clock," in which efficacy is determined by the kinetics of interferon signaling rather than its level. The IFN clock is structured into three phases: a fast-on activation phase, a fast-off recovery phase, and a persistent-on resistant phase. In this model, short-lived interferon signaling supports antitumor immunity, whereas prolonged signaling induces therapeutic resistance. In early signaling, Type I interferon drives immune cell coordination through myeloid priming and chemokine-guided T cell recruitment. Effective outcomes require attenuation of signaling; loss of feedback control promotes chronic interferon exposure. Persistent activation results in stable epigenetic remodeling that enforces immune evasion and progressive T cell dysfunction. We introduce the dISG ratio to operationalize this framework. It provides a transcriptomic metric that separates acute from chronic interferon states, enabling patient stratification. Emerging preclinical evidence suggests that modulating interferon kinetics may restore checkpoint blockade sensitivity, though clinical validation remains limited. The IFN clock reframes interferon biology as dynamic and time-dependent. It offers a framework for biomarker refinement and therapeutic design.

Immune checkpoint blockade targeting programmed death protein-1 (PD-1) and its ligand PD-L1 has shown substantial therapeutic benefit in only a subset of patients with solid tumors, highlighting the urgent need to improve cancer immunotherapy outcomes. Hypoxia, a hallmark of the tumor microenvironment (TME), plays a critical role in modulating antitumor immunity and contributes to resistance against PD-1/PD-L1 blockade. Hypoxia influences multiple microenvironmental factors, affecting disease progression, treatment responses, and prognosis in solid tumors. As a result, hypoxia has emerged as a pivotal therapeutic target in combination immunotherapy approaches. This review explores the molecular and cellular mechanisms by which the hypoxic TME induces resistance to PD-1/PD-L1 blockade across distinct tumor types. Furthermore, we examine current strategies that target hypoxia to enhance immunotherapy efficacy and discuss the potential benefits of combination regimens. Collectively, this study provides a comprehensive framework for the rational design and clinical optimization of PD-1/PD-L1 inhibitors, with the goal of expanding their therapeutic applicability in solid malignancies.

mRNA therapy offers precise control, scalability, and a favorable safety profile, making it a promising modality for disease treatment. However, its clinical success critically depends on effective mRNA delivery systems. Currently approved lipid nanoparticles (LNP) predominantly accumulate in the liver, leading to hepatotoxicity and restricting their application to liver-related diseases. Here, we synthesized a series of ionizable lipidoids with distinct linker structures-alkylated, hydroxylated, and esterified-and combined with DOPE and cholesterol to construct three-component lipid nanoparticles (tLNP). We demonstrate that the linker structure of ionizable lipidoids is a key determinant of both the mRNA delivery efficiency and organ selectivity of tLNP. Compared with alkylated or esterified counterparts, tLNP formulated with hydroxylated-linker ionizable lipidoids exhibit enhanced cellular uptake, superior mRNA delivery, and pronounced spleen-selective mRNA expression. Notably, these tLNP achieve a spleen-to-liver mRNA expression ratio of up to 85.88, effectively reducing liver toxicity and enabling extrahepatic mRNA therapies. Leveraging this platform, we developed a therapeutic mRNA vaccine (mLMP2A@BO10-tLNP) for EBV-associated cancers. The immunological role of the spleen and PEG-free tLNP design enhance vaccine safety and efficacy. This work provides design principles for efficient mRNA delivery and introduces strategies for treating EBV-associated cancers. STATEMENT OF SIGNIFICANCE: mRNA-based therapies have demonstrated transformative potential, with multiple applications advancing toward clinical translation. A critical factor in this process is the optimization of lipid nanoparticle (LNP) delivery systems. While current research has largely focused on the design of ionizable lipidoids, the regulatory role of their linker structures remains underexplored. This study reveals that linker structures critically govern LNP-mediated mRNA delivery efficiency and organ-selective expression, providing design principles for next-generation LNPs. Furthermore, by leveraging virus-derived antigens, this work presents a therapeutic mRNA tumor vaccine strategy that significantly reduces off-target cytotoxicity, addressing a key limitation of tumor antigen specificity. Together, these findings offer important insights for improving the safety and translational potential of therapeutic mRNA cancer vaccines.

Photothermal therapy (PTT) has attracted considerable attention in cancer treatment due to its noninvasive nature and lack of drug resistance. However, conventional photothermal agents often exhibit poor tumor accumulation, nonspecific thermal damage, and limited monotherapy efficacy, hindering their clinical translation. To overcome these constraints, we developed a tumor-microenvironment-adaptive peptide-based nano-photothermal agent (EIMSeO-IR783) founded on the principle of intermolecular coupling-induced supramolecular photothermal effects, and integrated it with a STING agonist for synergistic photothermal-immunotherapy. The nano-photothermal agent employs a redox-responsive peptide containing selenomethionine residues as a self-assembled skeleton, and realizing the transformation from nanoparticles to nanofibers through redox, thereby regulating the aggregation state of IR783 and inducing the formation of H-aggregates. These H-aggregates enhance non-radiative energy dissipation, leading to improved photothermal conversion efficiency and photostability. In the tumor microenvironment, overexpressed GSH drives adaptive morphological transformation, enabling precise in situ heat generation and improving tumor retention and ablation. Combined with the STING agonist, photothermal-induced immunogenic cell death promotes antigen release, dendritic cell maturation, and T-cell infiltration, eliciting a robust systemic immune response for synergistic photothermal-immunotherapy. This work proposes a tumor-microenvironment-guided strategy for constructing peptide-based photothermal systems, offering a new avenue for precise cancer therapy with controllable photothermal performance and immune synergy. STATEMENT OF SIGNIFICANCE: Photothermal therapy is a promising cancer treatment, but its effectiveness is limited by poor tumor accumulation, nonspecific heating, and insufficient therapeutic outcomes. This study presents a tumor-microenvironment-adaptive peptide-based nanophotothermal agent that undergoes structural transformation in response to intracellular glutathione, enabling localized heat generation and improved tumor retention. The adaptive assembly regulates H-aggregate formation of IR783, enhancing photothermal efficiency and stability. In combination with a STING agonist, the system induces immunogenic tumor cell death and stimulates systemic antitumor immunity, achieving synergistic photothermal-immunotherapy. This work introduces a peptide-guided strategy for controlling photothermal performance within tumors and provides a versatile platform for developing precise and multifunctional biomaterial-based cancer therapies.

Epitranscriptomic RNA marks have moved from molecular footnotes to a control layer deciding which transcripts are translated, stored or discarded. In solid tumors, this layer sits directly upstream of how the tumor microenvironment behaves: it links oncogenic signaling to antigen visibility, interferon tone, myeloid recruitment, stromal architecture and vascular fitness through three recurrent "conduit circuits" centered on NF-κB-driven inflammation, Wnt/β-catenin-mediated immune exclusion and Dll4-NOTCH-patterned angiogenesis. Yet translation into clinical practice has lagged: mechanistic work is fragmented, pharmacodynamic rules for short lead-in windows and axis-aligned combinations are rarely shared, and assays range from semi-quantitative bulk measures to non-standardized spatial maps. This review brings mechanistic and clinical observations together into a single map of how epitranscriptomic circuits can be used in trials. It describes how m⁶A and related marks shape these conduit circuits and clinical programs spanning stemness, perivascular niches, angiogenesis, metastasis and treatment resistance, and links them to a bench-to-bedside pharmacodynamic workflow that combines LC-MS, direct RNA sequencing, spatial profiling, perfusion imaging and tumor-informed ctDNA. Finally, four practical operating principles for clinical use of this biology are outlined: pharmacodynamic-gated axis alignment, perfusion-gated vascular normalization, lineage-aware spatial pharmacodynamics and reader-level, compartment-selective control, together with a proposed conduit-circuit response score to judge and combine epitranscriptomic interventions on the basis of tissue-level pharmacodynamic readouts.

Interleukins (ILs) are powerful regulators of anti-tumor immunity, yet their clinical impact in cancer has remained limited despite decades of therapeutic development. Although first-generation cytokines such as IL-2 established proof-of-principle for cytokine-driven cancer immunotherapy, their broader clinical translation was constrained not by insufficient immunostimulatory potency, but by imprecise deployment, including poor receptor selectivity, systemic exposure, and inadequate alignment with tumor immune context. Despite major advances in cytokine engineering, clinical benefit has continued to lag because cytokine deployment remains insufficiently matched to receptor biology, spatial pharmacology, and immune-state architecture. In this Review, we examine how next-generation interleukins, particularly IL-15, IL-21, and IL-10, are redefining cytokine therapy through functionally specialized roles in sustaining cytotoxic persistence, preserving effector competence, and rewiring suppressive immune states. We synthesize emerging strategies in receptor-biased engineering, spatially restricted delivery, and context-matched combination design, and propose a cytokine-guided framework that integrates receptor logic, delivery geometry, immune-state matching, and biomarker-informed deployment. We further argue that the principal barriers to clinical translation are no longer primarily molecular, but translational, including biomarker insufficiency, trial misalignment, tissue-level resistance, and patient heterogeneity. Overall, next-generation interleukins are best understood not as stronger cytokines, but as more precisely deployable immunotherapeutic modules for durable and rational cancer immunotherapy.

On-target, off-tumor toxicity presents a significant challenge in chimeric antigen receptor (CAR) T therapy for solid tumors. Traditional approaches to managing adverse reactions, such as suppressing in vivo CAR-T cell activity, risk impairing their antitumor efficacy, often resulting in treatment failure. Intravenously infused CAR-T cells initially traffic to the lungs, where they are activated by tumor-associated antigens (TAAs) expressed on pulmonary tissues, leading to acute lung injury. To address this, this study developed a strategy involving leukocyte function-associated antigen 1 (LFA-1) neutralization at the time of infusion. This approach modulates CAR-T cell pharmacokinetics to enhance efficacy while minimizing toxicity. Post-infusion, CAR-T cells preferentially sequester and activate in the lung, secreting tumor necrosis factor α (TNF-α), which upregulates intercellular adhesion molecule 1 (ICAM-1) expression on pulmonary endothelial cells. This triggers an "activation-adhesion" feedback loop via the LFA-1/ICAM-1 pathway, exacerbating lung injury. Neutralization of LFA-1 during infusion significantly reduces CAR-T cell adhesion to pulmonary endothelium, disrupts this feedback loop, and mitigates acute lung injury. By accelerating the pharmacokinetic progression of CAR-T cells beyond the lung, this strategy not only alleviates the acute toxicity associated with high-dose regimens but also enhances the antitumor efficacy of low-dose CAR-T cells, thus expanding the therapeutic window.

Claudin-6 (CLDN6) is an essential component of tight junctions (TJs) with a characteristic oncofetal expression pattern, being largely absent in normal adult tissues but re-expressed in multiple malignancies. This tumor-restricted expression profile provides a potential therapeutic window for CLDN6-directed targeting. Increasing evidence indicates that CLDN6 participates in diverse aspects of tumor biology, including regulation of tumor growth, metastasis, metabolic reprogramming, immune microenvironment remodeling, and therapeutic resistance. Consequently, CLDN6 has emerged as both a cancer biomarker and a promising therapeutic target. Furthermore, we focused on the expression landscape and biological functions of CLDN6 across cancer types and discussed its clinical relevance in tumor progression and prognosis. We further highlighted recent advances in CLDN6-targeted therapeutic strategies, including monoclonal antibodies (mAbs), T cell-engaging antibodies (TCEs), antibody-drug conjugates (ADCs), and adoptive cell therapy (ACT). Collectively, these developments provide important insights into the translational potential of CLDN6 and support its continued exploration as a precision therapeutic target in CLDN6-positive cancers.

Immune checkpoint blockade has reshaped lung cancer therapy, yet durable benefit remains confined to a minority of patients, and resistance often emerges even in initially responsive disease. An increasingly coherent explanation is that neural state-encoded through hierarchical neuroimmune interactions-can profoundly recalibrate anti-tumor immunity and influence treatment responsiveness. In this Review, we synthesize evidence across three integrated levels of neuro-immune regulation in lung cancer. First, lung tumors engage in local neural-immune-tumor interactions, including synapse-like coupling, neuronal mimicry, perineural niches shaped by injury signaling, tumor-induced neurogenesis and neuroendocrine transitions driven by therapy. These local interfaces concentrate catecholaminergic, cholinergic and sensory neuropeptide signaling within spatial microdomains, repeatedly targeting immune bottlenecks. Second, tumor-initiated peripheral-to-central neural circuits link vagal tumor sensing to brainstem autonomic integration, creating a feedback loop that reshapes peripheral immune status. Third, at the systemic level, psychological stress and circadian disruptions impose neuroendocrine constraints through the hypothalamic-pituitary-adrenal axis and sympathetic nervous system, further amplifying immune heterogeneity and clinical variability. By framing "neural state" as a measurable and actionable clinical variable, this unified three-layered neural-immune model clarifies opportunities for patient stratification, mechanism-informed combinations, and neuromodulatory interventions to optimize immunotherapy efficacy in lung cancer.

Adoptive T cell therapy has transformed the treatment of hematologic malignancies but fails to control solid tumors, where T cell dysfunction and an immunosuppressive tumor microenvironment (TME) remain the central barriers. Cytokines are essential regulators of both T cell fate and the TME, making cytokine engineering a key lever for overcoming these limitations. Here we review recent advances in intrinsic strategies that embed cytokine support directly into the engineered T cell product and extrinsic strategies that deliver cytokines to the tumor site to remodel the TME, providing a comprehensive analysis to guide rational strategy selection and combination. We further highlight synthetic cytokine and receptor designs that induce novel T cell states beyond the boundaries of natural T cell biology. Finally, we propose virtual cytokine networks as a framework for predicting patient-specific immune cascades triggered by any designed cytokine intervention, providing a path toward personalized cytokine-guided adoptive T cell therapy.

Redox imbalance is a defining feature of the tumor microenvironment (TME). Tumor cells adapt to this oxidative pressure through robust antioxidant pathways, whereas immune cells are easily impaired, leading to poor antigen presentation, compromised T-cell dysfunction, and an immunosuppressive milieu. These redox disturbances affect nearly every step of the cancer-immunity cycle. Targeting the redox balance is thus a critical objective. However, this therapeutic avenue is fraught with challenges. The complexity and dynamic characteristics of redox regulation pose major obstacles to achieving precise modulation in the TME. Nanomedicine offers practical strategies to modulate redox dynamics, either by amplifying oxidative stress or reinforcing antioxidant capacity. This review provides a critical overview of the biological mechanisms underlying redox dysregulation in the TME, highlights nanomedicine-based interventions for precise redox modulation, and evaluates their strategic applications across the cancer-immunity cycle. Furthermore, we synthesize essential design principles to navigate the redox paradox, ultimately discussing future perspectives for overcoming translational barriers in redox-based cancer immunotherapy.

Immunotherapy has emerged as a new pillar of cancer therapeutics that utilizes the patients own immune system to target tumor cells, a particular advantage in the context of hard-to-treat cancers. Virus-based immunotherapies have shown great potential as cancer treatments, including oncolytic viruses, viral vectors and virus-like particles. Here, we discuss the development, applications, challenges, and future opportunities of cowpea mosaic virus (CPMV) as a plant virus-based candidate for intratumoral immunotherapy, focusing on the treatment of aggressive, metastatic and refractory solid tumors. Evidence of antitumor efficacy has been gathered from mouse tumor models and veterinary clinical trials involving canine cancer patients. CPMV is promising as a monotherapy and as part of combination therapies, including chemotherapy, radiation, checkpoint inhibitors, and cytokine therapies. The putative mechanism of action is described, highlighting key features such as the capsid proteins and RNA acting as Toll-like receptor (TLR) agonists for TLR-2, TLR-4 and TLR-7, as well as the presentation of unique epitopes that prime a Th-1 balanced immune response. The nanoparticle structure of CPMV enhances efficacy by exerting multivalency and avidity effects. The biodistribution, toxicity, and agronomical safety profile of CPMV is also described, especially in relation to tumor retention, hematologic toxicity, allergenicity and adverse events, and viral shedding. This body of work provides a thorough exploration of a cancer immunotherapy candidate in development for more than 10 years, positioning CPMV as a promising intratumoral platform for hard-to-treat cancers.

In recent years, biologics-based modalities have increased significantly across various therapeutic areas. Monoclonal antibodies (mAbs) and emerging bioconjugates such as antibody-drug conjugates (ADCs) and antibody-oligonucleotide conjugate (AOCs) now span oncology and immunology areas. This growth has made delivery method and route of administration central to the drug product design. From a patient perspective there is a need for improved treatment accessibility and convenience. This need has driven a transition from intravenous (IV) infusion to subcutaneous (SC) delivery. The shift has accelerated the development and approval of biologic-device combination products, such as prefilled syringes, autoinjectors, and on-body delivery systems (OBDS) for SC administration of mAbs. Despite these advances, SC delivery is associated with pharmacokinetic and formulation challenges. These challenges are intrinsic to the molecule and can affect the dose delivery and frequency of administration. Short half-life, suboptimal bioavailability, and target-mediated drug disposition (TMDD) represent typical pharmacokinetic failure modes. Additionally, SC administration of large volumes (> 2 mL) and/or high concentrations (≥ 150 mg/mL) presents challenges, such as viscosity limitations, injection site reactions (ISRs), and injection related pain. For bioconjugates, the payload chemistry, linker stability, and drug-to-antibody ratio introduces further considerations that influence the route of administration. Together, these constraints define the boundaries within which SC delivery strategies must operate. Numerous strategies are currently being developed to address these challenges. At the molecular level, Fc engineering approaches such as YTE and LS mutations extend the half-life of antibodies and aim at reducing dosing frequency. On the other hand, enzyme-assisted delivery using recombinant hyaluronidase facilitates large volume SC administration by transiently increasing tissue permeability. At a formulation level, strategies such as viscosity-reducing excipients, computational modeling, and high throughput screening are increasingly employed to enable high-concentration drug products. On the device side, large volume autoinjectors and on-body delivery systems (OBDS) are being developed to break through 1-2 mL volume limitations with existing devices. This enables self-administration of higher volume drug products which was previously restricted to health care settings. In parallel, complementary delivery routes such as intradermal (ID) via hollow microneedles are being explored. ID delivery due to its proximity to dermal lymphatic offers potential uptake of drugs thereby improving pharmacokinetics. Additionally, due to its minimally invasive nature, this route of administration has the potential to address injection pain related issues. Collectively, convergence of routes of administration (SC or ID) with advancements in molecular engineering, formulation science, and device technology is essential for improved drug delivery. Emerging modalities will further continue to influence the development of devices, including considerations for dosing frequency. Achieving this integrated vision requires a deeper understanding of pharmacokinetic failure modes and regulatory frameworks governing biologic-device development combination products.

Adoptive cell therapy (ACT) against solid tumors is constrained by tumor heterogeneity, immunosuppressive microenvironments, and insufficient T cell potency and persistence. Although IL-12 has long been recognized as a potent enhancer of T-cell immunity, its clinical application has been hindered by systemic toxicity and the lack of a strategy to harness its benefits without chronic exposure. Here, we show that ex vivo IL-12 preconditioning programs tumor-specific T cells with increased antigen sensitivity, yielding superior antitumor activity compared with IL-7/IL-15. Systemic delivery of Pro-IL-12 further rejuvenates exhausted CD8+ tumor-infiltrating lymphocytes (TILs) by restoring IL-2 hypersensitivity, sustaining persistence, and strengthening effector machinery. When combined with ACT, a single dose of Pro-IL-12 reinstated IFN-γ production in exhausted TILs and doubled intratumoral T-cell accumulation via upregulation of IL-2Rα and increased IL-2 sensitivity. Pro-IL-12 markedly improved therapeutic outcomes, inducing durable complete tumor regression, promoting antigen spreading, and establishing long-term immune memory that prevented relapse from antigen-loss tumor. Across multiple xenograft models, including pancreatic cancer, lymphoma and triple negative breast cancer, distinct human CAR-T products paired with Pro-IL-12 enhanced tumor eradication without cytokine release syndrome. Together, these findings establish precision delivery of IL-12 as a translatable strategy that harnesses known IL-12 biology to achieve robust antitumor efficacy without systemic toxicity.

Immune checkpoint inhibitors (ICIs) demonstrate therapeutic benefits in advanced hepatocellular carcinoma (HCC), yet most patients exhibit limited responses to anti-PD-1 therapy and the molecular basis underlying this resistance remains largely undefined. Here, we identified the circular RNA SPECC1 (circSPECC1) as a regulator of antigen presentation and a crucial factor influencing the efficacy of anti-PD-1 therapy. circSPECC1 was significantly elevated in tumor tissues of HCC patients showing resistance to anti-PD-1 therapy and correlated with diminished intratumoral CD8+ T-cell infiltration. Elevated circSPECC1 levels facilitated tumor progression and impaired the therapeutic efficacy of anti-PD-1 treatment. Overexpression of circSPECC1 in cancer cells suppressed the proliferation and cytotoxic activity of CD8+ T cells both in vitro and in vivo. Mechanistically, circSPECC1 bound to the peptide-loading complex (PLC) chaperone ERP57 and disrupted its interaction with TAPBP (TAP binding Protein), thereby destabilizing PLC integrity. This aberrant interaction redirected MHC-I toward ER-associated degradation (ERAD), suppressing surface antigen presentation and impairing CD8+ T-cell activation. Importantly, targeted silencing of circSPECC1 using a PEG-PEI-folate nanosystem encapsulating si-circSPECC1 significantly restored sensitivity both murine and patient-derived xenograft HCC models to PD-1 blockade. In summary, these findings uncover a novel circRNA-guided ERAD of MHC-I mechanism that impairs antitumor immunity and highlight circSPECC1 as a promising therapeutic target to overcome immunotherapy resistance.

The rapid development of targeted protein degradation (TPD) has shown profound effects on disease treatment. Precise and effective targeted degradation tools that target endogenous proteins are essential to accelerate advances in treatment methods. Selective macroautophagy/autophagy relies on the activity of related receptors to achieve the degradation of specific intracellular components in lysosomes, but the methodology of selective autophagy for tumor therapy and chimeric antigen receptor (CAR)-T cell modification is yet unexplored. Here, we developed a peptide-based LC3-interacting region-targeting chimera (pLIRTAC) that accurately and efficiently targeted the degradation of AKT1 for glioma treatment. pLIRTAC could also inhibit the development of tumor cells by in vitro delivery after purification. For CAR-T cell therapy, pLIRTAC could significantly improve the efficacy of CAR-T cell-targeted lysis of tumor cells both in vitro and in vivo. pLIRTAC binds to autophagy-associated proteins through LC3-interacting region (LIR) motifs and to target proteins through protein-targeting short peptides, and targets the protein of interest (POI) based on the selective autophagy lysosomal pathway. pLIRTAC has been remarkably successful both in vivo and in vitro, providing a robust and effective tool for the control of endogenous abnormal proteins in cells, and can potentially further expand the therapeutic application of TPD technology.Abbreviation: ATG8s: mammalian Atg8 (autophagy related 8)-family proteins; Baf-A1: bafilomycin A1; CAR: chimeric antigen receptor; CQ: chloroquine; CRISPR: clustered regularly interspaced short palindromic repeats; EBSS: Earle's balanced salt solution; LIR: LC3-interacting region; 3 MA: 3-methyladenine; MFI: mean fluorescence intensity; pLIRTAC: peptide-based LC3-interacting region-targeting chimera; POI: protein of interest; PROTAC: proteolysis-targeting chimera; SARS: selective autophagy receptors; TPD: targeted protein degradation.

Systemic administration of glucocorticoids (GCs) has immunosuppressive effects that involve the upregulation of the transcription factor TSC22D3 in dendritic cells (DCs), thereby reducing their capacity for antigen presentation to T lymphocytes. Recently, we found that this effect is not mediated by direct action on the GC receptor in DCs but rather involves an indirect signaling circuitry. Indeed, GCs act on the GC receptor expressed by many cell types to cause the upregulation and release of the tissue hormone DBI/ACBP (diazepam binding inhibitor, acyl-CoA binding protein). DBI/ACBP, which is an inhibitor of macroautophagy/autophagy, then acts on the benzodiazepine-binding site of the gamma-aminobutyric acid type A receptor (GABAAR) to elicit the upregulation of TSC22D3. The indirect, DBI/ACBP-dependent upregulation of TSC22D3 by GCs is observed both in vivo (mice) and in vitro, in murine splenocytes and bone marrow-derived DCs, as well as in human peripheral blood mononuclear cells and monocyte-derived DCs. Inhibition of human mixed lymphocyte reactions (confronting DCs and lymphocytes from distinct donors) by DCs is reduced by DBI/ACBP neutralizing antibodies. Similarly, the suppression of antitumor immune responses (elicited by vaccination with dying cancer cells, immunogenic chemotherapy or PDCD1/PD-1 blockade) by GCs is reversed by DBI/ACBP neutralization. Epistatic experiments indicate that knockout of Tsc22d3 in DCs and inhibition of DBI/ACBP act on the pathway to reverse GC-mediated inhibition of cancer immunosurveillance. Of note, the benzodiazepine diazepam restores GC-induced immunosuppression when DBI/ACBP is inhibited. Altogether, these findings support a role for the DBI/ACBP-GABAAR system in immunosuppression by GCs.

5-Aminolevulinic acid (5-ALA)-induced protoporphyrin IX (PpIX) accumulation is widely used for fluorescence-guided surgery in malignant glioma. Beyond its diagnostic role, PpIX exhibits photo-, sono-, and radiosensitizing properties that enable locally applied tumor therapies. For these modalities, the cellular and subcellular localization of PpIX is a critical determinant of therapeutic efficacy, as it defines the site of reactive oxygen species generation and subsequent biological effects. In this comprehensive literature review, the cellular and subcellular distribution of 5-ALA-induced PpIX in gliomas and their tumor microenvironment (TME) was assessed. Studies were identified through a structured MEDLINE search and evaluated for evidence of PpIX localization in neoplastic and nonneoplastic cellular components. Accumulating data indicate that PpIX localizes not only within malignant glioma cells but also across multiple cellular components of the TME. Advanced imaging, single-cell, and spatial transcriptomic analyses demonstrate PpIX fluorescence frequently aligns with immunosuppressive myeloid populations and infiltrative tumor regions. Subcellularly, PpIX localizes in mitochondria, lysosomal-autophagic compartments, and extracellular structures, in a cell-type dependent manner. These distribution patterns provide a biological basis for the immunomodulatory, antiangiogenic, and cytotoxic effects observed in studies of 5-ALA-mediated local therapies. While macroscopic fluorescence remains clinically valuable for fluorescence-guided resection, the biologically relevant microscopic distribution of PpIX supports the concept that PpIX-based therapies may target not only tumor cells but also immunosuppressive and stromal compartments. A refined understanding of PpIX localization is therefore central to optimizing nonsurgical 5-ALA-based therapies and their integration into multimodal glioma treatment paradigms.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a leading cause of hepatocellular carcinoma (HCC) and confers resistance to immunotherapy. However, the underlying mechanisms remain unclear. We aimed to elucidate how the lipid-rich microenvironment of MASLD-HCC drives immune suppression and to identify actionable targets. Dendritic cell (DC)-CD8+ T cell interactions in HCC tissues were analyzed by multiplexed immunofluorescence staining. Mechanistic studies employed high-fat diet (HFD)-induced MASLD-HCC mouse models, genetic or pharmacological inhibition of Tim-3, and DC depletion or adoptive transfer. Lipid peroxidation, ferroptosis, and immune interactions were assessed using flow cytometry, transcriptomics, and functional assays. The therapeutic efficacy of Tim-3 blockade, alone or combined with anti-PD-1 therapy or lenvatinib, was evaluated in preclinical models. HFD reshaped the hepatic tumor immune microenvironment by inducing DC depletion and CD8+ T cell dysfunction, facilitating liver tumor progression. In human steatohepatitis-related HCC, DC infiltration and DC-CD8+ T cell interactions were markedly impaired, and high DC-specific Tim-3 expression correlated with poor prognosis. Mechanistically, the lipid-rich microenvironment induced DC depletion via Tim-3-dependent lipid peroxidation and ferroptosis. Genetic or pharmacological inhibition of Tim-3 in DCs attenuated lipid peroxidation, restored DC survival and CD8+ T cell activation, and suppressed tumor growth. Moreover, Tim-3 blockade synergized effectively with both anti-PD-1 and lenvatinib to achieve sustained tumor control. Our findings establish Tim-3 as a pivotal regulator of DC ferroptosis in metabolic liver cancer. Combining Tim-3 blockade with standard therapies represents a promising strategy to restore immune surveillance in MASLD-HCC. Our findings identify Tim-3 as a crucial metabolic immune checkpoint that governs dendritic cell ferroptosis and dendritic cell-mediated antitumor immunity in metabolic liver cancer. Targeted blockade of Tim-3 in dendritic cells holds great therapeutic potential for the treatment of steatohepatitis-related hepatocellular carcinoma, particularly for patients with metabolic dysfunction-associated steatotic liver disease-related hepatocellular carcinoma who exhibit resistance to anti-PD-1 therapy.

Dopamine (DA) has shown antitumor potential and is known to modulate immune responses under physiological conditions; however, its specific immunomodulatory role in cancer immunotherapy remains poorly defined. Triple-negative breast cancer (TNBC) is an aggressive malignancy with strong stemness properties and a limited response to immune checkpoint inhibitors (ICIs). Cancer stem cells (CSCs) contribute significantly to the suppression of the tumor immune microenvironment and immune evasion. However, the ability of DA to remodel the immune microenvironment by targeting CSCs and thereby increasing ICI efficacy in TNBC is not yet understood. To address this, we employed immunocompetent BALB/c mice bearing 4T1 TNBC tumors, performed RNA-seq analysis, and used flow cytometry to assess immune cells in tumors, the spleen, and tumor-draining lymph nodes. These analyses revealed that DA increases cytotoxic T lymphocyte (CTL) levels and activity, with a more pronounced effect observed in local tumor tissues, which indicates that T-cell activation is its primary antitumor mechanism. Using TNBC cell lines (MDA-MB-231, BT549, and 4T1) and CSCs enriched from these cell lines by suspension culture, PD-L1-knockout and D1DR-knockout 4T1 cells, and dopamine receptor (DR) agonists and antagonists, we revealed that the PD-1/PD-L1 pathway predominantly mediates CSC-driven immune evasion, growth, and metastasis in TNBC. Moreover, DA suppresses TNBC CSCs and downregulates PD-L1 expression in these cells specifically through dopamine receptor D1 (D1DR) activation. Furthermore, DA enhances both the antitumor efficacy of CTLs and the intratumoral CTL infiltration induced by a cytotoxic T-lymphocyte-associated protein 4 antibody (αCTLA-4) without compromising safety. DA also inhibits metastasis and prolongs the survival of 4T1 tumor-bearing mice. In conclusion, this study reveals a novel mechanism by which DA regulates tumor immunity in TNBC and demonstrates its novel potential for application in cancer immunotherapy. By investigating the CSC‒PD-L1‒immune axis, we also revealed that anti-CSC therapies are novel, promising ICI synergists for TNBC treatment.

Immune checkpoint inhibitors (ICIs) have revolutionized cancer therapy, yet their clinical application is constrained by a critical trade-off between potent antitumor efficacy and off-target cardiovascular toxicity. This adverse effect underscores a fundamental biological dilemma: how tumor immunotherapy disrupts cardiovascular homeostasis. Immune cells within the cardiovascular system undergo metabolic adaptations after immune therapy. This intricate crosstalk has brought metabolic checkpoints to the forefront of research. In this review, we systematically outline the features of tumor-associated metabolic remodeling and shared roles in cardiac metabolism and immunometabolism. We elucidate ICIs initiate a cascade of events leading to cardic dysfunction through metabolic signal pathways. Furthermore, we propose that the strategic integration of cutting-edge technologies including spatial metabolomics to enable precise reprogramming of metabolic networks will ultimately decouple the efficacy of cancer immunotherapy from its associated toxicities.

Nanozymes, engineered nanomaterials with enzyme-mimetic activities, have emerged as versatile platforms for ferroptosis-based cancer theranostics. Ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation, has emerged as a promising strategy to overcome resistance to conventional cancer therapies. By catalyzing redox reactions, nanozymes can generate reactive oxygen species (ROS) and promote ferroptotic lipid peroxidation, thereby triggering cell death in tumor cells that evade apoptosis-based treatments. In parallel, non-redox activities of nanozymes, including hydrolase- and phosphatase-like functions, enable them to remodel the tumor microenvironment (TME), modulate biomolecular signaling, and support targeted therapy. This review provides a systematic and design-oriented overview of nanozymes that interface with ferroptosis. We summarize how redox and non-redox nanozyme activities converge on key ferroptosis-related processes, such as ROS production, glutathione depletion, iron metabolism disruption, and TME regulation. We then highlight rational engineering strategies, including single-atom and multimetallic catalytic centers, biodegradable coordination frameworks, stimuli-responsive architectures, and protein corona engineering, that enhance catalytic specificity, tumor targeting, and biosafety. Theranostic implementations are discussed with emphasis on multimodal imaging-guided platforms and combination regimens that integrate chemotherapy, radiotherapy, phototherapy, and immunotherapy. Finally, we outline major translational challenges and future opportunities, including AI and computation-guided nanozyme design and adaptive, corona-informed systems tailored for personalized cancer therapy. This review aims to serve as a roadmap for developing clinically translatable nanozymes that unify diagnosis and treatment through ferroptosis-oriented precision oncology.

Advances in understanding tumor heterogeneity and the tumor microenvironment (TME) have ushered in a new era of targeted and immunotherapy. Both targeted and immunotherapy mainly rely on the use of specific antibodies against the tumor targets. Although monoclonal antibodies (mAbs) have achieved tremendous success, the full potential of these full-size antibodies is hampered by the large size, low tumor permeability, and uneven biodistribution, which will obstruct the clinical treatment outcome. Nanobodies (Nbs), derived from camelid heavy-chain antibodies, offer a revolutionary alternative due to their unique structural and functional properties. This review highlights the beneficial characteristics of Nbs, notably their ability to access hidden epitopes and penetrate dense TME, along with a favorable immunogenic profile. Their therapeutic applications in cancer are explored across multiple fronts: from monovalent and multivalent constructs targeting receptor tyrosine kinases and other cancer-related molecules to serving as delivery vehicles for drugs, radionuclides, and photosensitizers. In immunotherapy, Nbs play a pivotal role by activating immune cells, blocking immune checkpoints, and enhancing adoptive cell therapies. Despite challenges like rapid renal clearance, engineering and humanization strategies are advancing Nbs toward improved efficacy and safety in oncology. This review provides a comprehensive overview of current Nb-based therapies, their mechanisms, and future directions in oncology.

For peripheral immune cells to be effective against brain tumors, multiple coordinated steps are required, including migration and homing to the local microenvironment-collectively referred to as immune cell trafficking-followed by survival, proliferation, and persistence, all while remaining functional. This review focuses on the often-overlooked first step, trafficking, which is critical for initiating subsequent processes and driving antitumor responses. We summarize the migration cascade of immune cells, including T cells and macrophages, across the blood-brain barrier (BBB) and how primary brain tumors as well as brain metastases alter the molecular pathways and interactions associated with the immune cell migration cascade across the BBB and their functional homing. Subsequently, we highlight recent developments in engineering biomolecular, cellular, and physical methods to modulate the tumor microenvironment and improve the functional trafficking of immune cells in brain tumors. Our analysis reveals novel combinations to potentiate antitumor responses via feedforward mechanisms and modular therapeutic strategies designed to improve the migration and accumulation of functional immune cells in brain tumors. A deeper understanding of immune cell trafficking, distribution, and homing through the application of preclinical and clinical imaging techniques along with the development of in vitro/ex vivo platforms should enable new and more potent immunotherapy strategies against primary brain malignancies and brain metastases as well as accelerate their translation to the clinic.

Liver metastasis (LM) is a major cause of cancer-related mortality, driven largely by dynamic interactions between disseminated tumor cells (DTCs) and the liver microenvironment (LME). Liver fibrosis, a pathological condition characterized by the disruption of the LME and imposing a significant global health burden, is primarily orchestrated by activated hepatic stellate cells (aHSCs). However, the precise mechanisms through which liver fibrosis facilitates LM are poorly understood. Here, we demonstrated that liver fibrosis potently enhanced LM by promoting the early hepatic colonization of tumor cells in an aHSC-dependent manner. Mechanistically, we identified prostaglandin E2 (PGE2), secreted by aHSCs, as a key mediator that disrupted natural killer (NK) cell immune surveillance. Either the depletion of aHSCs or pharmacological inhibition of the PGE2-synthesizing enzyme Cyclooxygenase-2 (COX-2) with Celecoxib (CLX) restored NK cell function and suppressed LM. Notably, CLX treatment synergized with anti-NKG2A-based immunotherapy, significantly boosting its efficacy against LM in the fibrotic liver. Our findings unveil a critical "aHSC-PGE2-NK cell" axis in liver fibrosis-induced immunosuppression and provide a compelling therapeutic strategy for the clinical management of LM.

Chronic inflammation in intermediate/advanced tumors drives burdensome protumor immune cell communication, thereby weakening immunotherapy. Conventional anti-inflammatory therapies focus on alleviating chronic inflammation whereas ignore dysfunction and scarcity of myeloid and T cells, which hinder their intercellular communication restoration. To address the dilemma, an inflammatory condition-triggered protumor inflammation-immunosurveillance shift hydrogel (TRANS) is developed to initiate adaptive immune responses mediated by intercellular communication. Triggered by inflammatory conditions, TRANS releases celecoxib (CXB) to inhibit the COX-2/PGE2 pathway, thereby reprogramming tumor-associated macrophages and mitigating protumor inflammation. Furthermore, TRANS incorporates FMS-like tyrosine kinase 3 ligands (Flt-3L) and 4-1BB agonists (α-CD137) to respectively recruit type 1 conventional DC (cDC1) and revitalize tumor-infiltrating T cells, to rejuvenate immunosurveillance. TRANS inhibits 87.50% and 88.74% of primary and secondary colorectal tumors, generates antitumor immune memory to resist tumor rechallenge, and significantly reduces lung and liver metastases. Rechallenge model shows TRANS leads to antitumor immune memory formation. Single-cell RNA sequencing is preform to elucidate the mechanism of TRANS, which exhibits that TRANS exerts antitumor effects by optimizing the crosstalk between myeloid cells and T cells via CXCL9/10-CXCR3/DPP4. TRANS further gains better control of colorectal cancer when combined with immune checkpoints inhibitors. This study offers a novel perspective on immunotherapy by rebalancing inflammation-immunity dynamics.

Pathological angiogenesis and immunosuppression limit the efficacy of cancer therapies. Here, we report a biomimetic nanoparticle platform integrating a vascular endothelial growth factor receptor 2 (VEGFR2)-targeting aminated fullerene with tumor cell membrane coating. Phenotypic screening identified tetra [4-(amino)piperidin-1-yl]C60 epoxide (TAPC) as a potent inhibitor of angiogenesis. Transcriptomic analyses identified VEGFR2 as a highly expressed and clinically relevant target in colorectal cancer (CRC), prompting further mechanistic investigation. TAPC was found to target VEGFR2, reducing its expression and suppressing PI3K-AKT signaling in both tumor and endothelial cells. To enhance delivery, TAPC was encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles and coated with homologous tumor cell membranes to generate tumor cell membrane-coated nanoparticles (TAPC@CNPs). This formulation improved stability and supported systemic circulation, enabling tumor accumulation. In murine CRC models, TAPC@CNPs significantly inhibited tumor growth and reduced angiogenesis markers, including VEGFR2 and CD31. Furthermore, treatment decreased regulatory T-cell levels and increased T-cell infiltration and activation, indicating enhanced antitumor immunity. These findings establish TAPC as a fullerene-based VEGFR2 inhibitor and demonstrate that tumor membrane-coated nanoparticle delivery amplifies its anti-angiogenic and immune-modulating effects, offering a nanomaterial strategy to concurrently target angiogenesis and remodel the tumor immune microenvironment.

The minimal residual disease (MRD) following tumor resection remains a major challenge for preventing recurrence. Existing treatments usually exhibit poor specificity for scattered tumor cells at the surgical site. Moreover, few strategies successfully combine real-time MRD monitoring with sustained therapeutic intervention, further limiting their efficacy. To address these issues, we developed a PD-L1-targeted and lactate-responsive DNA hydrogel (Gel@FX11-SPNT). A key innovation lies in its dual-functional PD-L1 aptamers: they bind to PD-L1-positive tumor cells to facilitate in situ enrichment and block the PD-L1/PD-1 checkpoint to reactivate immunity. Structurally, the hydrogel network is crosslinked by lactate-responsive aptamers which are conjugated with fluorophore-quencher pairs. When exposed to lactate (a metabolite abundant in MRD microenvironment), the lactate-responsive aptamers undergo conformational changes which not only activates fluorescence for MRD monitoring but also triggers the hydrogel disassembly, allowing release of mitochondria-targeted FX11-SPNT. Under ultrasound irradiation, FX11-SPNT generates reactive oxygen species (ROS) and suppresses aerobic glycolysis, thereby inducing tumor cell apoptosis and immunogenic cell death, which was evidenced by the upregulation of calreticulin (CRT), high mobility group box 1 (HMGB1), and heat shock protein 70 (HSP70). This process promotes dendritic cell maturation and T-cell activation, thus establishing long-term immune memory that effectively eliminates residual tumor cells and inhibits metastasis.

High-fidelity imaging of pro-tumorigenic M2 macrophages reprogramming into anti-tumorigenic M1 phenotypes during tumor immunotherapy is crucial for guiding personalized treatment. Off-on probes, switching from low "off" state signals to high "on" signals upon biomarker activation, enable visualization of this process. However, most existing off-on probes fail to deliver high-fidelity imaging, as even low "off" state probe signals are amplified by extensive intratumoral accumulation without biomarker activation, producing a high probe background that is indistinguishable from biomarker-triggered signals and prone to false-positive diagnoses. Here we report semiconducting polymer intrachain donor-acceptor reconfiguration-tailored off-on NIR-II (1000-1700 nm) photoacoustic probes (BDPNP) with ultralow background for high-fidelity imaging of immunotherapy-associated macrophage reprogramming. BDPNP undergoes polymer intrachain donor-acceptor reconfiguration-induced large absorption redshift from a short wavelength (no NIR-II photoacoustic background) to a long wavelength (robust NIR-II photoacoustic signals) under the macrophage reprogramming biomarker nitric oxide. BDPNP shows ultralow background even after extensive tumor accumulation without immunotherapy, but signals amplify 21.4-fold upon immunotherapy. The design clearly distinguishes biomarker-triggered signals from probe background in vivo, enabling high-fidelity mapping of macrophage reprogramming in tumor immune microenvironment. This work establishes a general strategy for ultralow-background off-on NIR-II photoacoustic probes and provides a powerful tool for high-fidelity imaging of cancer immunotherapy response.

In vivo CAR-macrophage (CAR-M) therapy exploits macrophages' tumor-tropic homing, phagocytic capacity, and ability to remodel the immunosuppressive tumor microenvironment, with low risk of cytokine release syndrome and potential as an off-the-shelf therapy. However, clinical translation is currently constrained by the lack of efficient non-viral platforms for in situ macrophage engineering. To address this challenge, fluorinated ionizable lipids were rationally designed-capitalizing on fluorine's high electronegativity, low polarizability, and lipophobicity-to enhance lipid nanoparticle (LNP) delivery. Screening a library of 80 fluorinated lipids identified A1F5C5 as the lead candidate, demonstrating superior mRNA delivery to macrophages both in vitro and in vivo. Our investigation into the mechanism revealed that, while fluorination universally enhanced both cellular uptake and subsequent endosomal dissociation, the specific configuration with five fluorine atoms uniquely conferred a superior membrane fusion capability, which proved critical for achieving efficient endosomal escape. F5-LNPs encapsulating mRNA encoding an hPSMA-targeted CAR (F5-CAR) were formulated using this platform. Intravenous F5-CAR administration reprogrammed the tumor microenvironment, reducing M2-like macrophages and boosting granzyme B and perforin expression in CD8+ T cells. Notably, when combined with anti-PD-L1 blockade, this strategy achieved 100% complete and durable regression in MC38-hPSMA tumor-bearing mice. This work establishes fluorinated LNPs as a scalable platform for in vivo CAR-M engineering against solid tumors.

Chemodynamic therapy (CDT) based on overproduced reactive oxygen species (ROS), activates cytoprotective autophagy, an inexorable phenomenon-that enables tumor cell survival, therefore attenuating ROS-induced therapeutic efficacy. Herein, we develop a tumor microenvironment (TME)-responsive nanoplatform (MnOx-GOx-PM@Ma) composed of manganese oxide nanoflowers (MnOx NFs) co-loaded with glucose oxidase (GOx) and an activatable melittin pro-peptide (PM), and coated with macrophage membranes (Ma) for targeted delivery. Combined MnOx NFs and GOx trigger O2/H2O2 cyclic generation, thereby amplifying CDT in the acidic and glutathione (GSH)-rich TME. Meanwhile, the PM is selectively cleaved by lysosomal legumain to activate melittin, which disrupts lysosomal membranes and converts cytoprotective autophagy into a pro-death process. Additionally, the releasing Mn2+ exhibits excellent magnetic resonance imaging (MRI) contrast properties. Both in vitro and in vivo studies demonstrate that MnOx-GOx-PM@Ma effectively suppresses tumor growth through synergistic starvation therapy, enhanced CDT, and autophagy inhibition. Collectively, this work presents a strategy to overcome autophagy-mediated therapeutic resistance and optimize synergistic CDT-based antitumor therapy.

