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This multicohort, single-center, phase II trial explored the efficacy and safety of a biomarker-driven total neoadjuvant therapy (TNT) regimen in high-risk locally advanced rectal cancer (LARC) patients. The regimen included induction chemotherapy (4 cycles of mFOLFOX6) with targeted agents (bevacizumab for cohort B and cetuximab for cohort C), followed by short-course radiotherapy (25 Gy/5Fx) and consolidation chemotherapy (2 cycles of mFOLFOX6, with cetuximab in cohort C). The primary endpoint was complete response (CR; sustained clinical CR [cCR] + pathological CR [pCR]). Secondary endpoints included compliance, safety, pathological outcomes, and survival. Between April 2021 and October 2024, 90 of 96 patients completed the prescribed treatment. CR was achieved in 36 (18 in cohort B and 18 in cohort C) patients (40%). No grade IV-V toxicity was observed, although three and 41 patients experienced targeted agent-related adverse effects in cohort B and C, respectively. In the two cohorts, 43 and 38 patients underwent curative surgery with sphincter preservation in 39 (90.7%) and 34 (89.5%) patients, respectively. Postoperative complications occurred in seven (16.3%) and five (13.2%) patients, of whom one (2.6%) patient in cohort C received reoperation. This intensified TNT regimen with targeted agents for high-risk LARC patients achieved a satisfactory CR rate with acceptable toxicities and complications.

Minimal residual disease (MRD) status is widely used to assess depth of response and predict outcome in multiple myeloma, but bone marrow MRD testing is invasive and subject to sampling variability. Blood-based mass spectrometry (MS), which measures patient-specific monoclonal immunoglobulins, offers a non-invasive approach to monitoring residual disease. Using the structured assessment schedule of the CASSIOPEIA trial, we compared serum MS with bone marrow MRD by next-generation flow cytometry (NGF) and next-generation sequencing (NGS). This retrospective post-hoc analysis included 237 patients from the CASSIOPET companion study with paired serum and bone marrow samples prospectively collected at predefined CASSIOPEIA time points from post-induction through maintenance. MRD results by MS, NGF, and NGS were evaluated for associations with progression-free survival, agreement between methods, longitudinal changes, and sustained MRD status according to International Myeloma Working Group criteria. At all time points, MRD negativity by any method was associated with longer progression-free survival, with comparable landmark prognostic performance across platforms. Agreement between serum MS and bone marrow MRD improved during maintenance, with high positive predictive value and high negative predictive value relative to a 10⁻⁵ marrow reference, but more limited rule-out performance against NGS at 10⁻⁶. Sustained MRD negativity over 1 and 2 years identified patients with favorable outcomes irrespective of method. These findings support a complementary, phase-adapted role for serum MS alongside bone marrow MRD testing, enabling frequent non-invasive monitoring during maintenance while preserving bone marrow assessment when maximum sensitivity is required. The Cassiopeia trial is registered at www.clinicaltrials.gov as NCT02541383.

Extracellular vesicles (EVs)-derived miRNAs are valuable non-invasive biomarkers for early cancer diagnosis. However, most current methods detect either total EVs-miRNAs or those from a single subpopulation, overlooking subpopulation heterogeneity. Moreover, EVs isolation and miRNA analysis are often conducted in separate workflows, increasing sample handling and compromising reproducibility. Herein, we presented an integrated, single-tube platform for subpopulation-specific miRNA profiling, in which multivalent aptamer-based EVs capture and CRISPR/Cas12a-mediated signal amplification were co-localized on a rationally designed DNA scaffold. The multivalent aptamer system achieved a capture efficiency of 84.3%, significantly outperforming monovalent aptamers (56.4%). Upon in situ lysis of captured EVs subpopulation, the released miRNAs preferentially encountered adjacent recognition probes due to the increased local concentration, thereby efficiently initiating catalytic hairpin assembly (CHA). The resulting abundant duplex subsequently activated Cas12a, achieving a limit of detection (LOD) as low as 1.42 × 104 particles/μL. Critically, the modularity of the platform allowed for straightforward reconfiguration to target distinct EVs subpopulations by exchanging the capture aptamer. We demonstrated this flexibility by profiling miR-21 and miR-155 across two specific subpopulations (CD63+ and MUC1+). The results revealed both cell-line-specific expression patterns and marked heterogeneity across CD63/MUC1-captured EVs fractions. Meanwhile, MUC1+ subpopulation outperformed CD63+ in clinical diagnosis, with higher AUC values for both miR-21 (0.85 vs 0.75) and miR-155 (0.83 vs 0.73). Overall, our study highlighted the critical importance of EVs subpopulation heterogeneity in cancer diagnostics and provided more informative biomarker readouts.