Activated AKT and YAP signaling have been implicated in the pathogenesis of intrahepatic cholangiocarcinoma (iCCA), but their specific roles in tumor progression and regulation of the tumor microenvironment remain unclear. We developed two doxycycline-inducible iCCA mouse models, Akt/TRE-YAP and TRE-Akt/YAP, allowing selective inhibition of YAP or AKT in established tumors upon doxycycline withdrawal. Histological and molecular analyses were then performed to assess phenotypic changes and pathway regulation. YAP suppression in the Akt/TRE-YAP model initially induced tumor regression but ultimately led to transformation into steatosis-associated HCC due to persistent AKT signaling. Conversely, AKT suppression in the TRE-Akt/YAP model induced more profound iCCA regression with minimal tumor burden. Mechanistically, both AKT and YAP regulate tumor cell proliferation, while RNF125 acts as a tumor suppressor inhibited by YAP during iCCA pathogenesis. AKT uniquely regulates tumor metabolic pathways, whereas YAP controls iCCA differentiation and the immune microenvironment. YAP inhibition depleted neutrophils and increased CD4+ and CD8+ T-cell infiltration. The immunosuppressive role of YAP was confirmed by ectopic YAP activation in mouse iCCA and validated in human iCCA, where YAP/TAZ activation correlated with immunosuppressive features. Although YAP inhibition increased T-cell infiltration, these T cells expressed PD-1. Combined YAP suppression and anti-PD-L1 treatment further enhanced tumor regression. We define distinct molecular roles for AKT and YAP in iCCA progression and identify YAP as a key regulator of the tumor immune microenvironment. These findings support combining YAP inhibition with immune checkpoint blockade for iCCA treatment. Our studies elucidate the distinct contributions of AKT and YAP signaling to intrahepatic cholangiocarcinoma (iCCA) progression. Using innovative doxycycline-inducible models, we show that both pathways regulate tumor cell proliferation - YAP by repressing the tumor suppressor RNF125 - while AKT primarily governs metabolism and YAP controls differentiation and suppresses the immune microenvironment. Importantly, YAP inhibition reshapes the tumor immune landscape, and its combination with PD-L1 blockade induces profound tumor regression. These findings establish YAP as a key regulator of the iCCA immune microenvironment and provide a strong preclinical rationale for combining YAP pathway inhibition with immune checkpoint blockade in iCCA therapy.

Spatiotemporally-tailored activation of tumor specific immune-response remains a critical challenge for effective whole tumor cell vaccines (TCVs). Herein, we reported a TCV platform obtained by nanoparticle-mediated photo-chemotherapy to generate whole tumor cell vaccine under near-infrared (NIR) laser irradiation. The photo-chemo treatment can trigger the yielded IPL-TCV to activate stimulator of interferon genes (STING) pathways and immunogenic cell death (ICD), which broaden the antigen spectrum of TCV and enhance lymph node (LN) targeted delivery. The combination of TCV with NIR laser irradiation can synergistically activate both innate and adaptive immunity, leading to potent antitumor immune responses that suppress tumor growth. We further demonstrate that IPL-TCV strategy facilitates dendritic cell (DCs) recruitment, maturation, and subsequent T cell activation, which elicits remarkable efficacy in reducing tumor growth, preventing tumor occurrence. This study highlights photo-chemo coordination based personalized TCV strategy to augment cancer immunotherapy by spatiotemporally-tailored innate and adaptive immunity stimulation.

Conventional therapies for colorectal cancer (CRC) are often constrained by limited specificity, while the hypoxic, acidic, and immunosuppressive commonly contributes to immunotherapy resistance. Herein, a dual-responsive nanosystem with oxygen vacancies (AgFeO2-PVP, termed Vo-AFP) was engineered against CRC. This system is selectively activated within the tumor microenvironment (TME) by H2S and acidic conditions, triggering Fe2+ release to induce ferroptosis and forming a dual-vacancy structure composed of iron and oxygen vacancies. These defects introduce sub-bandgap (intermediate energy bands) that facilitate two-step single-photon absorption under 1064 nm laser irradiation and promote non-radiative electron-hole recombination, boosting photothermal conversion efficiency from 17.03% to 34. 15%. Density functional theory (DFT) calculations confirm that the dual-vacancy configuration induces charge redistribution, enhancing localized surface plasmon resonance (LSPR) in the Near-infrared II (NIR-II) window. The synergy between defect-assisted sub-bandgap absorption and LSPR enhancement enables effective photothermal ablation of deep tumors. In vivo results show that localized heating further accelerates Fe2+ release and suppresses the GPX4-mediated antioxidant pathway, augmenting ferroptotic death. This photothermal-enhanced ferroptosis significantly sensitizes tumors to aPD-L1 checkpoint blockade, remodels the immunosuppressive microenvironment, activates systemic antitumor immunity, and inhibits tumors and pulmonary lesions. Vo-AFP thus provides an integrated theranostic platform and a new paradigm for activatable, defect-engineered nanosystem in CRC immunotherapy.

In situ tumor vaccination activates host immunity through endogenous tumor-associated antigens (TAAs) and represents a promising strategy for tumor immunotherapy. However, insufficient immunogenicity and an immunosuppressive tumor microenvironment have limited the efficacy of in situ tumor vaccination in colorectal cancer (CRC). Here, we presented a versatile manganese-based nanoplatform (MnP@LNP) for chemodynamic therapy and in situ tumor vaccination to enable CRC immunotherapy. Upon encountering the acidic tumor microenvironment, MnP@LNPs underwent continuous release of Mn2+ accompanied by the generation of highly destructive hydroxyl radicals. When administered rectally to the primary tumor site, MnP@LNPs alone achieved 71.0% inhibition of primary tumors, while their combination with αPD-L1 increased the inhibition rate to 83.8% and concurrently suppressed distant tumors by 81.9%. The primary function of MnP@LNPs was to generate an in situ vaccine by inducing immunogenic cell death of tumor cells. The released TAAs, together with Mn2+ as an adjuvant, activated the cGAS-STING signaling pathway, driving a systemic cytotoxic T-lymphocyte response against metastatic tumors. Besides, this therapeutic strategy also reprogrammed the intestinal milieu by enriching beneficial microbial populations (e.g., Alistipes and Dubosiella) and upregulating the levels of immunostimulatory metabolites, such as indole-3-aldehyde and indole-3-lactic acid. Collectively, MnP@LNPs serve as a nanotherapeutic that orchestrates local-to-systemic antitumor immunity while fostering an immune-favorable intestinal ecosystem, representing a promising platform for CRC immunotherapy.

Cancer cells utilize tumor-derived exosomes to suppress antitumor immunity. Herein, we identify co-activator-associated arginine methyltransferase 1 (CARM1) as a key regulator of exosome biogenesis and metabolite sorting that inhibiting CD8+ T cell-mediated antitumor responses. Genetic ablation of CARM1 in breast cancer cells impairs immunosuppressive exosome secretion, enhancing CD8+ T cell infiltration, proliferation, and effector function. Mechanistically, CARM1 dimethylates apoptosis-linked gene-2 interacting protein X (ALIX) at arginine 757, facilitating its interaction with endosomal sorting complex required transport (ESCRT) components, and promoting tetraspanin-enriched exosome biogenesis. CARM1-dependent ALIX methylation enables selective packaging hypoxanthine into exosomes through direct binding to the ALIX F676 pocket. Exosomal hypoxanthine disrupts inosine metabolism in activated CD8+ T cells, inhibiting pentose phosphate pathway, glycolysis, nucleotide synthesis, and effector cytokine production. Co-administration of CARM1 inhibitor with inosine significantly enhances tumor-infiltrating CD8+ T cell cytotoxicity, reduces PD-1+TIM-3+ exhausted CD8+ T cells, and suppresses tumor growth. These findings establish the CARM1-ALIX-hypoxanthine axis as an immunosuppressive mechanism and suggest that combining CARM1 inhibition with inosine supplementation represent a promising therapeutic strategy for breast cancer.

Metabolic reprogramming is a hallmark of clear-cell renal cell carcinoma (ccRCC), driving tumor progression and altering the tumor microenvironment (TME), making it crucial to understand metabolic dysregulation in ccRCC and to identify new therapeutic targets for patients. In this study, metabolomic profiling identified elevated levels of methylmalonic acid (MMA) in ccRCC, attributed to downregulation of methylmalonyl-CoA mutase (MMUT). MMA produced by ccRCC accumulates in the TME and activates the suppressor of fused (SUFU)-regulated Hedgehog signaling pathway in a dose-dependent manner, promoting M2 polarization of macrophages and tumor progression. Mechanistically, MMA induces methylmalonylation at the K499 site of ubiquitin-specific peptidase 36 (USP36), inhibiting USP36-mediated deubiquitination and SUMOylation of SUFU, thereby promoting the expression of GLI family zinc finger 1 (GLI1) and its target genes. Both in vitro and in vivo experiments demonstrated that a low branched-chain amino acids (BCAAs) diet or treatment with the de-methylmalonylation agent MC3138 effectively inhibited M2 polarization of macrophages and tumor progression. These findings emphasize the critical role of MMA in ccRCC pathogenesis and suggest that combining a low-BCAAs diet with MC3138 therapy may offer a promising treatment strategy for ccRCC patients with elevated MMA levels.

Current PD-L1 degraders, whether antibody-based or small-molecule-mediated, are hindered by limitations in pharmacokinetics (e.g., poor tissue penetration) or pharmacodynamics (e.g., suboptimal degradation efficacy, immunogenicity concerns). These drawbacks highlight the necessity for novel PD-L1 degradation platforms using innovative technologies. This study aims to design and synthesize bifunctional small molecules as PD-L1 degraders by leveraging the unexplored E3 ligase SPOP, aiming to overcome the limitations of existing degraders and evaluate their potential in cancer immunotherapy. A series of SPOP-based bifunctional small molecules were designed and synthesized. Their PD-L1 inhibitory and degradation activities were assessed using HTRF and western blot assays, respectively. Mechanistic studies (His pull-down, bio-layer interferometry, western blot) were performed to verify ternary complex formation with PD-L1 and SPOP. In vivo pharmacokinetic properties and antitumor efficacy were evaluated in a B16-F10 tumor model, with analysis of tumor-infiltrating lymphocytes (TILs) to explore immune microenvironment effects. Compound SPOP9 exhibited potent PD-L1 inhibition (IC50 = 357.2 nM) and degradation (DC50 = 1.0 μM). Mechanistic studies confirmed its assembly into a stable ternary complex with PD-L1 and SPOP. SPOP9 showed favorable in vivo bioavailability (F = 74.8 %) and, at 10 mg/kg (i.p.), reduced tumor weight by 44 % in B16-F10 mice, superior to anti-PD-L1 antibody (TGI = 34.4 %). TIL analysis indicated SPOP9 activated the tumor immune microenvironment and downregulated PD-L1. SPOP9, as the first SPOP-binding bifunctional PD-L1 degrader, demonstrates promising preclinical efficacy and pharmacokinetic properties, addressing key limitations of existing degraders. It merits further investigation as a potential agent for cancer immunotherapy.

Intrahepatic cholangiocarcinoma (ICC) is a highly heterogeneous and aggressive malignancy with poor prognosis and limited treatment options. The lack of precise molecular classification and incomplete understanding of the tumor microenvironment (TME) impede therapeutic development. This study aims to dissect the cellular heterogeneity and intercellular interactions in ICC, with a particular focus on SPINK1-overexpressing epithelial subsets. We sought to determine the role of SPINK1 in modulating immune cell recruitment and tumor metabolism, and to evaluate its clinical relevance. We performed single-cell RNA sequencing (scRNA-seq) on human ICC tumors and adjacent normal tissues to construct a transcriptomic atlas. Integrative proteomic analysis, transwell assays, co-culture systems, and functional perturbation experiments (SPINK1 knockdown and ASCT2 inhibition) were conducted to explore epithelial-macrophage interactions and metabolic dynamics. ScRNA-seq identified a SPINK1-overexpressing epithelial subcluster enriched in ICC tumors. High SPINK1 expression correlated with significantly shorter patient survival. SPINK1-overexpressing epithelial subcluster secretes CCL20, which recruits lipid-associated macrophages (LAMs). This effect was reversed by CCL20 neutralizing antibodies. In co-culture, LAMs increased intracellular glutamine levels in SPINK1-overexpressing epithelial subcluster, promoting proliferation. Disruption of SPINK1 expression or glutamine transport abolished this metabolic support. Our findings uncover a novel SPINK1-CCL20-LAMs axis that orchestrates immune recruitment and metabolic reprogramming in ICC. SPINK1 facilitates tumor growth by establishing a glutamine-rich microenvironment via LAMs. These insights highlight SPINK1 as a potential therapeutic target and prognostic biomarker in ICC.

Gastric cancer (GC) has shown relatively poor responses to existing immune therapies. The extensive infiltration of tissue-resident memory T cells (TRM) in tumor microenvironment (TME) is associated with better Overall Survival (OS) in patients treated with immune checkpoint inhibitors (ICIs). However, the precise roles of TRM cells in cancer immunity and responses to ICIs in GC remain poorly understood. We found that the TRM cells representing the majority of tumor-infiltrating lymphocytes and expressing high level of immune checkpoint TIGIT in GC tissue. This study aims to elucidate the mechanisms through which GC cells modulate TRM cells by PVR/TIGIT axis to facilitate immune evasion. The immune status of GC was evaluated by quantifying T cell subsets in GC tissues through flow cytometry. The biological functions and molecular mechanisms by which the oncogenic factor PRDM15 mediates tumor immune evasion were investigated through the PVR/TIGIT axis using transcriptome sequencing, Chromatin immunoprecipitation (ChIP) assay and Co-immunoprecipitation assay. The therapeutic potential of PRDM15/PVR/TIGIT was analyzed using tumor-bearing models. We confirmed that PRDM15 promotes the transcription of PVR which is the ligand of TIGIT in GC cells. Aberrantly high expression of PRDM15 in GC tissues was associated with higher TNM stages and T cell immunosuppressive state in GC patients. PRDM15 was found to upregulate the proliferation, invasion and migration of GC cells. When co-cultured with TRM cells with high TIGIT expression, PRDM15 activated the PVR/TIGIT axis to inhibit TRM cells activation by upregulating PVR expression in GC cells. Mechanistically, PRDM15 recruited the histone methyltransferase complex PRMT5/Mep50/WDR5 to activate PVR transcription. This study demonstrated that PRDM15 in GC cells could recruits the histone methyltransferase complex PRMT5/Mep50/WDR5 to promote PVR transcription, thereby activating the PVR/TIGIT axis, which inhibits TRM cells activation and mediates immune escape and GC progression.

The aryl hydrocarbon receptor (AhR) plays a pivotal role in modulating immune responses and influencing tumor development by detecting metabolites derived from tryptophan breakdown. In patients suffering from pancreatic ductal adenocarcinoma (PDAC), elevated levels of AhR are strongly correlated with poor clinical outcomes. Despite this, the cell-autonomous functions of AhR in pancreatic tumor cells, particularly its role in modulating anti-tumor immunity within the tumor microenvironment, remain poorly characterized and require further investigation. CRISPR-Cas9 technology was used to generate Ahr-/-/AHR-/- Pan02, B16-F10 and Panc-1 cells. Subcutaneous tumor models in nude and C57BL/6J mice were used to evaluate the effect of tumor cell intrinsic Ahr on the anti-tumor immune response. Transcriptome sequencing, ATAC sequencing, CUT&Tag sequencing, ChIP and IP-MS were used to elucidate how AhR controls the expression and accessibility of major histocompatibility complex class I (MHC-I). Elevated AhR levels in PDAC patients correlate with worse outcomes. AhR protein is located in both cytoplasm and nucleus of tumor cells. Deletion of Ahr in Pan02 and B16-F10 leads to a significant upregulation of MHC-I and related gene expression. AhR drives tumor growth by suppressing T cell-mediated immunity. Mechanistically, AhR specifically suppressed MHC-I expression in pancreatic tumor cells by interacting with protein arginine methyltransferase 5 (PRMT5) to decrease chromatin accessibility, which led to impaired antigen presentation. We demonstrated that AhR inhibition improved the therapeutic efficacy of immune checkpoint blockade, chemotherapy, and PRMT5-targeted therapy in pancreatic cancer models. Tumor cell intrinsic AhR inhibits MHC-I expression in PDAC cells by epigenetic regulation mediated by PRMT5, resulting in reduced immunogenicity of pancreatic tumor cells. The discovery of the AhR-PRMT5-H4R3me2s-MHC-I axis provides critical mechanistic insights into how tumor-intrinsic epigenetic regulation of antigen presentation promotes the progression of PDAC.

Osteosarcoma remains a clinical challenge due to its high invasiveness, early metastasis, and intrinsic radioresistance, which collectively limit the efficacy of conventional treatments and immunotherapy. Here, we developed a pH-responsive hydrogel reservoir, referred to as VP/CaNPs@Gel, by integrating verteporfin (VP)-loaded CaCO3 nanoparticles (CaNPs) into a β-cyclodextrin-crosslinked polysaccharide hydrogel to achieve precise radiosensitization and immunomodulation. The hydrogel undergoes rapid in-situ crosslinking to form a robust scaffold, ensuring the sustained and pH-triggered release of VP and Ca2+ within the tumor microenvironment. In vitro assays demonstrated that VP/CaNPs@Gel significantly amplifies radiation-induced reactive oxygen species (ROS) production and triggers robust pyroptotic cell death. In an in vivo orthotopic osteosarcoma model, VP/CaNPs@Gel effectively suppressed tumor growth and markedly enhanced the efficacy of immune checkpoint blockade under X-ray. Mechanistically, treatment induced PD-L1 upregulation, elevated systemic IL-1β, IL-18, and IFN-γ levels, promoted dendritic cell maturation and CD8+ T cell infiltration, increased M1 macrophage polarization, and reduced regulatory T cells and M2 macrophages. These shifts effectively converted the immunologically "cold" tumor into a "hot" state responsive to immune checkpoint inhibitors. This carrier-within-a-carrier strategy provides a multifunctional platform that couples potent radiosensitization with immunomodulation, offering a promising approach to overcoming radioresistance and improving therapeutic outcomes in osteosarcoma.

Hepatocellular carcinoma (HCC) remains a major global health burden with limited effective therapeutic strategies. Although chimeric antigen receptor (CAR) T-cell therapy has shown encouraging potential, its efficacy in HCC is profoundly constrained by the immunosuppressive tumor microenvironment. In this study, we performed integrative analyses of HCC transcriptomic and clinical samples data to identify Copine-1 (CPNE1) as potential oncogenic drivers involved in tumor progression and immune suppression. Using functional and mechanistic assays in vitro with HCC cell line models and in vivo with mouse models, we examined the role of CPNE1 in regulating tumor proliferation, stemness, and its contribution to resistance against GPC3-targeted CAR-T cell therapy. We found that CPNE1 is significantly overexpressed in HCC and correlates with poor patient prognosis. Functional assays revealed that CPNE1 enhances tumor proliferation and stemness by activating the STAT3-transforming growth factor beta (TGF-β) signaling pathway. Moreover, CPNE1-induced secretion of TGF-β promotes T-cell exhaustion, which impairs the efficacy of CAR-T cells. Knockdown of CPNE1 restored CAR-T cell function, enhanced T-cell infiltration into tumors, reduced exhaustion, and improved antitumor efficacy without causing systemic toxicity. Clinically, high CPNE1 expression was associated with lower T-cell infiltration and poor outcomes in HCC patients treated with immune checkpoint inhibitors. These findings identify CPNE1 as a critical regulator of HCC progression and immune suppression, highlighting the CPNE1-STAT3-TGF-β axis as a promising therapeutic target in HCC.

Glioblastoma (GBM) as the most malicious primary brain tumor is infiltrative and often unresectable, which coupled with a profoundly immunosuppressive tumor microenvironment (TME) lead to a poor prognosis for patients. Here, we developed a golden vesicular nanoadjuvant (Au/TLR3-V) comprising gold nanoclusters and TLR3 agonist poly(I:C), formulated within an injectable hydrogel (Au/TLR3-V@gel) for locoregional photothermal-immunotherapy of large orthotopic GBM. Upon near-infrared (NIR) irradiation, Au/TLR3-V generated a potent photothermal response that induced immunogenic cell death and promoted dendritic cell (DC) activation. In TME‑mimicking systems, Au/TLR3-V pre-treated GL261 cells activated DCs accompanied by the upregulation of T‑cell‑recruiting chemokines. In murine models of advanced orthotopic GBM, a single intratumoral injection of Au/TLR3-V@gel followed by a brief NIR irradiation orchestrated a two-stage therapeutic process: localized thermal ablation of the tumor bulk (physical reduction) and subsequent release of Au/TLR3-V to facilitate DCs' capture of tumor-derived antigens and subsequently amplify DC activation (biological sweeping). When combined with anti‑CTLA‑4 blockade, this therapy achieved an 33% complete cure rate and established potent systemic anti-GBM immunity. By transforming an unresectable and immunologically "cold" tumor into a locally confined immunogenic "hotspot" using a soft, brain-compliant depot, this work provides a promising locoregional approach for unresectable GBM. STATEMENT OF SIGNIFICANCE: The treatment of unresectable glioblastoma is limited by the lack of biomaterials that can simultaneously provide safe intracranial retention, local tumor ablation, and immune activation. This study presents a brain-mimetic injectable hydrogel incorporating a golden vesicular nanoadjuvant (Au/TLR3‑V@gel) for locoregional photothermal-immunotherapy. The hydrogel offers mechanical compliance with brain tissue and sustained retention, while the released nanoadjuvant amplifies dendritic cell activation in response to tumor antigens generated by photothermal ablation. By coupling local cytoreduction with immune remodeling and further combination with anti-CTLA-4 blockade, this platform achieves durable survival benefits in orthotopic glioblastoma. These findings establish a promising biomaterials-based strategy for intracranial immunotherapy of unresectable brain tumors.

Transarterial chemoembolization (TACE) treatment for hepatocellular carcinoma (HCC) commonly results in residual disease; however, the angiogenic features of residual tumors remain poorly defined. This study aimed to characterize vascular niche remodeling after TACE and provide a translational rationale for optimizing combination therapy. We performed single-nucleus RNA sequencing on six treatment-naive and six post-TACE HCC tumor samples. The findings were subsequently validated across five independent cohorts using multiplex immunohistochemistry, plasma biomarker assays, public transcriptomic datasets, and retrospective clinical analyses. Multiple in vivo HCC models were further used to investigate the PDGFC-PDGFR axis and evaluate low-dose anti-VEGF-A plus anti-PD-L1 therapy. Residual tumor vessels following TACE exhibited heightened angiogenic activity while remaining structurally immature, with accompanying immunosuppressive features. Integrated analyses revealed that TACE markedly reprogrammed tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs) toward pro-angiogenic and immunosuppressive states. Notably, PDGFC+ TAMs acted on pericytes and CAFs to induce pericyte-to-CAF transition and activate stromal remodeling programs, thereby reinforcing a positive feedback loop linking aberrant angiogenesis and immunosuppression. More importantly, in preclinical HCC models, low-dose anti-VEGF-A plus anti-PD-L1 alleviated hypoxia, reduced PDGFC+ TAM accumulation, enhanced CD8+ T-cell immunity, and improved tumor control. Retrospective clinical analysis further supported the potential of low-dose bevacizumab plus atezolizumab to improve clinical outcomes while preserving hepatic function after TACE. This study demonstrates that TACE induces PDGFC+ TAM-mediated dysfunction of the vascular-immune microenvironment. Low-dose bevacizumab plus atezolizumab may counteract this pathological microenvironment by promoting vascular normalization and enhancing antitumor immunity, thereby representing a promising therapeutic strategy. Transarterial chemoembolization (TACE) commonly leaves residual tumors in hepatocellular carcinoma (HCC); however, tumor angiogenic features in this context remain poorly defined. By integrating single-nucleus transcriptomics with multi-cohort validation, this study shows that residual tumors exhibit a hypoxic, angiogenic, and structurally immature vascular niche accompanied by immunosuppression. PDGFC+ tumor-associated macrophages emerged as a central component of this niche and may promote pericyte-to-fibroblast transition, cancer-associated fibroblast activation, and pathological angiogenesis. In preclinical HCC models, low-dose anti-VEGF-A plus anti-PD-L1 therapy alleviated hypoxia, reduced PDGFC+ macrophage accumulation, increased pericyte coverage, enhanced CD8+ T-cell immunity, and improved tumor control. Retrospective clinical analysis further suggested that low-dose bevacizumab plus atezolizumab may improve clinical outcomes while preserving hepatic function after TACE. These findings provide a translational rationale for further evaluating dose-optimized anti-angiogenic therapy plus immune checkpoint blockade in patients with HCC undergoing TACE, and may help clinicians and researchers refine combination strategies.

Cytotoxic T lymphocytes (CTLs) play a pivotal role in antitumor immunity via inducing apoptosis in cancer cells. However, their effector functions can be compromised by the perturbations in the tumor microenvironment (TME). Among the diverse factors within the TME, the effects of stiffness of the extracellular matrix (ECM) on CTL-mediated responses require greater understanding. To address this gap, we three-dimensionally bioprinted CTLs and targeted cells in polyethylene glycol-based ECM mimics with tunable stiffness and uniform porosity. Our findings reveal that CTLs in stiffer ECMs showed reduced migration speed and impaired antigen-specific cytotoxicity. Synapse formation analysis revealed that stiffer ECMs impair CTL efficacy not by reducing the frequency of CTL-target cell contacts but by shortening the contact time between these cells, a consequence of disrupted immunological synapse formation. Live Ca2+ imaging and expression of phospho-ZAP70 confirmed these findings. Together, these results suggest that ECM stiffness modulates CTL function at multiple levels, from migration to cytotoxicity and synapse quality.

Mitochondria-ER contact sites (MERCs) are crucial signaling hubs, but their role in anti-tumor immunity is unclear. This study revealed that the mitophagy regulator PRKN ubiquitinated CD274 at MERCs in human cervical cancer cells, a key mechanism for anti-tumor immunity. CD274 expression inversely correlated with PRKN in cervical cancer. Upon mitophagy activation, CD274 was recruited from ER to MERCs by PINK1, enhancing its interaction with PRKN. PRKN then ubiquitinated CD274 at residues K89 and K105 within its extracellular domain. Functionally, a ubiquitination-deficient CD274 mutant promoted the anaerobic glycolysis and MTOR signaling, accelerating cancer cell growth. Coculture with ubiquitination-deficient CD274 mutant-expressing cancer cells increased the CD8+ T-cells' exhaustion. Single-cell RNA sequencing of mouse tumors showed the expansion of exhausted CD8+ T cells and myeloid-derived suppressor cells (MDSCs) with ubiquitination-deficient CD274 mutation. In vivo, ubiquitination-deficient CD274 mutant accelerated tumor growth and reduced the therapy efficacy of immune checkpoint inhibitors. Conversely, clinical sample analysis showed that CD274 localization at MERCs or its ubiquitination levels were closely associated with the improved immunotherapy efficacy. Thus, mitophagy-dependent recruitment of CD274 to MERCs for PRKN-mediated ubiquitination is a novel pathway that activates the anti-tumor immunity and improves the immunotherapy efficacy, presenting a promising strategic target for cervical cancer treatment.Abbreviations: CCCP, carbonyl cyanide m-chlorophenylhydrazone; CD, cluster of differentiation; CHX, cycloheximide; FCCP, carbonyl cyanide-p-trifluoromethoxyphenylhydrazone; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GZMB, granzyme B; IFNG, interferon gamma; LDHA, lactate dehygrogenase A; MAP1LC3, microtubule-associated protein 1 light chain 3; MFN2, mitofusin 2; MHC, major histocompatibility complex; MTOR, mechanistic target of rapamycin kinase; OCR, oxygen consumption rate; PBMC, peripheral blood mononuclear cell; PDCD1, programmed cell death 1; PI, propidium iodide; PINK1, PTEN induced putative kinase 1; PKM, pyruvate kinase, muscle; RPS6, ribosomal protein S6; TNF, tumor necrosis factor; TME, tumor microenvironment.

The human genome encodes ~1,900 secreted proteins, many of which mediate intercellular communication. Secreted proteins do not act cell-autonomously, limiting systematic approaches to characterize their functions. Here we introduce SecAct (Secreted Activity, https://secact.ccr.cancer.gov ), a computational framework that infers the signaling activities of 1,170 human secreted proteins from spatial, single-cell and bulk transcriptomic data. The inference model harnesses precomputed intercellular signaling signatures trained on 1,258 spatial transcriptomics samples spanning 37 cancer types. Transcriptomics data from antisecreted protein therapies validate SecAct's accuracy in predicting the repression of secreted protein activity following treatment. For spatial and single-cell transcriptomics data, SecAct provides interactive modules for analyzing secreted protein-mediated cell-cell communication. Applying SecAct to 54 cancer immunotherapy cohorts comprising 5,174 patients, we identified secreted proteins associated with tumor immunity. In vivo experiments validated lymphocyte antigen 86 (LY86), whose function in cancer was previously unknown, as an antitumor regulator.

Immune checkpoint inhibitors (ICIs) have revolutionized the treatment of advanced malignancies; however, their efficacy in solid tumors remains limited by therapy resistance. This resistance arises from the metabolic reprogramming of immune cells in the tumor microenvironment (TME), a metabolic 'cage' where immune and cancer cells compete for nutrients. Immune effector cells succumb to metabolic exhaustion amid nutrient competition, whereas immunosuppressive cells augment inhibitory functions via metabolic adaptation, collectively mediating tumor immune evasion. This review systematically delineates the metabolic reprogramming features of immune cells in the TME, dissects the molecular mechanisms governing ICI resistance, and summarizes combination strategies targeting metabolic pathways to reverse resistance, providing theoretical and translational insights for optimizing cancer immunotherapy.

The acidic tumor microenvironment (TME) is a major driver of immunosuppression and tumor progression in breast cancer. Although myosin VI (MYO6), an actin-dependent motor protein, is frequently upregulated in malignancies, its function in immune remodeling remains poorly understood. To identify MYO6 as a regulator of acidic TME remodeling and macrophage polarization in breast cancer, elucidate the underlying mechanism, and evaluate its therapeutic relevance for anti-programmed death-ligand 1 (anti-PD-L1) therapy. Integrated analyses of The Cancer Genome Atlas and Gene Expression Omnibus datasets were performed to evaluate the association of MYO6 with prognosis and M2 tumor-associated macrophage (TAM) infiltration in breast cancer. Transcriptomic, molecular, cellular, and pharmacological approaches were used to investigate MYO6-mediated acidic TME remodeling and macrophage polarization. In vivo studies assessed the effects of MYO6 inhibition by 2,4,6-triiodophenol (TIP) on tumor acidification, immunity, and anti-PD-L1 efficacy. MYO6 expression correlated with M2 TAM infiltration and poor prognosis in breast cancer. Mechanistically, nuclear MYO6 recruited SP1 and RNA polymerase II to the carbonic anhydrase 9 (CA9) promoter, upregulating CA9 and promoting acidification. The acidic TME activated the proton-sensing receptor GPR65 and the cAMP/PKA/CREB pathway in macrophages, driving M2 polarization. TIP-mediated MYO6 inhibition reduced acidification, restored antitumor immunity, and enhanced anti-PD-L1 efficacy in vivo. MYO6 is a key molecule regulating breast cancer immune suppression and immunotherapy responsiveness. By promoting CA9-dependent extracellular acidification, MYO6 drives macrophage M2 polarization through the GPR65/cAMP/PKA/CREB pathway. Targeting MYO6 with TIP may represent a promising strategy to overcome resistance to anti-PD-L1 therapy.

Cancer-associated mesothelial cells (CAMCs) are key modulators of the ovarian tumor microenvironment, contributing to tumor growth and immune evasion. Mesothelial cells (MCs) maintain peritoneal homeostasis and immune surveillance and represent the first point of contact during abdominal dissemination of ovarian cancers. Yet, their role in ovarian tumor immunity remains poorly understood. Lineage tracing, 3D models, and spatial transcriptomic profiling were used to characterize CAMC origin, localization, and phenotypic transitions during ovarian cancer progression. Multiplex cytokine panels were used to define the cytokine profiles associated with MC transformation into CAMCs. Functional studies were conducted in syngeneic ovarian cancer mouse models to assess the impact of CAMCs on tumor growth and response to immunotherapy. In parallel, CAMC-driven changes in immune cell phenotype and functional state within the tumor microenvironment were characterized. We demonstrate that CAMCs originate from peritoneal MCs, populate the tumor surface, and progressively infiltrate the tumor core while undergoing a phenotypic transition toward a fibroblast-like phenotype. We characterize the function of an unrecognized CAMC signature marked by SERPINB2+ expression and a combination of markers absent in normal MCs. CAMCSerpinb2+ cells have reduced expression of pro-inflammatory cytokines (IL-2, IL-7, IL-12, IL-15) and increased expression of IL-10, TGFβ1, and CCL17 compared with normal MCs. Functionally, the presence of CAMCSerpinb2+ cells correlates with accelerated tumor growth, reduced CD4+ T and B cell infiltration, an expanded Treg population, and ultimately resistance to combination immunotherapy in a syngeneic mouse model of ovarian cancer. These findings identify CAMCs as central regulators of immune suppression in ovarian cancer and reveal a distinct SERPINB2+ immunosuppressive CAMC state associated with tumor progression and immunotherapy resistance. Targeting CAMCs may represent a promising therapeutic strategy to restore antitumor immunity and improve responses to current ovarian cancer treatments and immunotherapies.

Targeted cancer therapies increasingly require platforms that can penetrate poorly perfused tumor regions while minimizing systemic toxicity. Bacteria, owing to their intrinsic tumor tropism, genetic programmability, and immunostimulatory properties, have re-emerged as versatile anticancer agents, ranging from attenuated tumor-colonizing strains to highly engineered "living therapeutics." In this review, we synthesize the mechanistic foundations and therapeutic advances of bacterial-based cancer therapy through four major themes. First, we examine foundational mechanisms, including tumor-selective colonization, direct oncolysis and cytotoxicity, activation of innate and adaptive immunity, and remodeling of the tumor microenvironment. Second, we discuss engineering strategies that enable controllable delivery of therapeutic payloads, such as cytokines, antibodies and nanobodies, enzyme-prodrug systems, toxins, and nucleic-acid therapeutics, while also improving biosafety and biocontainment. Third, we evaluate combination strategies integrating bacteria with chemotherapy, radiotherapy, phototherapy, and immunotherapy, with emphasis on how bacteria complement conventional modalities by targeting hypoxic, necrotic, and immunologically refractory tumor niches. Fourth, we summarize translational progress, including representative early-phase clinical experiences, manufacturing challenges, and major safety constraints. We also highlight emerging microbiome-disease databases and computational resources that may support target selection, biomarker discovery, and therapy-response stratification. Current evidence supports bacteria as a promising precision modality, particularly for immunologically "cold" or hypoxic tumors; however, major challenges remain in the predictability of intratumoral distribution, host clearance, genetic stability, and long-term safety. Addressing these barriers through rigorous engineering, standardized manufacturing, and clinically meaningful endpoints will be essential for the next generation of bacterial therapeutics in oncology.

Hepatocellular carcinoma (HCC) responds poorly to immune checkpoint blockade, largely because of an immunologically cold tumor microenvironment (TME) characterized by deficient antigen presentation and impaired cytotoxic T cell responses. Analysis of numerous HCC clinical cohorts, including our institutional datasets, reveals that stimulator of interferon genes (STING) pathway activity is positively correlated with patient survival, enhanced antigen presentation capacity, and an immune-activated TME. However, achieving effective and controllable STING activation in immune-cold HCC tumors remains challenging. Here, we develop a biomimetic nanomedicine that integrates photothermal therapy (PTT), STING pathway activation, and immune checkpoint blockade to treat refractory HCC. A coordination nanomedicine MCI-NP is engineered by co-encapsulating the STING agonist MSA-2 and indocyanine green (ICG) via Cu2+-mediated chelation, enabling stabilized PTT-induced immunogenic cell death together with robust STING-driven innate immune activation. Further cloaked with PD-1-overexpressing cell membranes, MCI-NP@mPD-1 achieves an approximately fivefold increase in tumor accumulation compared with uncoated MCI-NP and enables localized PD-1/PD-L1 axis blockade. Following a single treatment, MCI-NP@mPD-1-based photothermal immunotherapy effectively suppresses primary tumor growth and significantly prolongs survival by remodeling the TME toward an immune-active state, while inducing durable systemic immune memory that effectively limited postoperative lung metastasis. Without introducing additional nanocarriers or excipients, MCI-NP@mPD-1 offers a promising therapeutic paradigm for photothermal immunotherapy of immune-cold HCC.

The therapeutic efficacy of melanoma immunochemotherapy is severely limited by insufficient intratumoral drug retention, systemic toxicity, and immunosuppressive tumor microenvironment (TME). Thus, we developed a single-dose injectable localized chemo-immunotherapy platform that enables sustained tumor-restricted delivery and synergistic immune remodeling. This hybrid system integrates doxorubicin-loaded tumor-derived exosome-coated hollow mesoporous silica nanoparticles (DeH) with a mannose-modified β-cyclodextrin-polyethyleneimine carrier co-delivering dual Toll-like receptor agonists (RC-MCP) within a thermoresponsive polypeptide hydrogel (mPEG-b-PELG). The biomimetic exosome coating provides an exosome-sheathed nanostructure and amplifies doxorubicin-induced immunogenic cell death. Mannose-mediated targeting facilitates preferential uptake by antigen-presenting cells and enhances endosomal Toll-like receptor signaling, leading to robust dendritic cell maturation. Following intratumoral injection, the hydrogel undergoes in situ sol-gel transition to form a stable drug depot, enabling sustained and coordinated release while minimizing systemic exposure. As a result, a single intratumoral administration elicits potent antitumor immune responses, significantly suppresses tumor growth, alleviates TME immunosuppression, and exhibits favorable biocompatibility in murine melanoma model. Collectively, this work presents a rationally designed single-dose localized chemo-immunotherapy strategy for melanoma treatment.

Although calcium ion (Ca2+) metabolism modulation exerts potent antitumor effects by triggering programmed cell death, current pharmacological interventions are severely limited by suboptimal tumor tropism and stringent metabolic feedback control loops. Herein, we engineer a supramolecular nanomedicine based on reduced human serum albumin co-encapsulating fluvastatin and resveratrol as dual Ca2+ metabolic modulators (RH-Flu-Res). After internalization by high glutathione characteristics of tumor cells, fluvastatin and resveratrol are released, which then mobilize extracellular Ca2+ influx and trigger endoplasmic reticulum (ER) Ca2+ pool leakage via intracellular microenvironment acidification and ER stress, respectively, achieving spatiotemporal reprogramming of intracellular and extracellular Ca2+ metabolism. The Ca2+ is then chemotactically recruited to mitochondria lumen via the mitochondrial gap junction protein connexon 43 (mtCX43). This particular redistribution of Ca2+ initiates Ca2+ metabolism dysregulation and subsequently activates PANoptosis in cancer cells. Meanwhile, the redistribution of lactic acid by RH-Flu-Res reprograms the tumor microenvironment into an "immune hot" phenotype. Results demonstrate that RH-Flu-Res achieves exceptional tumor suppression in multiple oral models, accompanied by marked PANoptosis and robust antitumor immune responses. This study identifies mtCX43 as a Ca2+-dependent molecular switch regulating PANoptosis and establishes a novel paradigm for PANoptosis-based cancer therapy via supramolecular nanomedicine-mediated Ca2+ metabolic reprogramming.

This narrative review aims to critically summarize available data on the association between adult acromegaly and metabolic dysfunction-associated steatotic liver disease (MASLD), with a particular focus on clinical associations and treatment implications. From a pathophysiological perspective, growth hormone (GH) may improve hepatic steatosis, inflammation, and fibrosis by acting directly on hepatocytes and indirectly (through insulin-like growth factor-1) on hepatic stellate cells, thereby inducing their senescence. Nonetheless, GH excess may also favor the development of hepatocellular carcinoma, possibly through mechanisms unrelated to hepatic fibrosis. From a clinical perspective, individuals with acromegaly have low rates of hepatic steatosis and visceral adipose tissue (VAT), but high insulin resistance (IR), which contradicts the generally observed positive association between IR and VAT or hepatic steatosis. By contrast, limited data do not support lower rates of hepatic fibrosis in acromegaly, possibly because GH excess is controlled after diagnosis. From a therapeutic perspective, published evidence, derived mainly from case series, suggests that surgical or pharmacological management of acromegaly increases hepatic steatosis and VAT, despite decreasing IR. However, the long-term effect of acromegaly control on hepatic inflammation and fibrosis remains largely unknown. Acromegaly is associated with IR, type 2 diabetes mellitus, hypertension and cardiovascular disease; however, acromegaly is inversely associated with VAT and MASLD. Further mechanistic and clinical studies are warranted to better elucidate the intriguing association between acromegaly and MASLD.

Cancer type classification is challenging due to tumor heterogeneity and undefined tissue of origin (TOO), particularly in cancers of unknown primary (CUP) and multiple primary cancers (MPC). Accurate TOO identification is critical for guiding treatment and prognosis. We developed a stacked ensemble machine learning classifier that integrates 11 multidimensional cfDNA features spanning genomic, fragmentomic, methylation/repeat, and microbial signals. Base models were constructed using five algorithms, including Deep Learning, Distributed Random Forest, Gradient Boosting Machine, Generalized Linear Model, and XGBoost, within a five-fold cross-validation framework, and their predictions were aggregated into a final ensemble optimized for top-1 accuracy. The classifier achieved robust performance across 17 cancer types, with top-1 and top-2 accuracies of 78% and 89% in the training cohort (n = 1,814), and 80% and 90% in an independent validation cohort (n = 1,221). Notably, predictive performance was retained in samples with low tumor fraction (71% top-1, 85% top-2). Sensitivity varied across tumor types, with the highest performance observed in head and neck and colorectal cancers. Among CUP cases, 11 of 15 (73.3%) predictions matched clinically inferred primary sites based on multimodal diagnostics. Feature importance analysis identified nucleosome positioning, fragment size distribution, and repeat elements as key contributors to model performance. Collectively, this cfDNA-based classifier provides a robust and non-invasive approach for accurate cancer type identification and has the potential to support clinical decision-making.

Background With the advent of immune checkpoint inhibitor-based combination therapy, treatment sequencing for advanced hepatocellular carcinoma (HCC) has become increasingly complex. The Barcelona Clinic Liver Cancer (BCLC) 2026 recommendations emphasize individualized decision-making. Purpose To compare clinical outcomes of hepatic arterial infusion chemotherapy (HAIC) and tyrosine kinase inhibitors (TKIs) in patients with advanced HCC after atezolizumab-bevacizumab (AB) failure, within the BCLC 2026 treatment paradigm. Materials and Methods This multicenter retrospective study included patients who received AB and subsequently received either HAIC or a TKI between March 2022 and August 2025. HAIC used a cisplatin-fluorouracil regimen, and TKIs were given at standard doses. Tumor response and progression-free survival (PFS) were assessed using contrast-enhanced multiphase CT or liver MRI according to routine clinical practice. Imaging was reviewed by radiologists blinded to treatment allocation. Inverse probability of treatment weighting (IPTW) was applied for age, sex, Eastern Cooperative Oncology Group performance status, Child-Pugh class, portal vein tumor thrombosis, and tumor burden based on the up-to-seven criteria. Results This study included 90 patients (mean age, 62 years ± 10.9 [SD]; 73 men; HAIC group, n = 51; TKI group, n = 39). Baseline tumor burden was higher in the HAIC group than in the TKI group (percentage of patients with tumors beyond the up-to-seven criteria: 88% [45 of 51] vs 64% [25 of 39]; P = .01). In unweighted analyses, median overall survival (OS) was similar between the HAIC and TKI groups (10.4 vs 6.4 months; P = .62), whereas PFS favored HAIC over TKIs (median, 5.3 vs 3.6 months; P = .008). Objective response rate and disease control rate were higher with HAIC than with TKIs (objective response rate: 35% [18 of 51] vs 5% [two of 39], P = .002; disease control rate: 67% [34 of 51] vs 26% [10 of 39], P < .001). After IPTW, HAIC remained associated with longer PFS than TKIs (median, 7.1 vs 3.4 months; P < .001), and OS remained similar between groups (median, 10.5 vs 6.3 months; P = .32). Conclusion In patients with advanced HCC, after AB failure, second-line HAIC yielded higher response rates and longer PFS than TKIs. © RSNA, 2026 Supplemental material is available for this article. See also the editorial by Deyirmendjian and Tang in this issue.

Perfluorooctanoic acid (PFOA) has been classified as a human carcinogen based partly on limited epidemiologic evidence of increased kidney and testicular cancer risk. We conducted an updated analysis of cancer incidence in a high PFOA-exposed cohort of community members and workers in the mid-Ohio Valley with follow-up extended by approximately 10 years on average and linkage to the Virtual Pooled Registry-Cancer Linkage System to better characterize risk of malignancies of a priori interest and to explore risk of other cancers. Based on estimates of annual serum PFOA concentrations during 1952-2020, we evaluated associations between cumulative PFOA exposure and site-specific cancer incidence through 2020 among 32,050 participants (28,398 community members and 3,652 workers). Hazard ratios (HR) and 95% confidence intervals (CI) were estimated using Cox proportional hazards models adjusted for potential confounding factors. In the overall study population, we observed positive associations with kidney and testicular cancers. Associations with kidney cancer for high PFOA exposure were most pronounced among younger community members (≤60 years: HRQ4 = 1.82, 95% CI 1.02-3.26; Ptrend = 0.03) and those with localized disease (HRQ4 = 1.72, 95% CI 1.01-2.95; Ptrend = 0.17). High PFOA exposure was also associated with increased testicular cancer risk among community members (HRQ4 = 3.62, 95% CI 1.03-12.7; Ptrend = 0.12). We also observed positive associations with thyroid cancer, particularly among female participants, and with pancreatic cancer among older participants; notably, exposure-response relationships for both thyroid and pancreatic cancer were strongest among the more highly exposed workers (per 1-unit increase on the natural log scale, HRcont = 1.79, 95% CI 1.07-2.97; and HRcont = 1.70, 95% CI 1.08-2.67 for thyroid and pancreatic cancers, respectively). Our findings in this high PFOA-exposed cohort provide additional epidemiologic evidence of increased kidney and testicular cancer risk, as well as new evidence of associations with thyroid and pancreatic cancers. These findings have important implications regarding our understanding of the carcinogenic potential of PFOA and the cancer burden related to this ubiquitous environmental contaminant.