The purpose of this study was to compare, across readers with varying experience, the characterisation of prostate MRI lesions as grade group (GG) ≥ 2 cancer, by using the PI-RADS version 2.1 (PI-RADSv2.1) score alone and by combining prostate specific antigen density (PSAd), the PI-RADSv2.1 score and the output of a radiomics-based algorithm (Q-CAD). The MULTI database in which 21 readers (seven experienced seniors, seven less-experienced seniors, seven juniors) had assigned a PI-RADSv2.1 score to 240 prostate MRI lesions was retrospectively used. The lesions were outlined by two independent experts to compute their Q-CAD score. For each reader, four biopsy strategies were simulated. PI-RADS3 and PI-RADS4 strategies triggered biopsy in PI-RADSv2.1 ≥ 3 and PI-RADSv2.1 ≥ 4 lesions respectively. Combined3 and Combined4 strategies triggered biopsy when at least two of the following conditions were fulfilled: positive PI-RADSv2.1 score (≥ 3 for Combined3; ≥ 4 for Combined4), positive Q-CAD score (≥ 0.45 in peripheral zone; ≥ 0.79 in transition zone), PSAd ≥ 0.15 ng/mL/cm3. A total of 232 lesions were included. Using lesions' delineations by Expert 1 for the three readers' experience groups, the Combined3 strategy was significantly less sensitive for GG ≥ 2 cancers (87-88% vs. 91-96%; P = 0.026 to < 0.001), but significantly more specific (45%-55% vs. 15%-34%; P < 0.001) than the PI-RADS3 strategy. The Combined4 strategy was less sensitive than the PI-RADS4 strategy (84%-86% vs. 85%-91%) but the difference was significant only for less-experienced seniors (P = 0.023); it was significantly more specific (51%-63% vs. 27%-50%; P < 0.001) in all groups. The Combined4 strategy provided the highest net benefit for risk thresholds >12%-16%. Using lesions' delineations by Expert 2 yielded similar results. The combined strategies significantly increased specificity, at the cost of slightly reducing sensitivity for GG ≥ 2 cancers.

Early cancer diagnosis in patients with non-specific symptoms is limited by the lack of discriminatory tests. Within the Oxfordshire Suspected CANcer (SCAN) pathway, exploratory biomarker work showed that serum 1H NMR-based metabolomics can identify cancer with high accuracy. SCAN2 evaluated whether integrating metabolomics with glycomics provides complementary molecular information and improves discrimination in a clinically complex, real-world population. Serum from 369 SCAN patients (59 cancers) was analysed using AXINON® System-derived NMR metabolomics and HPLC-MS glycomics. Machine-learning models were trained to predict cancer status, with performance assessed by receiver operating characteristic (ROC) analysis of pooled cross-validated predictions. To place cancer risk in a broader clinical context, a second classifier modelling alternative non-cancer diagnosis was incorporated, and mean predicted probabilities from both models were jointly projected into a two-dimensional space, maintaining strict separation of training and test data. In the full cohort, integration of glycomics with metabolomics achieved an AUC of 0.814 (95% CI 0.808-0.820). In a refined sub-cohort excluding major comorbidities and selected cancer types (32 cancers, 277 non-cancers), performance improved to an AUC of 0.884 (95% CI 0.879-0.890). Discriminatory features included cancer-associated biantennary fucosylated glycans alongside amino acid metabolites (glutamate, histidine) and lipoprotein-related measures. A classifier distinguishing metastatic from non-metastatic disease (n = 29 vs. 30) achieved an AUC of 0.80. Joint probability analysis in the full cohort preserved cancer-associated signatures across comorbidity burden, with projection-based classification achieving an accuracy of 89.2% (95% CI 85.7-92.6). These findings validate the SCAN1 metabolomic signature in a more clinically complex cohort and indicate that integrating glycomics with metabolomics provides complementary biological information for cancer discrimination. Joint probability analysis provides an interpretable framework for cancer risk stratification within multimorbid diagnostic pathways, supporting the clinical potential of scalable multi-omics blood testing. EPSRC, EU Horizon 2020, Wellcome/MLSTF, Novo Nordisk Foundation.

Homologous recombination deficiency (HRD) is a critical biomarker for predicting response to poly (ADP-ribose) polymerase (PARP) inhibitors in high-grade serous ovarian cancer (HGSOC). Hence, genomic scar-based HRD testing must be implemented in routine clinical labs. This study aims to obtain a head-to-head comparison of some of the applications available for HRD status determination in a real-world clinical cohort to guide HRD testing standardization. HRD scores were obtained from HGSOC formalin-fixed, paraffin-embedded samples from the Molecular Prescreening Program at Vall d'Hebron Institute of Oncology (VHIO) using two approaches: VHIO-HRD (n = 229) and shallow sequencing-based HRD score (LSTsh-HRD; n = 123). Large-scale transitions (LST), telomeric allelic imbalance (TAI), and genomic loss of heterozygosity (LOH) were calculated using data from a custom hybrid-capture panel (VHIO-HRD score), and LST was determined from shallow whole-genome sequencing of a genomic library (LSTsh-HRD score). Validation was obtained by benchmarking against established and commercially available platforms. Tumor fraction (TF) was estimated using several computational tools. Optimized HRD status cutoff for VHIO-HRD (LSTVHIO-HRD + TAIVHIO-HRD + LOHVHIO-HRD) was set to ≥47 based on comparison with commercially available tools, whereas HRD status cutoff for LSTsh-HRD was predefined at ≥20. Despite differences in chemistry and scoring metrics, strong concordance in HRD classification was observed across assays. TF strongly influenced HRD performance, with optimal sensitivity at TF ≥0.4 for VHIO-HRD and ≥0.2 for LSTsh-HRD. We validated that HRD scar quantification can be implemented across testing laboratories using assays with different library preparation chemistries and biomarker calculation algorithms. TF is a critical determinant of accurate HRD scoring; LSTsh-HRD quantification offers improved sensitivity in low-cellularity contexts, and VHIO-HRD enables status classification across the continuum of score values in samples with TF ≥0.4. A multiplatform, standardized HRD testing strategy may enhance biomarker-driven patient selection for PARP inhibitor therapies in HGSOC and other HRD-associated tumors.