Nitrate and nitrite, present in ground and drinking water, as well as foods, may contribute to bladder cancer through the formation of carcinogenic N-nitrosamines. However, risk likely depends on both the source and endogenous nitrosating factors. Among participants of the Danish Diet, Cancer, and Health Cohort (N = 54,610) with no history of cancer, source-specific nitrate and nitrite intakes (drinking water, plant sources, naturally occurring animal sources, and additive-permitted meat sources) were estimated from national water monitoring data and food frequency questionnaires using food databases. Bladder cancer cases were ascertained through the Danish Cancer Registry. Multivariable-adjusted Cox proportional hazards models were used to examine associations, and analyses were stratified by established bladder cancer risk factors and factors hypothesized to influence endogenous N-nitrosamine formation to explore potential effect modification. During 27 years of follow-up, 1,058 bladder cancer cases were identified. No associations were observed for nitrate intake [HRlog (95% CI) per doubling of intake (mg/d): drinking water: 0.88 (0.73, 1.08), plant sources: 0.91 (0.74, 1.12), naturally occurring animal sources: 0.83 (0.67, 1.02), additive permitted meat sources 1.06 (0.84, 1.32)]. Similar estimates were observed for source-specific nitrite intakes. Associations did not appear to be modified by bladder cancer risk factors or endogenous nitrosating factors. The lack of associations between nitrate and nitrite exposure and bladder cancer may reflect either a true absence of effect or exposure levels that are too low be biologically relevant for bladder carcinogenesis. Future studies in populations with higher exposure levels are warranted.

Post-endoscopy upper gastrointestinal cancer (PEUGIC) is defined as cancer diagnosed 6-36 months after a prior index endoscopy during which cancer was not diagnosed. Few studies have examined PEUGIC survival. We describe survival in PEUGIC patients compared with upper gastrointestinal cancer (UGIC) patients who did not undergo endoscopy without a cancer diagnosis prior to their diagnosis. This was a population-based retrospective survival analysis of UGIC patients diagnosed between January 2009 and December 2018 in England. Patients were categorized into "detected UGIC" (without prior endoscopy) and "PEUGIC" diagnosed 6-12, 12-18, 18-24, or 24-36 months after index endoscopy. PEUGIC survival was analyzed according to pre-existing Barrett's esophagus (BE), esophageal cancer histological, and gastric cancer. Kaplan-Meier all-cause survival curves investigated survival from both diagnosis and index endoscopy date. 98710 UGIC patients (9068 PEUGIC [9.2%]) were studied. Pre-existing BE was recorded for 9975 UGIC patients (10.1%), including 2583 PEUGIC (25.9%). PEUGIC survival was better than detected UGIC survival but the difference decreased as time between index endoscopy and diagnosis increased. Patients with pre-existing BE and PEUGIC had much better survival, with more stage 1 cancer (39.5% versus 8.4% for PEUGIC without BE; P < 0.001). Survival in PEUGIC and detected UGIC patients was similar when BE patients were excluded. Survival in PEUGIC patients with squamous cell carcinoma and gastric cancer was the same as for detected UGIC. Better survival in patients with PEUGIC compared with detected UGIC appeared to be related to BE surveillance. Epidemiological biases, including selection, lead time, and immortal time, should be considered when reporting PEUGIC survival outcomes.

Intraoperative image guidance has become a central component of modern surgical oncology, with near-infrared (NIR) fluorescence imaging emerging as a powerful platform for real-time visualization and precision intervention. Owing to its favorable optical characteristics, including reduced tissue autofluorescence, improved penetration depth, and excellent safety profile, NIR fluorescence imaging has rapidly evolved from experimental validation to clinical integration in gastrointestinal surgery. This review summarizes recent advances in both nonspecific and molecularly targeted NIR fluorescence imaging strategies for gastric tumors, with a particular emphasis on their integration with targeted drug delivery and theranostic applications. Preclinical studies demonstrate that NIR fluorophores conjugated to drugs, antibodies, or tumor-specific ligands enable high-contrast visualization of primary and metastatic lesions while supporting image-guided resection and localized phototherapy in gastric cancer (GC), gastrointestinal stromal tumors (GISTs), and neuroendocrine tumors. Large-animal models further validate the translational feasibility of NIR imaging for intraoperative assessment of vascular perfusion, anastomotic integrity, and sentinel lymph node (SLN) mapping. Clinically, NIR fluorescence imaging, most commonly using indocyanine green, enhances SLN detection, identifies metastatic lymphatic pathways beyond conventional dissection fields, and enables objective evaluation of tissue perfusion, thereby reducing postoperative complications. Collectively, NIR fluorescence imaging represents a versatile platform that bridges molecular imaging, drug delivery, and surgical intervention. Its integration into gastric tumor management has the potential to improve surgical precision, optimize therapeutic delivery, and accelerate the development of personalized and minimally invasive treatment strategies.

Generative indigo carmine chromoendoscopy (generative chromoendoscopy), produced using deep learning-based synthetic image transformation, is a novel image-enhanced endoscopic approach, whose clinical feasibility has not been validated. This proof-of-concept study evaluated whether generative chromoendoscopy improves the visibility of gastric neoplasms compared with conventional white-light imaging (WLI) and real chromoendoscopy. This prospective multicenter study was conducted at three Japanese institutions between January and August 2025. A deep neural network-based program converted WLI into generative chromoendoscopy images. Patients with early gastric cancer or adenoma sequentially underwent endoscopy using WLI, generative chromoendoscopy, and real chromoendoscopy. During live endoscopy, two endoscopists independently assessed lesion visibility using a 7-point Likert-type scale (-3 to +3). The primary end point was visibility of gastric neoplasms with generative chromoendoscopy relative to WLI, with comparison to real chromoendoscopy. Secondary end points included clinicopathologic factors associated with improved visibility. 60 patients were included. Mean (SD) visibility scores were 0.67 (0.93) for generative chromoendoscopy and 0.51 (1.00) for real chromoendoscopy. The mean difference was 0.16 (95%CI 0.01 to 0.30). Differentiated-type histology (odds ratio [OR] 2.19, 95%CI 1.03 to 4.62), current Helicobacter pylori infection (OR 2.14, 95%CI 1.02 to 4.46), and expert endoscopist status (OR 2.14, 95%CI 1.22 to 3.76) were significantly associated with higher visibility scores using generative chromoendoscopy. Generative chromoendoscopy was feasible during live endoscopic procedures. It improved visibility of pre-identified gastric neoplasms compared with WLI and achieved visibility comparable to real chromoendoscopy.

The European Association of Urology (EAU) produces an annual guidelines document based on the most recent evidence for the diagnosis, treatment, and follow-up of testicular cancer (TC). To summarize the 2026 version of the EAU Guidelines on TC and highlight the main changes compared to the previous version. A multidisciplinary team of clinicians with specific expertise in the disease (urologists, medical oncologists, radiation oncologists, and pathologists) reviewed the results of a comprehensive appraisal of the published literature on the topic since the last Guidelines update paper in 2023. Recommendations based on the highest available level of evidence are presented across TC stages, histologies and prognostic categories regarding diagnosis, primary management, detection and treatment of relapse and survivorship care. Areas of lack of strong recommendation consensus are highlighted. The 2026 version of the EAU Guidelines on TC collates the highest available scientific evidence to standardize the management of patients with TC.

Defining precision postpolypectomy surveillance intervals is essential to optimize the balance between colonoscopy resource use and colorectal cancer (CRC) risk reduction within population-based fecal immunochemical test (FIT) screening programs. We sought to develop fecal hemoglobin (f-Hb) concentration-guided surveillance intervals and integrate them into the existing surveillance framework. Between 2010 and 2015, 3,929,387 individuals participated in FIT screening, of whom 89,771 participants aged 50 to 74 years with f-Hb ≥20 μg Hb/g and complete colonoscopy after polypectomy were enrolled and monitored through 2018 for ascertaining incident CRCs. CRC risks across different f-Hb concentrations were estimated using an accelerated failure time model, and surveillance intervals were proportionally adjusted-shortened at higher f-Hb and extended at lower levels. We compared pragmatic, precision-oriented surveillance intervals with current guidelines to evaluate the extent of colonoscopy reduction without compromising CRC risk. There were 89,771 postpolypectomy individuals stratified as high-risk (49.5%) and low-risk (50.5%) groups based on the United States surveillance guideline. During a mean 5.5-year follow-up, CRC incidence was 2.8/1000 person-years (95% confidence interval, 2.7-3.0), increasing stepwise from 2.2/1000 person-years at 20 to 49 μg/g to 4.0/1000 person-years at ≥450 μg/g. Hypothetical f-Hb-guided surveillance intervals were lengthened for low-risk individuals with low f-Hb and shortened for those with elevated f-Hb within both risk categories. If adopting this precision approach, colonoscopy demand could be reduced by 9.8% vs current United States guidelines, while maintaining comparable CRC risk. Precision surveillance intervals guided by f-Hb can complement current guideline-based strategies by optimizing colonoscopy resource use while preserving the benefits of FIT-based CRC screening programs.

Advanced primary liver cancers remain difficult to treat after failure of first-line therapy. The 2025 French Genomic Medicine Initiative aimed to identify actionable targets for personalized treatment. Patients with PLC progressing on systemic therapy were enrolled across eight centers. Tumor and blood samples were analyzed using whole-genome, whole-exome, and RNA sequencing to identify targetable alterations classified according to the ESCAT scale. Genomic results and potential therapies were reviewed by a molecular tumor board. A total of 120 patients were enrolled: 80 with hepatocellular carcinoma (HCC), 25 with cholangiocarcinoma (CCA), 9 with combined hepatocellular-cholagiocarcinoma (cHCC-CCA), 4 with fibrolamellar carcinoma, 1 with hepatic sarcoma, and 1 with hepatic epithelioid hemangioendothelioma (HEHE). Recurrent genomic alterations included TP53 (46%), TERT (44%), and CTNNB1 (20%) in HCC; TP53 (55%), ARID1A (20%), and BAP1 in CCA; and TP53 (67%), PIK3CA (22%), and ARID2 (11%) in cHCC-CCA. Among 103 interpretable genomes, 67 patients harbored at least one actionable alteration (HCC: 59%, CCA: 80%, cHCC-CCA: 78%). Thirty-one patients (22 HCC, 5 CCA, 3 cHCC-CCA, 1 HEHE, 1 hepatic sarcoma) received matched therapies: 1 ESCAT I, 2 ESCAT II, 21 ESCAT III, and 7 ESCAT IV. These patients had received prior systemic therapy, including ≥2 lines in 69% of cases. Disease control (DC; radiological response/stable disease) was achieved in 10 of 31 patients (32.3%), including 23% of HCC, 75% of CCA, and 67% of cHCC-CCA cases. DC was observed exclusively in patients treated for ESCAT I-III alterations (41.7%), with no clinical benefit in those treated for ESCAT IV alterations. Median progression-free survival was significantly longer in patients achieving DC compared with those with progressive disease (11.8 vs. 2.4 months; p = 0.009). Comprehensive genomic profiling in advanced PLC refractory to systemic treatment is feasible and associated with DC in a subset of pretreated patients with ESCAT I/II/III alterations. Our research demonstrates that comprehensive genomic profiling through the French Genomic Medicine 2025 (FGM2025) initiative is feasible and clinically impactful in advanced primary liver cancers, including rare subtypes. By integrating whole-genome, whole-exome, and RNA sequencing, actionable genomic alterations were identified in nearly two-thirds of patients - well beyond the reach of standard next-generation sequencing panels. These findings provide a strong rationale for implementing early, broad molecular profiling to optimize therapeutic matching, preserve liver function, and expand access to precision medicine. Ultimately, our work highlights the transformative potential of genomics-guided strategies in improving clinical outcomes and advancing personalized care for patients with hepatobiliary malignancies.

This study aims to assess the risks of primary and second primary keratinocyte cancers (KCs) in patients with rheumatoid arthritis (RA) and in relation to treatment with biologic disease-modifying antirheumatic drugs (bDMARDs) or targeted synthetic disease-modifying antirheumatic drugs. Nationwide cohort study of patients treated with Janus kinase inhibitors (JAKi), tumour necrosis factor inhibitor (TNFi), or non-TNFi bDMARDs, using data from the Swedish Rheumatology Quality Register linked to other registers including the National Cancer Register, 2012 through 2023. Adjusted hazard ratios (HRs) were estimated via Cox regression using TNFi as reference. We identified 21,756 unique patients with RA. Based on 155 incident KC with JAKi, 458 with non-TNFi and 766 with TNFi, the HR for JAKi vs TNFi was 1.39 (1.16-1.68), corresponding to 1 extra KC case per every 244 patients per year. For non-TNFi vs TNFi, the HR was 0.96 (0.86-1.08). By subtype, the HR for JAKi vs TNFi was 1.41 (1.13-1.75) for basal cell carcinoma and 1.49 (1.09-2.05) for squamous cell carcinoma (SCC). For abatacept vs etanercept, the HR for SCC was 1.48 (1.11-1.97). The HR for a second primary KC was 1.31 (0.94-1.82) for JAKi and 0.94 (0.75-1.17) for non-TNFi bDMARD vs TNFi. Patients treated with JAKi have an elevated risk of KC compared with patients treated with TNFi. Although the class of non-TNFi bDMARDs is not associated with increased KC risk, we repeated a drug-specific signal of increased risk for SCC with abatacept.

Gastrointestinal (GI) cancers accounted for 23.6% of new cancer cases and 34.2% of cancer-related deaths globally. A comprehensive understanding of GI cancer burden and mortality-attributable risk factors is urgently needed. We aim to identify high-risk regions and populations to inform targeted interventions and evidence-based policies for global GI cancer control. Cases, disability-adjusted life years (DALYs), and age-standardized rates (ASRs) were obtained from the Global Burden of Disease Study (GBD) 2021. The socio-demographic index (SDI) was employed to categorize regions. Joinpoint regression was used to assess temporal trends. Risk factor contributions were calculated via the comparative risk assessment framework. In 2021, GI cancers caused an estimated 5,255,409 (95% UI: 4,687,432-5,801,303) new cases, 3,698,636 (95% UI: 3,291,859-4,091,466) deaths, and 88,123,735 (95% UI: 79,029,696-97,844,204) DALYs. Regionally, nearly half of the cases and deaths occurred in East Asia and high-income Asia Pacific. Older adults (>65 years) and males bore a significantly greater burden across most GI cancer types. Colorectal cancer and stomach cancer were the major contributors to the burden of GI cancer. Risk-attributable mortality varied by cancer type, region, SDI level, sex, and age. Tobacco use and high body mass index (BMI) were pervasive cross-cancer risk factors. Tobacco use accounted for 11.29% (95% UI: 9.36-13.28) of stomach cancer deaths and 40.64% (95% UI: 33.64-47.05) of esophageal cancer deaths, whereas high BMI contributed to 9.51% (95% UI: 4.07-14.81) of colorectal cancer deaths and 11.69% (95% UI: 8.27-15.59) of gallbladder and biliary tract cancer deaths. GI cancers impose a major yet unequal global burden, disproportionately affecting males, older adults, and populations in East Asia. The varying risk patterns underscore the necessity of targeted, region-specific public health interventions.

Our aim was to evaluate the effect of addition of radical prostatectomy (RP) to best systemic therapy (BST) on cancer-specific mortality (CSM) in patients with oligometastatic prostate cancer (omPC). This randomised controlled trial included patients with omPC with a low metastatic burden (1-5 bone metastases with/without nodal involvement) on conventional or PET imaging. Patients were randomised to receive either RP with pelvic lymph-node dissection plus BST (RP + BST) or BST alone. The primary endpoint was CSM. Secondary endpoints included clinical progression and overall survival (OS). Study accrual was stopped early because of a change in medical practice. Statistical analyses included cumulative incidence plots, Gray's test, competing-risks regression, Kaplan-Meier estimates, and log-rank tests. Between May 2015 and December 2018, 132 patients were randomised. The median age was 67 yr (interquartile range 63-71) and median prostate-specific antigen was 20 ng/ml (interquartile range 10-39). The 5-yr CSM cumulative incidence was 13% for RP + BST and 23% for BST alone (p = 0.037), with a hazard ratio of 0.39 (95% confidence interval 0.16-0.98; p = 0.045). The 5-yr cumulative incidence of clinical progression including CSM was 59% for RP + BST and 60% for BST alone. The 5-yr OS rate was 81% for RP + BST and 74% for BST alone. Clavien-Dindo grade ≥III surgery-related complications occurred in nine of 66 (14%) patients in the RP + BST arm. Limitations include early discontinuation of study accrual and the lack of statistical significance for the OS benefit. While this trial has substantial limitations, the results support addition of RP as local therapy to BST in omPC. This trial is registered on ClinicalTrials.gov as NCT02454543.

In this issue of Cancer Cell, Liu et al. apply spatial multi-omics to map colorectal cancer micrometastases across primary tumors and matched liver and lung metastases, revealing liver micrometastases as an early evolved, stem-like, immune-suppressed residual disease state linked to a six-gene recurrence signature.

Although steatotic liver disease (SLD) is a dynamic condition, the prognostic impact of subtype transitions over time remains unclear. We investigated the association between SLD subtype transitions and the risk of long-term liver-related events (LREs). From the Korean National Health Insurance Service database, adults who underwent national health checkups in both 2015 and 2017 were recruited. SLD subtypes included metabolic dysfunction-associated SLD, metabolic and alcohol-related SLD, and alcohol-related liver disease. Transitions were classified into no transitions, transitions to more advanced subtypes, and transitions to less advanced subtypes. LREs included incident cirrhosis, hepatocellular carcinoma, liver transplantation, and liver-related mortality. Subdistribution hazard ratios (sHRs) were estimated using Fine-Gray competing-risk models. Among 1,988,878 individuals, 21.7% transitioned over two years: 11.3% transitioned to more advanced subtypes, whereas 10.4% transitioned to less advanced subtypes. In each SLD subtype, transitions to more advanced subtypes showed a higher risk of LREs, whereas transitions to less advanced subtypes showed a lower risk of LREs. In SLD, the transitions to more advanced subtypes showed a higher risk of LREs (sHR, 1.34; 95% confidence interval [CI], 1.28-1.40), whereas transitions to less advanced subtypes showed a lower risk (sHR, 0.86; 95% CI, 0.83-0.88). Sensitivity analyses stratified by sex, BMI, and a fatty liver index threshold of >60 demonstrated comparable findings. We found that transitions to more advanced SLD subtypes showed increased risks of LREs, whereas transitions to less advanced subtypes showed risk reduction. The findings highlight the need for management schemes considering SLD subtype changes over time.

Local ablation therapies (LATs), including radiofrequency (RFA) and microwave ablation (MWA), are established for small hepatocellular carcinoma (HCC) as standard treatment. External beam radiotherapy (EBRT), including stereotactic body radiotherapy (SBRT) and proton beam therapy (PBT), has emerged as a non-invasive alternative; however, direct comparative evidence remains limited. We conducted a systematic review and meta-analysis comparing the efficacy and safety of EBRT versus LAT in patients with HCC. PubMed/MEDLINE, Embase, Cochrane Library, and KoreaMed were searched through June 2025 per PRISMA guidelines. Eligible studies included retrospective or prospective comparisons of EBRT (SBRT/PBT) with LAT (RFA/MWA) reporting recurrence or survival. Hazard ratios (HRs) and odds ratios (ORs) were pooled using random-effects models. Twelve studies (2,768) patients were included. Compared with LAT, EBRT was associated with significantly lower local recurrence (OR 0.36, 95% confidence interval [CI] 0.25-0.50; P<0.001; I² = 0%) and improved local progression-free survival (HR 0.41, 95% CI 0.34-0.50; P <0.001), regardless of tumor size. Progression-free survival favored EBRT (HR 0.80, 95% CI 0.64-1.00; P = 0.049), whereas overall survival did not differ significantly between modalities (HR 1.24, 95% CI 0.83-1.84; P = 0.291). Severe toxicities (grade ≥3) were comparable between groups for acute (OR 0.78, 95% CI 0.27-2.24) and late events (OR 1.29, 95% CI 0.85-1.95). EBRT was associated with better local control without significant differences in survival or toxicity compared with LAT. These findings support the safety and efficacy of EBRT for local treatment of patients with liver-confined HCC.

In DESTINY-PanTumor02 Part 1, trastuzumab deruxtecan (T-DXd) demonstrated a clinically meaningful benefit in human epidermal growth factor receptor 2 (HER2)-expressing tumors. In this article, we report subgroup analyses in the biliary tract cancer (BTC) and pancreatic cancer (PC) cohorts from DESTINY-PanTumor02 Part 1. DESTINY-PanTumor02 (NCT04482309) Part 1 evaluated T-DXd 5.4 mg/kg in locally advanced/metastatic HER2 immunohistochemistry (IHC) 3+/2+ (local/central testing) tumors that had progressed after systemic treatment or without treatment options. The primary endpoint was objective response rate (ORR) by investigator assessment (INV). Secondary endpoints included progression-free survival, overall survival, and safety. Exploratory endpoints included outcomes by biomarker status and treatment history. Overall, 41 patients with BTC and 25 patients with PC had received T-DXd; median (range) follow-up duration was 6.01 (0.7-29.1) months and 4.99 (1.1-27.2) months, respectively. By INV, ORR was 22.0% [95% confidence interval (CI) 10.6%-37.6%] in the BTC cohort and 4.0% (95% CI 0.1%-20.4%) in the PC cohort; by independent central review, ORR was 26.8% (95% CI 14.2%-42.9%) and 12.0% (95% CI 2.5%-31.2%), respectively. Responses were seen across most patient subgroups in the BTC cohort; the highest ORR (56.3%, 95% CI 29.9%-80.2%) by INV was observed for those with HER2 IHC 3+ tumors (central test). Best objective response (by INV) of stable disease was observed in 61.0% of the BTC cohort and 68.0% of the PC cohort; median overall survival was 7.0 (95% CI 4.6-10.2) and 5.0 (95% CI 3.8-14.2) months, respectively. Adjudicated drug-related interstitial lung disease/pneumonitis occurred in 17.1% of patients with BTC and 4.0% of patients with PC. These data further support T-DXd as a recommended treatment in pretreated HER2 IHC 3+ tumors. Interpretation of PC cohort findings was limited by sample size and potentially confounded by the low number of patients with IHC 3+ PC. Safety was consistent with the known profile.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease worldwide and a major cause of cirrhosis, hepatocellular carcinoma, and liver-related mortality. Weight loss via positive health behavioral changes is the cornerstone of management of MASLD. However, this is a huge challenge for many patients, highlighting the need for effective pharmacological therapies. Two pharmacological agents have received accelerated approval for the treatment of metabolic dysfunction-associated steatohepatitis (MASH), the progressive inflammatory form of MASLD, namely resmetirom, an oral, liver-directed, selective thyroid hormone receptor-β agonist, and semaglutide, a glucagon-like peptide-1 receptor agonist. Despite recent advances, effective pharmacological therapy for MASH-related cirrhosis remains an unmet need, underscoring the importance of early identification and intervention. This narrative review summarizes the current pharmacological therapy landscape of MASLD, including emerging incretin-based therapies and fibroblast growth factor-21 analogues, as well as current guidance on the use of non-invasive tests for treatment selection and monitoring for treatment response. It also provides a background on recent advances in obesity and metabolic medicine, highlighting the evolving paradigm of complication-centric obesity management and the potential role of incretin-based therapies as the backbone of the management of obesity and obesity-related conditions, including MASLD.

Histone lactylation is an epigenetic modification triggered by lactate produced during glycolysis. In cancer, histone lactylation can coordinate metabolic and epigenetic states to promote tumor development and progression. Here, we identified circ2891 as a driver of metabolic reprogramming and histone lactylation in colorectal cancer (CRC). In patients, circ2891 was aberrantly upregulated in tumors and correlated with poor outcomes. Elevated circ2891 potentiated aerobic glycolysis and lactate production in CRC cells, supporting enhanced cell proliferation and tumor growth. Mechanistically, circ2891 specifically interacted with the m6A reader FXR1 and facilitated its phase separation, which stabilized PGK1 and ENO1 mRNAs by enhancing the recognition of their m6A modification sites. Consequently, elevated PGK1 and ENO1 led to increased aerobic glycolysis and drove histone lactylation, thereby transcriptionally activating the RNA helicase DDX21 to promote CRC progression. Simultaneously targeting circ2891 and histone lactylation suppressed patient-derived organoid and xenograft tumor growth. Together, these findings show that circ2891 promotes CRC by stabilizing PGK1/ENO1 via phase separation of FXR1, boosting glycolysis and histone lactylation to activate expression of DDX21. Targeting circ2891 offers a promising therapeutic strategy enhancing clinical outcomes.

Cancer-associated fibroblasts (CAFs) drive key aspects of tumor malignancy. However, the profound heterogeneity in cytokine secretion and cell-cell communication orchestrated by CAFs renders therapeutic strategies targeting these molecules largely ineffective against cancer. In contrast, exploring the metabolic reprogramming mechanisms of CAFs could be conducive to therapeutic intervention in colorectal cancer (CRC). Here, we conducted metabolomic and single-cell RNA-sequencing analyses that uncovered increased glycolysis, intracellular lactate, and histone lactylation in CAFs compared to normal fibroblasts (NFs). Elevated histone 3 lysine 18 lactylation (H3K18la) in CAFs promoted the transition of CAFs to myofibroblastic CAFs (myCAFs), increased collagen deposition, and enhanced organoid growth. Furthermore, fibroblast-specific conditional lactic dehydrogenase A (Ldha) knockout in a mouse model mitigated CRC tumorigenesis and progression in vivo. Cancer cell-derived TGF-β accelerated metabolic reprogramming and histone lactylation in CAFs. NPM1, a histone chaperone functioning as a cofactor of P300, enhanced the H3K18la level of the ROCK1 promoter and activated ROCK1 transcription. The RhoA/ROCK1/MLC2/MRTF-A pathway and extracellular matrix (ECM) remodeling were indispensable for the tumor-promoting effect of CAF histone lactylation in CRC. Furthermore, CAF histone lactylation limited CD8+ T cell infiltration and aggravated CD8+ T cell exhaustion via ECM remodeling. Stiripentol, a clinical drug that targets lactylation, potentiated the efficacy of immunotherapy in CRC models. Together, this study demonstrates that elevated lactate levels in CAFs reprogram the epigenetic landscape to promote tumor progression and immunosuppression, highlighting the potential of targeting CAF histone lactylation as a therapeutic strategy for CRC.

Cancer remains a major threat to global public health, highlighting the urgent need for precise and effective treatment strategies. Photothermal therapy (PTT) has gained increasing attention as a promising alternative owing to its non-invasiveness, deep tissue penetration, and oxygen-independent mechanism. However, the development of high-performance organic photothermal agents (PTAs) with pronounced near-infrared absorption, high photothermal conversion efficiency (PCE), and good photostability remains challenging. Herein, we propose a synergistic strategy combining π-bridge and donor engineering to design a series of aggregation-induced emission (AIE)-active small-molecule photosensitizers. Experimental and theoretical analyses reveal that stepwise introduction of a planar thiophene π-bridge and methoxy donor groups systematically regulates the excited-state energy dissipation pathways, enabling a continuous modulation from photodynamic therapy (IsoTPA), through balanced synergistic PDT/PTT (IsoTHTPA, PCE = 57.2%), to highly efficient PTT (IsoTHTO, PCE = 64.3%), which reveals the fundamental competitive interplay between intersystem crossing and non-radiative decay. Encapsulated into nanoparticles, IsoTHTO NPs demonstrate efficient photothermal ablation of 4T1 tumor cells and significant tumor growth inhibition under 660 nm laser irradiation both in vitro and in vivo. This work presents a reasonable molecular design approach for tunable access to PDT, synergistic PDT/PTT, and PTT modalities from a single molecular platform for cancer phototheranostics.

Colorectal cancer (CRC) cells accumulate iron to fuel proliferation yet paradoxically resist its toxicity. Jain et al. reveal that heme stabilizes succinate dehydrogenase subunit C, sustaining complex II-dependent coenzyme Q reduction and its redistribution to the plasma membrane, enabling CRC cells to buffer oxidative stress and iron-induced cell death.

Oral chemotherapy for colorectal cancer (CRC) is limited by poor tumor selectivity and microenvironment-driven resistance. Addressing these limitations demands materials that integrate tumor-selective targeting with immune microenvironment modulation. Here, clinical analysis of CRC specimens revealed pronounced transferrin (Tf) enrichment in CRC-associated intestinal regions. Guided by this finding, we engineered a gastrointestinal-stable cyclic Tf-binding peptide (cp) with high Tf affinity and constructed cp-modified ginger-derived extracellular vesicles (cp-GEVs) for in situ Tf-mediated sandwich-like targeting. By recruiting endogenous Tf, cp-GEVs established a Tf-mediated bridging interface that selectively engages Tf receptor-overexpressing intestinal epithelium and tumor cells, enabling efficient epithelial transcytosis, tumor-selective accumulation, and deep intratumoral penetration after oral administration. When loaded with irinotecan (CPT-11), CPT@cp-GEVs significantly enhanced intracellular drug delivery and reprogrammed immunosuppressive M2-like tumor-associated macrophages toward a pro-inflammatory phenotype, thereby disrupting cancer stem cell-enriched drug-resistant niches. In AOM/DSS-induced primary CRC models and patient-derived ex vivo systems, CPT@cp-GEVs significantly improved chemotherapeutic efficacy while attenuating resistance. Collectively, this work establishes a Tf-mediated sandwich-like targeting framework for oral cancer therapy, offering a conceptually distinct materials design paradigm that integrates endogenous ligand recruitment with immune microenvironment reprogramming.

Selenium deficiency is linked to gastric cancer, and p53 mutant tumors often acquire chemoresistance. To address this, we designed a camptothecin (CPT) dimeric prodrug with a hybrid -S-Se-S- linker (CPT-S-Se-S-CPT). It self-assembles into uniform nanoparticles (SSeSCPT NPs) with dual redox-responsive release triggered by glutathione and reactive oxygen species. Selenium plays dual synergistic roles: it enhances cellular uptake and pharmacokinetics while reducing systemic toxicity; it also acts as a redox-active center to amplify oxidative stress, activating p38/MAPK and unfolded protein response pathways, thereby inducing p53-independent apoptosis and overcoming drug resistance. In vivo, SSeSCPT NPs prolong plasma half-life, achieve efficient tumor accumulation via the EPR effect, and show potent antitumor activity in p53 mutant gastric cancer models. Surface selenium groups recognize Toll-like receptor 4 (TLR4) overexpressed on tumor cells, promoting clathrin/caveolin-mediated endocytosis and bypassing EPR size limitations. This nanoplatform integrates CPT chemotherapy, selenium intervention, and the p53 mutation background into a single system, establishing a toxicity-controlled, efficacy-enhanced strategy. It also reveals how selenium-based nanomaterials remodel redox homeostasis, bypass p53 deficiency, and reprogram apoptotic networks. This work provides mechanistic insights and translational directions for precision therapy of p53 mutant gastrointestinal malignancies.

Ultrasound-induced luminescence offers a light-free imaging modality with deep tissue penetration and spatiotemporal controllability; however, its broader application is hindered by weak luminescent signals that limit imaging depth and signal-to-noise ratios. Here, we report an organic-inorganic heterojunction sonosensitizer, TA@TiO2, formed by coupling a trianthracene derivative (TA) with titanium oxide (TiO2). Under ultrasound irradiation, the nanoscale charge-transfer interface promotes interfacial charge transfer, significantly enhancing reactive oxygen species (ROS) generation. This increased ROS triggers amplified chemical energy conversion, resulting in a markedly enhanced ultrasound-induced luminescence signal for deep-tissue optical imaging. Compared to TA nanoparticles, TA@TiO2 exhibits superior signal transmission in scattering media and maintains high luminescence at lower power densities. In vivo studies demonstrate that TA@TiO2 enables high-contrast imaging of deep-seated tumors, such as pancreatic cancer and glioma, while providing enhanced sonodynamic therapy efficacy. The positive correlation between ultrasound-induced luminescence intensity and ROS generation allows for dynamic, imaging-guided tumor therapy. These results establish heterojunction engineering as a potent strategy for advancing ultrasound-activated theranostics.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with poor outcomes. Obesity increases the risk of PDAC through metabolic dysregulation and inflammation. Ketogenic diet (KD) can alter metabolism and has been evaluated for its effects on tumor progression in nonobese PDAC using genetically engineered mouse models (GEMM). We hypothesized that KD may also prevent obesity-associated PDAC progression by altering body composition and cancer metabolism. Therefore, male PDAC GEMMs were subjected to diet-induced obesity (DIO) using high-fat diets or maintained on a low-fat diet (LFD) for 15 weeks. Mice were then randomized to continue the initial diets or switch to a KD or matched control diet for 6 weeks. Body weight and composition, glucose tolerance, ketone levels, pancreas histology, and tissue metabolomics were assessed. Furthermore, murine pancreas-derived organoids from DIO or LFD-fed GEMMs were treated with a ketone body and analyzed using untargeted metabolomics. In obese PDAC GEMMs, KD delayed cancer progression independent of weight loss, an effect not observed in nonobese LFD-fed mice. KD-mediated PDAC suppression was associated with enrichment of pancreatic metabolic pathways that support nonglucose energy production. Ketone-treated organoids recapitulated a subset of the KD-associated metabolic differences observed in vivo, suggesting a direct metabolic effect on cancer cells. These findings suggest potential benefits of a KD in preventing obesity-associated PDAC. The diet-cancer metabolic interactions highlight potential opportunities for dietary or metabolic interventions to prevent PDAC in high-risk obese populations. Ketogenic diet prevents pancreatic cancer progression in obese, but not in nonobese, genetically engineered mouse models and modifies pancreatic metabolic pathways, informing future dietary and therapeutic strategies for high-risk obese populations.

Despite the availability of RAS inhibitors and the dependence of >90% of pancreatic ductal adenocarcinomas (PDAC) on oncogenic KRAS mutations, resistance to KRAS inhibition remains a serious obstacle. We showed here that phosphoinositide 3-kinase (PI3K) plays a major role in this resistance through upstream activation of wild-type (WT) RAS signaling-beyond its known KRAS effector function. The combination of proximity labeling, CRISPR screening, live-cell imaging, and functional assays revealed that PI3K orchestrates phosphoinositide-mediated GAB1 recruitment to the plasma membrane, nucleating assembly of RAS signaling complexes that activate mitogen-activated protein kinase (MAPK) in an EGFR/SHP2/SOS1-dependent manner. Inhibiting PI3K enhanced sensitivity to mutant-specific KRAS inhibitors in PDAC cells, including in cells with clinically identified PIK3CA mutations. These findings refine RAS-PI3K signaling paradigms, reveal that PI3K-driven WT RAS activation drives resistance to KRAS inhibition, and illuminate avenues for augmenting KRAS-targeted therapies in PDAC. PI3K plays a functional upstream role in assembling wild-type RAS signaling complexes that confer KRAS inhibitor resistance, providing a rationale for combined targeting of PI3K and KRAS in pancreatic cancer.

Cancer tissue forms cancer stroma with a high elastic modulus due to increased type I collagen (Col I) production from fibroblasts and cross-linking of Col I fibers. Stiffened cancer stroma acts on cancer cells as a mechanical stress and promotes cancer progression. This highlights the need for culture scaffolds that recapitulate the structural, mechanical, and biological characteristics of the tumor stromal microenvironment. In this study, we attempted to create a scaffold material that mimics the fibrous structure and elastic modulus of in vivo cancer tissue using Col I. By gelation with collagen microfibers (CMFs) and collagen nanofibers (CNFs) at various concentrations, we were able to produce collagen gels with elastic moduli ranging from 1 to 18 kPa and 2 to 80 kPa, respectively. When cultured on CNF gels with a similar elasticity to in vivo colorectal cancer tissue, JC-011 exhibited proliferation comparable to that on Matrigel. Furthermore, JC-011 cells cultured on CNF gels exhibited cancer-related protein expression patterns highly similar to those observed on Matrigel. Together, these findings demonstrate that hierarchical CNF gels recapitulate a physiologically relevant tumor stromal microenvironment and provide a promising collagen-based platform for investigating cancer cell-matrix interactions.

Neuroendocrine prostate cancer (NEPC) persists in a profoundly hypoxic microenvironment, yet the mechanisms enabling tumor adaptation to this metabolically challenging niche remain undefined. Here, we identify the lipid kinase PIKfyve as overexpressed in NEPC, functioning as a central node in a stress-adaptive lipid kinase axis that supports adaptation to persistent endoplasmic reticulum (ER) stress. Mechanistically, NEPC requires PIKfyve-mediated lysosomal degradation and lipid recycling to maintain metabolic homeostasis under hypoxia. PIKfyve inhibition disrupts lysosomal function, exacerbates ER stress, and activates a compensatory sterol regulatory element-binding protein (SREBP)-dependent de novo lipogenesis program essential for NEPC survival. This stress-lipid axis creates a synthetic vulnerability between PIKfyve and fatty acid synthase (FASN), where dual inhibition synergistically amplifies ER stress, triggers the terminal unfolded protein response, and induces tumor cell death. These findings reveal a metabolic adaptation in NEPC and provide preclinical evidence that co-targeting PIKfyve and FASN can overcome hypoxia-associated stress adaptation.

Three-dimensional (3D) histology provides volumetric insights into tissue microarchitectures across entire specimens, holding great promise for more accurate prognostication. However, existing methods are too slow for intraoperative consultations. We present ULTRA (ultrarapid cleared stimulated Raman with AI), a label-free, stain-free, fixation-free, and section-free platform that leverages the chemical specificity of stimulated Raman scattering (SRS) microscopy for rapid 3D histological analysis. Through the synergistic development of a one-step tissue-clearing protocol and unsupervised learning algorithms, ULTRA delivers high-resolution, formalin-fixed, paraffin-embedded (FFPE)-grade deep 3D virtual histology within 30 min, covering orders of magnitude more tissue than slide-based methods. In human surgical glioma samples, ULTRA accurately resolves key histological features in 3D and delineates depth-dependent tumor infiltration margins at single-cell resolution. By compressing 3D histology from days or hours to an intraoperative timescale, ULTRA addresses a critical clinical gap and enables more informed surgical decision-making in the operating room.

Early diagnosis of HCC remains a clinical challenge, with survival strongly dependent on reliable biomarkers and translatable detection tools. Herein, we establish uracil-DNA glycosylase (UDG) and apurinic/apyrimidinic endonuclease 1 (APE1) as diagnostic biomarkers for early-stage HCC and develop a lock-open strategy based on an AND logic-gated, entropy-driven DNA nanowalker (ALEN) for highly sensitive detection of their enzymatic activities. ALEN integrates sequential UDG/APE1 processing with miR-21 gating to achieve on-track signal amplification. In the locked state, circuit leakage is minimized. Upon UDG/APE1 processing and miR-21 gating, ALEN is unlocked to initiate walker propagation, reactant regeneration, and spatially confined amplification selectively in tumor cells. This ability overcomes critical limitations of conventional entropy-driven DNA circuits, including low efficiency, limited turnover, and excessive leakage. When encapsulated in aptamer-functionalized lipid nanoparticles, ALEN gains robust biostability, biocompatibility, and HCC-specific accumulation. Using in vitro cellular models and in vivo mouse models of HCC, we confirmed that ALEN enables highly sensitive, cell type-specific discrimination of the activities of UDG and APE1, as well as timely and sensitive sensing of their dynamic fluctuations. Our results demonstrate that ALEN facilitates HCC diagnosis and monitoring of tumor progression, highlighting its promise as a clinical management tool.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, largely due to limitations in early diagnosis and effective treatment. To address these challenges, we developed a multifunctional probiotic nanoplatform, EcNA@Cur-Lip-FA, that integrates real-time diagnosis, targeted therapy, and immune modulation for enhanced CRC treatment. Specifically, this system leverages genetically engineered Escherichia coli Nissle 1917 (EcN) encoding acoustic reporter genes (ARGs) to produce ultrasound (US)-responsive gas vesicles (GVs), enabling real-time, noninvasive tumor imaging. Simultaneously, folic acid (FA)-modified curcumin-loaded liposomes (Cur-Lip-FA) are co-delivered with EcN, enhancing tumor-specific accumulation and therapeutic precision. After being taken up by tumor cells, curcumin is released locally upon exposure to US irradiation, generating reactive oxygen species (ROS), inducing immunogenic cell death (ICD), and activating the cGAS-STING pathway to promote dendritic cells (DCs) maturation and robust CD4+/CD8+ T-cell responses. Additionally, EcNA@Cur-Lip-FA downregulates angiogenesis-related factors (VEGF, VEGFR1, and ANGPT1), thereby inhibiting tumor neovascularization. In vitro and in vivo studies demonstrated that EcNA@Cur-Lip-FA enables precise tumor localization, dynamic imaging, and effective suppression of both primary CRC and metastatic liver lesions with favorable biocompatibility. This multifunctional platform offers a promising strategy for integrating diagnosis and treatment, paving the way for next-generation CRC theranostics.

Gastric cancer remains a leading cause of cancer mortality. The Correa cascade-stepwise progression from inflammation through atrophic gastritis, metaplasia and dysplasia to adenocarcinoma-has long defined its pathogenesis. Lineage tracing, single-cell multi-omics and organoid models now reveal marked cellular heterogeneity, plasticity and non-sequential trajectories. This review synthesises gastric stem-cell identity, niche architecture and precancerous progression. Homeostasis is sustained by cycling antral LGR5+ basal and IQGAP3+ isthmus progenitors and by quiescent corpus MIST1+/TROY+ chief-cell reserves, coordinated by Wnt/R-spondin, Notch, BMP, Hedgehog and EGFR signalling. Helicobacter pylori subverts this niche through type-IV-secretion-system delivery of cytotoxin-associated gene A and through BMP collapse, driving region-specific trajectories. In the antrum, hyperproliferation and CDX2-dependent intestinal metaplasia advance to TP53/KRAS/APC-mutant dysplasia. In the corpus, oxyntic atrophy triggers isthmus expansion via type-2 innate lymphoid cell (ILC2)-derived WNT5A and YAP signalling, alongside a tuft-cell-ILC2 interleukin-25/interleukin-13 circuit driving spasmolytic polypeptide-expressing metaplasia (SPEM). In lineage-tracing models, KrasG12D-mutant isthmus cells can also bypass SPEM to generate dysplasia directly. Further origins include ATOH1-deficient pit cells and bone-marrow-derived epithelial hybrids. Together, these findings support a regionally stratified, stem-cell-centred model in which metaplasia and dysplasia arise as parallel cellular fates. This refinement operates at the cellular level without displacing the tissue-level Correa sequence documented in long-term human cohorts. It nominates the remodelled stem-cell niche as a tractable pharmacological target and warrants molecular profiling of at-risk progenitor populations to complement, rather than replace, histopathological surveillance.

Aspartame, one of the most widely consumed artificial sweeteners globally, has been associated with increased cancer risk in epidemiological studies, yet the underlying mechanisms remain poorly understood. This study investigated the effects of subchronic aspartame exposure on intestinal tumorigenesis using the Apcmin/+ spontaneous intestinal tumor mouse model and elucidated the mechanistic pathways involved. Mice were exposed to physiologically relevant doses of aspartame (0.07 and 0.2 mg/mL) in drinking water for 3 months, followed by comprehensive analysis of tumor development, gut microbiota composition, and intestinal metabolomics. The results revealed that aspartame exposure significantly promoted tumor progression, as evidenced by increased proportions of medium-sized tumors in the small intestine. Microbiota analysis using 16S rRNA sequencing revealed selective enrichment of Akkermansia muciniphila (A. muciniphila) and increased microbial dysbiosis following aspartame treatment. Metabolomic profiling identified tryptophan metabolism as the most significantly altered pathway, with enhanced production of serotonin pathway metabolites. Mechanistic validation through in vitro experiments demonstrated that aspartame directly stimulated A. muciniphila growth and increased bacterial production of 5-hydroxytryptophan (5-HTP), which was subsequently converted to serotonin (5-HT) in tumor tissues. Culture supernatants from aspartame-treated A. muciniphila promoted colorectal cancer cell proliferation, an effect that was specifically blocked by carbidopa, an inhibitor of 5-HTP to 5-HT conversion. These findings reveal a novel aspartame-microbiota-serotonin axis that promotes intestinal tumor progression, challenging the perceived safety of long-term aspartame consumption and highlighting the need for reevaluation of current safety guidelines.