In glioblastoma studies, the presence of measurable disease is often required for trial eligibility. While response assessment according to RANO 2.0 relies on MRI, the recently introduced PET RANO 1.0 criteria allow standardized evaluation based on amino acid PET. This study compares the frequency of measurable disease according to PET RANO 1.0 vs. RANO 2.0 criteria at key enrollment timepoints of clinical trials. In this retrospective, single-center study, we included patients with IDH-wildtype glioblastoma who underwent both [¹⁸F]FET PET and MRI, after standard first-line radiotherapy (time-point T0) (defined per RANO 2.0) or at first progression (time-point T1). Two independent raters evaluated measurable disease using PET RANO 1.0 and RANO 2.0 criteria. Further, tumor size, target lesions and tracer uptake metrics were analyzed. In total 322 patients were included, 112 at T0 (median age: 59 years, IQR 54-69), and 210 at T1 (median age: 59, IQR 53-67). On MRI, measurable disease was identified in 57/112 patients (50.9%) at T0 and in 137/210 patients (65.2%) at T1 (median sum of products of cross-sectional diameters: 143mm2; 187mm2). On PET, significantly more cases with measurable disease were detected: 102/112 patients, (91.1%) at T0 and 201/210 patients (95.7%) at T1 (median volumes: 9.86cm³; 14.3cm³) (p = 0.001). PET RANO 1.0 detects a substantially larger subset of patients with measurable disease compared to RANO 2.0. These findings warrant prospective validation of PET-based measurable disease as an inclusion criterion for clinical trials in IDH-wildtype glioblastoma, with the potential to broaden trial eligibility.

Urinary extracellular vesicles (uEVs) provide noninvasive biomarkers for liquid biopsy owing to their ability to reflect disease-associated molecular alterations. However, the accurate analysis of low-abundance uEV surface proteins in complex biological matrices remains a major analytical challenge. Here, we report a proximity-activated dual-cascade (PADC) biosensing platform that integrates a target-responsive 3D DNA machine with a nanozyme-mediated ratiometric transduction circuit for high-fidelity multiplexed phenotyping of uEVs. Upon dual-recognition of target uEVs, the DNA machine undergoes proximity-induced spatial confinement on individual magnetic particles, creating a localized catalytic microenvironment that markedly enhances DNA walking kinetics and signal amplification efficiency. Coupled with an enzyme-nanozyme redox cascade, this confined catalytic process generates a robust ratiometric fluorescence output for sensitive and reliable molecular profiling. Benefiting from the spatially confined amplification architecture, the proposed platform enables ultrasensitive multiplexed analysis of CD63, EGFR, and PAK6 with detection limits down to 4.3 × 104, 2.2 × 104, and 1.7 × 103 particles/mL, respectively. Importantly, the target-uEV response was approximately six-fold higher than those generated by common urinary interferents or their mixture, demonstrating high analytical selectivity in complex urine matrices. To further evaluate its universality for multi-disease screening, multidimensional molecular profiling combined with machine learning algorithms was performed on clinical uEV samples collected from healthy controls, Diabetes Mellitus (DM) patients, and prostate cancer (PCa) patients. The optimal classification model achieved an overall multiclass diagnostic accuracy of 91% for distinguishing healthy, DM, and PCa groups. This work establishes a proximity-confined catalytic biosensing strategy for multiplexed uEV analysis and demonstrates its potential for uEV-based liquid biopsy and proof-of-concept disease stratification.