Glioblastoma (GBM) cells eliminate temozolomide (TMZ) through extracellular vesicles (EVs), a resistance mechanism independent of DNA damage repair that markedly reduces intracellular drug concentration. However, strategies to block this efflux pathway remain poorly explored. Levetiracetam (LEV), a first-line antiepileptic for GBM patients, suppresses seizures by inhibiting synaptic vesicle release, but its potential role in chemotherapeutic efflux has not been investigated. Chemical proteomics identified RAB5A and CD63 as novel LEV-binding targets, and analyses of TCGA datasets further supported their clinical relevance. Mechanistic studies combining co-immunoprecipitation, immunofluorescence, and in silico modeling revealed LEV-mediated disruption of endosomal trafficking and membrane fusion, thereby inhibiting TMZ efflux. Nanoparticle tracking analysis and electron microscopy were used to evaluate EV release, while LC-MS/MS was employed to quantify TMZ at the subcellular and intratumoral levels. Orthotopic GBM models were used to evaluate therapeutic efficacy. RAB5A and CD63 were identified as dual mediators of LEV activity. LEV competitively bound RAB5A, impairing endosomal maturation and TMZ trafficking. LEV also disrupted CD63-RAB35 interaction, promoting RAB35 proteasomal degradation and suppressing plasma membrane fusion. Collectively, LEV reduced TMZ efflux and synergistically enhanced TMZ cytotoxicity in vitro, while in orthotopic models the combination therapy inhibited tumor growth, was accompanied by immune microenvironment remodeling, and prolonged survival.

RNF43 is frequently inactivated by mutations in pancreatic ductal adenocarcinoma (PDAC), but the molecular mechanisms and therapeutic vulnerabilities associated with RNF43 loss remain poorly defined. Here, we demonstrate that RNF43 functions as an E3 ubiquitin ligase targeting YBX1 for degradation, thereby suppressing mitochondrial oxidative phosphorylation (OXPHOS). In RNF43-deficient PDAC models, stabilized YBX1 activates MYC through dual mechanisms-enhancing MYC mRNA stability via IGF2BP1 and physically interacting with c-Myc protein-leading to transcriptional upregulation of IDH2 and IDH3A and subsequent OXPHOS activation. Importantly, RNF43 loss conferred sensitivity to OXPHOS inhibition both in vitro and in vivo. Treatment with the OXPHOS inhibitor IACS-010759 suppressed the proliferation, migration, invasion, and metastasis of RNF43-mutant tumors. Our findings identify a RNF43-YBX1-MYC signaling axis associated with metabolic reprogramming in pancreatic cancer and suggest that OXPHOS inhibition may represent a potential therapeutic vulnerability in tumors with RNF43-inactivating mutations.

ARID1A is frequently mutated in non-tumorous human tissues and various cancers. However, its role in metabolic dysfunction-associated steatohepatitis (MASH) and hepatocellular carcinoma (HCC) onset and progression remains controversial. Given the critical role of the gut-liver axis in MASH and HCC, we hypothesized that the gut microbiota might contribute to ARID1A deficiency-induced liver diseases. Here, we demonstrate that liver-specific Arid1a deficiency drives MASH/HCC development through dysregulated bile acid metabolism and gut microbiota restructuring. Gut microbiota depletion using an antibiotic cocktail attenuates MASH progression and reduces HCC incidence in liver-specific Arid1a-deficient mice. While lipopolysaccharide (LPS) accumulation due to gut barrier dysfunction promotes HCC progression, it does not significantly affect MASH activity or tumor initiation in Arid1a-deficient mice. Mechanistically, ARID1A deficiency reduces chromatin accessibility and impairs SWI/SNF complex binding at the promoter of nuclear receptor FXR, which governs bile acid homeostasis, suppressing FXR transcription. Further analyses in Arid1a-deficient mice reveal altered gut microbiota composition enriched in bile salt hydrolase (BSH) genes and bile acid profiles showing elevated levels of the secondary bile acid taurodeoxycholic acid (TDCA). When administered to mice, TDCA exacerbates liver inflammation and fibrosis specifically in Arid1a-deficient mice by promoting neutrophil infiltration and hepatic stellate cell activation. Therapeutic interventions including vancomycin to eliminate bile acid-metabolizing bacteria, obeticholic acid to activate Fxr, and cholestyramine to excrete bile acids substantially ameliorate MASH pathology in Arid1a-deficient mice. Collectively, our findings establish the ARID1A-FXR-bile acid-gut microbiota axis as a pathogenic driver of MASH and HCC, offering mechanism-based therapeutic strategies for patients with ARID1A mutations.

Cancer remains a leading cause of death worldwide, and current systemic therapies are limited by poor tumor penetration and retention, systemic toxicities, and therapeutic resistance. Intratumoral (IT) immunotherapy offers a promising alternative. By delivering therapies directly into the tumor, IT delivery platforms can reduce systemic toxicity and improve efficacy. This review compares systemic and IT immunotherapy, emphasizing the ability of IT immunotherapy to bypass vascular and stromal barriers, reduce immune-related adverse events, and enable synergistic combinations that may be intolerable systemically. We examine IT delivery platforms including nanoparticles, bacterial vesicles, oncolytic viruses, device-assisted infusion systems, hydrogels, implantable scaffolds, drug-eluting seeds, and microneedle patches. For each, we discuss delivery strategies and translational considerations. We also highlight IT therapeutic cargos, including gene therapies, cell therapies, pattern recognition receptor agonists, cytokines, immune-checkpoint inhibitors, and metabolic modulators, and we discuss which delivery platforms are more suitable for particular therapeutic cargos. Finally, we explore synergy between IT immunotherapy and external energy modalities such as photothermal therapy, microwave ablation, magnetic hyperthermia, radiotherapy, sonodynamic therapy, photodynamic therapy, and cryotherapy. While challenges remain, the ability of IT immunotherapy to achieve durable systemic anti-tumor immune responses with minimal toxicity positions it as a transformative approach for cancer immunotherapy.

Testicular steroidogenesis is fundamental to male reproductive health, but its disruption by environmental and emerging chemicals remains insufficiently characterized due to limitations in existing test systems. Traditional animal models pose ethical and logistical challenges, while the validated H295R assay-based on a female adrenal carcinoma cell line-fails to reflect male gonadal steroidogenesis. This review uses a semi-systematic approach to evaluate over 1500 studies employing in vitro models, including primary Leydig cells, Leydig cell lines, stem cell-derived Leydig-like cells, and advanced 3D testicular systems. We assess species origin, developmental relevance, culture conditions, and the extent to which these models replicate key steroidogenic pathways. Most models rely on rodent-derived, cancerous cell lines cultured in two-dimensional monolayers, with limited representation of human and immature Leydig cells. A targeted full-text analysis examined the effects of 23 reference chemicals on testosterone, progesterone, androstenedione, and estrogen levels across the H295R assay and eight testicular in vitro models. Forskolin, genistein, prochloraz, and ketoconazole showed consistent effects and may serve as promising reference compounds. However, data for most chemicals in testicular models are scarce or inconsistent-particularly for androstenedione and progesterone-underscoring the need for improved model standardization. We propose future directions to enhance predictive power, including the development of hormone-responsive, species- and stage-specific models cultured under hormone-controlled conditions. Such advances are essential to improve chemical safety assessment and facilitate regulatory acceptance of alternative test methods.

Hepatoblastoma (HB) is the most common malignant liver tumor in children. One-fifth of patients exhibit a poor prognosis due to resistance to conventional chemotherapy, which typically includes doxorubicin. One of the underlying mechanisms is drug efflux through the MDR1 export pump. This study aims to explore pharmacological strategies for sensitizing HB by inhibiting MDR1. A panel of compounds, including established MDR1 inhibitors, natural products, clinically used drugs, and tyrosine kinase inhibitors (TKIs), was employed to inhibit MDR1 activity and enhance the response to doxorubicin. Wild-type (HepG2-WT) and doxorubicin-resistant (HepG2-DR) cells, with enhanced MDR1 expression, as well as murine xenograft models and patient-derived HB cells (HB-303) and organoids, were utilized. Curcumin did not sensitize HepG2-DR cells to doxorubicin, whereas verapamil and simvastatin enhanced doxorubicin cytotoxicity only at toxic concentrations. In contrast, several TKIs, including nilotinib, tivozanib, and, to a lesser extent, cabozantinib, exhibited synergistic effects with doxorubicin in HepG2-DR cells. These TKIs also improved the efficacy of doxorubicin in patient-derived HB organoids, a response that depended on MDR1 expression. Third-generation MDR1 inhibitors (tariquidar, elacridar, and zosuquidar) sensitized HepG2-DR and HB-303 cells at non-toxic nanomolar concentrations in vitro. Furthermore, the combination of doxorubicin and zosuquidar significantly reduced tumor growth even when these were generated from chemoresistant cells. In conclusion, we described pharmacological strategies to enhance HB response to chemotherapy. MDR1 inhibitors, such as zosuquidar, may enable dose reductions of chemotherapeutic agents, whereas the use of synergistic TKIs, such as tivozanib, may improve therapeutic outcomes and minimize adverse effects in children with HB. TG100-115, a TRPM7 kinase inhibitor, provides neuroprotection and attenuates NLRP3 inflammasome-mediated neuroinflammation in a neonatal mouse model of hypoxic-ischemic brain injury.

The early diagnosis of gastric cancer (GC) faces great challenges due to the lack of specific biomarkers. Therefore, it is crucial to discover effective markers and establish highly sensitive detection methods. This study validated the potential of BTN3A2 protein as an early GC biomarker by analyzing serum samples from patients with GC, benign gastritis, other digestive system cancers, and healthy controls across three clinical centers. Furthermore, a chemiluminescent (CL) imaging immunosensor based on copper-doped NiFe PBA (Cu-NiFe PBA) nanozyme probe was developed to achieve highly sensitive detection of BTN3A2 protein in serum of patients with GC and healthy controls. The high detection sensitivity for BTN3A2 was achieved by copper doping to enhance peroxidase-like activity of nanozyme for signal amplification. The proposed sensor exhibits a broad linear range from 0.5 to 5000 pg/mL, and achieves a detection limit of 0.15 pg/mL (S/N = 3). In addition, the established sensor is successfully utilized for BTN3A2 detection in serum samples from GC and healthy individuals across three clinical centers, and demonstrates perfect correlation with clinical outcomes when compared with traditional enzyme-linked immunosorbent assay. This study provides a candidate serum biomarker in the early stage of GC and offers a convenient, highly sensitive method for its detection.

Tumor necrosis factor-alpha (TNF-α) plays paradoxical roles in cancer, promoting tumor survival via NF-κB activation while also inducing necroptotic cell death. Here, we identify the intracellular pH (pHᵢ) as a decisive switch that reprograms TNF-α signaling from a prosurvival inflammatory response toward tumoricidal necroptosis. In both hepatocellular and cervical carcinoma models, alkalizing the pHᵢ with bicarbonate inhibits TNF-α-mediated IκB phosphorylation and NF-κB nuclear translocation while simultaneously activating the TNF-α-driven necroptotic cascade, which is characterized by the sequential phosphorylation of RIPK1, RIPK3, and MLKL. Mechanistically, bicarbonate induces the depolarization of the mitochondrial membrane potential, reactive oxygen species (ROS) generation, and a cyclophilin D-dependent permeability transition-effects that are synergistically amplified by TNF-α. Genetic knockdown of RIPK1 or RIPK3 abrogates necroptosis, whereas the pharmacological inhibition of ROS suppresses necroptosis and concurrently restores NF-κB nuclear translocation. ROS are identified as the key mediators determining the switch between these opposing TNF-α signaling pathways. In vivo, bicarbonate and TNF-α treatment suppresses tumor growth, enhances necroptotic signaling, and inhibits NF-κB activation. Thus, our work indicates that intracellular alkalization acts as a modifiable regulator to redirect TNF-α from a toxic, proinflammatory cytokine into a potent and selective tumoricidal agent. Graphical Abstract: Alkalization of the pHi by bicarbonate reprograms TNF-α signaling from a prosurvival NF-κB axis to a cytotoxic necroptotic pathway. a TNF-α alone activates NF-κB signaling, promoting inflammation and tumor growth. b Bicarbonate alkalizes the pHi and induces mitochondrial dysfunction and reactive oxygen species (ROS) production, which is further amplified by TNF-α. Elevated ROS levels inhibit NF-κB signaling and activate RIPK1-dependent necroptosis. This ROS-mediated switch redirects TNF-α signaling from a proinflammatory, tumor-promoting response to a cytotoxic, necroptotic pathway, resulting in the suppression of both tumor growth and inflammation. This diagram highlights the translational potential of combining bicarbonate with TNF-α for cancer therapy.

Biodegradable magnesium and its alloys have demonstrated ability of anti-tumor cells. However, it remains unclear whether they can regulate the tumor's immune-suppressive microenvironment required for effective treatment of tumor-associated esophageal stenosis. In this study, we investigate the effects of pure Mg, Mg-Cu, and Mg-Cu-Zn alloys on tumor growth and regulation of the tumor immune microenvironment. Our in vitro results show that all three materials effectively suppress the proliferation of human esophageal cancer cells (Eca109 and KYSE30) and mouse AKR esophageal cancer cells. These anti-tumor effects are further validated in vivo using subcutaneous allograft models in immunocompetent mice and xenograft models in immunodeficient mice, where each material inhibits tumor growth and promotes tumor necrosis. Despite these shared anti-tumor effects, only the Mg-Cu-Zn alloy significantly reduces the infiltration of CD163+ M2 tumor-associated macrophages within tumor tissues, indicating a distinct immunomodulatory function. RNA sequencing analysis reveals that the tumor suppression is associated with inhibition of proteasome activity and oxidative phosphorylation, whereas the improved immune microenvironment is linked to enhanced Fc gamma receptor (FcγR)-mediated phagocytosis-an effect not observed with pure Mg or Mg-Cu alloys. These findings demonstrate that the Mg-Cu-Zn alloy not only has anti-tumor properties but also plays a role in regulating the immune microenvironment, making it a promising candidate for biodegradable esophageal stents in tumor-associated stenosis.

The ketogenic diet (KD), an emerging nutritional intervention for cancer, reprograms cellular energy metabolism from glucose to ketone bodies, including acetoacetate (AcAc), acetone (Ac), and β-hydroxybutyrate (BHB). However, the mechanisms connecting ketone body signal sensing to tumor growth suppression remain elusive. Here, we show that RagC, a key component of mTORC1 pathway, senses BHB but not AcAc and Ac, to dictate tumor suppression. KD-derived BHB inhibits mTORC1 activity by promoting β-hydroxybutyrylation (Kbhb) of RagC at lysine 349 (K348 in mice). Mechanistically, RagC-K349bhb is dynamically catalyzed by p300 and erased by SIRT1, disrupting RagC interaction with Raptor/mTOR and blocking mTORC1 recruitment to lysosomes. Clinically, BHB-mediated RagC-K349bhb suppresses colorectal cancer (CRC) growth via mTORC1 inhibition in both RagC-K348R knockin mice and CRC patient-derived samples. Thus, we identify a BHB sensing mechanism by mTORC1 and highlight the potential role of RagC-K349bhb as a therapeutic target for BHB-based CRC treatment.

Metabolic dysfunction-associated steatohepatitis (MASH) serves as a primary contributor to liver fibrosis, cirrhosis, and hepatocellular carcinoma, yet specific diagnostic markers and therapeutic targets remain unavailable. This study elucidates the molecular mechanism by which UBQLN1 (ubiquilin 1) promotes MASH-related liver fibrosis by regulating small extracellular vesicles (sEVs) secretion and the functionality of the lysosome-mitochondria axis, as well as its clinical significance. Analysis of a multicenter cohort (n = 150) demonstrated significantly elevated UBQLN1 levels in both serum and serum-derived sEV from MASH patients, exhibiting diagnostic accuracies of 0.89 and 0.95, respectively. Furthermore, increased UBQLN1 was observed in mouse models of MASH, hiPSCs-derived human liver organoids, and oleic acid and palmitic acid injured hepatocytes (lipotoxic hepatocytes). Mechanistically, lipotoxic stress induces O-GlcNAcylation at the T277 site of UBQLN1 via OGT (O-GlcNAc transferase), which competitively inhibits its phosphorylation, consequently reducing ubiquitin-mediated degradation. Hepatocyte UBQLN1 facilitates the secretion of sEVs by regulating LAMP1-mediated fusion of multivesicular bodies (MVBs) with lysosomes. Subsequently, sEVs containing UBQLN1 regulate the activation of hepatic stellate cells by degrading the V-ATPase subunit ATP6V1B2 through E54D-dependent ubiquitin ligase activity, thereby inhibiting lysosomal acidification and mitophagy. Moreover, hepatic-specific knockdown of Ubqln1 or hepatocyte-specific knockdown of Ogt significantly alleviates fibrosis and metabolic disorders in MASH mice. This study elucidates the critical role of the post-translational modification regulatory network of UBQLN1 in the progression of MASH and proposes its translational potential as an integrated therapeutic target, providing a theoretical basis for the development of sEV-based intervention strategies.Abbreviations: ATP6V1B2 ATPase H+ transporting V1 subunit B2; Co-IP: co-immunoprecipitation; CCL4: carbon tetrachloride; ELISA: enzyme linked immunosorbent assay; GOT1/AST: glutamic-oxaloacetic transaminase; GPT/ALT: glutamic-pyruvic transaminase; hiPSCs: human induced pluripotent stem cells; HFD: high-fat diet; HFHC: high-fat and high-cholesterol diet; HSCs: hepatic stellate cells; LAMP1: lysosomal associated membrane protein 1; LTH-sEV: lipotoxic hepatocyte-derived small extracellular vesicles; LSECs: liver sinusoidal endothelial cells; MAP1LC3B/LC3: microtubule associated protein 1 light chain 3 beta; MVBs: multivesicular bodies; MASH: metabolic dysfunction-associated steatohepatitis; N-sEV: normal hepatocyte-derived sEV; OGT: O-linked N-acetylglucosamine (GlcNAc) transferase; O-GlcNAc: O-linked-β-D-N-acetylglucosamine; PAOA: oleic acid and palmitic acid; sEV: small extracellular vesicle; UBQLN1: ubiquilin 1.

Bone defect regeneration requires precise spatiotemporal orchestration of inflammatory response, blood vessel formation, and bone formation processes. A major unresolved issue is the fabrication of biomaterial scaffolds that can synergistically combine immunomodulation with angiogenic-osteogenic coupling to hierarchically regulate this regenerative cascade. This study developed a novel biomimetic composite scaffold that structurally and compositionally replicates natural bone by integrating methacrylic anhydride modified gelatin (GelMA) with pearl powder (PP) and dimethyloxalylglycine (DMOG) to enhance bone repair. Fabricated through controlled gradient cooling, ice-templating, and ultraviolet-photocrosslinking techniques, the scaffold exhibited a well-defined interconnected large porous structure with average pore sizes larger than 90 μm. The scaffold demonstrated optimal physicochemical properties, including high porosity (62.2%-81.5%), favorable swelling characteristic (476.4%-737.4%), exceptional mechanical performance, and sustained structural stability (180-day PBS immersion). In vitro assessments revealed the scaffold's superior cell adhesion capacity and significant enhancement of stem cell osteogenic differentiation, attributable to thecombined effects of PP and DMOG. Remarkably, the scaffold effectively modulated macrophage polarization, redirecting lipopolysaccharides (LPS)-activated macrophages toward the pro-regenerative M2 phenotype via suppression of tumor necrosis factor (TNF), mitogen-activated protein kinase (MAPK), and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathways. Furthermore, it substantially promoted endothelial cell migration and robust capillary network formation in tube formation assay. In a rat femoral condyle defect model, the scaffold demonstrated tri-modal therapeutic efficacy: accelerated bone regeneration, significant immune modulation, and enhanced angiogenesis. These multifunctional properties position this engineered scaffold as a clinically translatable solution for bone tissue regeneration.

Our previous studies demonstrated that the fat mass and obesity-associated protein (FTO) is upregulated in colorectal cancer (CRC). It demethylates G6PD/PARP1 and SLC7A11/GPX4 mRNAs, thereby protecting CRC from DNA damage and ferroptotic cell death. However, the mechanisms underlying FTO upregulation in CRC remain unclear. Unexpectedly, we show Ubiquitin-specific peptidase 30 (USP30) binds serine/glycine and senses their levels to protect FTO from proteosome degradation. Stabilized FTO demethylates 3-phosphoglycerate dehydrogenase (PHGDH) and phosphoserine aminotransferase 1 (PSAT1) mRNAs and inhibits their degradation in an m6A-YTHDF2-dependent manner, thereby promoting serine synthesis and CRC tumor growth. Furthermore, we identify sodium 2, 2-dichloroacetate (DCA) as a novel inhibitor of USP30, and DCA inhibits CRC serine synthesis and tumor growth. Clinically, USP30, FTO, PHGDH, and PSAT1 levels are highly correlated in CRC tissues. This study provides mechanistic insights into how USP30 senses serine/glycine levels to regulate serine synthesis via the FTO-PHGDH/PSAT1 axis, offering a potential therapeutic strategy for targeting serine/glycine metabolism in cancer.

eIF6 is overexpressed in multiple cancers. Previous work has showed that deficiency alters the gut microbiota. This study investigated the mechanism linking eIF6 deficiency, microbial dysbiosis, and colorectal cancer (CRC). eIF6 expression was assessed in human and mouse CRC samples. Functional assays were conducted in mice with AOM/DSS-induced CRC. Antibiotic treatment and faecal microbiota transplantation (FMT) were applied to evaluate microbiota-mediated effects. 16S rDNA sequencing and Dubosiella newyorkensis (D. newyorkensis) supplementation were used to identify key bacteria. Metabolites from the bacterial supernatant were analysed via targeted mass spectrometry. The effect of indole-3-carboxaldehyde (3-ICA) was tested in murine models. eIF6 expression was upregulated in CRC. Its deficiency reduced the tumour incidence and proliferation of tumours in mice and increased the abundance of beneficial bacteria such as Akkermansia and Dubosiella. FMT from eIF6 deficient mice and D. newyorkensis administration attenuated tumorigenesis and enhanced barrier function. 3-ICA, a metabolite of D. newyorkensis, also suppressed CRC progression. eIF6 deficiency exerts protective effects against CRC through the enrichment of D. newyorkensis and its metabolite 3-ICA, revealing a novel mechanism and potential therapeutic strategy for CRC.

Primary and metastatic lesions share similar biological properties despite existing in different microenvironments. A better understanding of the mechanisms underlying the maintenance of phenotypic homogeneity in heterogeneous microenvironments could help identify strategies for suppressing metastasis. Here, we found that mechanical memory enabled tumor cells to resist biomechanical stress and sustain malignant traits. Matrix stiffness activated the RhoA-ROCK1 signaling pathway, leading to actin cytoskeletal remodeling and suppression of mitochondrial fission. The dysregulation of mitochondrial dynamics enhanced fatty acid β-oxidation, leading to the accumulation of acetyl-CoA and subsequent histone hyperacetylation. The stiffness-mediated epigenetic reprogramming was mitotically heritable, allowing progeny cells to retain a proliferative advantage after detachment from stiff environments. Inhibition of RhoA-ROCK1 disrupted mechanical memory, thereby reducing metastasis. These findings suggest that targeting mechanical memory may be a strategy for preventing metastasis.

Cholangiocarcinoma (CCA) is a biliary cancer with a poor prognosis and marked chemoresistance. The transcription factor SOX17, which is essential for cholangiocyte differentiation, is frequently downregulated in CCA, as is the case with several tumor suppressor genes. This study provides a proof-of-concept for further development of cellular therapeutic strategies to restore SOX17 expression in CCA cells. For this aim, we used EGI-1 cells as the target cell model derived from extrahepatic CCA. Recombinant Tat-SOX17 protein was first produced in bacteria and purified using Ni-NTA affinity columns and asymmetric flow-field-flow fractionation. Tat-SOX17 entered EGI-1 cells and reached their nucleus. The addition of the α-fetoprotein signal peptide (AFPsp) to the chimeric protein enabled the efficient secretion of the fusion protein formed by AFPsp, SOX17, and reverse Tat (TatR) by donor cells. For the production of secretome enriched in AFPsp-SOX17-TatR protein, HEK293T cells, derived from embryonic kidney, were used. In EGI-1 cells cultured in the presence of this secretome, target gene expression, cell cycle progression, apoptosis, proliferation, colony formation, and cell migration were assessed. Protein expression and localization were analyzed by Western blotting and immunofluorescence. AFPsp-SOX17-TatR enters EGI-1 cells, reaches their nucleus, and modulates the expression of SOX17-dependent genes, such as ABCB1 and ABCG2. Moreover, a marked reduction in proliferation and colony-forming ability was found. In contrast, no significant effect on cell cycle progression, apoptosis, or cell migration was observed. Similar treatment of immortalized human cholangiocytes also increased their SOX17 content, resulting in upregulation of the cholangiolar marker cytokeratin 7 (CK7), but did not affect their proliferation rate. In conclusion, using chimeric proteins such as AFPsp-SOX17-TatR, which contain components for secretion from donor cells and entry into target cancer cells, can provide a promising approach for treating tumors such as CCA, which are characterized by reduced expression of tumor suppressor genes, including SOX17.

Ferroptosis, a form of oxidative cell death, represents a therapeutic vulnerability for treating apoptosis-resistant cancers. Here, we identify leucine zipper transcription factor-like 1 (LZTFL1) as a key regulator of ferroptosis that rewires glutathione (GSH) metabolism. Mechanistically, LZTFL1 promotes oxidation of glucose-6-phosphate dehydrogenase (G6PD), thereby limiting NADPH production and impairing GSH regeneration. GSH depletion in turn enhances LZTFL1 translation via an AKT-mammalian target of rapamycin (mTOR)-eukaryotic initiation factor 4E (eIF4E) pathway, establishing a feedforward loop that amplifies ferroptosis. In vivo, the LZTFL1-formin homology 2 domain-containing 1 (FHOD1)-G6PD axis sensitizes multiple tumor models, including patient-derived xenografts, to ferroptosis, leading to enhanced lipid peroxidation, reduced GSH levels, suppressed tumor growth, and prolonged survival. LZTFL1 expression restores cisplatin sensitivity in resistant lung and ovarian cancer cells and predicts improved survival outcomes in patients with lung adenocarcinoma. Moreover, FDA-approved agents upregulate LZTFL1 and re-sensitize resistant tumors to cisplatin. These findings highlight LZTFL1 as a potential biomarker and a therapeutic target for enhancing ferroptosis-based cancer therapy.

Platinum (Pt)-based anticancer drugs have been a cornerstone of chemotherapy for decades, yet their clinical application remains constrained by dose-limiting systemic toxicity and drug resistance. In this Account, we summarize our systematic efforts to address these challenges through two complementary strategies: 1) functionalization of Pt(IV) prodrugs and 2) spatially controlled targeted delivery. The kinetic inertness and octahedral geometry of Pt(IV) complexes offer a versatile platform for axial functionalization, allowing the integration of diverse bioactive ligands that are released upon intracellular reduction. Exploiting this feature, we have developed multifunctional Pt(IV) prodrugs that co-target DNA damage repair and apoptotic pathways, rewire cholesterol and energy metabolism, induce nonapoptotic cell death including PANoptosis and autophagy-associated death, and epigenetically silence resistance-associated gene networks via chromatin compaction. To engage the tumor immune microenvironment, we have incorporated immunomodulators─including STING agonists, TREM2/CD33 inhibitors, and STAT3 blockers─to amplify innate and adaptive antitumor immunity. Furthermore, we have developed radiotherapy-responsive Pt(IV) prodrugs that undergo rapid, X-ray-triggered reduction mediated by hydrated electrons, enabling spatiotemporally precise drug activation with markedly attenuated systemic toxicity. This strategy is currently advancing toward clinical translation through IND-enabling studies. In parallel, we have established targeted delivery platforms to improve the spatial precision of Pt agents. Mitochondria-targeted complexes redirect cytotoxicity to an organelle lacking efficient DNA repair, disrupting bioenergetics and triggering intrinsic apoptosis. At the tissue level, biotin-mediated targeting exploits overexpressed vitamin transporters for tumor-selective accumulation, while Pt(IV)-antibody conjugates (Pt-ADCs) achieve antigen-specific delivery, upregulate tumor MHC-I expression, expand TCR clonotypes, and synergize with PD-1 blockade. Additionally, a stimuli-responsive in situ self-assembly strategy enables enzyme-triggered nanostructure formation and intracellular disassembly for enhanced tumor accumulation and burst drug release. An immunocompetent patient-derived organoid platform has been established to screen these agents in a clinically relevant setting. The integration of multifunctional modulation, targeted delivery, and externally controlled activation within single Pt-based systems creates a synergistic framework that simultaneously addresses resistance and toxicity. Moving forward, our research will focus on optimizing pharmaceutical properties, advancing radiotherapy-responsive Pt(IV) prodrugs and Pt-ADCs toward clinical evaluation, and refining predictive screening platforms. These programmable Pt therapeutics hold considerable promise for delivering safer and more effective precision chemotherapy to cancer patients.

The cellular response to hypoxia is driven by hypoxia-inducible factors (HIF), which regulate genes involved in glycolysis, angiogenesis, and cell proliferation, as well as inflammation and tumor progression. HIF activation is well characterized and is primarily regulated by oxygen-dependent prolyl hydroxylation and subsequent degradation. SET1B, a histone H3 lysine 4 methyltransferase, has recently emerged as a key modulator of HIF target gene transcription, but evidence suggests that it plays a broader role in modulating HIF transcriptional activity beyond histone methylation. In this study, we revealed that SET1B interacts with RNA polymerase II to coordinate sustained HIF-mediated transcriptional activity through multiple functional domains. In clear-cell renal cell carcinoma (ccRCC), SET1B is critical for sustained HIF activity, and SET1B expression correlated with disease progression and metastasis in patient samples. Moreover, SET1B depletion enhanced the efficacy of HIF2 inhibitors. These findings establish SET1B as a driver of tumor progression and potential therapeutic target in ccRCC. SET1B functions as a key regulator of HIF-dependent transcription and cancer growth under low-oxygen conditions, revealing a therapeutic target to enhance treatment efficacy and potentially slow disease progression in kidney cancer.

Transposable elements (TE), mobile DNA sequences that can move independently about the genome, make up about half of the human genome and are mostly silenced by epigenetic modifications in healthy differentiated cells. Global epigenetic dysregulation during oncogenic transformation causes TE expression, contributing to transcriptional rewiring and TE activity as insertional mutagens. TE nucleic acids are sensed in the cytoplasm, initiating inflammation that can be therapeutically manipulated to activate antitumor immune responses. In this study, we detail how TEs contribute to six of the classic hallmarks of cancer and propose widespread TE expression and activation as a new hallmark of cancer. TEs are expressed at significantly higher levels in cancer cells compared with normal cells as a consequence of epigenetic dysregulation. These elements contribute to transcriptional rewiring, genomic instability, and innate and adaptive immune responses to cancer. Altered TE expression in cancers is a therapeutic vulnerability that scientists are beginning to harness for novel diagnostics and anticancer therapies. See related commentary by Sun and Greenbaum, p. 1489.

Synergistic photodynamic and photothermal therapy (PDT/PTT) holds immense promise for cancer treatment, yet its efficacy is often compromised by limited photosensitizer performance, glutathione (GSH)-mediated ROS quenching, and tumor-protective autophagy. Herein, we developed a GSH-responsive hydrophilic porphyrin copolymer (PPS) that covalently integrates porphyrin into the polymer backbone, ensuring high loading capacity and minimal carrier-related toxicity. PPS exhibits robust absorption around 800 nm, enabling simultaneous PDT and PTT under a single NIR laser. By incorporating the autophagy inhibitor chloroquine (CQ), the resulting CQ@DPPS nanosystem achieves multi-modal synergy: intracellularly, CQ not only inhibits phototherapy-induced autophagic flux to suppress cell survival but also facilitates the lysosomal escape of photosensitizers, heightening programmed cell death via Bcl-2 inhibition and Caspase-3 activation. Crucially, CQ@DPPS significantly activates NK cell-mediated innate immune surveillance and enhances the secretion of granzyme B (GZMB) in NK cells by downregulating MHC-I expression on the surface of tumor cells. In vivo, CQ@DPPS combined with 808 nm irradiation achieved complete tumor regression in HCT116-bearing mice and significantly attenuated hepatic metastasis. This work provides a potent strategy to overcome phototherapy resistance through the integration of autophagy inhibition, dual-mode phototherapy, and innate immune activation. STATEMENT OF SIGNIFICANCE: Clinical translation of PDT/PTT therapy is hindered by inefficient photosensitizer delivery, GSH-mediated ROS depletion, and cytoprotective autophagy in tumors. Here, we report a GSH-responsive hydrophilic porphyrin copolymer with porphyrin covalently integrated into the polymer backbone, enabling high loading, reduced carrier burden, and effective 808 nm-triggered PDT/PTT. Co-delivery of CQ in the CQ@DPPS nanosystem not only suppresses protective autophagic flux, but also promotes lysosomal escape, mitochondrial apoptosis, and NK cell-mediated antitumor immunity through reduced MHC-I expression and increased GZMB secretion. This multi-mechanistic platform achieved complete tumor regression and reduced hepatic metastasis in vivo, offering a promising biomaterial strategy to overcome major barriers in phototherapy and facilitate clinical translation.

Cancer stem cells (CSCs) play a pivotal role in the initiation, metastasis, and recurrence of pancreatic tumors, significantly contributing to drug resistance and reducing the efficacy of traditional chemotherapy. To address these challenges, we developed an amphipathic self-assembled nano-prodrug to co-deliver all-trans retinoic acid (ATRA), gemcitabine (GEM), and ferroptosis inducer (RSL3) for the combined therapy of pancreatic ductal adenocarcinoma (PDAC). The amphipathic prodrug (ATRA-GEM) was synthesized by conjugating ATRA and GEM via an ester bond, enabling good self-assembly and the simultaneous loading of the ferroptosis inducer RSL3. In tumor microenvironment, this combinational nano-prodrug disassembles in response to high level of esterase, triggering the controlled release of ATRA, RSL3, and GEM. The released drugs exhibited a combinational therapeutic effect in vitro by combining apoptosis, ferroptosis, and differentiation therapy. For in vivo application, the nano-prodrug was modified with DSPE-PEG-RGD to enhance tumor-targeting capability and prolong blood circulation. This nano-prodrug demonstrated significant anti-PDAC efficacy and good biosafety in both cell-derived xenograft and patient-derived xenograft models. STATEMENT OF SIGNIFICANCE: Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest cancers due to drug resistance and tumor relapse driven by cancer stem cells (CSCs). Here, we report an amphipathic self-assembled nano-prodrug that simultaneously delivers a differentiation agent (ATRA), a chemotherapeutic (gemcitabine), and a ferroptosis inducer (RSL3). Unlike existing therapies that target only one tumor population, this strategy integrates CSC differentiation, ferroptosis-induced cell death, and chemotherapy into a single platform, enabling coordinated elimination of both CSCs and bulk tumor cells. This nano-prodrug also features high drug-loading capacity and tumor-targeted, stimulus-responsive release. This work provides a new strategy for overcoming resistance and achieving more durable therapeutic outcomes in PDAC, with broad implications for treating other refractory cancers.

Dysregulated Hippo signaling critically promotes malignant proliferation, invasion and distant metastasis across multiple cancers, while precise and coordinated modulation of this core pathway remains a major unmet therapeutic hurdle in clinical cancer treatment. Here, we report for the first time the albumin-based, ultrasound-responsive nanodroplets (FLU-NDs) encapsulating fluvastatin (FLU) to achieve accurate and targeted Hippo pathway reprogramming for efficient tumor suppression. FLU-NDs possess satisfactory drug-loading capability, excellent biocompatibility, and robust contrast-enhanced ultrasound performance. Notably, FLU-NDs achieve efficient tumor accumulation through a dual-delivery strategy that integrates albumin-mediated enrichment with ultrasound-triggered spatiotemporal precise release. Mechanistically, we provide evidence for the first time that ultrasound-targeted microbubble destruction (UTMD) combined with FLU-NDs enables synergistic modulation of Hippo pathway through a reactive oxygen species (ROS)-mammalian sterile 20-like kinase 1 (MST1)-yes-associated protein (YAP) regulatory axis. Specifically, cavitation-induced ROS effectively activates upstream kinase MST1, while ultrasound-triggered release of FLU suppresses the downstream YAP, thereby facilitating coordinated upstream activation and downstream inhibition. Using this previously unrecognized UTMD-associated Hippo modulation strategy, FLU-NDs induce robust apoptosis, reverse EMT, and significantly inhibit tumor growth and metastasis in vitro and in vivo. Collectively, this study establishes a novel, high-efficacy and low-toxicity theranostic platform for precise Hippo-targeted cancer therapy.

Chronic liver disease represents a major global public health challenge, and its malignant progression to hepatocellular carcinoma is the leading cause of death among affected patients. Gut microbiota dysbiosis is a critical driver of this process. As the central hub of the "gut-liver axis," the gut microbiota, when disrupted, compromises the integrity of the intestinal mucosal barrier, promoting the translocation of microbial metabolites, such as lipopolysaccharides and aberrant secondary bile acids, to the liver. In turn, key signaling pathways become activated, including TLR4/NF-κB, Wnt/β-catenin, and PI3K/Akt, sustaining persistent hepatic inflammation and oxidative stress. These pathological processes accelerate the progression from liver fibrosis to cirrhosis, promote genomic instability, and suppress tumor suppressor gene expression, paving the way for the malignant transformation of hepatocytes. Leveraging its holistic regulatory properties, characterized by multi-component, multi-target, and multi-pathway actions, Chinese medicine can intervene at multiple stages of this inflammation-to-cancer cascade by modulating both the structure and function of the gut microbiota. It does so first by enriching beneficial short-chain fatty acid-producing bacteria, such as Lactobacillus and members of the phylum Firmicutes, which helps restore the intestinal mucosal barrier, limit endotoxin translocation, and alleviate hepatic inflammation and fibrosis. In parallel, by normalizing bile acid metabolism and reestablishing gut microbial homeostasis, Chinese medicine counteracts the development of a tumor-permissive microenvironment marked by immune suppression and DNA damage in hepatocytes induced by microbial metabolites. At the same time, it enhances anti-tumor immune responses mediated by CD8+ T cells and other immune effectors. Drawing on evidence from multi-omics analyses and clinical studies, this review examines the core mechanisms and recent advances regarding how Chinese medicine monomers and formulations modulate the gut microbiota to impede the progression of chronic liver disease to HCC. It highlights gut microbiota dysbiosis as a key driver of hepatocarcinogenesis and highlights the therapeutic potential of targeted microbiota regulation by Chinese medicine, providing a conceptual foundation and strategic approaches for the precision prevention and treatment of hepatocellular carcinoma.

Chronic stress has emerged as a critical factor that exacerbates cancer progression, yet its underlying mechanisms and targeted therapeutic strategies remain elusive. Herein, we demonstrate that restraint-induced stress significantly enhances breast tumor growth and proliferation in a murine model. To counteract this effect, we engineered a piezoelectric nanomaterial based on Cu/BaTiO₃ (CBTO) and show that ultrasound activation of CBTO (CBTO-US) effectively suppresses stress-driven tumor growth without adverse effects on body weight or food intake. At the molecular level, transcriptomic profiling (RNA-seq) revealed that restraint stress downregulates key pathways associated with apoptosis, MAPK signaling, and glycolysis, whereas CBTO-US treatment reverses these stress-induced transcriptional changes. Functional analyses demonstrated that CBTO-US markedly reduces lactate accumulation in both tumor tissues in vivo and in E0771 breast cancer cells in vitro under stress conditions. This reduction occurs independently of lactate dehydrogenase activity; consistent with this, Seahorse metabolic flux analysis confirmed that CBTO-US suppresses glycolytic activity upstream of lactate production. Mechanistically, CBTO-US increases the intracellular NAD⁺/NADH ratio and elevates cytosolic Ca²⁺ levels, leading to attenuation of stress- and lactate-induced ERK phosphorylation and consequent inhibition of MAPK signaling. Collectively, these results establish a piezoelectric-catalyzed metabolic regulation strategy that counteracts stress-driven tumor progression, highlighting CBTO-US as a potential therapeutic modality for stress-driven breast cancer. STATEMENT OF SIGNIFICANCE: Chronic stress facilitates breast cancer progression, but effective therapies targeting this link are lacking. Here, we show that ultrasound-activated piezoelectric nanoparticles (Cu/BaTiO₃) suppress stress-driven tumor growth by reprogramming cancer metabolism. These nanoparticles generate electrical signals under ultrasound, reducing lactate production and restoring cellular redox and calcium balance to block pro-cancer MAPK signaling. This work introduces a wireless, non-invasive strategy that converts mechanical energy into anti-cancer metabolic control-offering a new paradigm for treating stress-associated malignancies and broadening the therapeutic potential of piezoelectric nanomaterials.

Artesunate is an antimalarial agent widely recognized for its high efficacy, low toxicity, and favorable tolerability profile. Beyond its primary activity, artesunate has demonstrated promising anti-tumor properties with a well-established safety record. Emerging scientific evidence indicates that its anticancer effects are mediated by multiple mechanisms, including the inhibition of tumor cell proliferation, migration, invasion, and angiogenesis, induction of apoptosis and autophagy in tumor cells, modulation of the tumor microenvironment, regulation of immune responses and circumvention of drug resistance. Collectively, these findings underscore the considerable potential of artesunate as a therapeutic candidate in cancer treatment. In this review, we highlight the effects of artesunate on various types of cancer and explore its mechanisms of action, aiming to suggest potential research directions and support the expansion of its clinical use. In addition, we have compiled the latest in vitro, preclinical and clinical trials and we summarize the positive effects of its combination with other chemotherapies, especially in cases where tumors develop resistance. We also explore strategies to enhance its anticancer efficacy through novel drug delivery methods, and finally, we emphasize recent findings on pharmacology, toxicity, and the emerging issue of artesunate resistance in cancer treatment, and provide a critical perspective.

Despite advances in emerging therapies, colorectal cancer (CRC) incidence continues to rise and clinical outcomes remain suboptimal, highlighting the need for improved management strategies. One potential therapeutic target is osteoprotegerin (OPG), a glycoprotein which exhibits context-dependent functions by interacting with both Receptor Activator of Nuclear Factor Kappa-B Ligand (RANKL) to produce anti-resorption effects and TNF-related apoptosis inducing ligand (TRAIL) to promote cell survival, potentially modulating key hallmarks of CRC progression. However, its translational potential remains underexplored. This review summarizes OPG's structure, expression, and physiological roles, followed by a comprehensive analysis of its mechanistic contributions to CRC progression. We then examine therapeutic strategies targeting OPG and its potential as a diagnostic biomarker, before concluding with perspectives on future research. Present evidence suggests that circulating OPG levels are associated with prognostic factors in CRC such as overall survival and metastasis rates. However, unresolved mechanistic questions remain, particularly regarding dominant cellular sources of OPG and whether circulating OPG levels directly reflect inflammatory and/or tumor dysfunction. Clinically, OPG holds promise as a biomarker for CRC, though it may be more effectively utilized within multi-marker panels rather than as a standalone biomarker. Current research has predominantly focused on the RANKL signaling pathway, leading to development of anti-RANKL therapeutics such as Denosumab, while OPG-specific therapeutics remain underdeveloped. Collectively, future research could investigate whether compartment-specific OPG levels could serve as a biomarker for CRC and predict responses to immunotherapeutics, alongside drug repurposing of existing anti-RANKL therapeutics, and non-pharmacological strategies targeting OPG.

The expanding landscape of regulated cell death (RCD)-encompassing ferroptosis, cuproptosis, pyroptosis, and apoptosis-has revealed a highly interconnected signaling network rather than strictly isolated pathways. A critical missing link in decoding this mechanistic crosstalk is the dysregulation of subcellular ion homeostasis. Here, we systematically review how the spatiotemporal interplay between transition metals (e.g., Fe²⁺, Cu⁺) and classical ions (e.g., Ca²⁺, K⁺, Cl⁻) dictates cell fate. Moving beyond single-ion paradigms, we highlight lipid peroxidation as the primary biochemical convergence hub where metal-catalyzed oxidative stress and channel-mediated organelle dysfunction intersect. Furthermore, we contextualize these ionic derangements within specific pathophysiological microenvironments, particularly focusing on how hypoxia, acidosis, and metabolic exhaustion re-wire ion channel repertoires in cardiovascular ischemia/reperfusion injury and the tumor immune microenvironment. By interrogating the dual role of ion-driven RCD in both tumor cells and anti-tumor immunity, we expose novel therapeutic vulnerabilities. Finally, we outline emerging precision strategies-including multiplexed in vivo biosensors, spatial transcriptomics, and stimuli-responsive nanomedicine-designed to spatiotemporally manipulate ion fluxes. This review provides an actionable conceptual framework for targeting the ion homeostasis-lipid peroxidation axis, ultimately guiding the development of disease-modifying therapies across oncology, cardiology, and neurology.

The clinical prognosis for osteosarcoma (OS) remains bottlenecked by chemoresistance and pulmonary metastasis. OS features dense infiltration by tumor‑associated macrophages (TAMs) that are hijacked into an immunosuppressive, pro-tumorigenic M2-like phenotype. Overcoming this therapeutic plateau requires a paradigm shift toward active remodeling of the tumor immune microenvironment. This review evaluates the trajectory of TAM-targeted interventions in OS, emphasizing the critical transition from monotypic phagocytosis checkpoint blockade (e.g., CD47, GD2) to multimodal synergistic regimens. We systematically dissect how next-generation nanomedicine, targeted metabolic stressors (ferroptosis, cuproptosis), and pharmacological rewiring can forcibly induce immunogenic cell death and reverse M2 polarization. Addressing the unique reconstructive demands of OS, we spotlight the development of immuno-regenerative scaffolds-bifunctional biomaterials engineered to synchronize post-resection tumor clearance with active osteogenesis. Finally, we highlight how spatial transcriptomics and biomimetic platforms are mapping physical immune-exclusion barriers and novel therapeutic subpopulations. Breaking the OS therapeutic stalemate ultimately demands interventions that breach these spatial architectures and fundamentally reprogram TAMs, guided by real-time functional imaging (e.g., ferumoxytol MRI) and high-resolution biomarkers (e.g., PSME2).

Metallothioneins (MTs), a family of cysteine-rich, low-molecular-weight proteins with high metal-binding affinity, are promising candidates for preventing and treating oxidative stress-related diseases. MTs confer protection against diverse pathologies, including diabetes and its complications, aging-related disorders, and cancer. Their mechanisms of action encompass scavenging reactive oxygen and nitrogen species (ROS/RNS), regulating metal ion homeostasis, and suppressing inflammatory responses and apoptosis. The expression of MTs is tightly regulated by multiple cis-acting elements, such as metal, antioxidant, and glucocorticoid response elements, as well as by epigenetic mechanisms. Furthermore, MTs modulate critical signaling pathways-including MAPK, NF-κB, and JAK/STAT-thereby participating in cellular stress adaptation. Recent advances have introduced novel strategies for inducing MT expression, including nanomaterial-based delivery systems and molecular chaperones, thereby enabling targeted therapeutic applications. This review aims to synthesize available evidence, elucidate the central role of MT in oxidative stress-related diseases, and highlight its translational potential as a therapeutic target.

Breast cancer (BC) is estimated to be around 2.3 million new cases and 670 000 deaths in 2020 resulting it into the most frequent cancer globally and ranks fifth among mortalities. Numerous studies have established the link between BC and microRNAs (miRNAs), which are endogenous non-protein-coding master regulators that modulate genetic expressions and influences many physiological and pathological cellular processes. In addition, hypoxia, which also consequently leads to cancer, is a pivotal driver of biological behaviour and malignant traits of cancer cells and eventually tumour cell survival. Hypoxia and hypoxia-inducible factors (HIFs) further regulates the miRNAs expression and promotes invasion and metastasis via increased invasive, migratory, and intravasation properties of the cancer cell, which initiates the metastatic process. In subsequent stages of metastasis, cancer cells navigate immune escape, vascular extravasation, metabolic reprogramming, and tumour-stromal crosstalk at the distant tissue site. Hypoxia dysregulates cancer metabolism post-transcriptionally through miRNAs, promoting glycolysis and fatty acid metabolism by inducing the expression of genes such as GLUT1, SCD, lipidome, and glycolytic enzyme biosynthesis. Hypoxia, on the other hand, also regulates immunogenic reprogramming by circumventing immunosurveillance, perpetuating cell survival and progression via hampering CD-47, PD-L1 markers and T-cell activation. Another process of cancer known as programmed cell death types I and II also involves a complex intricate protein mediated by HIF-1α. Hypoxia-mediated drug resistance influence drug therapeutic effect via altering transporter flux, drug targets, and inactivate of apoptotic pathways. In this review, we are focusing on understanding the clinical implications of hypoxia-driven miRNAs in BC diagnosis and prognosis. Therapeutic landscape for these molecules remains largely untapped, resulting into conducting more research in this field for harbouring potential translational discoveries in the diagnosis, prognosis and therapeutics for BC.

The widespread use of synthetic food additives, such as Tertiary-butylhydroquinone (TBHQ), a common preservative in food and pharmaceuticals, poses health risks, including DNA damage, genotoxicity, and cancer, necessitating sensitive monitoring methods. This study introduces a highly sensitive electrochemical sensor using screen-printed gold electrodes (SPGEs) modified with nanostructured gadolinium orthovanadate (GdVO4). GdVO4, a stable narrow-bandgap semiconductor, enhances sensor performance by promoting electron transfer and increasing the electroactive surface area. Differential pulse voltammetry (DPV) revealed two linear TBHQ detection ranges (2-22.66 μM and 24.88-32.32 μM) with sensitivities of 2.397 μAμM-1 cm-2 and 39.24 μAμM-1 cm-2, respectively. The sensor achieved a low detection limit of 0.19 nM and a quantification limit of 6.43 nM. Reproducibility testing showed a relative standard deviation of 2.05% at 100 μM TBHQ. Real-sample analysis of commercial oil and butter samples confirmed accurate TBHQ detection with high recovery rates, demonstrating the sensor's reliability and suitability for routine food-safety monitoring.

Pancreatic ductal adenocarcinoma (PDAC) exemplifies early systemic dissemination, with circulating tumor cells (CTCs) at its core. We advance a unified conceptual framework that positions CTCs as the systemic execution hub of PDAC metastasis, dynamic entity that coordinates the metastatic cascade via four cardinal functions: Seeding, Adapting, Engineering, and Signaling. Integrating eco-evolutionary dynamics, this hub actively drives phenotypic selection, niche remodeling, and immune evasion, while providing real-time biologic intelligence through liquid biopsy. Robust clinical correlation has not yet translated into routine practice because of technical variability, biological complexity, and a lack of interventional evidence. We therefore propose an evidence-driven, phased roadmap: grounded in prospective clinical cohort data, progressing from immediate multi-center technical standardization and pragmatic trials, such as minimal residual disease (MRD)-triggered salvage therapy, to mid-term biomarker-driven adjuvant trials and long-term integration into multimodal liquid biopsy ecosystems, aimed at intercepting this execution hub. By reframing CTCs from correlative indicators to actionable therapeutic targets and dynamic sentinels, this framework charts a path toward transforming the management of this recalcitrant systemic disease.

Epigenetic dysregulation drives tumor progression and therapeutic resistance, creating a critical need for targeted modulators. Protopanaxadiol (PPD) type ginsenosides, triterpenoid saponins from Panax ginseng, have emerged as promising natural epigenetic regulators, but their clinical translation is limited by poor bioavailability and context dependent activity. Preclinical and early clinical evidence is synthesized here to elucidate the mechanistic underpinnings and translational potential of these agents.Structurally distinct ginsenosides converge on conserved epigenetic enzyme families with tumor specific isoform selectivity: one ginsenoside modulates DNA methyltransferases (DNMTs) across ovarian, renal, and liver cancers; another selectively targets DNMTs and histone deacetylases (HDACs) in colorectal cancer (CRC); a third modulates HDACs and RNA modifiers in leukemia and breast cancer. These interventions reactivate silenced tumor suppressors, suppress epithelial mesenchymal transition (EMT), reverse metabolic reprogramming, and remodel the tumor immune microenvironment. Highly-glycosylated ginsenosides act indirectly via upstream signaling, while low-glycosylated metabolites exhibit superior bioavailability. Key mechanistic principles include family level epigenetic convergence, multi-enzyme co-regulation, and functional complementarity. Clinical translation is hindered by subtherapeutic intratumoral accumulation, lack of biomarker guided studies, and RNA modifier dependency variability. Ginsenosides represent a pharmacologically unique class of epigenetic modulators, with precision oncology potential when paired with delivery optimization and epigenetic biomarker stratification.

Macroautophagy/autophagy is a critical process for maintaining cellular homeostasis and has emerging implications in cancer biology. DRAM2 (DNA damage regulated autophagy modulator 2), a transmembrane protein enriched at lysosomal membranes, has been implicated in autophagy regulation; however, the upstream mechanisms governing its trafficking and function remain unclear. In this study, we identified RPS6KA3/RSK2, a stress-responsive kinase downstream of the MAPK pathway, as a novel upstream kinase of DRAM2. RPS6KA3/RSK2 interacted with and phosphorylated DRAM2 at Ser263 within its cytosolic tail. This phosphorylation was required for AP3D1/AP-3-dependent trafficking of DRAM2 to the late endosomal-lysosomal pathway, thereby facilitating autolysosome formation and sustaining autophagic flux. In contrast, the non-phosphorylatable DRAM2S263A mutant failed to bind AP3D1/AP-3, exhibited defective lysosomal trafficking, and was partially redistributed toward plasma membrane-proximal compartments, where it enhanced exosome secretion. Bioinformatic analyses revealed a strong positive correlation between RPS6KA3/RSK2 and DRAM2 expression in melanoma tissues, and elevated DRAM2 expression was associated with poor patient prognosis. Depletion of RPS6KA3/RSK2 or DRAM2 impaired autophagic flux and inhibited melanoma cell proliferation. Similarly, expression of the DRAM2S263A mutant suppressed melanoma progression in vitro and in vivo by disrupting autophagy. Moreover, DRAM2 protein levels were elevated in skin cancer tissues compared to normal tissues. Collectively, our findings uncover a phosphorylation-dependent trafficking switch that bifurcates DRAM2 function between autophagy and exosome secretion, and establish the RPS6KA3/RSK2-DRAM2 axis as a critical regulator of melanoma progression. This signaling pathway may represent a promising therapeutic target for autophagy-associated malignancies.Abbreviations: AGC: protein kinase A, G, and C families; AP-3: adaptor protein 3; CD: cytosolic domain; CSNK2/CK2: casein kinase 2; co-IP: co-immunoprecipitation; CQ: chloroquine; CREB: cAMP responsive element binding protein; CTKD: C-terminal kinase domain; DRAM2: DNA damage regulated autophagy modulator 2; EBSS: Earle's balanced salt solution; ESCRT: endosomal sorting complexes required for transport; GEPIA: gene expression profiling interactive analysis; GPS: global positioning system; GRK7: G protein-coupled receptor kinase 7; GSK3B: glycogen synthase kinase 3 beta; IP-MS: immunoprecipitation-mass spectrometry; LAMP1: lysosome associated membrane protein 1; LAMP2: lysosome associated membrane protein 2; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MAPK: mitogen-activated protein kinase; MVB: multivesicular body; NTA: nanoparticle tracking analysis; NTKD: N-terminal kinase domain; PI4K2: phosphatidylinositol 4-kinase type 2 alpha; PRKAA2: protein kinase AMP-activated catalytic subunit alpha 2; RPS6KA3/RSK2: ribosomal protein S6 kinase A3; SKCM: skin cutaneous melanoma; SQSTM1: sequestosome 1.

Natural products are biologically active compounds used for therapeutic interventions for various diseases, particularly infections. Autophagy is an intracellular catabolic pathway involving lysosomal degradation and is closely associated with immunological pathways, effectively combating bacterial, viral, fungal, and parasitic infections. Accumulating evidence suggests that autophagy activation or inhibition by natural products promotes antimicrobial responses against various pathogens. Numerous natural products can modulate autophagy through diverse signaling pathways, suggesting their potential as a host-directed therapeutic strategy that may complement conventional drug regimens or help mitigate drug resistance in various infectious diseases. However, it remains largely unclear whether these effects are mediated by direct modulation of autophagy or indirectly through associated mechanisms, including enhanced immune defense, attenuation of pathological inflammation, or crosstalk with other organelle functions. Additionally, multiple pathogens can evade host responses; thus, autophagy activation may inadvertently create favorable conditions for certain pathogens. This review discusses the current knowledge of natural products in terms of their antimicrobial actions through autophagy regulation, particularly the roles of distinct natural product classes, such as polyphenols, alkaloids, terpenoids, quinones, peptides, and macrolides in modulating autophagy for potentially contributing to control various infectious diseases. Exploring the intricate molecular interplay between natural products and autophagy in limiting infections may provide valuable insights that could inform the development of innovative host-directed antimicrobial treatments based on autophagy regulation.Abbreviations: 3-MA: 3-methyladenine; AM: alveolar macrophages; AMP: antimicrobial peptides; AMPK: 5' adenosine monophosphate-activated protein kinase; ARDS: acute respiratory distress syndrome; ART: artemisinin; ASFV: African swine fever virus; ATG: autophagy related; AZM: azithromycin; BafA1: bafilomycin A1; BECN1: beclin 1; BMDM: bone marrow-derived macrophage; BNIP3: BCL2 interacting protein 3; BNIP3L: BCL2 interacting protein 3 like; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CAMKK2: calcium/calmodulin-dependent protein kinase kinase 2; CBD: cannabidiol; CF: cystic fibrosis; CGA: chlorogenic acid; CGAS: cyclic GMP-AMP synthase; CHUK/IKKα: component of inhibitor of nuclear factor kappa B kinase complex; CLP: cecal ligation and puncture; CLR: clarithromycin; CMA: chaperone-mediated autophagy; CoV: coronavirus; DHT: dihydrotanshinone I; EGCG: epigallocatechin-3-gallate; EIF2A: eukaryotic translation initiation factor 2A; EIF2AK2: eukaryotic translation initiation factor 2 alpha kinase 2; ESKAPE: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.; ESRRA: estrogen related receptor alpha; FOXO1: forkhead box O1; FUNDC1: FUN14 domain containing 1; HBV: hepatitis B virus; HCV: hepatitis C virus; HDT: host-directed therapy; HIV: human immunodeficiency virus; HMGB1: high mobility group box 1; HSV: herpes simplex virus; IAV: influenza A virus; ICT: isocryptotanshinone; IFN: interferon; IKBKB/IKKβ: inhibitor of nuclear factor kappa B kinase subunit beta; IL: interleukin; INH: isoniazid; IRF3: IFN regulatory factor 3; KEAP1: kelch like ECH associated protein 1; LAMP: lysosomal associated membrane protein; LAP: LC3-associated phagocytosis; LPS: lipopolysaccharide; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MAPK: mitogen-activated protein kinase; MDM: monocyte-derived macrophage; MDR: multidrug-resistant; MON: monotropein; Mtb: Mycobacterium tuberculosis; MTOR: mechanistic target of rapamycin kinase; mtROS: mitochondrial ROS; NET: neutrophil extracellular trap; NFE2L2/Nrf2: NFE2 like bZIP transcription factor 2; NFKB/NF-κB: nuclear factor kappa B; NLRP3: NLR family pyrin domain containing 3; NLRX1: NLR family member X1; NOTCH1: notch receptor 1; NTM: nontuberculous mycobacteria; OMS: ohmyungsamycin; PAK1: p21 (RAC1) activated kinase 1; PINK1: PTEN induced kinase 1; PKM/PKM2: pyruvate kinase M1/2; PLD: phospholipase D; PM: peritoneal macrophage; PPM1A: protein phosphatase, Mg2+/Mn2+ dependent 1A; PRKN/parkin: parkin RBR E3 ubiquitin protein ligase; PtdIns3K: phosphatidylinositol 3-kinase; PtdIns3P: phosphatidylinositol-3-phosphate; PTEN: phosphatase and tensin homolog; RB1CC1/FIP200: RB1 inducible coiled-coil 1; RELA/p65: RELA proto-oncogene, NF-kB subunit; RIF: rifampicin; ROS: reactive oxygen species; RSV: resveratrol; RUBCN/rubicon: rubicon autophagy regulator; SAR: selective autophagy receptor; SIRT: sirtuin; STING1: stimulator of interferon response cGAMP interactor 1; STX17: syntaxin 17; Tat: trans-activator of transcription; TB: tuberculosis; TBK1: TANK binding kinase 1; TFEB: transcription factor EB; TLR: toll like receptor; TNA: tanshinone IIA; TNF: tumor necrosis factor; UA: ursolic acid; ULK1/Atg1: unc-51 like autophagy activating kinase 1; UPR: unfolded protein response; UVRAG: UV radiation resistance associated; VAMP8: vesicle associated membrane protein 8; VDR: vitamin D receptor; WIPI2: WD repeat domain, phosphoinositide interacting 2; ZFYVE1/DFCP1: zinc finger FYVE-type containing 1; ZIKV: Zika virus.

Iron is vital for life but can be toxic in excess by forming reactive oxygen species. Ferroptosis, a type of regulated cell death, relies on iron-dependent lipid peroxidation and requires a labile iron pool (LIP) in cells. Ferritin stores iron safely, and its degradation increases the LIP. Ferritinophagy, the autophagic breakdown of ferritin, is crucial for releasing stored iron to trigger ferroptosis. This review examines ferritinophagy's molecular mechanisms, highlighting NCOA4 (nuclear receptor coactivator 4) as the main receptor targeting ferritin for lysosomal degradation. It also discusses the regulatory network controlling NCOA4, including transcriptional factors like TP53/p53 and MYC/c-Myc, RNA-binding proteins, and post-translational modifications such as ubiquitination. We explore ferritinophagy-induced ferroptosis as a promising anti-cancer approach. Research shows that various natural compounds, repurposed drugs, and new metal complexes can induce tumor cell death by activating the NCOA4-ferritinophagy pathway, which is crucial for overcoming therapeutic resistance in many cancers. Understanding this pathway highlights the relationship between iron metabolism, macroautophagy/autophagy, and cell death, offering a foundation for new treatments for cancer and iron-related diseases.Abbreviation: FTH1: ferritin heavy chain 1; GPX4: glutathione peroxidase 4; GSH: glutathione; HIF: hypoxia-inducible factor; LIP: labile iron pool; MAPK/JNK: mitogen-activated protein kinase; NCOA4: nuclear receptor coactivator 4; PUFAs: polyunsaturated fatty acids; SLC7A11: solute carrier family 7 member 11; TFRC: transferrin receptor; TFEB: transcription factor EB.

Colorectal cancer (CRC) is a leading cause of cancer-related mortality, often diagnosed at advanced stages when treatment options are limited. Histone deacetylases (HDACs) are frequently overexpressed in CRC, promoting tumor cell proliferation and survival through epigenetic regulation. In the current study, we identified piroctone olamine (PO), an antifungal agent, as a novel inhibitor of CRC cell proliferation which simultaneously activates autophagy to promote CRC cell death with minimal cytotoxicity to normal cells. Inhibition of autophagy completely abolished PO-induced cell death, confirming that autophagy is the primary pathway for cell death. Transcriptomic analysis revealed that PO significantly upregulated the expression of beclin-1 (BECN1), an essential autophagy-related gene. Structural analysis indicated that PO contains an oxime group, structurally similar to hydroxamate-based HDAC inhibitors. We also found that PO specifically targeted and inhibited HDAC6, leading to increased acetylation of histone H3 at lysine 9 (H3K9ac). The increase in H3K9ac promoted the binding of transcription factors at the BECN1 promoter, enhancing BECN1 transcription, as confirmed by ChIP-qPCR and dual-luciferase assays. In vivo, PO effectively reduced HDAC6 expression, elevated BECN1 levels, promoted autophagy, suppressed tumor growth in a syngeneic mouse CRC model. In conclusion, PO selectively inhibits HDAC6, promotes histone acetylation at H3K9, upregulates BECN1 expression, and induces autophagy-dependent cell death in CRC cells. These findings position PO as a promising candidate for the development of HDAC6-targeted therapies in CRC.

Biliary tract cancers (BTCs) are a group of rare but highly lethal malignancies that include intrahepatic cholangiocarcinoma (iCCA), extrahepatic cholangiocarcinoma (eCCA), and gallbladder carcinoma (GBC). A major challenge in modeling and treating these cancers is their highly heterogeneous mutational landscapes, both within and across subtypes, as well as across geographic and etiological contexts. This heterogeneity necessitates large cohorts to enable robust statistical analyses. Here, we collate and analyze data from more than 30 next-generation sequencing studies to provide a comprehensive overview of BTC mutational patterns and their potential clinical implications. Mutation data from 5,123 BTC samples from 13 countries were standardized, focusing on a set of 29 bona fide oncogenes and tumor suppressor genes. We performed statistical analyses to evaluate hypotheses related to mutation prevalence, etiology, co-mutation patterns, and recurrent mutations. This analysis provides robust estimates of mutation prevalence across geographic regions, BTC subtypes, and hepatitis status, highlighting genes whose alteration frequencies vary by anatomical location and etiology. or example, iCCA in Eastern vs. Western hemispheres showed large differences in alteration prevalence for FGFR2, IDH1, KRAS, TP53, CDKN2A, and BAP1, whereas eCCA and GBC exhibited only modest hemispheric differences. We further show that these differences, as well as those observed between iCCA and eCCA, are partly explained by variation in the mutation profiles and prevalences of small- vs. large-duct subtypes. The large sample size also enabled systematic characterization of gene-gene co-occurrence and mutual exclusivity across BTC subtypes, along with a clinically oriented catalogue of recurrent mutations in oncogenes and tumor suppressor genes. This integrative cross-study analysis provides a global view of BTC genomics, clarifying how geography, etiology, and anatomical subtype collectively shape driver mutation landscapes. Our results provide statistical depth, uncover new mutational relationships, and suggest high-priority avenues for basic and translational research in biliary tract cancer. By integrating data across diverse geographic and etiological contexts, this study highlights how mutation prevalence and co-mutation patterns vary systematically with anatomical subtype, hepatitis status, and region. These findings offer a more nuanced framework for interpreting BTC genomics and underscore the importance of stratified approaches to research and treatment. Collectively, this work establishes a robust foundation for future biological studies, biomarker development, and the design of clinical trials that are tailored to geographic origin, underlying etiology, and specific mutational and co-mutation landscapes, ultimately supporting more precise and effective therapeutic strategies in BTC.

Cancer stem cells (CSCs) are a critical subpopulation within heterogeneous tumors which are largely responsible for therapeutic resistance, recurrence and metastasis. Herein, we report an ultrasound-mediated strategy based on a sono-responsive core-shell nanoplatform (MTCA) that concurrently disrupts physical barriers and reprograms intracellular homeostasis to precisely regulate and eradicate CSCs. The MTCA was engineered and surface-modified with a carbonic anhydrase IX (CAIX) inhibitor to enable selective targeting of CAIX-overexpressing CSCs and perturb their pH homeostasis. The copper-titanium dioxide shell depletes intracellular glutathione and amplifies reactive oxygen species generation under ultrasound activation, thereby overcoming CSC-associated antioxidant defenses. Meanwhile, ultrasound-induced mechanical effect transiently disrupts extracellular integrity, significantly enhancing nanoplatform penetration into CSC-enriched tumor region. This synergistic approach was found to effectively downregulate stemness-associated gene expression and suppresses CSC viability in CSC-enriched 3D tumors spheroids, which also inhibited tumor growth and metastasis in in vivo tumor models. Collectively, this study establishes an ultrasound-enabled paradigm for coordinated physical and biochemical modulation of CSC niches, offering a promising strategy to overcome CSC-driven therapeutic resistance.

SMAC is a mitochondrial intermembrane space protein, which is released during apoptosis and whose known function is antagonism of inhibitor of apoptosis proteins in the cytosol, to facilitate caspase activation. Recent data suggest that SMAC can also be released by sub-lethal signals in the apoptosis pathway, in the absence of cell death. We here explored potential functions of SMAC in non-apoptotic cells. We found that a portion of SMAC is spontaneously released into the cytosol in the absence of apoptosis, regulated by the BCL-2-family proteins BAX and BAK and the fission GTPase DRP1. In cancer cell lines, SMAC was required for the activation of caspases in lethal and non-lethal conditions, while this contribution to caspase-activation was much smaller in non-malignant fibroblast lines. In cells with high levels of cytosolic SMAC, SMAC deficiency reduced in vitro migration, invasion and anchorage-independent growth as well as metastasis in a xenograft model in zebrafish. SMAC-deficient cells further showed a reduced activity in interferon signaling, associated with reduced cytosolic presence of mitochondrial DNA and activation of the stimulator of interferon genes (STING), and SMAC expression levels correlated with interferon-induced genes in cancer data sets. We further found that SMAC can regulate mitochondrial morphology and integrity. Finally, high gene-expression of SMAC was associated with poor prognosis in patients of several cancer types. These results identify SMAC as a regulator of inflammation and growth behavior of cancer cells. They further report a mitochondrial function of SMAC and demonstrate a role of SMAC in human cancer biology across several cancer entities.

Platinum resistance in breast cancer remains a major therapeutic challenge due to convergent escape mechanisms. Here, we identify the PRMT6/USP7 complex as a dual epigenetic orchestrator of chemoresistance. Mechanistically, USP7-mediated deubiquitination of H2BK120ub facilitates PRMT6-dependent deposition of the repressive mark H3R2me2a, concomitant with the exclusion of activating H3K4me3. This coordinated histone crosstalk reprograms the chromatin landscape, leading to transcriptional silencing of the ferroptosis regulator TAZ (WWTR1). Consequently, TAZ suppression disrupts lipid peroxidation and blocks cisplatin-induced ferroptosis, a critical cell death pathway. Simultaneously, the PRMT6/USP7 complex recruits the E3 ubiquitin ligase RNF168 to sites of DNA damage, promoting H2AX monoubiquitination and robust activation of both homologous recombination (HR) and non-homologous end-joining (NHEJ) repair pathways, thereby enhancing DNA damage tolerance. To translate this mechanism, we engineered an injectable hydrogel for the sequential co-delivery of cisplatin and PRMT6/USP7 inhibitors, demonstrating significantly enhanced therapeutic efficacy. Our study unveils a previously unrecognized bifunctional role for the PRMT6-USP7 axis in orchestrating epigenetic reprogramming and DNA repair to confer platinum resistance, providing profound mechanistic insights and a compelling co-targeting strategy for overcoming chemoresistance in breast cancer.

Both alpha and beta cells are dysfunctional in type 1 diabetes (T1D), but beta cells die while alpha cells survive the immune attack. Understanding the mechanisms underlying alpha-cell resistance could identify new approaches to protect beta cells. Herein, we analysed single-cell datasets from human alpha and beta cells under basal/unstimulated conditions and under immune-mediated stress. Alpha cells exhibit enhanced immune-like gene expression compared to beta cells. We also found that the tumor suppressor Maternally Expressed Gene 3 (MEG3), a T1D risk gene, is highly expressed in beta cells while almost undetectable in alpha cells. These observations were confirmed by analysing bulk RNA-sequencing data from fluorescence-activated cell-sorted alpha and beta cells isolated from primary human islets from non-diabetic donors. Additionally, MEG3 knockdown in human insulin-producing EndoC-βH1 cells and human islets microtissues decreased cytokine-induced damage and apoptosis, preserving beta-cell function under inflammatory conditions. The fact that alpha cells exhibit increased immune-like and anti-apoptotic activity as compared to beta cells suggests that they are better equipped to endure the autoimmune assault in T1D. In addition, the marked difference in the expression of the pro-apoptotic factor MEG3 in beta cells compared to alpha cells may explain, at least in part, why beta cells are more susceptible to damage and cell death in a diabetogenic environment than neighbor alpha cells within the same islet.

The quiescence-activation transition of cancer stem cells regulated by environmental stimuli has been shown to potentially contribute to cancer maintenance and regrowth after therapy. However, little is known about how the neoplastic niche couples with neighboring signals to control the activation of quiescent rectal cancer stem cells during radiotherapy. Lineage-tracing experiments were performed usingHopxCreERT2;RosatdTomato mice and organoids to visualize the dynamics and radioresistance ofHopx-expressing cells in vivo. BrdU pulse-chase assays and cell cycle analysis were used to identify whether Hopx+ stem cells were label-retaining cells (LRCs). The paracrine pro-survival effect of radiotherapy-induced dying cancer cells on neighboring Hopx+ quiescent stem cells was analyzed in the context of both apoptosis and necroptosis blockade. Human rectal cancer organoids and patient-derived xenografts (PDXs) were used to assess the radiotherapy-enhanced efficacy of Hopx targeting. Lineage tracing experiments revealed that Hopx+ quiescent stem cell subpopulation exhibited an enhanced regeneration, which functionally drove the recurrence of rectal cancer after irradiation. Mechanistically, iron released from radiotherapy-induced tumor cell death triggered a Stat3-dependent pro-survival program in neighbor-surviving Hopx+ quiescent stem cells. Interestingly, we demonstrated activated Hopx+ cancer stem cells antagonized ferroptosis that should be caused by iron-overload via the inhibition of de novo lipid synthesis. Collectively, quiescent cancer stem cells could establish a new dependency on anti-apoptotic programs in their dying neighbors. This study highlights targeting and regulating Hopx+ quiescent stem cells could be a promising therapeutic approach to overcome the refractoriness of human rectal cancer.

Depression, chronic stress, fear, and anxiety have been implicated as major risk factors in tumor growth, invasion, and metastasis. These psychological conditions stimulate the secretion of neurotransmitters and stress hormones through prolonged activation of the fight-or-flight response. The arousal of stress-related systems, such as the hypothalamic-pituitary-adrenocortical axis and the sympathetic nervous system, induces physiological alterations. Consequently, stress-associated mediators suppress antitumor immune responses, enhance the release of inflammatory cytokines, and promote tumor cell migration and survival within an altered tumor microenvironment through diverse signaling pathways. This review highlights recent advances in understanding how depression and chronic stress regulate tumor growth, angiogenesis, invasion, and metastasis. We further discuss how stress response systems remodel the tumor microenvironment to promote cancer progression, providing insights into the interface between behavioral and biological mechanisms and identifying potential therapeutic targets. We emphasize the rationale and importance of developing pharmacological interventions to inhibit tumor progression and enhance the efficacy of conventional therapies and immunotherapies in patients with cancer-associated depression. Moreover, we discuss emerging pharmacological strategies with the potential to prevent stress-related cancer progression.

Acetate is the metabolic precursor of acetyl-coenzyme A (CoA), fuelling histone acetylation. We aimed to investigate whether the acetate-acetylation axis is hijacked in tumour endothelial cells (TECs) to govern hepatocellular carcinoma (HCC) progression. The endothelial acetate-acetylation axis and its impact on malignant phenotypes and anticancer therapy were systematically dissected using clinical specimens, primary endothelial cells (ECs) from HCC (tumour endothelial cells, TECs) or non-tumour liver tissues (non-tumour endothelial cells, NECs), EC lines and diverse mouse models. Compared with NECs, TECs showed elevation of acetate transporter (monocarboxylate transporter 1, MCT1), metabolic enzyme ACSS2 and H3K27ac. Acetate was highly enriched within tumour and surrounding parenchyma, and correlated positively with tumour angiogenesis. Functionally, acetate or hepatoma-conditioned media increased endothelial H3K27ac and EC migration, which were attenuated by inhibiting MCT1 or ACSS2. Notably, acetate-treated ECs, but not acetate alone, drived CD8+ T cell exhaustion and regulatory T cell (Treg) expansion. In mouse hepatoma allograft models, acetate administration increased H3K27ac levels in TECs, driving angiogenesis, tumour growth and metastasis, while reducing CD8+ T cells and expanding Tregs. Mechanistically, acetate orchestrated pro-angiogenic and immunosuppressive transcriptional programmes in ECs via histone acetylation. Therapeutically, pharmacological ACSS2 inhibition, EC-targeting simACSS2-liposomes and adeno-associated virus (AAV)-TIE2-shmACSS2 all decreased H3K27ac levels in TECs, inhibited angiogenesis, increased CD8+ T cells and reduced Tregs. Crucially, ACSS2 inhibition synergised with anti-programmed cell death protein 1 (PD-1) to alleviate immunosuppression, curb angiogenesis and suppress tumour progression. Hepatic acetate accumulation and concomitant MCT1/ACSS2 upregulation in TECs drives endothelial epigenetic remodelling, thus fuelling angiogenesis, immunosuppression and HCC progression. Targeting this metabolic-epigenetic axis represents a novel approach to potentiate HCC therapy and sensitise immunotherapy.

Fanconi anemia (FA) is characterized by defective DNA repair and chronic p53 activation, predisposing to acute myeloid leukemia through persistent genomic instability. The molecular adaptations enabling cell survival under chronic stress remain poorly understood. This study investigates the role of ZMAT3, a p53 responsive RNA binding protein overexpressed in FA deficient cells, to elucidate its role in cellular adaptation. Using FA patient fibroblasts and Fancg KO mouse models, we launched transcriptomic profiling, lipidomic, and functional analyses. RNA-seq and lipidomic analyses revealed unexpected alterations in sphingolipid metabolism upon ZMAT3 depletion, linked to ceramide accumulation and downregulation of ASAH1 (acid ceramidase). Functionally, ZMAT3 depletion increased DNA damage and enhanced ferroptosis susceptibility, an iron dependent cell death mechanism. Mechanistically, our results showed that ZMAT3 promotes ASAH1 expression, limits ceramide accumulation and protects cells from ferroptosis induced death which could participate to preleukemic clonal evolution and pointing to areas for further exploration in FA-associated malignancies.

SWI/SNF chromatin remodelers are represented by three biochemically distinct subcomplexes, the abundant cBAF and the less abundant PBAF and GBAF. Genetics have identified important roles for PBAF in development and disease; however, relating PBAF-mediated phenotypes to biochemical function in chromatin regulation and gene activation has been challenging. Here, we show that the PBRM1 subunit of PBAF is critical for the completion of TGFβ1-mediated epithelial-mesenchymal transition (EMT) of mammary cells in vitro as well as the metastasis of murine breast cancers in vivo. Using epigenomics to profile different stages of EMT, we find that PBRM1 is necessary for targeting PBAF to inducible promoters marked by H3K14ac. We further find that PBRM1 facilitates DNA accessibility at sites bound by TGFβ1-inducible transcription factors, such as Atf3, for the induction of genes involved in migration, cell survival, and inflammation, providing evidence that PBAF is a vulnerability in late-stage metastatic cancers.

Extrachromosomal DNA (ecDNA), as a dynamic genetic vector, enables tumor cells to exhibit enhanced adaptability under chemotherapeutic stress. However, its functional role in tumor malignancy and the development of drug resistance in gastric cancer (GC) remains unclear. This study aims to investigate the relationship between chemotherapy and ecDNA in GC, and explore its potential as a novel therapeutic target. We first analyzed our clinical data to reveal associations between ecDNA induction by chemotherapy and metastatic progression. Then we investigated the role of ecDNA in mediating chemoresistance and malignant phenotypes in cisplatin-resistant cell models using WGS, SEM, western blot, qPCR, flow cytometry, and immunofluorescence. We found that neoadjuvant chemotherapy (NAC) promoted ecDNA formation from linear amplicons, which was associated with increased tumor mutational burden, poorer survival, and higher distant metastasis risk. The generation of ecDNA in GC was more likely to originate from chromosomes 5, 7, 8, 9, and 12. In cisplatin-resistant models, ecDNA emergence promoted various malignant phenotypes, including DNA damage repair, G1 arrest, autophagy, proliferation, migration, and invasion, which were attenuated by ecDNA inhibition. EcDNA inhibitors delayed acquired resistance in wild-type cells and showed combinatorial activity with cisplatin in resistant GC, supporting clinical potential. Our study revealed that cisplatin could promote ecDNA generation, which enhanced malignant phenotypes, including cell proliferation, motility and drug resistance, and was associated with poorer patient prognosis. Targeting ecDNA with inhibitors could reverse these effects, supporting further preclinical evaluation.

Triple-negative breast cancer (TNBC) is a clinically aggressive and molecularly heterogeneous subtype defined by the absence of estrogen receptor, progesterone receptor, and HER2 expression. Chemotherapy remains the mainstay of treatment, but recent advances in immune checkpoint inhibitors, DNA damage response-targeted therapies, antibody-drug conjugates, and small-molecule inhibitors have expanded therapeutic options. Despite these developments, clinical outcomes are still limited by tumor heterogeneity, suboptimal biomarker selection, resistance, and treatment-related toxicity. In this review, we summarize current and emerging therapeutic strategies for TNBC with a focus on their underlying biological rationale. We highlight key approaches targeting DNA repair deficiency, immune regulation, oncogenic signaling pathways, and antigen-directed therapies, and discuss their clinical development and application. We also address the role of molecular subtyping and biomarkers in guiding treatment selection, as well as the challenges posed by resistance and limited durability of response. Overall, this review provides an updated overview of TNBC treatment strategies and discusses ongoing challenges and future directions toward more effective and individualized therapies.

The emergence of resistance to CDK4/6 inhibitors (CDK4/6i) is a major barrier to long-term survival in metastatic hormone receptor-positive breast cancer. Mechanisms of CDK4/6i resistance are highly diverse and are currently unpredictable. In a recent issue of Nature, Safonov and colleagues report that germline BRCA2 (gBRCA2) alterations predispose tumors toward loss-of-function alterations in RB1, a key mechanism of tumor escape from CDK4/6is. Leveraging large-scale clinical genomics, the authors show that gBRCA2-altered tumors are enriched for RB1 alterations and derive less benefit from CDK4/6i-based therapy while retaining sensitivity to PARP inhibition. Mechanistically, they demonstrate that the shared location of BRCA2 and RB1 on chromosome 13q leads to RB1 hemizygosity in gBRCA2 tumors and that homologous recombination deficiency-associated mutagenesis facilitates acquisition of a second inactivating hit. Furthermore, baseline RB1 hemizygosity predicts inferior outcomes on CDK4/6is independent of germline status, supporting its role as a predictive biomarker. These findings have immediate clinical implications with regard to the sequencing of PARP inhibitors and CDK4/6is in patients with gBRCA2 mutations and highlight a potential opportunity to anticipate and intercept resistance, leading to more durable clinical benefit.

Black women have twice the endometrial cancer mortality of White women, and studies suggest that differences in ultrasound diagnostic accuracy partially contribute to this disparity. To evaluate racial and ethnic differences in the diagnostic performance of transvaginal ultrasound for detecting endometrial neoplasia. A prospective cohort study of 1833 women aged 50 years or older at risk of endometrial neoplasia, conducted at a large urban academic medical center from February 2014 to August 2022, with follow-up through March 2023. The statistical analyses were conducted from March 2024 to December 2025. Women underwent endometrial assessment via transvaginal ultrasound followed by sonohysterogram-directed biopsy and follow-up observation to establish final outcome. Assessment of transvaginal ultrasound diagnostic performance for detecting endometrial cancer and hyperplasia across racial and ethnic groups, measured by: (1) completion rate (endometrial accessibility) and (2) accuracy of completed scans. To assess any association with fibroids, analyses were repeated after excluding women with fibroids. Subgroup analyses were conducted among women with endometrial cancer, those with postmenopausal bleeding, and those without prior exposure to estrogen or tamoxifen. A total of 1833 women met inclusion criteria (mean [SD] age, 60.3 [8.1] years). Most were postmenopausal (1218 women [87.4%]), and 832 (45.4%) were Black, 705 (38.5%) were Hispanic, and 253 (13.8%) were White. Black women had significantly lower completion rates (adequate endometrial visibility) for transvaginal ultrasound compared with White women (75.7% vs 88.9%; relative risk, 0.85; 95% CI, 0.80-0.90; P < .001). Accuracy of completed transvaginal ultrasound was slightly lower among Black women compared with White women (sensitivity, 96.3%; 95% CI, 91.3%-100% vs 100%; negative predictive value, 97.9%; 95% CI, 95.0%-100% vs 100%). After excluding women with fibroids, differences in completion and accuracy between Black and White women disappeared. Similar findings were observed in subgroup analyses of women with a final diagnosis of endometrial cancer, those presenting with postmenopausal bleeding, and those without prior exposure to estrogen or tamoxifen. In this cohort study of a diverse population of women, ultrasound completion rates were lower among Black women, primarily due to fibroids impairing endometrial visibility. Nevertheless, transvaginal sonography demonstrated a 76% completion rate and retained excellent ability to exclude cancer in Black women, regardless of fibroid presence.

Background Foundation models show promise in medical imaging but remain underexplored in three-dimensional modalities. Despite the adoption of digital breast tomosynthesis (DBT) in breast cancer screening, no dedicated foundation model currently exists for this modality. Purpose To develop and evaluate a foundation model for DBT (DBT-DINO) and assess the impact of domain-specific pretraining across multiple clinical tasks. Materials and Methods This retrospective study used DBT images from Mass General Brigham acquired between March 2011 and February 2024. Self-supervised pretraining was performed using Meta AI's DINOv2 methodology on more than 25 million two-dimensional sections from 487 975 DBT volumes from 27 990 patients. Three downstream tasks were evaluated: (a) breast density classification using 5000 screening examinations, (b) 5-year risk of developing biopsy-proven breast cancer using 106 417 screening examinations, and (c) lesion detection using 393 annotated volumes. The performance of DBT-DINO was compared with that of ImageNet-pretrained DINOv2 baselines using McNemar and DeLong tests. Results A total of 4981 patients (mean age, 57.76 years ± 11.40 [SD]; 4855 female) were included for density classification, 31 561 patients (mean age, 60.09 years ± 10.47; 31 559 female) were included for risk prediction, and 199 female patients were included for lesion detection. For breast density classification, DBT-DINO achieved 79% (786 of 997 examinations) accuracy, outperforming the DINOv2 baseline (73% [728 of 997 examinations]; P < .001). For 5-year breast cancer risk prediction, DBT-DINO had an area under the receiver operating characteristic curve (AUC) of 0.78 and DINOv2 had an AUC of 0.76 (P = .057), showing no evidence of a difference. In lesion detection, DINOv2 had an average sensitivity of 67% (91 of 136 lesions), whereas DBT-DINO had a sensitivity of 62% (84 of 136 lesions) (P = .60), again with no evidence of a difference. Conclusion DBT-DINO demonstrated strong performance in breast density classification; however, there was no evidence of a difference compared with the ImageNet baseline in 5-year breast cancer risk prediction or lesion detection, suggesting that domain-specific pretraining for localized detection tasks required further refinement. © RSNA, 2026 Supplemental material is available for this article. See also the editorial by Wu in this issue.

Long-acting granulocyte colony-stimulating factors (G-CSFs) are the standard of care for primary prophylaxis against chemotherapy-induced neutropenia in breast cancer, but comparative data remain limited. This prospective observational cohort study was conducted at a single center in Shanghai, China. Patients with breast cancer scheduled for high-febrile-neutropenia-risk chemotherapy and planned for long-acting G-CSF primary prophylaxis were eligible. Patients received subcutaneous pegfilgrastim, mecapegfilgrastim, telpegfilgrastim, or efbemalenograstim alfa once per cycle per clinical practice. The primary outcome was the incidence of grade 3/4 neutropenia (leukopenia), assessed during chemotherapy cycles and at follow-up visits. Safety was assessed via bone pain-related treatment-emergent adverse events (BPR TEAEs). Between March 25 and September 30, 2025, 407 patients were analyzed. Grade 3/4 neutropenia (leukopenia) occurred in 24.1%, varying numerically across agents (telpegfilgrastim 30.2%, mecapegfilgrastim 16.4%; overall P = 0.061). After multivariable adjustment, telpegfilgrastim was associated with a higher risk of grade 3/4 neutropenia (leukopenia) versus mecapegfilgrastim (OR = 2.46, P = 0.011). BPR TEAEs occurred in 62.4%, with similar incidence across groups (P = 0.32). Lower BPR TEAE risk was associated with ddEC chemotherapy (OR = 0.32, P = 0.0010) and postmenopausal status (OR = 0.53, P = 0.031). In exploratory analyses, administration at 24-48 h (versus <24 h) was associated with lower grade 4 neutropenia (leukopenia) (9.9% versus 27.3%, P = 0.011) and severe BPR TEAEs (0.5% versus 4.5%, P = 0.032), and a 3 mg pegfilgrastim dose was associated with comparable myeloprotection to 6 mg (29.4% versus 26.5%, P = 0.81) but fewer BPR TEAEs (29.4% versus 60.3%, P = 0.022). All four long-acting G-CSFs demonstrated clinical efficacy. Numerical variations and adjusted analyses indicated distinct clinical profiles, supporting tailored prophylaxis. Optimizing administration timing and considering dose reduction could improve benefit-risk balance. Larger prospective studies are needed to confirm these exploratory findings. None.

Postmastectomy radiation therapy (PMRT) improves locoregional control and survival in patients with early breast cancer, but it is unclear whether these benefits extend to human epidermal growth factor receptor 2 (HER2)-positive patients in the anti-HER2 era. This was a retrospective cohort analysis of prospectively collected data from the phase III ALTTO trial and includes all ALTTO participants who underwent mastectomy regardless of randomisation arm. The phase III ALTTO trial enrolled patients between 2007 and 2011 from 946 centers from 44 countries in four different continents. Eligible patients had histologically confirmed HER2-positive breast cancer with either node-positive disease or node-negative disease with pathologic tumour size ≥1 cm. PMRT was non-randomised and delivered at the investigator's discretion. Primary efficacy analyses were conducted in the intention-to-treat population; safety analyses included all patients who received at least 1 dose of anti-HER2 therapy. Endpoints were locoregional recurrence (LRR), distant recurrence (TTDR), disease-free (DFS), and overall survival (OS). DFS and OS were analysed using Cox models; LRR and TTDR competing risk models. All models included the interaction between number of metastatic nodes, 0 (pN0), 1-3 (pN1) or ≥4 (pN2) and PMRT. Multivariable models accounted for patient and tumour characteristics. Hazard ratios (HR) and their 95% confidence interval (CI) are reported. Overall, 4154 patients treated with mastectomy were analysed: 1987 (47.8%) with and 2167 (52.2%) without PMRT. Baseline characteristics were worse in the PMRT group, whereas adjuvant endocrine and chemotherapy use/type was similar between groups. At a median follow-up of 9.2 years (IQR, 6.4-10), 129 LRR events occurred: 43 (2.2%) in the PMRT group and 86 (4.0%) in the no-PMRT group. In patients with pN2, PMRT was associated with improved LRR control (HR 0.31; 95% CI 0.16-0.59), TTDR (HR 0.59; 95% CI 0.44-0.79), DFS (HR 0.58; 95% CI 0.45-0.75), and OS (HR 0.61; 95% CI 0.44-0.86). In patients with pN1, PMRT showed marginal benefit in DFS (HR 0.82; 95% CI 0.64-1.05) but not in LRR (HR not calculated), TTDR (HR 0.89; 95% CI 0.65-1.23), nor OS (HR 0.88; 95% CI 0.62-1.24). Only 229 (15%) patients with pN0 received PMRT, with no evidence of benefit in any endpoints. In this large HER2-positive, predominantly node-positive, adjuvant cohort, PMRT improved locoregional control and conferred a clinically meaningful survival benefit in pN2. In the landscape of personalized treatment and growing interest in axillary de-escalation, future prospective studies are warranted to better define which HER2-positive patients with low nodal burden (particularly pN1 disease) derive meaningful benefit from PMRT in the context of modern systemic therapy. The publication of this analysis was funded by the Italian Ministry of Health through Ricerca Corrente funds.