Metabolic syndrome (MetS) includes obesity, insulin resistance, hypertension, and dyslipidemia. While androgen deprivation therapy (ADT) is associated with an increased risk of dyslipidemia, adiposity, and MetS, evidence is limited on the occurrence and timing of metabolic dysfunction among men receiving concurrent androgen receptor pathway inhibitor (ARPI) therapy and whether patterns vary by age. To characterize the occurrence and rate of MetS during the first year following initiation of ADT-ARPI, examine associations with age and ARPI type, and evaluate component metabolic outcomes (secondary end points). This retrospective cohort study of adult patients from January 2014 to September 2025 with up to 12 months of follow-up per person used data from a national, deidentified health record dataset (Epic Cosmos) and included patients with prostate cancer who initiated treatment with ADT-ARPI (abiraterone acetate, enzalutamide, apalutamide, and darolutamide) without evidence of MetS or its components before treatment. Data were analyzed between October 2025 and January 2026 (further analyses were done with revisions through May 2026). Concurrent ADT and ARPI use, with index date defined as the first date of overlap between therapies. New-onset MetS during the 12 months following the index date. Secondary outcomes included individual metabolic abnormalities. The cohort included 16 924 men with prostate cancer (mean [SD] age, 73.1 [9.1] y; 509 [3.0%] Asian individuals, 912 [5.4%] Hispanic individuals, 3562 [21.0%] non-Hispanic Black individuals, and 11 083 [65.5%] non-Hispanic White individuals). Medical ADT use predominated, and enzalutamide was the most frequently used ARPI. During the first year following initiation of concurrent ADT and ARPI therapy, the cumulative incidence of metabolic syndrome increased steadily, reaching nearly 40%, and varied by age group. Hypertension was the most frequently documented component outcome. Metabolic associations varied by age, with the highest incidence of metabolic syndrome among patients aged 70 to 79 years (51.7 events per 1000 person-months; 95% CI, 50.7-53.9). Heterogeneity in metabolic outcomes by ARPI type was observed in exploratory analyses. This study found that while ARPIs are standard of care for advanced prostate cancer, metabolic abnormalities were frequently documented shortly after initiation of concurrent ADT and ARPI therapy. This suggests that monitoring should extend beyond cancer-specific outcomes to include early detection of metabolic dysfunction, ideally through multidisciplinary care. The early burden of metabolic abnormalities highlights the need to evaluate scalable interventions addressing cardiometabolic risk in men with prostate cancer.

In this issue of Blood Cancer Discovery, Merz and colleagues study more than 600 recipients of chimeric antigen receptor T-cell (CAR T) and/or bispecific antibody therapy in multiple myeloma with an emphasis on patients who received both modalities sequentially. They find a clear progression-free survival advantage when CAR T therapy was used first; importantly, because outcomes with both modalities were similar when following progression after the converse modality, this advantage was driven by CAR T therapy's up-front benefits. See related article by Merz et al, p. XX .

FLT3-internal tandem duplication measurable residual disease testing for patients in remission from acute myeloid leukemia is now recommended by the recently updated clinical standard-of-care guidelines. This companion technical note provides important laboratory and clinical recommendations regarding such testing.

Patients with relapsed or refractory multiple myeloma (RRMM) with extraosseous extramedullary disease (EMD) have inferior outcomes and lack effective therapies. We developed anti-B-cell maturation antigen (anti-BCMA)/G protein-coupled receptor, class C group 5 member D (GPRC5D) bispecific chimeric antigen receptors (CARs) to investigate the activity and safety of the CAR T cells in patients with extraosseous EMD. In this single-arm, open-label, phase 2 trial, we enrolled 37 patients with RRMM with extraosseous EMD, and anti-BCMA/GPRC5D bispecific CAR T cells were administered at 2.0 × 106 CAR T cells per kg. At a median follow-up of 10.1 months (interquartile range, 6.4-19.1), 36 of 37 patients (97%) obtained an overall response and measurable residual disease negativity, including 16 (43%) with stringent complete response. The median progression-free survival was 5.8 months (95% confidence interval, 2.2-9.4), and the median overall survival was not reached. The most common grade 3 or worse adverse events were hematologic toxicities (except lymphopenia; 37/37). Twenty-seven patients (73%) experienced cytokine release syndrome, all cases of which were grade 1 or 2. Two patients (5%) had grade 1 or 3 immune effector cell-associated neurotoxicity syndrome. These findings support that anti-BCMA/GPRC5D bispecific CAR T cells induced a high response rate in patients with RRMM with extraosseous EMD, and the safety profile was manageable. This ongoing trial is registered at www.clinicaltrials.gov as NCT05509530.

Teclistamab (Tec) and talquetamab (Tal) are bispecific CD3 T-cell engagers targeting B-cell maturation antigen and G protein-coupled receptor, class C, group 5, member D, respectively, and have transformed how we treat relapsed/refractory multiple myeloma (RRMM). Early onset toxicities, such as cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS), require utilization of pre-medications, step-up dosing (SUD) schemas, and close monitoring while late onset toxicities, such as hypogammaglobulinemia, myelosuppression, and notably infections, can lead to significant complications and treatment interruptions, which may require extending bispecific antibody (BsAb) dosing intervals to mitigate toxicities. While current prescribing information for Tec and Tal recommend repeat SUD for prolonged dose delays, we report a series of six patients (pts) who continued therapy with extended dosing intervals (EDI) (8-12 weeks) without repeating SUD. Notably, all pts had achieved a  ≥  complete response (CR) prior to transitioning to extended interval dosing, which was chosen either to improve tolerability or per provider preference based on depth of disease response. Despite experiencing CRS (83% of pts) and ICANS (17% of pts) during their initial SUD, no CRS or ICANS events occurred after 23 doses of Tec/Tal given at EDI, despite a median duration between doses of 77 days. At the time of this report, all patients in this cohort have sustained, deep disease response and remain on BsAb therapy. Despite small sample size, this report adds to limited available data supporting safe and feasible omission of SUD in RRMM pts slated for Tec/Tal therapy resumption via EDI.

Population-based genomic newborn screening identifying newborns at risk for early-onset cancers has not been evaluated. Here, within a Michigan birth cohort (1987-2020), we identify all children developing a solid or central nervous system malignancy by age 8 years (n = 1948). We perform targeted sequencing of 11 cancer predisposition genes using archived newborn dried blood spot DNA. We find pathogenic or likely-pathogenic germline variants (P/LP) in 6.8% of cases (n = 132): RB1 (n = 69), TP53 (n = 24), SMARCB1 (n = 8), WT1 (n = 7), RET (n = 6), SUFU (n = 6), PTCH1 (n = 4), DICER1 (n = 4), APC (n = 3) and PHOX2B (n = 1). We show approximately 1/27,000 newborns develop an early-onset malignancy with an associated pathogenic or likely-pathogenic variant. Germline variant prevalence is 100% in medullary thyroid carcinoma, 40% in retinoblastoma, and 11-30% across five additional diagnoses, with strong gene-tumor specificity (p < 0.001). P/LP variants are rare in comparison datasets from healthy newborns and gnomAD. Our data support newborn screening for selected cancer-risk genes.

Confirmatory prostate-specific antigen measurement reclassifies many younger men below the biopsy threshold, offering a simple, low-cost way to reduce magnetic resonance imaging and biopsy. For most, reclassification means deferral rather than exclusion, and follow-up remains insufficient to rule out missed clinically significant cancers."

In health care, artificial intelligence (AI) is emerging as a powerful tool to improve the quality of care, support health care professionals, and build a more streamlined and efficient health care system while also enhancing the patient experience. In particular, integrating AI into clinical practice through an innovative management approach like lean and change management could optimize diagnostic and therapeutic pathways, increase operational efficiency within hospitals, and promote more personalized, patient-centered care. At the same time, uncertainty remains about how these new technologies should be implemented in order to become broadly accessible in clinical practice. This expert opinion explores the potential role of AI in breast cancer patients' care from the perspective of a team of Italian opinion leaders. The manuscript identifies two main areas of application: (i) AI solutions designed for direct patient interaction (i.e. conversational agents interacting with patients, providing informational and organizational support during the care journey and contributing to personalized care and improved patient quality of life) and (ii) AI solutions for clinical support to health care providers (i.e. tools or algorithms aimed at supporting physicians, reducing workload, and optimizing operational flows). When implemented alongside lean and change management strategies, these solutions have the potential to streamline care pathways and improve the satisfaction and interaction of patients and health care professionals, ultimately benefiting all stakeholders. The document also addresses the ethical, regulatory, methodological, and educational challenges associated with the implementation of AI solutions in clinical practice.

Acute myeloid leukemia/myelodysplastic syndromes (AML/MDSs) carrying p53 mutations are refractory to various standard therapies. Arsenic trioxide (ATO) may be effective in restoring function to p53 structural mutants. Here, we report that mutant p53 rescued by ATO treatment strengthened interferon responses triggered by the DNA hypomethylating agent decitabine by transactivating interferon regulatory factor 7 (IRF7) directly. Decitabine also increased the transactivation activity of ATO-rescued mutant p53 by inducing p53-serine-20 phosphorylation and blocking p53-inhibitory mouse double minute 2 homolog (MDM2). ATO and decitabine together killed p53-mutant AML cells and suppressed tumor growth in cell line-derived xenografts. In a first-in-human pilot clinical trial for testing the combination of ATO and decitabine (PANDA-T0 trial, NCT03855371), which enrolled five patients with AML/MDS harboring p53 structural mutations, the ATO and decitabine regimen produced manageable adverse events, and four of the five treated patients achieved complete remission at the level of the bone marrow, associated with p53 activation and interferon response. In 103 p53-mutant patients whose samples were deposited in Ruijin AML/MDS sample repository, 48 distinct p53 missense mutants were identified, 21 of which were classified as ATO and decitabine regimen applicable because of their competencies in activating p53 and interferon responses upon cotreatment. This study establishes an alternative treatment regimen for patients with p53-mutant AML/MDS and provides a proof-of-concept framework for p53-targeted therapy that differentiates between p53 mutations.