Culmerciclib (®) is an oral, small molecule CDK2/4/6 inhibitor developed by Chia Tai Tianqing Pharmaceutical for the treatment of breast cancer. By simultaneously inhibiting CDK2, CDK4 and CDK6, culmerciclib may help overcome resistance mechanisms in HR+/HER2- breast cancer. The drug received its first approval in China on 9 December 2025 for use in combination with fulvestrant in adults with HR+/HER2- advanced or metastatic breast cancer whose disease has progressed after endocrine therapy. This article summarizes the milestones in the development of culmerciclib leading to this first approval.

Brain metastases in patients with breast cancer (BCBM) are common. While the Breast Graded Prognostic Assessment (Breast GPA) predicts overall survival (OS), its ability to predict intracranial progression-free survival (icPFS) has not been established. We conducted a retrospective, multi-institutional cohort study to evaluate the association between Breast GPA and icPFS among BCBM patients treated initially with stereotactic radiosurgery (SRS) or whole-brain radiation therapy (WBRT). icPFS was defined as the time from initial local BCBM treatment to the first local salvage treatment or death. Among 2263 patients, 49% received SRS and 51% WBRT. Subtypes included hormone receptor (HR)+/HER2- (31%), HR+/HER2+ (21%), HR-/HER2+ (17%), and triple-negative breast cancer (TNBC) (24%). The median icPFS ranged from 6 months (TNBC) to 12 months (HR+/HER2+). By Breast GPA, the median icPFS ranged from 5 months (GPA 0.0-1.0) to 13 months (GPA 3.5-4.0), and the median OS ranged from 6 to 37 months, respectively. The bias-corrected c-index for Breast GPA was 0.606 for icPFS and 0.648 for OS. icPFS and OS were strongly correlated (r = 0.783; 95% confidence interval, 0.739-0.836). In multivariable analysis, breast cancer subtype, number of BCBM, and Karnofsky Performance Status were the strongest icPFS predictors. The Breast GPA predicts OS and icPFS in BCBM patients, providing valuable benchmarks for clinical trial design. These data establish historical controls for icPFS by subtype and Breast GPA and may guide systemic therapy prioritization in patients at greatest risk of early intracranial progression.

Accurate intraoperative margin assessment and reliable detection of metastatic sentinel lymph nodes (SLNs) remain critical challenges in breast cancer surgery. We developed sa-FCA, a novel XIAP-targeting near-infrared fluorescent probe that achieves enhanced tumor specificity and sensitivity, along with a prolonged imaging window, via tumor-selective cascade activation and in situ self-assembled nanostructures. In a mouse model of breast cancer, sa-FCA demonstrated precise tumor localization, sensitive detection of metastatic SLNs, and identification of spontaneous pulmonary metastases. Clinical evaluation using human specimens showed excellent discrimination between tumor and benign tissues such as fibroadenoma (n = 252 patients, 99.59% sensitivity and 97.37% specificity), with clear lesion delineation. Furthermore, it accurately distinguished between metastatic and non-metastatic SLNs, with a sensitivity of 98.18% and a specificity of 92.31% (n = 69). By significantly improving metastatic SLN detection, sa-FCA overcomes the intraoperative diagnostic delays inherent in conventional pathology and facilitates immediate surgical decision-making. The ability of sa-FCA to precisely target tumors, differentiate fibroadenoma, and identify metastatic nodes allows for complete tumor resection with maximal normal tissue preservation, thereby enhancing both oncological safety and cosmetic outcomes. These findings establish a new paradigm for image-guided breast cancer surgery and promote the clinical translation of fluorescence-guided interventions.

FeDeriCa (NCT03493854) showed that the fixed-dose combination of pertuzumab and trastuzumab for subcutaneous injection (PH FDC SC) was noninferior to intravenous (IV) P and H, with comparable total pathological response rates and safety profiles when given as neoadjuvant therapy for human epidermal growth factor receptor 2 (HER2)-positive early breast cancer (BC). We report long-term efficacy and safety. Patients received eight neoadjuvant chemotherapy + P IV + H IV cycles or PH FDC SC every 3 weeks (cycles 5-8; 1:1 randomisation). Patients continued adjuvant HER2-targeted treatment to complete 18 cycles. Data cut-off was 2 June 2023, with a median follow-up of 51.4 (P + H IV arm) and 51.2 months (PH FDC SC arm). Four-year event-free rates (invasive disease-free survival, event-free survival, distant recurrence-free interval, and overall survival) were 89.6 [95% confidence interval (CI) 85.5-93.7], 88.5 (95% CI 84.4-92.5), 92.5 (95% CI 89.2-95.8) and 95.5 (95% CI 92.9-98.1) in the P + H IV arm and 88.5 (95% CI 84.3-92.7), 86.6 (95% CI 82.2-90.9), 91.9 (95% CI 88.4-95.4) and 94.1 (95% CI 91.1-97.1) in the PH FDC SC arm, respectively. No new safety signals (including cardiac safety) were observed. Long-term efficacy of PH FDC SC and P + H IV was comparable. Safety remained similar and consistent with P + H + chemotherapy. PH FDC SC is established as a faster, more convenient, less invasive patient- and health care professional-preferred alternative to P + H IV for HER2-positive BC.

Current treatment guidelines for patients diagnosed with stage II to III triple-negative breast cancer (TNBC) rely on the use of neoadjuvant therapy (NAT). A critical unmet need is a sensitive test that can inform the presence of distant micrometastases to help personalize postneoadjuvant systemic therapy beyond pathological complete response (pCR). In this study, 286 plasma samples from 86 patients from the SCANDARE study were retrospectively analyzed for circulating tumor DNA (ctDNA) at three presurgical timepoints: baseline, during and post-NAT, and during postsurgical follow-up, when available, using an ultrasensitive, tumor-informed, whole-genome-based assay. After a median follow-up of 5.1 years, 15 patients (17.4%) had developed distant metastases, and 19 patients (22.1%) died; 100% of pretreatment baseline plasma samples were positive for detected ctDNA, and the majority of ctDNA detections during and post-NAT, presurgery, were at ultrasensitive (<100 parts per million) levels. Ultrasensitive ctDNA detection post-NAT, presurgery was highly prognostic of distant recurrence-free interval, disease-free survival, and overall survival. Notably, ultrasensitive ctDNA detection status improved the prognostication of patients without pCR at surgery, with 94% of non-pCR patients with undetectable ctDNA post-NAT, presurgery remaining distant recurrence-free. These findings highlight the potential of ctDNA to refine risk assessment in patients with early-stage TNBC and support its investigation in interventional settings to guide systemic treatment decisions.

Intratumor heterogeneity for human epidermal growth factor receptor 2 (HER2) in HER2-positive breast cancer is a driver of resistance to HER2-targeted therapies. The advancement of treatments for HER2 heterogeneous (HET) tumors has been hindered by the lack of preclinical models that accurately mimic the human disease. In this study, we describe human HER2 HET breast cancer models composed of ERBB2-amplified (HER2hi) and nonamplified (HER2lo) cell populations derived from the same tumor. Utilizing these models, together with cellular barcoding, we demonstrate subclonal cooperation between HER2hi and HER2lo subpopulations. Furthermore, HER2lo cells drive resistance to HER2-targeting antibody-drug conjugates (ADC) like trastuzumab deruxtecan (T-DXd) but are sensitive to HER2 kinase inhibitors. CRISPR screens in HET cocultures identified sensitizers of HER2lo cells to T-DXd, including ATP-binding cassette subfamily C member 1 and ubiquitin-specific peptidase 9 X (USP9X). USP9X inhibition enhances the lysosomal targeting of HER2, thereby potentiating ADC payload release and reducing tumor recurrence after T-DXd treatment. Our results elucidate the functional relevance of HER2 heterogeneity and propose improved therapies for these tumors. Studies of HER2 HET breast cancer models demonstrated that HER2lo cells drive HER2-targeting ADC resistance and accelerate recurrence by cooperating with HER2hi cells. We identified novel therapeutic strategies to sensitize HER2lo cells to T-DXd, providing mechanistic insight and offering promising avenues to overcome resistance and improve patient outcomes.

To address the long-standing inconsistency in Perfluorooctanoic Acid (PFOA)-related carcinogenic evidence, a stepwise analytical framework was developed that integrated CiteSpace-based global hotspot mapping, organ-specific meta-analysis, and nonlinear restricted cubic spline (RCS) dose-response modeling to move from research landscape identification to quantitative heterogeneity synthesis and internal dose-based risk estimation. Bibliometric results revealed a recent frontier shift from organ-specific cancer risks toward serum PFOA biomonitoring, mixed exposure assessment, and endocrine-disruptive mechanisms, providing the conceptual basis for organ-oriented evaluation. Meta-analysis of 34 datasets across the four organ-specific cancers most associated with PFOA exposure demonstrated a heterogeneous and divergent risk profile, with modest yet measurable cumulative elevations in cancer risk of kidney (odds ratio (OR) = 1.27; 95% confidence interval (CI), 1.01-1.59; P = 0.04), borderline statistical significance in breast cancer risk (OR = 1.13; 95% CI: 1.00-1.27; p = 0.04), while liver cancers (OR = 1.02; 95% CI: 0.95-1.09; p = 0.64) and thyroid cancers (OR = 0.90; 95% CI: 0.76-1.05; p = 0.19) were statistically non-significant. The dose-response risk estimation with the RCS model further uncovered distinct nonlinear carcinogenic architectures, as characterized by Odds Ratios varying with serum/plasma PFOA concentration, including cumulative amplification in kidney, borderline statistical accumulation in breast cancers, and adaptive, non-monotonic modulation in liver and thyroid cancers. These findings highlighted organ-specific susceptibility rather than uniform carcinogenicity and advance a biomarker-driven, nonlinear, and prediction-oriented paradigm for global environmental carcinogenic risk assessment, providing mechanistically informed evidence to refine PFAS regulatory prioritization and precision public health decision-making.

Drug-drug cocrystal technology especially for natural products, can optimize the physicochemical properties of drugs without altering their chemical structures, thereby enabling synergistic and dual-drug therapeutic effects. In this study, a novel drug-drug cocrystal composed of emodin (EM) and matrine (MA) was successfully prepared using ultrasound-assisted crystallization in a benchtop ultrasonic bath operating at a fixed frequency of 53  kHz. The optimal preparation conditions were a nominal ultrasonic power of 500  W, a crystallization temperature of 35°C, a sonication time of 3 h, and an EM:MA molar ratio of 1:1.5. The cavitation effects generated by ultrasound disrupted the pre-existing intermolecular hydrogen bonds (e.g., drug-solvent or drug self-association) and accelerated molecular diffusion, thereby facilitating the formation of desired drug-drug hydrogen bonds for rapid co-nucleation, shortened co-crystallization time, and high product uniformity. The process was systematically optimized using response surface methodology (RSM) and artificial neural networks (ANNs). The resulting EM-MA cocrystal was characterized by PXRD, DSC, FT-IR, SCXRD, and SEM, which confirmed the formation of O-H···O=C heterosynthons and the stabilizing role of hydrogen-bonding. Cellular assays showed that the cocrystal exhibited stronger inhibitory activity against 4 T1 breast cancer cells than the physical mixture of EM and MA. These results indicate that the novel cocrystal may have potential for further development as a combination drug product, which could improve the bioactivity of natural products.

Women who have full-term pregnancies have a reduced risk of breast cancer, although the underlying mechanisms are not well understood. Here we show that tissue-resident memory-like T (TRM-like) cells are enriched in the breasts of parous women and mice compared to nulliparous individuals. These cells develop during mid-gestation, persist after lactation and are closely associated with mammary epithelial cells, suggesting dependence on epithelial-derived signals. Impaired alveolar differentiation or a deficiency in epithelial cell-derived cytokines interleukin (IL)-15 and transforming growth factor (TGF)β diminished the expansion of mammary TRM-like cells. Single-cell transcriptomics revealed the expanded TRM-like cells were heterogeneous, comprising CD8αβ+ and CD8αα+ subsets. Lineage tracing showed that these TRM-like cells acquire effector functions and contribute to tumor control. Depletion of TRM-like cells abrogated breast tumor protection in parous mice, while enhanced IL-2Rβ signaling-induced TRM-like cells and conferred protection in nulliparous mice. Together, we show a mechanism by which pregnancy can induce anticipatory breast cancer protection in a tissue-specific manner.

Cells dynamically regulate their morphology, contractility, and metabolism in response to the mechano-chemical properties of their microenvironment. Here, we show matrix stiffness and ligand density jointly govern the bioenergetics of contractile cells through a nonequilibrium active chemo-mechanical model built around a newly introduced cellular metabolic potential. This concept links ATP hydrolysis to mechanosensitive signaling, quantifies the energetic cost of stress fiber assembly, and determines mechanically stable states. The metabolic potential enables quantitative prediction of cell contractility, morphology, and ATP consumption in different stiffness 2D and 3D environments, and we find quantitative agreement with experimental measurements in MDA-MB-231 breast cancer cells. The model further predicts activation of AMPK accompanies increased energetic demands in stiffer microenvironments which we experimentally validate and correlate with increased mitochondrial membrane potential, glucose uptake, and intracellular ATP levels. Together, these findings establish a predictive quantitative framework linking mechanosensitive control of cell shape and contractility to actomyosin-dependent ATP consumption and AMPK-mediated energy replenishment. STATEMENT OF SIGNIFICANCE: While it is well established that extracellular matrix stiffness alters cell shape, contractility, and signaling, how these cues quantitatively integrate with cellular energy homeostasis in diseases such as cancer remains unclear. In this work, we establish a framework connecting extracellular mechanics, actomyosin organization, and intracellular energy regulation into a single energetic principle governing cell behavior. We show that mechanical environments reshape the cellular energy budget, driving cells toward morphologies and contractile states that balance energetic demand with metabolic supply. This perspective reframes mechanotransduction not simply as a signaling cascade, but as a process fundamentally constrained by energy economics, providing new insight into how cancer cells navigate mechanically heterogeneous tissues, with implications for invasion, metastasis, and therapeutic targeting of mechano-metabolic vulnerabilities.

Resistance to therapeutic agents represents a critical barrier in the clinical management of triple-negative breast cancer (TNBC), necessitating novel therapeutic strategies. We discovered that mammalian target of rapamycin (mTOR) and glutaminase 1 (GLS1) mediated glutamine metabolism-a TNBC-addicted process-critically drives aggressive proliferation and confers therapeutic resistance. The combined treatment with rapamycin (an mTOR inhibitor) and CB839 (a GLS1 inhibitor) elicited synergistic anti-tumor effects in diverse TNBC cell lines. This synergy arose from‌ inhibition of metabolic rewiring, ‌characterized by‌ perturbations in adenosine triphosphate (ATP) homeostasis, redox equilibrium, metabolite pools, and signaling cascades, ‌ultimately triggering multimodal‌ cell death mechanisms. Furthermore, ‌synergistic‌ pharmacological inhibition of mTOR and GLS1 ‌significantly suppressed‌ in vivo tumor growth and metastasis ‌in murine models‌, ‌with no overt toxic effects observed‌. Overall, our findings indicate dual inhibition of mTOR and GLS1 as a promising clinical strategy to counteract glutamine-driven resistance, enhancing therapeutic efficacy in TNBC.

Brain metastasis (BM) is a leading cause of mortality in breast cancer patients. This study utilizes single-cell RNA sequencing (scRNA-seq) to identify high-risk malignant subclusters and uncover potential therapeutic targets driving BM. Five pairs of scRNA-seq data of primary breast tumors (PT) and BM were retrieved from the Gene Expression Omnibus (GEO) database. Following unsupervised clustering and cell-type annotation, specialized bioinformatic pipelines-including inferCNV and pseudotime trajectory analysis-were implemented to delineate malignant subpopulations and evolutionary states associated with high brain metastatic potential. Molecular mechanisms were investigated using ChIP-qPCR, nascent RNA labeling assay, and dual-luciferase assays. Clinical significance was evaluated in a 124 paired-patient cohort using propensity score matching (PSM), while in vivo metastatic potential was assessed via a murine carotid artery injection model. We identified a specific malignant epithelial subcluster (cluster 3) characterized by high lactate levels and superior BM potential and subsequently established CREB1 lactylation as a biomarker of this cluster. Mechanistically, lactate dehydrogenase A (LDHA)-mediated lactylation of CREB1 at lysine 136 (K136) enhances its transcriptional activity, upregulating cytoskeletal genes including CALML5, CNN2, and PDLIM1. This axis facilitates pseudopodia formation, cellular migration. In vivo, LDHA knockdown significantly reduced intracranial tumor burden. Clinically, high lactylation scores and high CALML5 expression are independent predictors of BM and overall survival. The LDHA-lactylation-CREB1-cytoskeleton axis is a novel driver of BM, serving as a promising therapeutic target and prognostic biomarker for breast cancer.

Triple-negative breast cancer (TNBC) exhibits robust self-regulatory mechanisms against harsh microenvironmental stress, yet the precise molecular drivers remain elusive. Here, we identified circSCLT1 as a novel circular RNA with energy stress-responsive properties. Systematic in vitro and in vivo assays demonstrated that circSCLT1 promotes TNBC metastasis and radioresistance. Mechanistically, energy stress triggers HNRNPA1-dependent circSCLT1 biogenesis. Cytoplasmic circSCLT1 scaffolds a ternary complex with HNRNPA1 and CSDE1, blocking HNRNPA1 nuclear translocation and stabilizing IL1B via binding to its coding sequence (CDS). Additionally, the circSCLT1-containing complex displaces the E3 ubiquitin ligase TRIM25 from CSDE1 and redirects it toward TIAL1, thereby inducing K48-linked polyubiquitination and proteasomal degradation of TIAL1. Furthermore, circSCLT1 suppresses arginine dimethylation of HNRNPA1 by reinforcing the binding between CSDE1 and HNRNPA1, which in turn facilitates stress granule (SG) assembly under severe energy stress. These circSCLT1/HNRNPA1/CSDE1-containing SGs shield IL1B from autophagy-mediated degradation. Collectively, these actions amplify IL1B/NF-κB signaling, sustaining pro-survival networks that drive metastasis and radioresistance. Importantly, pharmacological inhibition of IL-1β secretion using disulfiram (DSF) synergizes with circSCLT1 silencing to markedly suppress tumor progression and extend survival in preclinical TNBC models. Our findings elucidate the oncogenic role of circSCLT1 in TNBC and underscore its potential as a novel therapeutic target, providing a rationale for combining IL-1β inhibition with circSCLT1-targeted therapy.

PIWI-interacting RNAs (piRNAs) are a class of endogenous noncoding RNAs that perform pivotal regulatory functions in numerous physiological activities and pathological progression. Dysregulated piRNA expression is closely associated with the progression and prognosis of cancers. Entropy-driven circuit have been widely designed for the imaging and therapy, but their imaging efficiency is always constrained by the uncontrolled off-site signal leakage and the inefficient circuitry activation that may result in low imaging contrast. Herein, we construct a programmable deoxyribozyme (DNAzyme)-activated entropy-driven logic circuit for the high-contrast imaging of piRNAs and the discrimination of cancer cells. We design a methylation-modified DNAzyme as a fuel strand generator and a hairpin probe that confines the fuel strand in the hairpin structure to suppress the signal leakage from the entropy-driven reaction circuit. The highly expressed fat mass and obesity-associated protein (FTO) in cancer cells can catalyze the demethylation of DNAzyme to activate its cleavage activity, inducing the cleavage of hairpin probes and the release of abundant fuel strands to drive entropy-driven reaction circuit. In the presence of target piRNAs, the fuel strands initiate toehold-assisted branch migration reaction to release signal probes and produce enhanced fluorescence signals. The proposed logic circuit system can profile endogenous piRNAs at the single-cell level, discriminate breast cancer tissues from healthy counterparts, and achieve in situ high-contrast imaging of piRNA in living cells. Importantly, the proposed logic circuit system can distinguish different cell subtypes with high accuracy and good specificity, providing a potential platform for clinical diagnosis.

Triple-negative breast cancer (TNBC) is characterized by aggressive biological behavior, including rapid post-treatment recurrence propensity, heightened metastatic dissemination, and significantly diminished survival outcomes. These features emphasize the necessity for innovative therapeutic strategies in TNBC treatment. Herein, we developed selenium nanoparticles modified with a mushroom polysaccharide-protein complex (PTR-SeNPs) and evaluated them in vitro anti-tumor efficacy across 17 human TNBC cell lines, followed by elucidation of the mechanism underlying PTR-SeNPs-induced apoptosis. In vitro evaluation across TNBC cell models revealed that PTR-SeNPs exhibit promising anti-tumor efficacy with preferential induction of mitochondrial-dependent apoptosis, demonstrating significant cytotoxic efficacy through MAPKs/Bcl2 pathway. Notably, further conjugation of PTR-SeNPs with anti-human MUC1 antibodies generated dual-modified nanoparticles (MUC1@PTR-SeNPs), which significantly enhanced anti-tumor activity in five TNBC cell lines with high/medium MUC1 expression. Furthermore, oral administration of MUC1@PTR-SeNPs for 30 days markedly inhibited tumor growth in mice bearing MDA-MB-468 xenografts via induction of mitochondria-mediated apoptosis. This work highlights the therapeutic potential of PTR-SeNPs against human TNBC, elucidates their molecular mechanisms, metabolic profile, and toxicity, thereby advancing this novel nano-mineral as a future treatment strategy for TNBC.

Entosis is a non-apoptotic cell death process implicated in various important biological processes, such as tumorigenesis. Entotic death is preceded with the formation of cell-in-cell structures that are well known to be controlled by two spatially separated core elements: adherens junction and actomyosin. However, the molecular mechanism underlying their coordination remains a longstanding open question. In this study, by profiling isogenic breast cancer cells, ARHGAP36 was identified as a potent inducer of entotic cell-in-cell formation, consistent with multiple lines of tumor-suppressive evidence both in vitro and in vivo. This effect is attributed to the concomitant promotion of P-cadherin-mediated cell-cell adhesion and RhoA-regulated actomyosin contraction. Mechanistically, ARHGAP36, through the arginine-rich domain at the N-terminal, binds to β-catenin to stabilize P-cadherin expression in a way accompanying with, and mutually exclusive from, its interaction with PKAc to activate RhoA signaling. Thus, this study unveiled a heretofore unrecognized coordination mechanism for entosis, where ARHGAP36 engages both adherens junction and actomyosin to drive cell-in-cell formation, providing a promising cancer therapeutic target.

Olaparib resistance limits its therapeutic efficacy in triple-negative breast cancer (TNBC). Exploring the mechanisms underlying olaparib resistance and developing combination strategies are of great clinical significance for improving the long-term therapeutic benefit in TNBC. This study aimed to determine whether ZUP1 is associated with olaparib resistance in TNBC patients and to elucidate the underlying mechanisms involved. We established an olaparib-resistant TNBC cell model and integrated transcriptomic profiling with public datasets to DUBs linked to resistance. Candidate function was assessed by drug sensitivity assay, clonogenic assay, apoptosis assay, HR/NHEJ reporter assays and comet assays. Mechanistic studies used co-immunoprecipitation, chromatin fractionation, and PARylation assays. In vitro co-culture assays and flow cytometry analysis were used to evaluated immune infiltration. High-throughput virtual screening (HTVS), biolayer interferometry (BLI), and deubiquitination assays nominated small molecule inhibitors and assessed therapeutic synergy with olaparib. We identified ZUP1, a recently uncovered deubiquitinase, as significantly upregulated in TNBC patients who did not respond to olaparib. Mechanistically, ZUP1 stabilized PARP1 by removing its polyubiquitin chains at lysine 425, resulting in increased PARylation and enhanced chromatin retention of SSRP1 and SPT16, thereby promoting DNA repair. ZUP1 deficiency significantly increased olaparib-induced DNA damage, facilitates cytosolic dsDNA release to activate STING signalling, and enhanced CD8+T cell infiltration into tumors. High-throughput virtual screening identified procyanidin C1 as a potential ZUP1 inhibitor. Combination treatment with procyanidin C1 and olaparib significantly suppressed tumor growth in olaparib -resistant TNBC models. ZUP1 facilitates olaparib resistance in TNBC by stabilizing PARP1 and enhancing DNA damage repair. Pharmacological inhibition of ZUP1 with procyanidin C1 represents a promising therapeutic strategy to overcome olaparib resistance in TNBC.

This perspective brings together multiple observations concerning the regulatory architecture and functional interactions of the TP53 gene and its isoforms. The TP53 gene encompasses an internal enhancer-promoter region (P2) located between exons 2 and 5, which is transactivated by full-length TAp53α, to drive the expression of truncated TP53 isoforms ΔN133p53α, β or γ. ΔN133p53α homo-monomers or dimers very likely form hetero-tetramers with homo-dimers of full-length TAp53α. The TAp53α-ΔN133p53 tetramer then acts as a transcription factor augmenting mitochondrial efficiency, DNA repair, telomere restoration and reversing cellular senescence, thus opposing many outputs of tumor suppressive TAp53α homo-tetramers. Other transcription factors, such as estrogen receptor (ER), OCT1, and SOX9 also may bind the P2 promoter-enhancer to promote transcription of oncogenic TP53 isoforms. These transcription factors, together with full-length TAp53α, appear to cooperate to maintain an open chromatin state in the P2 region in a manner reminiscent of a neural "associative memory" network.

Tumor progression and drug resistance in breast cancer are fundamentally governed by a complex microenvironment, characterized by extreme cell densities and associated nutrient/oxygen gradients. Conventional in vitro models often fail to replicate these features due to mass transport limitations in non-perfused systems. Here, we present a next-generation tumor-on-a-chip platform engineered via high-resolution projection micro-stereolithography (PµSL) to sustain exceptionally high-density breast cancer tissues (up to 400 million cells/mL) under continuous perfusion. By integrating a vascular-inspired perfusion architecture with precision-engineered micropores (∼7 µm), our system enhances nutrient and drug transport compared with traditional non-perfused 3D cultures. This high-density microenvironment drives density-dependent molecular reprogramming of ERα, Ki67, and CTSL2, closely mirroring the gene expression signatures observed in high-grade patient tumors. Notably, the drug resistance to Elacestrant observed in our high-density tissues uniquely aligns with clinically reported Cmax concentrations, effectively bridging the gap between in vitro screening and in vivo outcomes. This platform provides a physiologically relevant tool for studying aggressive tumor biology and evaluating personalized therapeutic strategies with high predictive power.

Understanding gene spatial expression and the organization of multicellular systems is vital for disease diagnosis and studying biological processes. However, existing models often struggle to integrate gene expression data with cellular spatial information effectively. Here we introduce SpatialFormer, a hybrid framework combining convolutional networks and transformers to learn single-cell multimodal and multiscale information in the niche context, including expression data and subcellular gene spatial distribution. Pretrained on 700 million cell pairs from 17 million spatially resolved single cells across 71 Xenium slides, SpatialFormer merges gene spatial expression profiles with cell niche information via the pairwise training strategy. Our findings demonstrate that SpatialFormer distills biological signals across various tasks, including single-cell batch correction, cell-type annotation and co-localization detection. The perturbation analysis identified gene pairs essential for the immune cell-cell communication in pulmonary fibrosis, epithelial-myoepithelial co-localization and tumor transition signals in breast cancer. These advancements enhance our understanding of cellular dynamics and offer additional pathways for applications in biomedical research.

Daedaleopsis confragosa, a medicinal mushroom, produces endoperoxide-containing steroids with anticancer activity, but their mechanisms remain unclear. This study investigated the anticancer potential of steroidal compounds from D. confragosa as food-derived bioactive compounds. Network pharmacology identified Src and PI3K-Akt pathway as key targets. Among seven isolated compounds, ergosterol peroxide (compound 1), 9,11-dehydroergosterol peroxide (compound 2), 5α,8α-epidioxyergosta-6-en-3-ol (compound 3), and (3β,5α,8α)-5,8-epidioxyergost-6-en-3-ol (compound 5) significantly inhibited MDA-MB-231 breast cancer cell viability, with compound 2 exhibiting the highest potency. These compounds induced cell death and reduced phosphorylated Src levels, with similar effects in 3D spheroid models. Molecular docking and dynamics simulations revealed that compound 2 and its metabolites (2-1,2-2) interact with Src near the ATP-binding site. Notably, metabolite 2-2 exhibited stronger binding affinity (-56.93 kcal/mol) than ATP (-40.29 kcal/mol). These findings suggest that steroidal compounds from D. confragosa inhibit Src kinase by blocking ATP binding, demonstrating significant therapeutic potential.

Patients with gastric cancer face substantial risk of recurrence after surgical resection. Molecular profiling of primary tumors may improve risk stratification to guide postoperative management. To identify genomic features of primary gastric tumors associated with disease recurrence and patterns of metastatic spread. This single-center cohort study took place at an academic quaternary referral center and included patients who underwent curative-intent resection of gastric adenocarcinoma from 2010 to 2024. Patients were classified by recurrence status and pattern of metastatic spread. Patients with gastric cancer (stages I to III) who underwent a margin-negative resection and had genomic sequencing of their primary tumor were included. Primary analysis excluded patients who had no evidence of disease with less than 2 years of follow-up. These data were analyzed from June 2025 through March 2026. Primary tumor specimens were sequenced using a targeted panel of cancer-associated genes (MSK-IMPACT). Correlation of disease-free survival and patterns of metastatic spread (hematogenous, peritoneal, or lymphatic) with primary tumor genomic profile. Among 438 patients who underwent complete oncologic resection, 377 had sufficient clinical follow-up or developed recurrence (median [IQR] age, 64 [55-71] years; 113 female [30%] and 264 male [70%]). Recurrence was identified in 179 patients, whereas 198 patients had no evidence of disease. In a multivariable analysis, alterations in KRAS (hazard ratio [HR], 1.54; 95% CI, 1.04-2.28; P = .03) and PIK3CA (HR, 2.15; 95% CI, 1.25-3.69; P = .006) were independently associated with worse disease-free survival. Tumors of patients with hematogenous recurrence had greater chromosomal instability (fraction genome altered, 0.11 vs 0.03; P = .001), whole-genome duplication (47% vs 15%; P = .02), and more frequent alterations of genes modulating cell cycle regulation (39% vs 6%; P < .001) compared with peritoneal recurrence. Bone metastasis arose from more genomically stable primary tumors (fraction genome altered, 0.005 vs 0.114; P < .001) and tumors with Lauren diffuse-type histology (36% vs 3%; P < .001) compared with other sites of hematogenous spread. This study identifies genomic alterations associated with disease-free survival and patterns of recurrence after resection of gastric cancer. These clinicogenomic risk factors may personalize postoperative surveillance strategies and inform perioperative treatment decisions.

The incidence of cancer in people with cardiovascular disease (CVD) is poorly understood. This study aimed to understand the sex differences in incident cancer and cancer-related mortality in people with and without CVD. Participants were drawn from the European Prospective Investigation into Cancer (EPIC)-Norfolk study and followed up for a median of 26.2 years. Participants were stratified by sex and incident CVD during follow-up. Cumulative cancer incidence was calculated for each group. Sex-specific lifetime risks of cancer were determined, accounting for competing risks of death. Cause-specific hazard ratios (HRs) were determined using Royston-Parmar models, adjusted for ethnicity and time-updated covariates, including deprivation index, cardiovascular risk factors, comorbidities, and medication. Participants' CVD status was updated throughout follow-up. A total of 22 534 participants were cancer-free at baseline (57% women; mean age 58.6 ± 9.3 and 59.5 ± 9.3 for women and men, respectively); 10 889 participants either had prevalent CVD at baseline or developed CVD during follow-up. Men had more comorbidities. The adjusted lifetime risk of incident cancer in CVD participants at age 75 was higher in men (17.3%) than in women (12.8%), with a significant cause-specific HR of 1.39 (99% confidence interval [CI] 1.25-1.53, P < .0001). Corresponding data for participants with no CVD were 10.2% for men and 8.2% for women, with an HR of 1.27 (99% CI 1.15-1.41, P < .0001 for men vs women). A similar trend exists for cancer-related mortality (HR 1.51, 99% CI 1.31-1.75, P < .0001 for men with CVD and HR 1.31, 99% CI 1.14-1.50, P < .0001 for men without CVD). This study showed an increased lifetime cancer risk and mortality in men compared with women, which was mediated by comorbid CVD.

Treatment strategies for patients with metastatic breast cancer (MBC) are becoming increasingly complex, underscoring the need for reliable biomarkers to inform clinical decision making. Circulating tumor cells (CTCs) are well-established prognostic biomarkers in MBC, while tumor-derived extracellular vesicles (tdEVs) have recently demonstrated complementary significance in patients undergoing first-line chemotherapy. This study seeks to validate the complementary role of tdEVs in a larger patient population and establish a clinically relevant cut-off. Archived baseline CellSearch® images from 355 patients with MBC (NU16B06 study) were reanalyzed using the Automated CTC Classification, Enumeration and PhenoTyping (ACCEPT) software to quantify CTCs and tdEVs. Multiple tdEV cut-offs were evaluated for association with overall survival (OS), alone and with CTCs (≥5, CTChigh). The selected cut-off was validated in a subcohort of 385 patients from the phase III STIC CTC trial. In the NU16B06 cohort, higher tdEV levels were consistently associated with shorter OS, regardless of cut-off. As models performed similarly (Brier Score difference <0.005), a cut-off of >20 tdEVs/7.5 ml (tdEVhigh) was selected based on prior literature and clinical relevance. Patients with tdEVhigh had nearly doubled risk of death compared with tdEVlow. Combining tdEVs with CTCs identified distinct prognostic subgroups, with CTChigh/tdEVhigh patients showing the shortest OS. Among CTClow patients, tdEVhigh was associated with increased mortality risk. In the STIC CTC cohort, the cut-off was validated for OS and progression-free survival, with tdEVhigh consistently associated with poorer surivial. When combined, patients with CTChigh/tdEVhigh had the poorest outcomes, while tdEVhigh remained associated with worse prognosis among CTClow patients. These findings support the use of tdEVs as complementary prognostic biomarkers to CTCs, potentially enhancing clinical trial design and informing treatment decisions in MBC.

The unprecedented expansion of approved oncology therapies has prolonged survival and transformed the prognosis for many patients diagnosed with cancer. However, cancer treatments may be associated with cardiovascular toxicities that manifest through vascular, myocardial, or metabolic pathways, potentially limiting the use of cancer therapeutics and adversely affecting outcomes. Oncology clinical trials provide an important opportunity to evaluate cardiovascular safety signals by generating data on the incidence, timing, and spectrum of toxicities. However, progress has been limited by inconsistent definitions and variable approaches to event characterization. This scientific statement aligns the advances in cardiovascular medicine and cardiovascular clinical trials to provide criteria for systematic selection, rigorous characterization, and adjudication of cardiovascular endpoints in contemporary oncology trials. The proposed framework links drug-specific mechanisms to endpoint selection and standardizes the approach to definitions of adverse cardiovascular events, including heart failure, arrhythmias, myocarditis, and thrombotic events. Definitions of major adverse cardiac events, clinical events, and surrogate endpoints are discussed, along with strategies for alignment with the Common Terminology Criteria for Adverse Events and patient-reported outcomes. Practical guidance is provided for prospective surveillance, decentralized and hybrid clinical trial designs, independent endpoint adjudication, and statistical approaches to competing risks and late-emerging toxicities. By harmonizing cardiovascular endpoint assessment across oncology trials, this scientific statement aims to enhance risk stratification, facilitate regulatory acceptance, and inform clinical decision-making, ultimately improving patient safety while supporting innovation in cancer therapeutics.

Colorectal cancer (CRC) is a leading cause of cancer-related mortality worldwide. Organized screening programs using the fecal immunochemical test (FIT) have been implemented to reduce CRC mortality through early detection. To evaluate the association between an organized FIT-based screening program and CRC-specific mortality in a population-based setting. This retrospective, population-based cohort study included all residents of the Basque Country, Spain (approximately 2.2 million inhabitants) between 2004 and 2024. The FIT-based CRC screening program targeting individuals aged 50 to 69 years was initiated in 2009. Individuals aged less than 50 years were included in the study as a nonscreening reference group. Data analysis was performed in April 2026. Biennial screening of asymptomatic individuals aged 50 to 69 years using single-sample FIT. Age-standardized mortality rates (ASMRs) for CRC per 100 000 population (based on the European Standard Population 2013) and annual mortality trends from 2004 through 2024 were calculated. Segmented regression and counterfactual analyses estimated changes in CRC mortality after screening implementation. Screening participation, FIT positivity, colonoscopy adherence, CRC detection rates, and cancer stage distribution were also evaluated. Among 438 134 total deaths (50.9% in men) in a population of approximately 2.2. million, 16 298 (3.7%) were attributable to CRC. Age-standardized CRC mortality declined from 32.8 to 23.1 deaths per 100 000 population between 2004 and 2024 (reduction of 29.6%; P = .002). In individuals aged 50 to 69 years, CRC mortality declined by 50.1% (from 39.7 to 19.8 deaths per 100 000 population). Segmented regression identified a significant inflection point in the year 2012 (95% CI, 2011-2014; P < .001), 3 years after screening implementation. Counterfactual analyses estimated a 46.2% relative reduction in CRC mortality in the group aged 50 to 69 years compared with projected trends without screening. Among individuals aged 70 years or older, CRC mortality declined by 19.6% (from 165.4 to 132.9 deaths per 100 000 population), corresponding to a 45.4% relative reduction compared with projected rates. Overall, 3464 of 4892 screen-detected CRCs (70.8%) were diagnosed at stage I or II throughout the study period. In this retrospective, population-based, descriptive cohort study, organized FIT-based screening was associated with a sustained decline in CRC mortality, particularly among individuals aged 50 to 69 years. These findings support the long-term benefit of organized FIT-based screening programs as a public health strategy to reduce CRC-related mortality.

Colorectal cancer (CRC) incidence is increasing in sub-Saharan Africa, including Nigeria, where more than half of patients die within 1 year due to late diagnosis and limited treatment. Without ready access to screening, identifying risk factors is critical. However, regional data are scarce, and it is unclear whether risk factors from high-income populations apply to Nigerians, who may have distinct clinicodemographic patterns and tumor characteristics. To assess whether 13 CRC risk factors identified in high-income populations are associated with CRC in adults in Nigeria. This multicenter case-control study was conducted in 6 hospitals across 3 of Nigeria's 6 geopolitical zones, with recruitment from April 2020 to September 2024. Participants were patients newly diagnosed with CRC and cancer-free controls matched on age, sex, location, education, and recruitment site. Data were analyzed from April 2023 to November 2025. Family history of cancer, anthropometric factors (adult height, body mass index [BMI], and somatotype or body size over the life course), physical activity, and dietary and lifestyle factors (processed meat, red meat, fruit, vegetables, fiber, alcohol, smoking, and diabetes). The associations of risk factors with CRC were assessed using odds ratios (ORs) and 95% CIs, estimated using multivariable unconditional logistic regression. Among 2063 participants (1103 [53.5%] male; median [IQR] age, 54 [42-64] years) enrolled, most had secondary education or higher (1756 participants [85.2%]), lived in urban areas (1817 participants [88.1%]), and were Yoruba (1524 participants [73.9%]). Cases were older than controls (median [IQR] age, 56 [45-66] vs 52 [41-63] years) but were otherwise similar. Family history of cancer (OR, 1.55; 95% CI, 1.06-2.26), adult height (OR per 5-cm increase, 1.09; 95% CI, 1.00-1.20), body size or type over the life course (tertile 3 vs tertile 1: OR, 1.88; 95% CI, 1.30-2.72), and processed meat intake (lowest vs highest tertile of intake: OR, 1.43; 95% CI, 1.07-1.91) were positively associated with CRC risk, and an inverse U-shaped association was identified between physical activity and CRC risk (OR vs first quartile of activity, quartile 2: 0.63; 95% CI, 0.44-0.88; quartile 3: 0.52; 95% CI, 0.34-0.79; quartile 4: 0.77; 95% CI, 0.58-1.02), consistent with data from high-income populations. BMI was inversely associated with CRC risk (OR vs BMI 18.5-24.9, BMI <18.5: 2.63; 95% CI, 1.79-3.86; BMI ≥30: 0.42; 95% CI, 0.28-0.63), likely reflecting prediagnostic weight loss. No associations with diabetes, smoking, or intakes of red meat, fruit, vegetables, fiber, or alcohol were observed, likely due to low prevalence or variability. This case-control study of CRC risk factors in adults in Nigeria found CRC risk patterns similar to those observed in high-income populations. These insights offer immediate opportunity to tailor education, prevention, and early detection in the region, especially as economic development leads to lifestyles more similar to high-income populations. These data provide a foundation for prospective research and investigations in other African populations.

Raised inflammatory biomarkers predict an increased short-term risk of recurrent vascular events after stroke or transient ischemic attack, but it is uncertain whether the hazard is maintained thereafter. In the absence of previous population-based studies, we aimed to determine the predictive value of biomarkers for the long-term risk of recurrent stroke in intensively treated patients following transient ischemic attack or stroke. We studied 12 blood biomarkers related to inflammation, neuronal cell damage, and thrombosis soon after transient ischemic attack or stroke in a population-based study (OXVASC [Oxford Vascular Study]; 2002-2011) with follow-up to 2025. We used Cox and negative binomial regressions to assess associations between each log-transformed biomarker (per SD increase) and recurrent stroke (first event and total number), adjusted for age, sex, and baseline vascular risk factors. Of 1292 consecutive patients (mean/SD age, 73.1/13.2 years; 47.8% male), 365 recurrent strokes occurred in 280 patients during 11 336 patient-years. All inflammatory biomarkers were associated with 10-year risk of recurrence, with little impact of adjustment for other risk factors (adjusted incidence rate ratio: IL-6 [interleukin-6], 1.21 [95% CI, 1.04-1.42]; CRP [C-reactive protein], 1.29 [95% CI, 1.11-1.49]; TNFR-1 [tumor necrosis factor receptor-1], 1.24 [95% CI, 1.05-1.46]; and NGAL [neutrophil gelatinase-associated lipocalin], 1.15 [95% CI, 0.99-1.33]). Moreover, number of elevated (top quartile) inflammatory markers was associated with higher burden of recurrence (incidence rate ratio, 1.31 [95% CI, 1.15-1.50]; P<0.001), with no diminution over time (90 days: hazard ratio, 1.24 [95% CI, 0.98-1.57]; 90 days-1 year: 1.24 [95% CI, 0.95-1.62]; 1-5 years: 1.22 [95% CI, 1.01-1.47]; and 5-10 years: 1.34 [95% CI, 1.06-1.68]), and similar associations for transient ischemic attack and minor stroke. Biomarkers related to neuronal cell damage or thrombosis were generally less predictive of recurrent stroke. Inflammatory biomarkers were more predictive of long-term recurrent stroke than neuronal or thrombotic biomarkers, particularly when several inflammatory markers were raised. Predictive value was independent of other vascular risk factors and did not diminish with duration of follow-up, supporting trials of anti-inflammatory strategies for long-term secondary prevention.

The incidence of early-onset prostate cancer (EOPC) is rising, and by 2045 a 24.5% increase in cases and a 50% rise in mortality are projected. Accumulating evidence indicates that EOPC represents a distinct disease entity, characterized by unique molecular features, risk factor profiles, and clinical behavior that differ from standard-onset prostate cancer (SOPC). Nevertheless, research in this field remains nascent, and no consensus exists regarding the optimal management of EOPC. We synthesize current evidence on the epidemiology, molecular pathology, clinicopathological characteristics, survival, management, and early detection of EOPC. EOPC exhibits a distinctive molecular landscape, with TMPRSS2-ERG fusions occurring in 63-90% of cases as a hallmark alteration, whereas mutations in PTEN, SPOP, and CHD1 are significantly less frequent. Notably, the prevailing focus on hereditary EOPC has inadvertently led to the neglect of sporadic cases, which dominate clinical practice. Although localized EOPC confers no significant prognostic advantage over SOPC, high-risk or metastatic early-onset disease substantially elevates prostate-cancer-specific mortality. By critically appraising the existing evidence, we identify key knowledge gaps, such as the understudied sporadic EOPC subgroup and the lack of dedicated clinical trials, and propose future research directions to inform early detection and optimize therapeutic strategies for this unique patient population.

Radiotherapy for head and neck cancer commonly induces salivary gland dysfunction, resulting in xerostomia that substantially impairs quality of life after treatment. To identify protective strategies, we established the irradiation-induced submandibular glands (SMG) injury mice model and performed integrated proteomic and metabolomic profiling of gland tissue and saliva. Spermidine levels were significantly altered in irradiated SMG during the postradiotherapy recovery phase. Oral spermidine supplementation restored salivary flow, preserved acinar aquaporin-5 expression, and attenuated vacuolization, apoptosis, and fibrosis in vivo; these effects were validated in SMG organoids. Single-cell RNA transcriptomic analysis further showed that spermidine increased the proportion of acinar cells, enhanced Golgi-mediated secretory gene expression, and suppressed inflammatory cytokine signaling. In the randomized, double-blind, proof-of-concept clinical trial, oral spermidine supplementation during peri-radiotherapy significantly reduced the incidence of xerostomia at 3 months compared with placebo (48.28 vs. 79.31%, p = 0.0391), while increasing salivary output and improving quality of life without serious adverse events. These findings highlight the potential of spermidine as a safe, metabolite-based strategy for preserving salivary gland function and alleviating radiation-induced xerostomia. Clinical trial registration: The RCT has been registered in Clinical. gov (NCT07035626).