Emerging evidence suggests the Portfolio diet may confer benefits beyond managing hyperlipidemia. This systematic review and meta-analysis evaluated the association between adherence to the Portfolio diet and disease outcomes in observational studies. Four databases were searched through September 2025, and 10 studies consisting of 513,284 participants were included. Data from 4 prospective cohort studies (n = 352,396) were pooled using random-effects models for disease outcomes. Each 1-standard deviation increase in the Portfolio Diet Score (PDS) was associated with a 6% lower risk of cardiovascular disease (CVD) (RR = 0.94, 95% CI: 0.92 to 0.96) and a 10% lower risk of all-cause mortality (RR = 0.90, 95% CI: 0.86 to 0.95). Qualitative synthesis suggested that higher adherence to the Portfolio diet was associated with lower type 2 diabetes (T2D) risk and lower odds of cancer. In conclusion, higher adherence to the Portfolio diet is inversely associated with risks of CVD and mortality, while the favorable associations with T2D and cancer remain preliminary and hypothesis-generating.

Blood or marrow transplant (BMT) recipients are at increased risk of developing subsequent cutaneous malignant neoplasms. There is a need to develop risk stratification tools to identify those at highest risk to inform dermatologic surveillance and evaluate risk mitigation strategies. To develop and validate a risk stratification tool for subsequent cutaneous malignant neoplasms among BMT recipients that incorporates readily available patient and treatment information. This prognostic study included participants in the multi-institutional, longitudinal BMT Survivor Study (BMTSS), which included individuals who underwent autologous or allogeneic BMT from January 1, 1974, to December 31, 2014, at 1 of 3 participating sites in California, Minnesota, and Alabama and survived 2 or more years after BMT. The present cohort was restricted to individuals who underwent a single transplant and completed at least 1 BMTSS survey. Data were analyzed from December 3, 2024, to June 3, 2026. Risk stratification models included the following candidate variables: age at BMT, sex, race, ethnicity, prior cutaneous malignant neoplasm, pretransplant exposure to monoclonal antibodies, total body irradiation, chronic graft vs host disease, and posttransplant immunosuppression. The main outcomes were incident basal cell carcinoma (BCC), squamous cell carcinoma (SCC), or melanoma after BMT, ascertained based on participant report with medical record validation when available. Models for risk stratification were developed and validated using time-dependent area under the receiver operating characteristic (AUC) curve with repeated random partitioning into training and testing cohorts. Results were averaged to make a final model. Individuals were classified into low or high risk for subsequent cutaneous malignant neoplasm based on clinically meaningful risk stratification score thresholds. Of the 3448 included BMTSS participants (1917 [55.6%] male; mean [SD] age at BMT, 41.9 [19.2] years) who were alive without a cutaneous malignant neoplasm event 2 years after BMT, 282 (8.2%) were diagnosed with BCC, 183 (5.3%) with SCC, and 73 (2.1%) with melanoma. At 15 years after BMT, the test model AUCs were 0.81 (95% CI, 0.76-0.85) for BCC, 0.90 (95% CI, 0.86-0.93) for SCC, and 0.83 (95% CI, 0.74-0.90) for melanoma. The 15-year cumulative incidence of BCC, SCC, and melanoma in the low-risk group was 2.4% (95% CI, 1.5%-3.9%), 3.2% (95% CI, 2.4%-4.2%), and 0.8% (95% CI, 0.4%-1.5%), respectively. In the high-risk group, the 15-year cumulative incidence of BCC, SCC, and melanoma was 13.3% (95% CI, 11.6%-15.4%), 13.6% (95% CI, 11.2%-16.5%), and 4.4% (95% CI, 3.2%-6.1%), respectively. In this prognostic study, risk stratification models demonstrated excellent performance, highlighting potential utility for identifying BMT recipients who are at greatest risk of subsequent cutaneous malignant neoplasms.

We conducted a prospective, multicenter, Phase II study to evaluate the safety and efficacy of venetoclax/azacitidine/low-dose cytarabine/aclarubicin/granulocyte colony-stimulating factor (G-CSF) (VA-CAG) in young patients with newly diagnosed acute myeloid leukemia (ND-AML). The VA-CAG regimen included venetoclax (100 mg/day, Days 1-2; 200 mg, Day 3; 400 mg/day, Days 4-21), azacitidine (75 mg/m2, Days 1-7), cytarabine (10 mg/m2/12 h, Days 1-7), aclarubicin (12 mg/m2, Days 1, 3, 5, and 7), and G-CSF (5 μg/kg/day, Days 0-8). The primary endpoint was the complete remission (CR) rate after Cycle 1; the secondary endpoints included measurable residual disease (MRD)-negative remission rate, adverse events, and duration of remission (DOR). A total of 120 subjects were enrolled. The median age was 50 years (interquartile range, IQR, 36-57). The CR and composite complete response (CRc) rates were 91% (95% CI 86%-96%) and 95% (95% CI 91%-99%), respectively; 84% (95% CI 81%-88%) of the CRc patients achieved MRD-negative remission. Common Grade ≥4 adverse events included neutropenia (96%), thrombocytopenia (86%), febrile neutropenia (24%), pneumonia (4%), and sepsis (4%). The median times to recovery of absolute neutrophil count (ANC) ≥ 0.5 × 109/L and platelet count ≥ 20 × 109/L for responding patients were 14 days (IQR, 9-19) and 10 days (IQR, 4-16), and the 60-day mortality rate was 0%. With a median follow-up of 22.1 months (95% CI 20.4-23.7 months), 34 subjects subsequently received hematopoietic stem cell transplantation (HSCT). The median DOR values of all patients and the patients who did not receive HSCT were not reached. Hence, the VA-CAG regimen is a safe and effective first-line induction chemotherapy for ND-AML patients.