Industrial volatile chemical products (VCPs), particularly those used in industrial coating applications, have emerged as major sources of reactive organic carbon (ROC) emissions with implications for atmospheric chemistry and human health. However, coating-related ROC emissions remain poorly constrained, limiting assessment of their environmental and health impacts. Here, we characterized full-volatility ROC emissions from representative industrial coating processes across gas and particle phases and estimated China's national ROC emissions from downstream coating processes at approximately 5.31 Tg yr⁻¹ in 2019. Volatile organic compounds (VOCs) dominated the characterized ROC mass in the high-volatility range, with aromatics (35.6%) and cyclic alkanes (20.4%) as major hydrocarbon groups, whereas oxygenated and functionalized organics increased at lower volatility ranges. However, emission mass alone did not reflect health relevance. Species prioritized by quantitative lifetime cancer risk (LCR) and hazard index (HI) assessment represented approximately 22.6% of the characterized ROC mass but dominated the calculated risk indicators. The top two cancer-risk contributors, 1,2-dichloroethane and trichloroethylene, accounted for 96.3% of the summed LCR while representing only 5.07% of the characterized ROC mass. In addition, a recurrently detected emerging-pollutant subset, including polycyclic aromatic hydrocarbons (PAHs), phthalates, phenolic antioxidants, and cyclic siloxanes, represented only ∼0.22% of the characterized ROC mass but was prioritized by complementary toxicity screening. These findings indicate a pronounced mismatch between emission mass and health hazard in coating-related ROC emissions, suggesting that conventional mass-based VOC metrics may underestimate health-relevant risks. A hazard-prioritized, tiered management framework is therefore proposed to support risk-oriented mitigation and more comprehensive full-volatility ROC regulation.

Long-term side effect surveillance after immunotherapy clinical trial participation in individuals with or at risk of type 1 diabetes is needed. Participants from 14 randomized controlled trials (47%) joined the Long-term Investigative Follow-up in Type 1 Diabetes TrialNet or Immune Tolerance Network Type 1 Diabetes Extension Study, and 98% answered at least one health-related outcomes question. Participants were observed for a median of 1.78 (interquartile range 0.89-4.04) years after original trial completion and, in some, up to 18 years. There were no differences in the frequency of self-reported health outcomes, including moderate to severe hypoglycemia events, hospitalizations, serious infections (including COVID-19 infection), new allergic reactions, autoimmune disease, and cancer incidence, between those receiving active and placebo therapy (n = 209-473 responses per question; P > 0.05 for all). We observed no differences in self-reported health outcomes between those who received active immunotherapy versus placebo. Continued long-term follow-up of immunotherapy trial participants is essential.

Breast density impacts cancer detection by masking tumors within fibroglandular tissue and there are disparities in screening outcomes across racial groups. However, it remains unclear whether these differences reflect inherent tissue characteristics or systemic bias. To isolate the effect of breast density on lesion detectability across racial subgroups, we conducted a retrospective case-control study using raw tomosynthesis projections from 902 women (453 cases, 451 matched controls) across BI-RADS density categories and self-reported race. Identical in-silico spiculated masses (8-15 mm) and microcalcification clusters (10-14 mm) were inserted into the projections using a calibrated lesion model. Images were reconstructed in the same manner to avoid proprietary processing in lesion detection. Lesion detectability was assessed with Channelized Hotelling Observers. Regression and causal mediation analyses examined the relationships between race, density, and detectability. As result, detectability decreased with increasing density; for masses, the area under the receiver operating characteristic curve (ROC AUC) reduced significantly from 0.93 to 0.85 (BIRADS A to D), whereas for microcalcifications AUC decreased from 0.85 to 0.78 across the same density range. Discrimination remained higher for masses than calcifications (AUC = 0.89 vs. AUC = 0.82). Stratified analyses showed slightly higher detectability in Non-Hispanic Black women compared with Non-Hispanic White and Asian American women, largely reflecting differences in density. Mediation analysis revealed that breast density accounted for 38-55% of the observed race-associated detectability differences. Mediation analyses have shown that density is the dominant factor in detectability. These findings support the development of calibrated detection models and personalized screening strategies that account for breast density.

Inflammation in acute respiratory distress syndrome (ARDS) and COVID-19 causes disruption of the alveolar-capillary barrier and tissue damage, which has been associated with increased mortality. Elevated pro-inflammatory plasma biomarker (specifically IL-6) levels reflect the degree of systemic inflammation and are linked to severity of lung injury. Understanding the causal pathway between inflammation, interventions, and patient outcomes could improve clinical care. To assess IL-6 as a mediator of effective interventions in ARDS and COVID-19. We leveraged individual patient data from 5 large randomized controlled trials. Interventions were imatinib, anakinra, low tidal volume ventilation, high positive end-expiratory pressure (PEEP), and simvastatin. We evaluated IL-6 as a mediator for the effectiveness of interventions on 28-day survival with joint modeling. Additionally, we analyzed the role of other pro-inflammatory markers (IL-8, tumor necrosis factor receptor 1, and C-reactive protein). The association between IL-6 and mortality was meta-analyzed using a random effects model. For 2563 patients, IL-6 measures were available at baseline and at least 1 other timepoint. Interleukin-6 mediated the effects of imatinib, anakinra, and low tidal volume on mortality. Higher PEEP and simvastatin did not alter IL-6 trajectories. In all studies, there were associations between higher IL-6 concentrations and increased mortality over 28 days (pooled hazard ratio [HR] for a log10-unit increase = 4.83, 95% CI, 3.50-6.66). The mortality benefit by anakinra and imatinib in COVID-19 and low tidal volume ventilation in acute lung injury may operate through resolution of inflammation defined by a reduction in plasma IL-6 levels.

To comprehensively investigate clinical, molecular, radiological, and surgical factors for prediction of progression-free survival (PFS) in patients with isocitrate dehydrogenase (IDH)-mutant astrocytomas. A total of 210 patients with newly diagnosed World Health Organization (WHO) grade 2-4 IDH-mutant astrocytomas between 2005 and 2023 were included. Clinical, molecular, radiological, and surgical factors were evaluated. Total, contrast-enhancing, nonenhancing, and necrotic tumor volumes were quantified via automatic volumetric segmentation (cm3). Significant predictors of PFS were identified using univariable and multivariable Cox analyses. The median PFS was 106.8 months, with a 5-year PFS rate of 67.5%. On multivariable analysis, higher initial KPS (hazard ratio [HR] =0.76, P = .029) remained as a favorable predictor of PFS, while higher WHO grade (grade 3, HR = 1.01, P = .997; grade 4, HR = 2.91, P = .013; reference standard as grade 2, overall P = .032), larger total tumor volume (HR = 1.03, P = .013), and lesser extent of resection (EOR) (subtotal resection, HR = 1.80, P = .043; partial resection, HR = 2.14, P = .014; biopsy, HR = 3.79, P = .008; reference standard as gross total resection, overall P = .015) remained as unfavorable predictors of PFS. Age and O6-methylguanine-DNA methyltransferase promoter methylation were not significant predictors, while the adverse prognostic impact of WHO grade was primarily driven by WHO grade 4 tumors. Our work presents a detailed analysis of a large series of IDH-mutant astrocytomas, underscoring the prognostic importance of WHO grade 4, tumor volume, and EOR.

Perioperative bleeding is common. Evidence for tranexamic acid (TXA) in urology remains limited and conflicting, and major urologic guidelines provide no recommendations. In an a priori planned analysis, we evaluated TXA among patients undergoing urologic surgery in POISE-3. POISE-3 was an international randomized trial of patients undergoing surgery with bleeding and cardiovascular risk factors. Participants received TXA (1 g intravenous bolus at surgery start and end) or placebo. The primary efficacy outcome was a 30-day bleeding composite (life-threatening/major/critical organ), and the primary safety outcome was a 30-day thrombosis composite (myocardial injury after noncardiac surgery [MINS], nonhemorrhagic stroke, peripheral arterial thrombosis, or symptomatic proximal venous thromboembolism [VTE]). Among 1124 urologic participants (556 TXA and 568 placebo), 489 had laparoscopic/robotic, 360 open, 244 transurethral, and 31 percutaneous surgery. The composite bleeding outcome occurred in 45 (8.1%) with TXA and in 62 (10.9%) with placebo (hazard ratio [HR] = 0.73, 95% confidence interval [CI] = 0.50-1.07). Major bleeding occurred in 34 (6.1%) with TXA and in 54 (9.5%) with placebo (HR = 0.63, 95% CI = 0.41-0.97). The composite safety outcome occurred in 67 (12.1%) with TXA and in 62 (10.9%) with placebo (HR = 1.12, 95% CI = 0.79-1.58; MINS: 62 vs 58; strokes: 2 vs 2; VTEs: 3 vs 3). We identified no interactions by surgical approach, cancer status, or recent antithrombotic usage. Patient-important thrombotic events were few, limiting precision for stroke and VTE estimates. TXA reduced major bleeding and supports perioperative use in urologic surgery, especially when the bleeding risk is high.

The burden of chronic liver disease is rapidly increasing worldwide, driven primarily by metabolic dysfunction-associated steatotic liver disease, metabolic and alcohol-associated liver disease (MetALD), and alcohol-associated liver disease (ALD). Genetic predisposition contributes substantially to variability in disease onset, progression, and outcomes, and recent advances in genomic discovery have brought polygenic risk scores (PRSs) and targeted sequencing closer to clinical relevance. This review summarizes the role of genetic testing in clinical hepatology, including monogenic drivers of disease and the growing role of common variants and PRS. Specific populations, including patients with cryptogenic cirrhosis and lean metabolic dysfunction-associated steatotic liver disease, may be enriched for monogenic drivers of disease. In addition, patients with chronic liver disease may benefit from incorporation of genetic risk scores, including PNPLA3, TM6SF2, HSD17B13, and other key variants in determining risk for fibrosis progression and cirrhosis. Across metabolic dysfunction-associated steatotic liver disease and alcohol-associated liver disease, PRSs demonstrate modest improvements in predicting fibrosis progression and liver-related events, especially when integrated with clinical risk factors and comorbidities. However, their performance remains limited for population-level screening. Similarly, PRS alone has limited diagnostic accuracy for hepatocellular carcinoma and more complex models with clinical features and multiomic biomarkers are likely needed. Emerging therapies targeting PNPLA3 and HSD17B13 variants represent a paradigm shift toward genetically informed treatment. Yet challenges remain, including limited ancestral diversity in genomic datasets, pleiotropic effects of variants, cost-effectiveness, and the need for integration with other omics and electronic medical records. As evidence matures, combining genetic risk with clinical and environmental factors may enable more personalized approaches to prognostication and therapy in liver disease.

Studies reported to date indicate that local therapy may improve outcomes in men with de novo metastatic (M1) prostate cancer (PCa). The aim of this study was to evaluate the effect of local therapy to the primary tumor on progression-free survival (PFS) in men with M1 PCa. In a multicenter, randomized phase 2 trial, men with de novo M1 PCa, after receiving 6 mo of best systemic therapy (BST), were randomly assigned (1:1) to continue BST alone (arm 1) or BST with the addition of either radiotherapy or surgery to their primary tumor (arm 2). The primary endpoint was PFS, defined as the time from randomization to progression (Prostate Cancer Working Group 2) by prostate-specific antigen, radiographic or symptomatic progression, or the time to change systemic therapy as per physician discretion and/or clinical decision, or death, whichever occurred first. Immunohistochemistry for the tumor suppressors p53, RB1, and PTEN was performed on available prostate biopsies at baseline and 6 mo. The aggressive variant PCa molecular signature (AVPC-MS) was assigned if two or more of these tumor suppressors were abnormal. In the current intent-to-treat analysis, the Kaplan-Meier product-limit method was used to estimate the median PFS. Between March 2013 and April 2018, 119 patients were randomized (arm 1: 59 and arm 2: 60). The median follow-up for patients who survived was 66 mo (64 mo for BST alone and 67 mo for BST plus local therapy groups). BST included androgen deprivation therapy (n = 119), with docetaxel (n = 37) or with androgen receptor pathway inhibitor agents (n = 9). Local therapy included surgery (n = 45), radiation (n = 13), or none (n = 2). At data analysis, 88 patients met Prostate Cancer Working Group 2 progression, and 53 patients had died. The median PFS was 17.9 mo (95% confidence interval [CI] 11.7-36.4) in arm 1 and 14.8 mo (95% CI 11.4-42.9) in arm 2 (hazard ratio [HR] 0.89, 95% CI 0.59-1.34, p = 0.6). Toxicity was limited in both arms, with grade 3 toxicities in four patients (6.7%) in arm 2 and zero patients in arm 1. Three patients required palliative intervention for symptomatic local progression in arm 1, while an additional six patients crossed over to receive local therapy after meeting castration-resistant PCa progression criteria. Predictors of worse overall survival (OS) for all comers included CHAARTED high-volume (HR 1.84, 95% CI 1.06-3.19) and clinical cT3b/T4 (HR 1.97, 95% CI 0.88-4.41) disease. Having the AVPC molecular profile (AVPC-MS) at baseline or 6 mo was significantly associated with worse PFS (HR 1.74, 95% CI 1.02-2.98, p = 0.04). However, there was no statistically significant association with OS (HR 1.83, 95% CI 0.94-3.56, p = 0.08). This phase 2 randomized study failed to demonstrate improved PFS in men with de novo M1 PCa treated with BST with the addition of local therapy to their primary tumor. Its effect on OS is being tested in an ongoing phase 3 trial (SWOG 1802). We identified biomarkers of potential prognostic value (CHAARTED volume status, cT3b/cT4 disease, and the AVPC-MS) that may serve to optimize therapy selection and stratification in this population, but these require further evaluation.

The interplay between somatic mutations and copy number alterations influences tumor evolution and prognosis. These alterations are often treated independently, overlooking gene mutant dosage (GMD)-a key property of their interaction. Here we develop a computational framework that infers mutation copy number and multiplicity from targeted sequencing panels without requiring matched normal samples. We derive GMD for over 500,000 mutations across 60,000 pan-cancer samples. By stratifying more than 20,000 patients according to GMD across multiple genes, we identify 46 tumor-type-specific biomarkers predictive of survival, 13 of which were undetectable using binary mutant/wild-type models, 26 were associated with metastatic spread and 20 predicted metastatic tropism. Our method reveals GMD patterns as independent predictors of disease prognosis, metastatic potential and site-specific dissemination across diverse tumor types. This augmented insight into genomic drivers enhances our understanding of cancer progression and metastasis and holds the potential to substantially enhance biomarker discovery.

MYC rearrangement (MYC-R) is a known adverse prognostic factor in multiple myeloma (MM), yet its role in the double-hit model remains undefined. Meanwhile, the prognostic value of MYC gain/amp has not been clearly characterized. This study aimed to evaluate the prognostic impact of MYC-R and/or gain/amp to assess their potential role in refining risk stratification. A prospective cohort of 227 patients with newly diagnosed MM (enrolled in the NICHE clinical trial, NCT04645199) received bortezomib-based induction therapy and, if eligible, underwent upfront autologous stem cell transplantation. The FISH panel included del(13q), del(17p), del(1p), 1q21 gain/amp, IgH rearrangement, t(4;14), t(11;14), t(14;16), t(14;20), MYC break-apart probe, and IgH/MYC fusion probe. Median progression-free survival (PFS) was significantly shorter in patients with MYC-R (34.0 months) or MYC gain/amp (34.3 months), compared with those without MYC abnormalities (49.2 months; P = 0.003 and 0.016, respectively). IgH/MYC was notably more common in cases with undefined-partner IgH translocations (16.6%) than those with defined partners (5.7%), and was associated with poorer outcomes. In multivariate analysis, MYC-R remained an independent predictor of PFS (hazard ratio 1.96, P = 0.007) and improved the discriminative performance of the double-hit model by increasing the concordance index for PFS from 0.549 to 0.581. Both MYC-R and MYC gain/amp indicate adverse prognosis in newly diagnosed MM. The independent prognostic value of MYC-R supports its integration into routine FISH panels and future refinements of the double-hit risk model.

Transarterial chemoembolization (TACE) is the standard of care for liver-confined hepatocellular carcinoma (HCC) that is not amenable to curative treatment; however, TACE has demonstrated unsatisfactory survival benefits. To evaluate whether combining TACE with subsequent thermal ablation improves clinical outcomes compared with TACE alone in patients with liver-confined unresectable HCC. The open-label, phase 3 TORCH randomized clinical trial was conducted from May 2015 to August 2024 at 2 tertiary medical centers in China. Patients with Barcelona Clinic Liver Cancer stage B HCC were enrolled. The data cutoff was October 31, 2025. Patients were randomly assigned (1:1) to receive either TACE combined with subsequent selective radiofrequency ablation (TACE-ablation) or TACE alone. The primary end point was progression-free survival (PFS), assessed per Response Evaluation Criteria in Solid Tumors (RECIST), version 1.1. Secondary end points included overall survival (OS), treatment response and PFS per modified RECIST, untreatable PFS, and safety. Among 241 patients included in the intention-to-treat population, 121 received TACE-ablation (median [IQR] age, 59.0 [51.0-66.0] years; 108 [89.3%] male), and 120 received TACE alone (mean [IQR] age, 58.0 [50.0-64.0]; 106 [88.3] male). The number of patients with 6-and-12 tumor burden scores of lower than 6, 6 to 12, and more than 12 points were 34 (28.1%), 79 (65.3%), and 8 (6.6%) in the TACE-ablation group and 31 (25.8%), 76 (63.3%), and 13 (10.8%) in the TACE alone group, respectively. At the data cutoff, the median PFS per RECIST, version 1.1, was 17.7 months (95% CI, 11.4-23.1 months) in the TACE-ablation group vs 7.3 months (95% CI, 6.4-10.4 months) in the TACE alone group (hazard ratio [HR], 0.47; 95% CI, 0.34-0.65; P < .001). TACE-ablation also resulted in statistically significant prolonged untreatable PFS compared with TACE (35.1 months vs 12.3 months; HR, 0.40; 95% CI, 0.27-0.58; P < .001). Median OS was 88.6 months (95% CI, 43.1 months to not estimable) with TACE-ablation and 35.1 months (95% CI, 25.4-45.5 months) with TACE alone (HR, 0.50; 95% CI, 0.34-0.73; P < .001). Clinically meaningful improvements in both PFS and OS were observed in patients with low to moderate tumor burden scores (≤6 and 6-12 points). Grade 3 and 4 treatment-related adverse events occurred in 23 patients (23.2%) in the TACE-ablation group and 24 (18.3%) in the TACE alone group. In this phase 3 randomized clinical trial, TACE combined with subsequent thermal ablation demonstrated superior survival outcomes than TACE alone in patients with liver-confined unresectable HCC. Sequential TACE-ablation could serve as a feasible treatment option for such patients. ClinicalTrials.gov Identifier: NCT02435953.

Colorectal cancer (CRC) cells are addicted to iron, which fuels nucleotide synthesis, mitochondrial respiration, and proliferation. Yet paradoxically, high intracellular iron is cytotoxic to most cells, raising the question of how CRC cells tolerate and exploit iron-rich environments. Ferroptosis, an iron-dependent form of cell death, is thought to mediate iron toxicity. However, whether most ferroptosis regulators, identified through synthetic chemical screens or small molecule activators, play a role in modulating iron toxicity, particularly in vivo, remains unclear. Here, using multi-omics profiling, CRISPR screening, and in vivo models, we uncover a heme-succinate dehydrogenase (SDH)-coenzyme Q (CoQ) axis that enables CRC cells to buffer iron-induced oxidative stress. Heme-dependent SDH reduces CoQ, which redistributes to mitochondrial and plasma membranes to detoxify lipid reactive oxygen species (ROS) as a radical-trapping antioxidant. These findings reveal that CRCs co-opt metabolic cofactors both for growth and for survival under physiologically toxic iron levels, uncovering new vulnerabilities for therapy.

Steatotic liver disease (SLD) comprises a heterogeneous group of liver diseases defined by pathological hepatic lipid accumulation with varying degrees of steatohepatitis and progressive fibrosis, which may culminate in cirrhosis and/or hepatocellular carcinoma. SLD encompasses metabolic dysfunction-associated steatotic liver disease (MASLD), formerly called non-alcoholic fatty liver disease (NAFLD); MetALD, describing patients with MASLD consuming moderately high amounts of alcohol; and alcohol-associated/related liver disease (ALD). In addition, SLD encompasses less common aetiologies, including drug-induced and monogenic causes, as well as cryptogenic disease, which lacks metabolic risk factors or an identifiable cause. As the leading cause of chronic liver disease worldwide, MASLD, including metabolic dysfunction-associated steatohepatitis (MASH), is a complex multisystem disorder associated with extrahepatic organ dysfunction. Consequently, MASLD has become a major focus of multidisciplinary research, driving innovation across hepatology, immunology, cardiometabolism, addiction medicine, nutrition, prevention, and beyond. Over the past two decades, a broad array of in vivo, in vitro, and ex vivo experimental models have been developed to recapitulate diverse aspects of SLD pathophysiology, genetics, inflammation, treatment response, and multi-organ crosstalk, substantially advancing mechanistic understanding and therapeutic development. However, the rapid expansion and heterogeneity of available models have also highlighted the need for improved categorisation, standardisation, and harmonisation in line with evolving disease definitions and clinical concepts. In this EASL position paper, we critically review and classify currently available experimental SLD models and propose key criteria required for their appropriate use across distinct SLD subtypes. These criteria encompass systemic and hepatic metabolism, cardiometabolic comorbidities, histopathology, immunopathology, and molecular features relevant to disease stage and aetiology. This position paper aims to guide informed model selection, promote consistent nomenclature, and enhance rigour and translational relevance in preclinical and experimental SLD research.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal human cancers, mostly due to its insidious onset that consequently leads to diagnosis at advanced stages. The complex biology of PDAC limits effective therapeutic options, yet over 90% of cases harbor KRAS mutations, making it a compelling therapeutic target. KRAS had a notorious history of being "undruggable." Emerging novel inhibitors, however, have fundamentally shifted this paradigm by demonstrating the possibility of direct KRAS targeting. Although promising in preclinical efficacy, the clinical benefit of these inhibitors was frequently curtailed by rapid and heterogeneous resistance due to the intrinsic complexity of PDAC. Concurrently, building on structure-based drug discovery, artificial intelligence (AI) is now infusing this traditional workflow with dynamic and extensive enhancements. AI-enabled approaches in molecular biology-including high-accuracy protein structure prediction, large-scale virtual screening, and the uprising of generative molecular design-have accelerated the discovery of potent and selective molecular structures. Comprehensive hybrid models and quantum-level data processing enabled the exploration of previously latent chemical spaces, thereby creating more opportunities for the search for effective PDAC therapies. Discussions of progress in KRAS inhibitors and in AI drug discovery are rising but remain largely in parallel. As a result, this review aims to bridge these two exciting fields together. Through discussions of novel strategies to target KRAS in PDAC, emerging paradigms to address KRAS resistance in clinical trials, and current innovations in AI algorithms for molecule screening and design, we hope to underscore the unique challenges and prospects of leveraging artificial intelligence to accelerate the discovery of effective therapies for PDAC.

Radiotherapy, while a cornerstone treatment for esophageal squamous cell carcinoma (ESCC), is paradoxically associated with significant weight loss that portends poor patient outcomes. The mechanisms driving this metabolic complication remain elusive. Here, we identified adipose depletion - rather than muscle atrophy - as the primary contributor to radiotherapy-induced weight loss in ESCC. We demonstrated that secretory autophagosomes (SAPs) released post-irradiation mediate systemic fat loss through integrated in vitro and in vivo studies. Proteomic profiling revealed enrichment of PBK (PDZ binding kinase) in radiation-induced SAPs, with functional studies establishing PBK as the master regulator of adipocyte lipolysis. Mechanistically, SAP-delivered PBK activated MAPK1/ERK2 (mitogen-activated protein kinase 1), triggering a downstream PRKA/PKA-LIPE/HSL signaling cascade that increases lipolytic rate. Clinically, elevated circulating SAPs levels predicted severe fat loss and reduced median survival in a ESCC cohort. Critically, pharmacological inhibition of PBK with OTS-514 rescued adipose mass in preclinical models while enhancing tumor radiosensitivity. Our work redefines radiotherapy-induced cachexia as an adipose-centric process orchestrated by SAPs, unveils PBK as a therapeutic target, and provides actionable biomarkers for early intervention. These findings bridge the gap between localized radiotherapy and systemic metabolic sequelae, offering a dual-strategy approach to improve both survival and quality of life in ESCC patients.Abbreviations: ADSCs: adipose-derived stem cells; CM: conditioned media; ESCC: esophageal squamous cell carcinoma; EVs: extracellular vesicles; eWAT: epididymal white adipose tissue; GA: gastrocnemius muscle; iWAT: inguinal white adipose tissue; LIPE/HSL: lipase E, hormone sensitive type; LIR: LC3-interacting region; MAP2K1/MEK1: mitogen-activated protein kinase kinase 1; MAPK/ERK: mitogen-activated protein kinase; OS: overall survival; PBK: PDZ binding kinase; PRKA/PKA: protein kinase cAMP-dependent; RT: radiotherapy; SAPs: secretory autophagosomes; sEVs: small extracellular vesicles.

Glioblastoma (GBM) is highly heterogeneous, complicating effective tumor elimination. Among GBM subtypes, the mesenchymal (MES) variant is associated with the poorest prognosis. Notably, macrophage infiltration is significantly higher in MES GBM. However, the regulatory networks underlying this distinct MES GBM microenvironment remain poorly understood. Here, we demonstrate that MES glioblastoma stem cells (GSCs) exhibit a preferential expression and secretion of the protein LY96. Mechanistically, MES GSCs engage an autocrine LY96-TNFRSF1B signaling axis that activates NF-κB, thereby promoting their proliferation, self-renewal, and maintenance of the MES state. Inhibition of LY96 in vivo suppresses GBM growth and prolongs survival in animal models. Additionally, LY96 acts in a paracrine manner on tumor-associated macrophages (TAMs), driving their polarization toward a MES-like state. Collectively, our findings identify LY96 as a key regulator of the MES state in GSCs and a driver of MES-like polarization in TAMs. Therapeutic strategies targeting LY96 and its downstream effectors may improve GBM treatment outcomes.

Beyond established rare fusions, such as ALK and ROS1, emerging ultrarare fusions involving receptor tyrosine kinases or their ligands, including EGFR-SHC1, further guide us to uncover novel mechanisms of oncogenic activation and corresponding treatment strategies. Collectively, rare and ultrarare genomic events are driving precision oncology toward an increasingly individualized era of "ultraprecision" cancer therapy. See related article by Zheng et al., p. 1573.

The FDA has launched a pilot program to establish real-time clinical trials that includes two proof-of-concept clinical trials that will report endpoints and data signals in real time. AstraZeneca is conducting TrAVeRse, a phase II trial in treatment-naïve mantle cell lymphoma, and Amgen is planning the phase Ib STREAM-SCLC study in limited-stage small cell lung carcinoma.

At this year's ASCO Annual Meeting, investigators also presented encouraging early clinical data for several KRASG12D-selective inhibitors, including RNK08594, GFH375, and DN022150, suggesting that this type of drug could play a role in the treatment of several solid tumors, such as pancreatic ductal adenocarcinoma and non-small cell lung cancer.

A 14-protein blood test can identify people at high risk of lung cancer more than 5 years before a tumor becomes detectable on imaging, including never-smokers who fall outside current screening criteria. In a retrospective analysis, the test also identified individuals most likely to benefit from the anti-IL-1β drug canakinumab, which roughly halved lung cancer incidence in the high-risk group but not in low-risk participants.

The treatment arsenal for small cell lung cancer could soon include antibody-drug conjugates, with early data from three phase I studies indicating robust response rates to CD56-targeting DXC006, DLL-3-targeting BL-M14D1, and B7-H3-targeting SYS6043. The latter also appears active in other tumor types, including ovarian and breast cancers. As well, phase II findings on a fourth ADC candidate, HER2-targeting trastuzumab brengitecan, suggest its strong efficacy in platinum-resistant ovarian cancer.

Stromal immunosuppressive pathways are key modulators of response to immune checkpoint inhibitors, but the tumor-intrinsic consequences of blocking these pathways remain incompletely defined. We conducted a clinical trial of bintrafusp alfa, a bifunctional PD-L1/TGFβ inhibitor, in small cell lung cancer (SCLC). Among 34 evaluable patients, 18% had partial responses, 20% stable disease, and 62% progressive disease; 38% of progressors met the criteria for hyperprogressive disease (HPD). HPD was also observed across other tumor types (n = 450), in higher frequencies with bintrafusp alfa than PD-(L)1 blockade alone. Blood and tumor profiling showed that HPD correlated with systemic immune suppression and elevated TGFβ signaling. Functional studies demonstrated that tumor-intrinsic TGFβ signaling restrains proliferation in a subset of SCLC; pathway blockade triggers hyperproliferation. External validation across cell lines and tumor samples confirmed a tumor-intrinsic TGFβ-high transcriptional state associated with inferior survival. These findings identify a context-dependent, growth-constraining function of TGFβ and support tumor-intrinsic biomarker guidance while targeting stromal immunosuppressive pathways. This study identifies tumor-intrinsic TGFβ signaling as a context-dependent growth restraint in SCLC and a driver of HPD following TGFβ blockade. A reproducible TGFβ-high mesenchymal state is linked to inferior survival, supporting biomarker-guided use of TGFβ-targeted immunotherapy.

Although epidermal growth factor receptor (EGFR) fusions in non-small cell lung cancer (NSCLC) typically show sensitivity to tyrosine kinase inhibitors (TKI), we identified an EGFR-SHC1 fusion subtype that exhibits intrinsic resistance to EGFR-TKI monotherapy through a dual-activation mechanism in the preclinical and clinical settings. EGFR-SHC1 fusion protein comprises of N-terminal EGFR and C-terminal SHC1. We demonstrated that EGFR-SHC1 simultaneously activates the EGFR kinase domain (KD) and SRC-mediated phosphorylation of the SHC1 fusion partner, thereby driving ERK/AKT pathway activation and tumorigenesis independent of KD inhibition. Structural modeling coupled with domain-specific mutagenesis revealed that SHC1 phosphorylation establishes a kinase-independent bypass mechanism. Notably, dual-targeted inhibition using afatinib (EGFR-TKI) in combination with dasatinib (SRC-TKI) induced marked tumor regression in a TKI-refractory patient with NSCLC with EGFR-SHC1. This study illustrates a cooperative oncogenesis between kinases and scaffold proteins in fusions, providing a clinically actionable strategy for overcoming TKI resistance in patients with these oncogenic fusions. This study identifies a previously unrecognized mode of oncogenic signaling in receptor tyrosine kinase (RTK) fusions, in which a non-kinase fusion partner actively drives tumorigenesis through kinase-independent mechanisms. By challenging the classical kinase-centric model of RTK fusions, it provides a framework for understanding intrinsic resistance to TKIs and for developing rational combination therapies. See related commentary by Le and Wolf, p. 1480.

Lung adenosquamous carcinoma (ASC) is a rare histological subtype of non-small cell lung cancer (NSCLC), and prospective data regarding the efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) in this patient population remain scarce. This prospective, multicenter, single-arm phase II trial (NCT04354961) evaluated the safety and efficacy of aumolertinib as first-line treatment for treatment-naïve patients with stage IV EGFR-mutant ASC. Eligible patients received aumolertinib 110 mg orally once daily, with progression-free survival (PFS) as the primary endpoint and objective response rate (ORR), disease control rate (DCR), overall survival (OS), and safety as secondary endpoints. The study was terminated early due to slow accrual, with 12 patients enrolled (median age: 66 years; 58.3% male; 58.3% with EGFR exon 19 deletion). After a median follow-up of 29.0 months, the median PFS and OS were 11.1 months (95% CI, 4.27-NA) and 16.7 months (95% CI, 11.3-NA), respectively. Confirmed ORR and DCR were 58.3% and 83.3%. Treatment-related adverse events (TRAEs) occurred in 75% of patients, with 33.3% experiencing grade 3-4 TRAEs; no treatment-related deaths were observed. Aumolertinib exhibits clinical activity with a manageable safety profile in EGFR-mutant ASC patients. However, its reduced efficacy compared with EGFR-mutant adenocarcinoma highlights the need for histology-specific therapeutic strategies. Trial Registration: NCT04354961.

Stereotactic body radiation therapy (SBRT) is effective for early-stage non-small cell lung cancer (NSCLC), but the treatment of ultra-central NSCLC (UCNLC) near critical structures remains challenging. This retrospective study analyzed 66 patients with stage I-II unresectable UCNLC who underwent SBRT at Shanghai Pulmonary Hospital (2020-2023). UCNLC was defined by planning target volumes (PTVs) abutting/overlapping vital structures (proximal bronchial tree, esophagus, heart, great vessels, pulmonary vessels) or internal target volumes (ITVs) within 1 cm of these organs. Tailored SBRT plans with non-uniform PTV margins were used in 45.5% of cases, enabling 53.3% of patients to receive a biologically effective dose ≥ 100 Gy. With a median follow-up of 32.2 months, the median progression-free survival (PFS) was 42.9 months; 1- and 3-year PFS rates were 93.9% and 68.8%, respectively. Median local control (LC) and overall survival (OS) were not reached, with 1- and 3-year LC rates of 98.5% and 84.6%, and OS rates of 98.5% and 88.7%, respectively. Toxicities were predominantly grade 1-2, with only one grade 3 pneumonitis. Larger ITV predicted poorer LC and PFS, whereas T stage independently predicted PFS and OS. Therefore, a non-uniform PTV strategy maintained clinical efficacy without compromising safety, supporting its application in UCNLC management.

Liquid biopsy is a promising, noninvasive approach for cancer detection, but current methods often trade off accuracy, operability, and cost. To address these limitations, we introduce an artificial perception system (APS) for liquid biopsy that combines a structurally defined DNA-carbon nanotube sensor array with machine learning (ML) models. The array produces multichannel fluorescence fingerprints from serum, which are decoded by ML models to classify disease state. In a total of 253 serum samples spanning liver, lung, and ovarian cancers and noncancer controls, the APS achieved mean sensitivity of 89% and specificity of 96%. Notably, early-stage lung cancer was detected with 92% sensitivity and 95% specificity at an estimated cost of ∼$4 USD per test. Insights from SHAP analysis and Mantel test revealed the detection mechanisms of APS, supporting biological plausibility and clinical translation. These results highlight a path toward accurate, scalable, and affordable multicancer detection and early cancer screening.

Circulating tumor antigens (ctA; tumor markers) are blood-based proteins that can offer prognostic value in non-small cell lung cancer (NSCLC) and may serve as potential early surrogates for survival. Given the significantly reduced testing time and cost of ctA compared with circulating tumor DNA (ctDNA), we further explored the utility of ctA using samples collected from over 2,300 patients participating in five clinical trials (IMpower130, 131, 132, 150, and 110). We analyzed a panel of six ctA (CA125 (cancer antigen 125), CEA (carcinoembryonic antigen), Cyfra21-1 (cytokeratin 19 fragment 21-1; CYFRA), NSE (neuron-specific enolase), SCC (squamous cell carcinoma antigen), and ProGRP (progastrin-releasing peptide)) and CRP (C-reactive protein) from the serum of patients with metastatic NSCLC in these trials, which investigated combinations of atezolizumab (anti-programmed death-ligand 1)±bevacizumab±chemotherapy. Previous work showed that an optimized cut-off using two ctA or a machine learning (ML) model of ctDNA features, both taken at 6 weeks, can stratify patients with stable disease (SD) for survival risk in IMpower150. Building on this approach, we applied an ML model combining ctA features at baseline and at 6 weeks, trained across a much larger aggregate dataset from multiple clinical studies. Previous findings from ctA analysis of IMpower150 were confirmed and found to be applicable to several other trials analyzed in this study. We found that ML model predictions provided similar prognostic performance (c-index of 0.73 and 0.71 in squamous and non-squamous test datasets, respectively), with CYFRA being the top feature for both histologies. While ctA demonstrated limited potential in differentiating treatment effects to inform early drug development, deriving an optimal prediction cut-off for 1-year overall survival (OS) showed that ctA model predictions could effectively stratify patients by radiographic response with 61% sensitivity and 78% specificity, adding significant prognostic value to radiographic imaging. Patients with partial response (PR), progressive disease (PD), or stable disease (SD) at either 6 weeks of treatment or best confirmed overall response could be separated into low-risk or high-risk groups for OS.

Therapeutic resistance remains a major challenge in cancer treatment, driven by compensatory signaling and stress response pathways that sustain tumor survival. Topoisomerase IIβ-binding protein 1 (TopBP1), a multifunctional scaffold protein with nine BRCT domains, integrates replication stress signaling with oncogenic networks and is frequently overexpressed in aggressive cancers. Its BRCT7/8 domains mediate critical interactions with E2F1, mutant p53, MIZ1, PLK1, and CIP2A, making TopBP1-BRCT7/8 an attractive therapeutic target. Using docking-guided screening and structure-activity relationship-driven optimization, we developed CS18 as a potent and selective BRCT7/8 inhibitor that disrupts oncogenic TopBP1 complexes without interfering with DNA replication. CS18 suppresses MYC transcriptional programs, restores E2F1-mediated apoptosis, and induces mitotic catastrophe. It exhibits broad-spectrum anticancer activity and synergizes with poly(ADP-ribose) polymerase (PARP) inhibitors in multiple cancer types and enhances osimertinib sensitivity in EGFR-mutated non-small cell lung cancer (NSCLC) cells. CS18 demonstrates efficacy in patient-derived breast cancer xenografts and overcomes osimertinib resistance in refractory NSCLC in vivo. These findings establish CS18 as a chemically distinct TopBP1 inhibitor with translational potential to overcome therapeutic resistance and advance precision oncology.

Hypothesis generation in biomedicine is constrained by human cognitive limitations in synthesizing insights from fragmented biomedical knowledge and multimodal data sources. Here we introduce XunZi, an AI biologist that integrates logical reasoning and multimodal data fusion to autonomously generate de novo therapeutic target hypotheses with testable mechanisms. XunZi has been trained on 24.4 million publications and 613.6 TB of multisource data spanning 21,008 human genes and 5,850 diseases, and outperforms existing methods in both accuracy and interpretability across diverse disease contexts. In Parkinson's disease (PD), where complex mechanisms and limited targets hamper therapy development, XunZi identifies aberrant activation of CHK2 and IRAK4 kinases across multiple models. Pharmacological or genetic inhibition of Chk2 rescues dopaminergic neuron loss and motor deficits in PD mice. We further demonstrate XunZi's broad versatility in diseases such as non-small-cell lung cancer. XunZi establishes a paradigm-shifting framework to translate fragmented biomedical knowledge and data into actionable therapeutics.

Mitochondria are semi-autonomous organelles whose functions critically depend on nucleus-encoded proteins. TOM40 is the core β-barrel protein of the translocase of the outer mitochondrial membrane (TOM) complex that mediates the import of most nucleus-encoded mitochondrial proteins. Here, we show that the small GTPase RAB32 facilitates the mitochondrial localization of TOM40 and the mitochondrial protein homeostasis in non-small cell lung cancer (NSCLC) cells. Accordingly, knockout of RAB32 results in mitochondrial dysfunction and inhibits NSCLC progression in xenograft and autochthonous NSCLC mouse models. We further identify ubiquitin-specific peptidase 13 (USP13) that removes the K48-linked polyubiquitin chains from RAB32 to prevent its proteasomal degradation. Consistently, knockout of USP13 causes destabilization of RAB32, impairs TOM40 mitochondrial localization and mitochondrial function, and inhibits NSCLC progression, which are restored by reconstitution of wild-type USP13 or RAB32, but not the catalytically inactive USP13C345A/M664/739E. Our study has revealed a previously uncharacterized RAB32-USP13 axis for mitochondrial functions and NSCLC progression.

Although non-small cell lung cancer (NSCLC) is among the most prevalent malignancies, currently available treatments remain largely ineffective, highlighting the need to elucidate the precise mechanisms of tumorigenesis. The Mpr1/Pad1 N-terminal domain-containing protein (MPND) is a member of the JAMM family of deubiquitinases. In this study, we found that MPND is frequently deleted, and its expression is significantly downregulated in NSCLC tumor tissues. Low MPND levels correlated with poor clinical outcomes, and depletion of MPND enhanced cell migration, invasion, and cancer stem cell-like characteristics in NSCLC cells. Mechanistically, MPND interacted with histones, removing the ubiquitin moiety from monoubiquitinated histone H2A K119 and H2B K120 (H2B K120ub). Loss of MPND elevated the ubiquitination marks, leading to alterations in chromatin architecture. Furthermore, MPND modulated gene transcription, and its depletion activated the transforming growth factor-beta (TGFβ)/SMAD3 signaling pathway. Specifically, the knockout of MPND increased H2B K120ub and chromatin accessibility at the SMAD3 locus, facilitating transcriptional activation. In mouse tumor models and clinical samples from patients with NSCLC, the loss of MPND consistently triggered activation of the TGFβ/SMAD3 axis, thereby promoting tumor growth and metastasis. Together, these data reveal an epigenetic pathway underlying NSCLC progression and suggest that targeting the TGFβ/SMAD3 axis may be particularly effective for the treatment of MPND-deficient NSCLC tumors. Loss of MPND in lung cancer promotes tumor growth and metastasis by increasing histone ubiquitination to epigenetically activate TGFβ/SMAD3 signaling, suggesting that targeting this axis may provide therapeutic benefit in MPND-deficient tumors.

Macroautophagy/autophagy is a well-established homeostatic mechanism that contributes to the integrity of multiple regulatory biological activities including but not limited to the gastro-intestinal tract and cognitive integrity. Autophagy also plays a central role in tissue regeneration, metamorphosis and development whereas defects in autophagy are associated with a wide range of disorders including metabolic diseases such as diabetes, organ pathophysiologies including liver, lung and heart disease, cancer, and microbial infection. In the field of cancer therapy, most research efforts have focused on cytoprotective autophagy, with substantial preclinical and clinical studies designed to interrogate the outcomes of pharmacologically (or genetically in preclinical work) inhibiting autophagy to enhance the efficacy of chemotherapeutic agents. There is lesser but nevertheless robust evidence for the cytotoxic function of autophagy while our laboratory and a few others have identified the nonprotective form of this cellular response. However, cytostatic autophagy, a distinct functional outcome of autophagy characterized by sustained proliferative arrest, has remained relatively underexplored. Cytostatic autophagy can be defined as a cellular condition in which autophagy activation coincides with durable proliferative arrest, and in which genetic or pharmacological inhibition of autophagy relieves the growth-arrest phenotype without inducing overt cytotoxicity. In this review, we provide the first comprehensive synthesis of the scientific literature addressing cytostatic autophagy, tracing its historical development and consolidating the experimental evidence that led to its current conceptual definition. We further discuss the molecular mechanisms underlying cytostatic autophagy, including the selective degradation of key cell-cycle regulators and the interplay between autophagy and senescence-associated signaling pathways.Abbreviations: CQ, chloroquine GBM, glioblastoma; HCQ, hydroxychloroquine; LV, left ventricular; NRVMs, neonatal rat ventricular cardiomyocytes; NSCLC, non-small cell lung cancer; OIS, oncogene-induced senescence; PI3K, phosphoinositide 3-kinase; PtdIns3K, phosphatidylinositol 3-kinase; PtdIns3P, phosphatidylinositol-3-phosphate; SASP, senescence-associated secretory phenotype; TASCC, TOR-autophagy spatial coupling compartment; TMZ, temozolomide.

Precise segmentation of medical images plays a crucial role in modern clinical practice, providing important foundations for the quantitative analysis of medical images and clinical decision making. However, although deep learning techniques have achieved significant success in conventional medical image segmentation, they still exhibit obvious limitations when faced with complex structure segmentation tasks such as colorectal cancer: high-quality medical data acquisition is not only difficult, but also even when data is relatively sufficient, the diverse morphological features of lesions cannot be adequately represented due to their high variability, which severely limits the generalization capability of traditional data-driven segmentation methods. To address these challenges, we propose a feature-driven segmentation model that improves performance by deeply mining intrinsic data information rather than relying on parameter stacking. Specifically, we introduce a spectrum-prior-boundary triple modeling paradigm, where frequency domain reconstruction and modulation is employed to establish mappings between different frequency bands and heterogeneous signals to identify ambiguous signals, a level set-based segmentation algorithm is used to construct abdominal anatomical distance fields to incorporate morphological priors, and an auxiliary edge branch is designed by integrating deep semantic and shallow detail features to strengthen boundary awareness. Extensive experiments on colorectal cancer segmentation demonstrate that the proposed method achieves significant improvements in both segmentation accuracy and boundary perception over other state-of-the-art methods. Furthermore, evaluations on lung cancer and breast cancer segmentation tasks validate its strong generalization capability across diverse lesion types.

Leptomeningeal metastasis from a solid tumor carries a dismal prognosis with median survival of 3-5 months after diagnosis and treatment with external beam radiation therapy. It targets the whole-brain or the entire central nervous system depending on disease extent, and is the current standard therapeutic approach, however, its effectiveness is limited by insufficient cytotoxicity and severe off target radiotoxicity. Radiopharmaceutical therapy is a promising strategy to deliver therapeutic radiation doses directly to the leptomeningeal space while minimizing damage to the normal central nervous system tissue. In this study, the tumor targeting alkylphosphocholine (NM600) was radiolabeled with 177Lu for SPECT/CT imaging and 225Ac for therapeutic efficacy in the context of solid tumor leptomeningeal metastasis. In in vitro experiments 225Ac-NM600 induced significantly greater apoptosis, reduced colony formation, and increased expression of DNA damage markers, compared to 177Lu or external beam radiation. In vivo therapeutic and toxicity profiles were assessed in leptomeningeal metastasis models of triple negative breast cancer and non-small cell lung cancer in NSG mice. Intrathecal administration of 177Lu- or 225Ac-NM600 demonstrated excellent tumor targeting with minimal off target effect, confirmed by SPECT/CT imaging and ex vivo biodistribution. 225Ac-NM600 was well tolerated, inhibited tumor growth, and significantly improved overall survival in both models. Moreover, central nervous system radiotoxicity was markedly lower with 225Ac-NM600 RPT compared to external beam radiation therapy. Compared with whole brain external beam radiation therapy, 225Ac-NM600 provides superior anti-tumor efficacy and reduced off target toxicity supporting its further evaluation in clinical trial.