We report the results of a prospective trial aimed at describing the feasibility and accuracy of prostate-specific membrane antigen (PSMA) radio-guided surgery (RGS) during robot-assisted radical prostatectomy (RARP) with extended pelvic lymph node dissection (ePLND) in patients with prostate cancer (PCa). This was a phase 2 study (NCT04832958) that enrolled 82 patients with localized PCa and a lymph node invasion (LNI) risk >5%. All patients underwent preoperative PSMA positron emission tomography (PET). [99mTc]Tc-PSMA-I&S was administered intravenously the day before surgery, followed by single-photon emission computed tomography/computed tomography. Side effects, perioperative outcomes, and the performance characteristics of PSMA-RGS for LNI detection were measured. A total of 62 patients completed all study procedures and were included in the final analyses. Median blood loss and length of stay were 50 ml and 4 d, respectively. No adverse events after tracer administration or intraoperative complications were recorded. Four patients experienced a Clavien-Dindo grade 3 complication within 30 d. Overall, PSMA-RGS identified two additional patients with high-risk PCa and pathologically node-positive (pN1) disease compared with preoperative PSMA PET, exhibiting a lower positive predictive value (PPV) and a similar negative predictive value (NPV) in the per-patient analysis (PPV: 50% vs 70%; NPV: 83% vs 83%). The PPV and NPV at per-region analysis in patients with molecular imaging node-positive (miN1) disease were higher for PSMA-RGS than for PSMA PET (PPV: 55% vs 42%; NPV: 85% vs 81%). In conclusion, PSMA-RGS identifies additional patients with pN1 disease missed by preoperative PSMA-PET among men with high-risk PCa, and extends the ePLND template in patients with miN1 disease, in whom PSMA PET underestimates the nodal burden. However, its NPV is suboptimal for avoiding ePLND in patients with PCa who have a LNI risk >5% and negative PSMA-RGS findings during RARP.

The purpose of this study was to compare the image quality and lesion detection between ultra-low dose (ULD) chest-abdomen-pelvis computed tomography (CAP-CT) reconstructed with a deep-learning image reconstruction (DLR) algorithm, and standard-dose CT (STD-CT) in cancer follow-up. A total of 106 patients undergoing CAP-CT for the follow-up of cancer were prospectively included. Each patient underwent both STD-CT and ULD-CT acquisitions. ULD-CT images were reconstructed using two DLR levels (Smooth/Smoother). Dosimetric indicators, objective image quality, subjective image quality, and lesion detection were compared. Agreement between protocols and readers was assessed using Gwet's AC1 or AC2 coefficients. ULD-CT significantly reduced radiation exposure, with a mean CTDIvol reduction of -71.5% and dose-length product reduction of -71.5% (P < 0.05). Minor but statistically significant differences in HU values were observed between STD-CT and ULD-CT across most tissues. For all organs or tissues, image noise was significantly higher with ULD-CT-Smooth than with STD-CT (P < 0.001), and with ULD-CT-Smoother than with STD-CT, except for dorsal vertebra (P = 0.39) and trachea (P = 0.26). For all organs or tissues, image noise was significantly lower with the Smoother DLR level than with Smooth DLR level (P < 0.001). Agreement between STD-CT and ULD-CT regarding lesion detection was almost perfect for thoracic, abdominal, and bone lesions. Detection of infracentimetric hepatic was lower with ULD-CT, whereas detection of larger lesions (≥ 10 mm) remained similar. Subjective image quality was lower with ULD-CT, with moderate inter-reader agreement, and lower diagnostic confidence than with STD-CT. One of the two readers considered that 19%-24% of ULD-CT examinations were uninterpretable. ULD-CT with DLR offers substantial radiation dose reduction but resulted in poorer image quality and lower detection of small low-contrast abdominal lesions compared with STD-CT. Although lesion detection remained equivalent for thoracic and skeletal lesions, the high proportion of suboptimal or uninterpretable examinations may limit routine use of ULD protocols in cancer follow-up.