Rapid and reliable detection of EGFR exon 19 deletion (19del) is important for precision treatment of non-small cell lung cancer, yet current genotyping methods remain poorly suited for point-of-care testing (POCT). This limitation is particularly significant in signal-off assays, where reaction failure can be misinterpreted as a true negative result. Here, we present a dual-site personal glucose meter (PGM)-readout platform for minimal-instrumentation analysis of EGFR 19del. In this system, target-dependent flap endonuclease 1 (FEN1) cleavage generates adenosine monophosphate (AMP), which triggers a glucose-consuming kinase cascade and enables direct quantitative readout using a commercial PGM without optical instrumentation or nucleic acid amplification. To improve interpretability, the platform incorporates an endogenous internal control through a dual-site design consisting of a mutation-discriminating detection site and a conserved control site, thereby enabling reliable interpretation of signal-off readouts while confirming sample adequacy. The platform achieved sensitive detection with a limit of detection of approximately 27 pM and discriminated mutation fractions down to ∼0.44% in wild-type/mutant mixtures. Its feasibility was further demonstrated using cell-derived genomic DNA and serum-containing samples. These results indicate that the proposed platform provides an accessible and practical strategy for decentralized EGFR 19del testing.

Tumor-associated macrophages (TAMs) have emerged as a promising immunotherapeutic target in non-small cell lung cancer (NSCLC). However, the indiscriminate cytotoxicity of chemotherapies and the immunosuppressive tumor microenvironment paradoxically impede this potential. To overcome these limitations, we engineered β-selenoester-crosslinked nanocapsules delivering Gefitinib, designed to induce opposite cell fates in cancer cells and macrophages. In cancer cells, the Se-C bond in β-selenoester is ultrasensitive under the intrinsic reactive oxygen species (ROS) level. It generates acrylates through selenoxide elimination reaction, which further depletes intracellular GSH to regenerate cytotoxic ROS. The ROS-triggered positive-feedback induces nanocapsule disassembly, enabling rapid Gefitinib release and apoptosis induction. The released Gefitinib also disrupts the CD47-SIRPα "don't eat me" axis to enhance macrophage phagocytic activity. In macrophages, low ROS level limits Gefitinib exposure, but the selenium metabolites generated from the elimination reaction are sufficient to promote macrophage activation. This selective cell fate programming yielded no macrophage toxicity at cancer-cell IC50 levels and a 91.1 % tumor suppression in vivo. Collectively, this work demonstrates a divergent cell fate induction strategy based on β-selenoester-crosslinking for integrated TAM-mediated immunotherapy.

Mutations in tumor suppressor p53 that gain oncogenic functions (Onc-p53) are frequent in lungs and many other solid tumors often associated with chromosome aberrations. Why cells or tumors with Onc-p53 develop chromosomal aberrations and whether the abnormalities contribute to tumor growth remain elusive. Evidence in this communication demonstrate for the first time that replication stress induced by Onc-p53 triggers re-copying of DNA replication forks, which generates replication intermediates that cause persistent mitotic aberration and DNA segregation errors. Replication intermediates from re-copied replication forks induced by Onc-p53 activate ATM signaling, which stabilizes Onc-p53, reinforces its ability to upregulate replication factors for sustaining replication stress, thus generating a feedforward cycle accelerating tumor formation. In agreement with this observation our time lapse video microscopy show in real time that persistent mitotic aberration and DNA segregation errors induced by Onc-p53 confer selective growth advantage. Accordingly, human lung tumors with Onc-p53 show selection of cells with mitotic aberration during serial passages. Knock down of active replication forks reduces re-copied fork generation by Onc-p53 and specifically induces apoptotic death of lung cancer cells expressing Onc-p53 in xenograft lung tumors in cooperation with inhibitors of ATM activation, deselecting cells with Onc-p53 with mitotic errors. This communication reveals a novel mechanism which interconnects replication stress induced by Onc-p53 to its stabilization and ability to generate chromosomal aberration in lung cancer cells that both accelerate tumor growth and serve as a targetable therapeutic vulnerability. These findings will be extremely valuable for tumor-specific treatment of a high percentage of cancer patients with p53 mutation.

Glioblastoma (GBM) is an aggressive and highly lethal brain tumor. Secondary glioblastoma (sGBM), which arises through malignant progression from lower-grade diffuse glioma, represents a clinically and biologically distinct subset of GBM. In this disease context, the protein tyrosine phosphatase receptor type Z1-mesenchymal-epithelial transition factor (PTPRZ1-MET; ZM) fusion has emerged as a recurrent oncogenic driver associated with adverse clinical outcomes. In this study, we analyzed 159 patients with sGBM, including 15 ZM-positive and 144 ZM-negative cases, and confirmed that ZM-positive tumors were associated with significantly shorter overall and progression-free survival. Transcriptomic profiling identified 359 genes upregulated in ZM-positive tumors, with enrichment in cell-cycle regulation and mitotic spindle-related pathways. To explore surrogate biomarkers associated with ZM fusion status, we benchmarked eight machine-learning classifiers and retained XGBoost as the primary feature-prioritization model. MET, PCDHGA3, and FAM3C emerged as the most informative biomarkers, and the fixed three-gene panel showed stable discriminative performance across cross-validation, nested evaluation, feature-pool sensitivity analyses, repeated random seeds, and class-weighted modeling. Protein-level validation in an independent formalin-fixed, paraffin-embedded (FFPE) cohort using multiplex and conventional chromogenic immunohistochemistry supported the pathology-compatible detection of elevated MET, PCDHGA3, and FAM3C expression in ZM-positive tumors. Collectively, these findings support a robust three-gene molecular signature that may facilitate the identification and stratification of ZM fusion-positive sGBM and provide biological insights with potential translational relevance for precision glioma diagnosis.

Diffuse midline glioma (DMG) remains one of the most aggressive and uniformly fatal brain tumors, with a median overall survival (OS) of only 11 months and no systemic therapy yet demonstrating a meaningful survival benefit. Primarily affecting children and young adults, DMG represents an area of profound unmet clinical need where families and oncologists have faced decades of therapeutic stagnation. The recent accelerated approval by the United States (US) Food and Drug Administration (FDA) of dordaviprone (ONC201, ModeysoTM) for patients with progressive H3 K27M-altered DMG marks the first regulatory recognition of a systemic therapy for this devastating disease. The decision has generated both optimism and debate within the neuro-oncology community. For many, the approval symbolizes long-overdue progress and affirms that systemic agents can demonstrate activity in DMG. For others, it raises concerns over whether the current evidence base is sufficient to justify widespread clinical adoption. The accelerated approval pathways allow for earlier access to therapy for serious life-threatening conditions based on surrogate endpoints, such as durable objective response rates (ORRs), while mandating post-approval studies to confirm benefit. However, while approvals using data from single-arm trials can make promising therapies available to patients expeditiously, such exceptional approvals present challenges that must be carefully considered. In the case of dordaviprone, this mechanism was applied following an integrated analysis of early-phase studies (ONC006 (NCT02525692), ONC013 (NCT03295396), ONC014 (NCT03416530),1 ONC016 (NCT05392374), and ONC018 (NCT03134131))2 that demonstrated measurable radiographic and biological responses in a subset of patients. Here, we examine the scientific, clinical, and regulatory context of the dordaviprone approval through contrasting perspectives. We consider both the landmark nature of this decision and the potential risks of overinterpreting preliminary evidence, aiming to clarify what this approval means for current practice and for the future development of therapies in DMG and other brain tumors.

Glioblastoma recurrence is driven by diffuse microscopic infiltration beyond the contrast-enhancing tumor margin. GlioMap is an open-access AI model predicting voxelwise infiltration and recurrence risk from multiparametric MRI. This prospective study aimed to validate GlioMap's biological accuracy and prognostic relevance through histopathological assessment, transcriptomic profiling, and survival analysis within the SupraGlio trial (NCT05735171). Patients with newly diagnosed glioblastoma underwent neuronavigated biopsies targeting AI-predicted high-risk (HRoR) and low-risk of recurrence (LRoR) regions beyond the contrast-enhancing tumor. Histopathological infiltration served as the ground truth, and transcriptomic profiling characterised each region's molecular phenotype. Model performance was evaluated using accuracy and area under the receiver operating characteristic curve (AUC). Survival analyses assessed the prognostic value of postoperative HRoR volume. Fifty-eight biopsies from 27 patients were analyzed. GlioMap achieved 0.81 accuracy (95% confidence interval [CI], 0.71-0.91) and 0.84 AUC (95% CI, 0.73-0.93) for histologically confirmed infiltration. Transcriptomic analysis of 48 samples from 16 patients revealed progressive upregulation of invasion- and angiogenesis-related genes (CD44, CHI3L1, STAT3, VEGFA) and downregulation of neuronal markers (MBP, GABRA1) from LRoR to HRoR regions and the tumor core, confirming a neural-to-mesenchymal gradient. Postoperative HRoR volume >1.6 cm³ predicted shorter overall survival (P = .04) and progression-free survival (P  =  .008). To our knowledge, this study provides the first prospective, biopsy-controlled, molecular validation of an AI model for mapping glioblastoma infiltration. By accurately identifying histologically and transcriptionally infiltrated regions, GlioMap offers a biologically grounded imaging biomarker that could guide extended resection and personalized radiotherapy planning, potentially improving tumor control and patient outcomes.

Clinical and radiographic models predict incidental meningioma growth, but their molecular architecture is unknown. We analyzed serial magnetic resonance imaging, IMPACT groups (Incidental Meningioma: Prognostic Analysis Using Patient Comorbidity and MRI Tests), targeted gene expression, DNA methylation, and copy number (CNA) profiling of 238 consecutive incidental meningiomas from a single neurosurgical center. The cohort was 81% female, with a median age of 59 years at detection and median tumor volume of 3.83 cm³. Symptoms developed in 15.5%; 93.7% were treated (median time-to-treatment 1.06 years), with 5% recurrence. IMPACT groups stratified treatment-free (P < .0001) and symptom-free survival (P = .0007). Most meningiomas were molecularly low risk, although those from the hypermitotic DNA methylation group had higher IMPACT scores (P = .0020). Incidental meningiomas had distinct CNA and gene expression patterns and favorable outcomes compared to 1434 nonincidental meningiomas. These findings support surveillance for most incidental meningiomas, but early treatment may improve outcomes for molecularly higher-risk cases.

Single-cell-level resolution tumor therapy represents an advanced strategy against glioblastoma but lacks suitable theranostic agents. Here, we developed a spatiotemporal-switchable, two-dimensional (2D), bismuthene-based second near-infrared window (NIR-II) nanozyme. In this platform, the bismuthene scaffold simultaneously directed the assembly of indocyanine green (ICG) into ordered J-aggregates and anchored monodispersed platinum (Pt) atoms. The resulting J-aggregates acted as an optical antenna with a long-wavelength absorption peak at 895 nanometers and high photobleaching resistance of 78.0%, enabling the identification of single tumor cells with a resolution of 44.3 micrometers at 1350 nanometers for precise glioma resection. Postoperatively, the spatiotemporal-switchable function was activated for therapeutic intervention, in which the photothermal effect amplified the original efficiency of the catalase-like activity of Pt atoms by threefold, driving a surge in intracellular oxygen to combat tumor hypoxia. Upon 808-nanometer irradiation, the induced oxygen release in the tumor microenvironment amplified ICG-mediated photodynamic therapy, and combined with bismuthene-mediated photothermal therapy, it effectively inhibited residual tumors. In an orthotopic glioma mouse model, this approach minimized recurrence and achieved increased survival without inducing neurological or motor deficits. This work provides an atomic-level and molecular-level design blueprint for NIR-II nanotheranostic agents, paving the way toward clinical translation of single-cell-level precision medicine for brain malignancies.

With low five-year survival estimates, poor prognosis, and high recurrence probabilities, glioma is considered one of the most intractable malignant tumors. Despite the discovery of lymphatic vascular system and immune system in the central nervous system (CNS), immune checkpoint blockade therapeutics, such as programmed death ligand 1 (PDL1, also called B7H1 or CD274) antibodies, are prevented from the CNS and glioma sites due to the existence of biological barriers including the blood-brain barrier (BBB) and blood-brain tumor barrier (BBTB). Herein, we constructed a BBB/BBTB-crossing recombinant antibody by fusing the PDL1 antibody (αPDL1) and the targeting moiety RAP22 peptide (RAP22) through a matrix metalloproteinase 2 (MMP2)-responsive cleavable linker, abbreviated as αPDL1-mRAP22. Not only was αPDL1-mRAP22 able to block PD1/PDL1 pathway, reduce T cell apoptosis, enhance T cell killing ability towards glioma cells in vitro, but also it showed higher accumulation in the glioma site, prolonged survival time, and potent immune responses as well as synergistic effects with temozolomide (TMZ) in vivo, offering a novel strategy for glioma immunotherapy.

Glioblastoma (GBM) remains highly lethal, and standard temozolomide (TMZ) therapy is undermined by O6-methylguanine-DNA-methyltransferase (MGMT)-driven resistance. Herein, we present a Fe3+-responsive aggregable gold nanoplatform (siMGMT@P-T D-Q Au NP) for systemic delivery of MGMT-targeting siRNA (siMGMT) and enables on-demand photothermal therapy (PTT). The platform reconciles the size-performance trade-off of siMGMT@P-T D-Q Au NP: it preserves a small hydrodynamic size during circulation to promote blood-brain barrier (BBB) penetration and tumor access. When in the tumor microenvironment, Fe3+-induced siMGMT@P-T D-Q Au NP rapid aggregation promotes plasmonic coupling, red-shifts near-infrared absorption, and markedly amplifies 808-nm photothermal ablation. Concurrently, Furin triggers siMGMT release to suppress MGMT and resensitize tumor cells to TMZ. The proposed sequential regimen comprises intravenous administration of siMGMT@P-T D-Q Au NP, Fe3+-triggered siMGMT@P-T D-Q Au NP aggregation to enhance PTT to kill tumors, and subsequent oral TMZ as maintenance therapy. This strategy integrates gene silencing to reverse chemoresistance, Fe3+-responsive aggregable enhancement of PTT, and standard-of-care chemotherapy, addressing key delivery barriers while aiming to improve therapeutic breadth with minimal invasiveness. STATEMENT OF SIGNIFICANCE: Our research covers the following highlights: 1. The constructed size-adaptive Au NP first crosses the blood-brain barrier as small particles and then accumulates as larger ones to enhance therapy. 2. The enhanced photothermal conversion efficiency of Au NP enables synergistic photothermal therapy against glioma. 3. Downregulation of drug-resistance gene MGMT maximizes the therapeutic efficacy of temozolomide. 4. The combined strategy of gene silencing and photothermal-chemotherapy achieves enhanced glioma treatment. Over all, this platform represents an innovative size-adaptive gold nanoplatform that achieves siRNA-mediated reversal of TMZ chemoresistance and enables synergistic photothermal therapy against glioblastoma, offering a paradigm for the future development of integrated, multimodal glioma treatment.

Mitophagy, the selective autophagic degradation of mitochondria, often acts as a pro-survival mechanism in tumor cells, including Glioblastoma (GBM), by clearing damaged mitochondria and mitigating oxidative stress. GBM is a highly aggressive brain tumor characterized by profound resistance to conventional therapies. Our recent study identified Molephantin (EM-5), a natural small molecule capable of crossing the blood-brain barrier, as a potent anti-GBM agent. Mechanistically, EM-5 triggers severe mitochondrial dysfunction and massive reactive oxygen species (ROS) production in GBM. Crucially, we discovered that EM-5 acts as a novel late-stage mitophagy inhibitor. It specifically blocks the fusion of mitophagosomes with lysosomes without affecting early autophagosome formation or lysosomal acidification. This ROS-driven fusion defect leads to the toxic accumulation of damaged mitochondria, thereby amplifying oxidative stress and driving GBM cells into apoptosis. Collectively, our work establishes that targeting late-stage mitophagy flux via ROS modulation is a valuable paradigm for the discovery and development of therapeutic agents against GBM.

Glioblastoma harbors frequent alterations in the retinoblastoma pathway, providing a genetic rationale for therapeutic targeting with cyclin-dependent kinase 4/6 (CDK4/6) inhibitors. The NOA-20 trial did not reveal a progression-free survival benefit of CDK4/6 inhibition plus radiation therapy in newly diagnosed, O6-methylguanine DNA methyltransferase (MGMT)-unmethylated glioblastoma. In fact, CDK4/6 inhibitor monotherapy has not demonstrated efficacy in solid tumors. We aimed at discovering response modulators to CDK4/6 inhibition, paving the way for rational combination therapies. We conducted genome-wide CRISPR-Cas9 screens in human glioma cell lines and stem-like cells (LN229, LN18, LNZ308, T98G, and GS-9) under CDK4/6 inhibition, employing knockout (Brunello library) and activation strategies (Calabrese library), followed by genetic and pharmacological validation of selected candidate genes in vitro and ex vivo (primary cultures) as well as the investigation of 1 functionally instructed combination therapy in vivo. Loss of AMBRA1 and gain of function of CCNE1 reduced sensitivity to CDK4/6 inhibition in glioma cells, whereas disruption of checkpoint kinase 1 (CHEK1) or FAM122A resulted in synthetic lethality in combination with CDK4/6 inhibition. AMBRA1-deficient glioma cells exhibited increased sensitivity to CHK1 inhibition, revealing a context-specific vulnerability. Combined inhibition of CHK1 and CDK4/6 led to synergistic antiglioma activity in vitro, ex vivo, and in vivo. Our data identify AMBRA1, CCNE1, CHEK1, and FAM122A as potential molecular modifiers of CDK4/6 inhibition response in experimental glioma and provide a biological rationale for combinatorial targeting with CDK4/6 inhibition in glioblastoma.

Glioblastoma (GBM) depends on selenoproteins, yet the mechanisms underlying their dysregulation remain poorly understood. The tRNA-binding protein TRNAU1AP also remains largely uncharacterized, with its mechanistic functions in in vivo tumorigenesis undefined. Additionally, while IGF2BP3 is the most dysregulated N6-methyladenosine (m6A) reader in GBM, the pathways through which it promotes glioblastoma stem cell (GSC) self-renewal have yet to be elucidated. TRNAU1AP expression in human GBMs and public datasets was analyzed by Western blotting, immunohistochemistry, and gene expression profiling. Functions of TRNAU1AP in GSCs were evaluated through gain- and loss-of-function assays assessing proliferation, self-renewal, and tumorigenicity. Proteomics, spatial transcriptomics, RNA immunoprecipitation, and polysome profiling were employed to investigate TRNAU1AP-mediated regulation of selenoprotein synthesis. RNA immunoprecipitation and phase-separation assays were used to characterize the TRNAU1AP-EEFSEC interaction. Multiomics and RNA stability analyses were performed to elucidate IGF2BP3-dependent regulation of TRNAU1AP expression. TRNAU1AP is essential for the proliferation, stemness, and tumorigenesis of GSCs and is associated with poor patient survival. TRNAU1AP interacts with EEFSEC to form a phase-separated complex that enhances EEFSEC binding to sec-tRNAsec, thereby promoting the translation of several selenoproteins. These selenoproteins act as key effectors mediating the oncogenic functions of TRNAU1AP. Furthermore, IGF2BP3 upregulates TRNAU1AP expression through m6A-dependent transcript stabilization, leading to increased selenoprotein synthesis and enhanced GSC stemness and tumorigenic potential. These findings establish novel oncogenic roles of TRNAU1AP, and reveal that IGF2BP3-TRNAU1AP coupling constitutes an important mechanism for the selenoprotein synthesis and for gliomagenesis, thereby advancing the understanding of RNA-binding proteins in cancer biology.

Accurate MRI-based brain tumor analysis requires not only tumor subtype classification but also localization at an anatomical granularity that is consistent with radiology reports. Most vision-only methods address localization and classification as separate label-prediction tasks, and therefore provide limited alignment with the fine-grained anatomical semantics used in routine reporting. To address this limitation, we propose LIGHT (Learning Image-text Grounding for Hierarchical Tumor analysis), a 3D vision-language framework that formulates brain tumor localization and subtype classification as image-text retrieval in a shared embedding space. Given multimodal MRI inputs, LIGHT retrieves coarse anatomical regions, fine-grained subregions, and tumor subtypes in a unified coarse-to-fine procedure. The framework has three main components: (1) a hierarchical retrieval vocabulary containing 21 coarse-grained regions, 563 fine-grained subregions, and 5 tumor subtypes; (2) large-scale foundation pretraining on a curated patient-disjoint in-house dataset of 99,813 MRI-report pairs; and (3) grounded task fine-tuning that uses segmentation-guided tumor crops and multi-template prompt supervision to improve tumor-prompt alignment. Across 11,034 annotated 3D brain tumor MRI cases from in-house, external clinical, and public datasets, LIGHT achieved 72.1% accuracy for coarse-grained localization, 70.1% top-1 accuracy for fine-grained localization, and 76.0%, 83.7%, and 84.6% accuracy for subtype classification on three 3D datasets, respectively. Additional public 2D subtype-classification experiments provide further evidence of cross-setting subtype recognition. These results suggest that report-grounded image-text retrieval can provide interpretable anatomical and diagnostic outputs for brain tumor MRI analysis. Code: https://github.com/qiuzhaoyu/LIGHT.

Diffuse intrinsic pontine glioma (DIPG) is a highly lethal and therapeutically refractory pediatric brain tumor, and the effects of distinct disease backgrounds on treatment response and residual cell-state plasticity remain poorly understood. We evaluated abemaciclib, an FDA-approved CDK4/6 inhibitor, combined with temozolomide (TMZ) and radiation (XRT) in two patient-derived orthotopic xenograft (PDOX) models established from a treatment-naïve biopsy (IBs-9119DIPG) and a previously treated autopsy tumor (IBs-A0317DIPG). Treatment activity was assessed in PDOX-derived 3D tumor organoids and in randomized DIPG PDOX studies, followed by survival analysis, immunohistochemistry, and endpoint single-cell RNA sequencing (scRNA-seq). The triple therapy generated synergistic antitumor effects in PDOX-derived organoids and significantly prolonged survival in both PDOX models (P < 0.05) despite their distinct baseline molecular and cell-state differences. Endpoint scRNA-seq revealed reduced oligodendrocyte-progenitor-like (OPC-like) cells in both models and decreased astrocyte-like cells in the IBs-A0317DIPG model as cell-state changes associated with treatment response. In contrast, neural progenitor-like (NPC-like) cells expanded in IBs-A0317DIPG, whereas mesenchymal-like and mitotic-like populations persisted in IBs-9119DIPG as candidate therapy-tolerant states. Pseudotime trajectory analysis uncovered a resistance-associated trajectory characterized by an exit from stemness toward differentiation in OPC-like cells in treatment-naïve IBs-9119DIPG, in contrast to the enrichment of stem-like OPC-like and NPC-like cells in therapy-resistant IBs-A0317DIPG. A transcriptionally defined radiation-resistance-associated subpopulation with candidate radiosensitization target genes (NPAS3, TBC1D15, and INPP4B) was also identified. Overall, the triple therapy improved survival in clinically distinct DIPG PDOX models and revealed therapy-associated residual cell-state changes that may inform future strategies to improve durable DIPG tumor control.

High dimension spatial technologies are broadly being used to map the cellular and molecular organization of human cancers including tumor antigens and drug targets. This has revealed extensive spatial heterogeneity of target proteins, but does not quantify drug distribution itself. Exposing additional complexities for drug targeting and delivery, Lu and colleagues recently applied highly multiplexed spatial proteomics to tumors from patients treated with fluorescently labeled antibody therapeutics. Quantifying drug penetration and distribution on cells of specific phenotypes, this approach identified compounding challenges that would limit therapeutic success as both target and therapeutic distribution are highly variable between patients and within individual tumors. Macrophage uptake, stromal barriers, vascularization differences, and epithelial permeability were all associated with altered antibody distribution. As expected, highly desmoplastic pancreatic cancers had poor drug delivery, and similar fibroblast and extracellular matrix (ECM) rich microenvironments were also identified in head and neck squamous cell carcinomas (HNSCCs) as broadly prevalent barriers to drug delivery. Providing a measurement strategy for single cell drug distribution, this work points towards opportunities to predict these barriers in precision medicine and design drugs that overcome them as a clinically impactful frontier.

Forty percent of U.S. adults have obesity, a complex condition determined by genetics, behavior, environment, and other modifiable and nonmodifiable factors. Obesity is associated with certain comorbid conditions, including type 2 diabetes, high cholesterol, heart disease, and some cancer types, and may increase the severity of communicable illnesses, such as influenza. Treatments of obesity include behavioral modification, pharmacotherapy, and surgical procedures, but high cost and coverage restrictions are a substantial barrier for many patients. In this position paper, the American College of Physicians (ACP) offers recommendations on how policymakers can increase the affordability and accessibility of obesity treatment for adults and address modifiable factors that contribute to obesity, like diet and physical activity. ACP also includes recommendations on reducing stigma associated with obesity, improving understanding of obesity among physicians, increasing access to healthy food, and curbing the proliferation of unhealthy, highly processed fare. Population health strategies for addressing obesity are also discussed.

Cardiovascular disease is the leading cause of death in women, yet significant disparities persist in diagnosis, treatment, and research representation. This clinical consensus statement outlines the rationale and framework for establishing women's heart centres (WHCs) in Europe. Women's heart centres are proposed as hub-and-spoke reference networks embedded within existing cardiovascular systems, delivering multidisciplinary, sex-sensitive care across the life course. The document defines referral pathways, operational standards, and core and advanced training competencies in women's cardiovascular health. Key domains include ischaemia/myocardial infarction with non-obstructive coronary arteries, cardio-obstetrics, cardio-oncology, autoimmune disease, mental health, and cardiac rehabilitation. Implementation strategies emphasize scalable models, integration with primary care, telemedicine, quality improvement, and research engagement. Although long-term outcome data remain limited, available evidence suggests improved diagnostic precision, risk factor control, and patient-reported outcomes. Establishing WHC offers a structured approach to reduce inequities and strengthen cardiovascular care for women across Europe.

Over the past decade, the development and implementation of novel circulating tumor "liquid biopsies" in clinical practice has revolutionized cancer care. These tools provide sensitive and specific analysis of tumor status which enable non-invasive alternatives to tissue biopsy for real-time tumor monitoring, guided targeted therapy, predicted treatment response, and assessing therapeutic resistance and residual disease. The FDA has already approved several assays to detect genetic alterations in plasma cell-free DNA for use as companion diagnostics matched to specific molecularly targeted therapies for cancer, setting the stage for future use in early relapse detection and additional applications which are expected to be incorporated into standard-of-care practices for aggressive cutaneous malignancies. It will therefore be important for the dermatology community to have a working knowledge of liquid biopsy techniques and their relevance to the management of cutaneous malignancies. This review highlights state-of-the-art liquid biopsy tools currently in practice with an emphasis on the development and implementation of circulating biomarkers in skin cancers. It is anticipated that liquid biopsy tools will soon become integrated into evolving standards-of-care for improved management of patients with aggressive cutaneous malignancies.

Targeted kinase inhibitors induce marked tumor regressions but are limited by the emergence of drug-tolerant residual disease and resistance. Although tumor-intrinsic and adaptive immune mechanisms have been extensively studied, a better understanding of the contribution of innate immunity to targeted therapy durability is needed to devise optimal treatment strategies. Here, we showed that natural killer (NK) cells constrain targeted therapy resistance and identified a regression-associated macrophage program that promotes NK cell recruitment. In immunocompetent melanoma models that recapitulate patient treatment trajectories, tumor regression was characterized by robust NK infiltration, which transitioned to an NK-excluded residual state preceding resistance. A regression-associated macrophage subset (F4/80hiCCL5⁺MHCII⁺CD63⁺) contributed to NK cell recruitment via CCR2/5 signaling. Genetic depletion of macrophages using LysM-cre;iDTR mice impaired NK cell infiltration. Pharmacologic inhibition of PTPN22, a negative regulator of immune activation, reprogrammed the residual tumor microenvironment, restored NK cell recruitment, and delayed resistance onset. Analysis of patient tumor datasets from melanoma and lung cancer revealed concordant NK dynamics during therapy, linking this innate immune program to clinical outcomes. These findings identify innate immune remodeling as a key contributor to targeted therapy durability and identify NK cell recruitment, promoted in part by regression-associated macrophages, as a therapeutically actionable node for extending durability of response across oncogene-driven cancers.

Uveal melanoma is a highly aggressive intraocular malignancy with limited therapeutic options for metastatic disease. Existing transgenic uveal melanoma mouse models inadequately recapitulate human disease progression, whereas transplant models lack immune competence for studying the tumor immune microenvironment and therapeutic interventions. To address these limitations, we developed a genetically engineered mouse model incorporating stepwise genetic alterations implicated in human uveal melanoma progression. Spatiotemporally controlled expression of mutant GNAQQ209L from the endogenous locus induced choroidal nevi with limited penetrance. Concomitant BAP1 deletion enhanced nevus formation, whereas further MYC activation led to fully penetrant intraocular tumors with the potential to disseminate. Single-cell RNA sequencing revealed that malignant cells segregated into melanocytic and neural crest-like subpopulations characterized by distinct transcriptional and biosynthetic programs. Trajectory analyses inferred dedifferentiation from the melanocytic toward the neural crest-like state during tumor progression. Comparison with human uveal melanoma revealed commonalities with highly aggressive class 2 uveal melanoma, including gene expression signatures and copy-number gains affecting genes that map to human chromosome 8q beyond the activated MYC allele, suggesting cooperative effects of multiple drivers in this chromosomal region. The tumor microenvironment featured immunosuppressive macrophage populations and exhausted T cells, closely resembling human uveal melanoma. This physiologically relevant, immune-competent model provides a platform for investigating uveal melanoma biology, functionally characterizing candidate driver genes, and developing immune-based therapeutic strategies. Development of a mouse model resembling the genetic progression and phenotypic plasticity of human uveal melanoma offers an immune-competent preclinical platform for understanding disease immunogenomics and testing immune and targeted treatments.

The T-cell receptor (TCR) repertoire records an individual's immunological history, but most unique CDR3 sequences in any one person are private and uninformative about anyone else. A small subset, however, recurs predictably across unrelated donors. These public TCRβ sequences are independently generated by multiple mechanisms: enrichment by thymic positive selection on a largely shared self-peptide-MHC ligandome, further amplified by selection on common foreign antigens and convergent recombination, together, they are what makes a personal repertoire computationally legible: they provide the shared coordinate system on which otherwise incommensurable repertoires can be aligned and compared. This review takes public TCR sequences as its protagonist. We trace the biology of TCR publicity through new measurements on a 1.5-billion-sequence meta-repertoire (7943 samples, 41 studies) that quantify five interrelated properties: the universe is finite, with Chao2-bounded ceilings (i.e., a lower estimate) of ≈1.97 × 109 amino acid and ≈8.25 × 109 nucleotide CDR3β sequences of which ≈22.5% and ≈12% have already been observed; the rarefaction curve already bends within [0, 7943] by a factor of ×3.3 (amino acid) and ×1.6 (nucleotide) relative to a linear-at-initial-rate extrapolation; publicness is a cohort-scale statistic, with the heavy tail of the publicness distribution growing predictably from N = 200 to N = 7943; super-public sequences are ≈0.014% of unique amino acid CDR3s but carry 8.45% of the total observation mass; and the recapture rate of a CDR3 already observed in another donor reaches 73.3% (amino acid) and 36.2% (nucleotide) at N ≈ 7900. We then trace the family of computational methods that exploit public sequences as features: frequency vectors, sequence-similarity networks, self-supervised transformer embeddings (CVC, Beaker, TCR-BERT, SCEPTR), and our own anchor-based graph neural networks (GraffiTee). We summarize their performance across cancer detection, autoimmunity, infectious disease, immunotherapy monitoring, and immunological aging, alongside the structural confounders (HLA, age, sex, sequencing platform) that bound generalizability. We connect repertoire-level classification to TCR-pMHC binding prediction and antigen identity, and ask what stands between current capabilities and a universal TCR-based diagnostic.

The transcriptional coactivators p300 and CREB-binding protein (CBP) have emerged as central regulators of enhancer-mediated transcription and key epigenetic integrators of oncogenic signaling in cancer. Through their histone acetyltransferase activity, particularly the deposition of histone H3 lysine 27 acetylation (H3K27ac), p300/CBP control enhancer activation and coordinate transcriptional programs that govern cell identity, tumor progression, and therapeutic response. Dysregulation of enhancer activity is increasingly recognized as a fundamental mechanism driving oncogenic transcription across diverse malignancies. Recent advances in chemical biology have enabled pharmacological targeting of p300/CBP through several complementary strategies, including catalytic histone acetyltransferase (HAT) inhibitors, bromodomain (BRD) inhibitors, dual BET-p300 inhibitors, and emerging targeted protein degradation approaches. In this review, we summarize the molecular functions of p300/CBP, recent progress in the development of pharmacological agents targeting these coactivators, and the current landscape of clinical translation. Catalytic inhibition of p300/CBP directly suppresses H3K27ac and provides a measurable tumor pharmacodynamic biomarker, whereas alternative targeting modalities modulate enhancer-driven transcription through distinct molecular mechanisms. Early-phase clinical studies indicate that p300-directed agents can achieve pharmacodynamic modulation in patients. We further discuss the translational relevance of p300 targeting in osteosarcoma, a malignancy characterized by genomic instability and heterogeneous transcriptional states rather than dominant kinase mutations. Collectively, these findings position p300 catalytic inhibition as a mechanistically defined and pharmacologically tractable strategy for targeting enhancer-driven oncogenic transcription and support continued clinical development of p300-directed therapies.

Nucleocytoplasmic transport is a central but underappreciated component of the proteostasis network as it controls the trafficking and partitioning of proteins between the nucleus and cytoplasm through the nuclear pore complex (NPC). Transport of large proteins across the NPC is mediated by karyopherins, a conserved family of importins and exportins that function through a Ran GTPase-dependent cycle. Beyond their canonical transport activities, karyopherins can directly contribute to proteostasis by acting as chaperone-like factors that prevent aberrant phase separation and protein aggregation. Dysregulation of karyopherin-mediated transport emerges as a convergent contributor underlying aging and diverse age-associated diseases, including neurodegeneration, cancer, cardiovascular dysfunction, chronic inflammation, and progeroid syndromes. Here, we highlight how age-related alterations in nucleocytoplasmic transport reshape proteome organization and intracellular signaling, and discuss emerging therapeutic strategies targeting karyopherins to restore proteostasis and cellular homeostasis.

CD8+ T cell exhaustion is increasingly recognized as a regulated adaptation to chronic antigenic stimulation rather than a simple immune failure. Indeed, recent studies reveal that exhaustion is imprinted early after T cell activation, integrating transcriptional and epigenetic cues to balance effector function with long-term persistence. Key regulators, including Inhibitor of DNA binding 3 (ID3), Thymocyte selection high mobility box protein (TOX), MYB, Krüppel-like factor 2 (KLF2), and Special AT-rich sequencing binding protein 1 (SATB1), orchestrate this process, preserving stem-like precursor populations that sustain immunity during chronic infection and cancer. This emerging view frames exhaustion as a context-dependent extension of the memory program rather than its collapse. By defining the molecular and functional logic of exhaustion, we highlight how these insights can inform new approaches to manipulate T cell fate for therapeutic benefit.

The extracellular matrix (ECM) plays a pivotal role in shaping tumor behavior by providing biochemical and biophysical cues to cancer cells. Traditional 2D culture systems fail to recapitulate this complexity, and in vivo systems do not readily allow for interrogation of how individual ECM components influence tumor cell behavior. Here, we introduce a fully synthetic, tunable three-dimensional (3D) hydrogel that mimics a soft tissue tumor microenvironment (TME) to enable mechanistic studies of ECM-tumor interactions. The hydrogel features a proteolytically degradable poly(ethylene glycol) base network functionalized with integrin-binding peptides derived from ECM components collagen I and fibronectin. To model metastatic ECM and further tune the hydrogel, we incorporated a tenascin-C (TNC)-derived peptide. To study the impact of these tunable ECM parameters on cancer cell behavior, we encapsulated Ewing sarcoma (EwS) cells within the hydrogels. EwS is an aggressive bone and soft tissue tumor that commonly metastasizes to lung. Our studies demonstrated matrix-dependent growth and phenotypic variation of EwS cells. Specifically, the TNC peptide drove divergent tumor behaviors and induced cell state changes consistent with alterations induced by the native TNC protein. To facilitate downstream functional assays, the hydrogel incorporates sortase-degradable crosslinkers that enable non-perturbative recovery of encapsulated cells. This platform provides a reductionist and reproducible model for studying the ECM's role in cancer cell biology while addressing long-standing challenges in PEG hydrogel degradation and cell retrieval. Collectively, this work establishes a biomaterials-based framework to dissect EwS tumor-ECM interactions in a controllable 3D microenvironment. STATEMENT OF SIGNIFICANCE: Engineered biomaterials to model Ewing sarcoma (EwS) have been previously employed to study metastasis to the bone. While important efforts, platforms to study EwS metastasis in lung - the most common site of metastasis - have not been developed. Here, we introduce a user programmable biomaterial designed to mimic the environment of soft tissue metastases. The platform features several attributes: 1) it is mechanically matched to lung tissue; 2) it is readily functionalized with extracellular matrix protein-derived peptides (e.g., Tenascin-C); 3) encapsulated cells can be retrieved in a "biologically invisible" manner via sortase-mediated gel degradation, enabling expanded downstream analysis. This platform provides a powerful, reproducible tool to dissect tumor behavior and identify new targets for cancer therapy.

For people with localized, high-risk prostate cancer, treatment with 6 months of apalutamide and androgen deprivation therapy (ADT) both before and after surgery reduces the risk of metastases and death compared with perioperative ADT alone, according to results of the phase III PROTEUS trial. At 5 years, 78.2% of patients who received apalutamide plus ADT were alive and free from metastasis, compared with 73.5% in the ADT only group.

Arsenic is a known carcinogen that may contaminate human drinking water. Since 2000, regulations in the United States (US) have limited arsenic in the water to 10 parts per billion (ppb). However, evidence suggests that even low levels of arsenic may still increase the risks of bladder, prostate and kidney cancer. We performed an ecological study in Washington State (WA) to examine the relationship between low levels of arsenic in drinking water and incident urologic cancers. Average county-level arsenic levels in drinking water were estimated from public water systems in WA (2000-2018). Incident bladder, prostate and kidney cancer cases in WA (2014-2018) were ascertained from the Department of Health. Multivariable Poisson regression was conducted to estimate adjusted incidence rate ratios (aIRRs) for the associations between arsenic exposure and cancer risk. We found that average arsenic levels at the county-level were 3.8 ppb (SD 1.1). When comparing the highest (4.16-7.50 ppb) versus lowest (1.65-3.14 ppb) quartiles for arsenic levels in drinking water, the highest quartile was associated with an increased risk of bladder cancer (aIRR: 1.21, 95%CI 1.09-1.35), prostate cancer (aIRR: 1.20, 95%CI 1.12-1.29), with a suggestive positive association with kidney cancer (aIRR: 1.10, 95%CI 0.97-1.26). These findings demonstrate that low levels (<10ppb) of arsenic were associated with an increased risk of incident bladder and prostate cancer. Additionally, these data suggest that US drinking water standards may not adequately reduce the risk of cancer associated with low levels of arsenic exposure.

Researchers have discovered that black splotches on brown bullhead catfish in lakes in New England and Canada are likely caused by a transmissible cancer, the first one found in fish. The discovery raises questions about the cancer's origin, how it might spread, and how widespread the phenomenon might be-and could offer insights into cancer biology more broadly.

RNA-triggered chromatin shredding may offer a new way to attack cancers driven by mutations that have resisted conventional drugs, two new studies show. In mouse models, upon activation by a target transcript, the CRISPR enzyme Cas12a2 can selectively eliminate tumor cells carrying mutations in TP53, MYC, and other hard-to-drug cancer genes.

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by β-amyloid (Aβ) deposition, neuroinflammation, and cognitive decline. Microglia, the brain's primary immune cells, play a central role in AD pathogenesis by driving neuroinflammatory responses. Deubiquitinating enzymes (DUBs) regulate microglial activation, but the role of the ovarian tumor domain-containing DUB OTUD6A in AD remains unclear. In this study, we demonstrate that OTUD6A is upregulated in microglia across multiple AD models, including Aβ-infused mice, 3 ×Tg mice, and APP/PS1 mice. Otud6a KO or microglia-specific knockdown Otud6a ameliorated cognitive deficits and reduced neuroinflammation in AD mice. Besides, OTUD6A binds to C/EBPβ by removing K48-linked ubiquitin chains at lysine 253 (K253), thereby leading to C/EBPβ accumulation and enhancing NF-κB signaling and proinflammatory cytokine production. Moreover, mutation of OTUD6A catalytic residue (C157A) abolished its deubiquitination activity, confirming its role in C/EBPβ stabilization. Furthermore, C/EBPβ knockdown reversed OTUD6A-mediated neuroinflammation, validating the microglial OTUD6A-C/EBPβ-NF-κB axis as a critical pathway in AD pathogenesis. Therefore, our findings highlight OTUD6A as a novel regulator of microglial activation and suggest that targeting this DUB could provide a therapeutic strategy to mitigate neuroinflammation in AD.

The ubiquitin-proteasome system (UPS) is a fundamental regulatory mechanism governing protein homeostasis via ubiquitin-mediated manner. In cell cycle progression, the UPS plays a pivotal role in regulating key cell cycle regulators, including cyclins, cyclin-dependent kinase (CDK) inhibitors, and checkpoint proteins. Through the orchestrated actions of E3 ubiquitin ligases, such as the anaphase-promoting complex/cyclosome (APC/C) and Skp1-Cullin1-F-box (SCF) complexes, timely degradation of cell cycle proteins ensures proper cell cycle transitions, DNA replication, and mitotic fidelity. Dysregulation of UPS components has been implicated in various pathological conditions, including cancer, where aberrant stabilization or premature degradation of key cell cycle regulators disrupts normal cellular proliferation. This review provides a comprehensive overview of the UPS's role in cell cycle control, highlighting the molecular mechanisms of ubiquitination and degradation, key regulatory E3 ligases and deubiquitinating enzymes (DUBs), and the therapeutic potential of targeting the UPS in cell cycle-associated disorders.

Cell-cell communications involve signal transmission from sending cells to receiving cells expressing specific receptors. Extracellular vesicles (EVs) mediate this process by transporting diverse biomolecules. G-protein-coupled receptors (GPCRs) are canonical membrane receptors that integrate various extracellular signals into intracellular responses. However, whether and how GPCRs engage in EV-mediated communications remain elusive. Here, we report that adhesion GPCRs (aGPCRs) induce the formation of migrasomes and retractosomes, two newly identified EV subtypes, through their extracellular adhesion-like domains and G12/13-protein signaling. Remarkably, activated receptors undergo ectocytosis into these EVs and are subsequently internalized by receiving cells, eliciting de novo G-protein activation. We further demonstrate that cancer-cell-derived migrasomes transfer aGPCRs such as GPR56 to endothelial cells in vitro and in vivo, thereby enhancing angiogenic potential. Together, our findings uncover that aGPCRs promote migrasome formation and provide a novel mechanism of cell-cell communications through EV-mediated intercellular spread of active GPCRs.

Manipulation of ligand-receptor interactions (LRIs) in a user-defined manner is one of the effective ways to regulate cell motility and disease progression. Molecular engines orchestrate LRIs to regulate cell motility by harnessing various forms of energy within physiological surroundings. However, engineering chemical engines remains largely unexplored, primarily due to the insufficient chemical cue density, uncertain conformational changes of scaffolds, and a limited repertoire of environment-responsive switchable scaffolds. Enzymatic biofuel cells (EBFCs) are electrochemical devices that convert biofuels into other accessible energy through electrochemical reaction for the construction of self-powered molecular tools, offering a viable strategy to overcome the above limitations. Here, we present an enzyme biofuel cell-based self-powered molecular engine (EBFC-SME) for rewiring chemical cues-based LRIs and regulating cell motility. We employ EBFC-SME as a proof-of-concept platform for reprogramming proton-responsive HGF/c-Met interaction. The system comprises a glucose dehydrogenase (GDH)-incorporated bioanode and a bilirubin oxidase (BOD)/functional nucleic acids (FNAs)-encapsulated iron-alginate (IA) hydrogel-coated biocathode. Through glucose-initiated redox reactions, this configuration enables the EBFC-SME to generate abundant protons and release FNAs. These products subsequently mediate the in-situ assembly on tumor cell membrane, which blocks c-Met pathway activation and ultimately suppresses tumor cell migration. Different from conventional chemical engines, the EBFC-SME efficiently regenerates chemical cues via intrinsic energy conversion reactions for the manipulation of scaffold-mediated LRIs. This EBFC-SME platform provides a robust "sensing-conversion-initiation" tool for the chemical regulation of cellular motility, holding significant promise in precision biomedicine.