The rapid expansion of immuno-oncology (I-O) and other advanced therapies is reshaping the complexity of early-phase clinical development. While core Phase I principles remain fundamental across oncology, emerging modalities introduce additional requirements for translational integration, specialized safety management, and therapeutic-specific infrastructure. However, the organizational functions of Phase I units have not evolved at the same pace and remain largely centered on conventional operational responsibilities. Drawing on the practices of leading Phase I units across the USA, Europe, and China, and comparing evolving regulatory frameworks of the US Food and Drug Administration, European Medicines Agency/Medicines and Healthcare products Regulatory Agency, and China's National Medical Products Administration, we propose a global perspective on the future development of Phase I units. We present a three-layer framework encompassing core Phase I foundations applicable across oncology, advanced translational capabilities, and ecosystem-level functions supporting emerging therapeutic development. Beyond ensuring patient safety and high-quality trial conduct, modern Phase I units should integrate translational research, artificial intelligence-enabled and model-informed drug development, regulatory science, and public engagement, while specialized centers may additionally support advanced therapy infrastructure, such as point-of-care manufacturing. Recognizing the diversity of institutional resources, we further propose a collaborative network model to facilitate scalable implementation and international harmonization. This framework positions Phase I units as integrated platforms connecting clinical investigation, translational science, regulatory innovation, and emerging therapy development, providing practical guidance for building future-ready early-phase clinical research infrastructure worldwide.

Relapsed or refractory CNS lymphoma (R/R CNSL) has limited treatment options. This multicenter retrospective study enrolled 84 consecutive R/R CNSL patients (median age 59 years; 53 PCNSL, 31 SCNSL) to evaluate efficacy and safety profiles of glofitamab therapy. With a median of 2.5 prior lines of therapy, the objective response rates (ORR) and complete response rates (CRR) for PCNSL were 88% (CRR 59%) with glofitamab monotherapy (n=32) and 100% (CRR 81%) with combination therapy (n=21), respectively; for SCNSL, ORR and CRR were 88% (CRR 75%) with glofitamab monotherapy (n=8) and 91% (CRR 65%) with combination therapy (n=23). At 14.7 months follow-up, median progression-free survival was 19.5 months for PCNSL and 13.5 months for SCNSL. Cytokine release syndrome occurred in 40% (all grade 1-2) of patients, and ICANS in 8%. Glofitamab-based therapy demonstrated substantial activity with manageable toxicity in this study, offering a promising treatment paradigm for R/R CNSL patients.

Relapsed and/or refractory disease remains the leading cause of death in AML, highlighting the need for broadly applicable, high-sensitivity approaches to MRD detection. We developed AML-CAPP-Seq (Cancer Personalized Profiling by Deep Sequencing), a personalized hybrid-capture assay that tracks both canonical AML drivers and patient-specific variants identified by whole-exome sequencing. In 56 patients with longitudinal plasma and matched peripheral blood and bone marrow samples, AML-CAPP-Seq enabled universal MRD assessment and resolution of clonal dynamics using a median of 30.5 variants per patient. Plasma ctDNA outperformed cellular compartments for MRD detection and more strongly predicted relapse-free (HR 17.8, p<0.0001) and overall survival (HR 17.0, p<0.0001) than standard-of-care MRD methods. Among 29 allogeneic transplant recipients, peri-transplant ctDNA-MRD dynamics markedly improved relapse risk stratification (HR 36.0, p=0.0009). Together, these results establish personalized ctDNA profiling as a minimally invasive, highly sensitive, and generalizable platform for enhanced clinical MRD detection and clonal surveillance in AML.

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

Global Vhl knockout results in vascular defects and early lethality, limiting our knowledge of VHL (von Hippel-Lindau)/HIF (hypoxia-inducible factor) signaling in coronary vessel formation and homeostasis. The hypoxia pathway has been implicated in cardiovascular diseases (CVDs) characterized by inflammation and vascular remodeling, such as atherosclerosis, but its involvement in Kawasaki disease (KD) remains unknown. Coronary artery dilation and vessel rupture are the most serious complications of KD. However, the molecular mechanisms underlying these cardiac events are not fully understood. We investigated the role of the VHL/HIF pathway in cardiovascular pathology and its relevance to KD. We generated a novel mouse model with genetic hyperactivation of the hypoxia pathway in progenitors contributing to coronary vessels and cardiac fibroblasts. We characterized the model using echocardiography, magnetic resonance imaging, histology, and molecular profiling. In parallel, we examined cardiac tissues from patients with KD with fatal coronary aneurysms for evidence of HIF signaling and inflammation using immunohistochemistry. Mice with conditional deletion of Vhl in the Wt1 (Wilms tumor 1) lineage developed normally but exhibited cardiomegaly, vascular abnormalities, progressive coronary artery dilation, pericardial hemorrhage, and systemic inflammation shortly after birth. Histologic analysis revealed coronary arteritis, elastin breaks, vascular remodeling, smooth muscle cell loss, perivascular fibrosis, and frequent intracoronary thrombus formation. In addition, vascular calcification, severe cardiac inflammation, and interstitial hemorrhages were observed, culminating in sudden death between 15 and 20 weeks of age, likely due to vessel rupture. Cardiac transcriptomic profiling identified dysregulated expression of genes involved in extracellular matrix organization, epithelial-mesenchymal transition, angiogenesis, inflammation, coagulation, and calcification, indicating compromised vascular stability and increased remodeling in Vhl conditional knockout mice. Simultaneous deletion of Hif2a rescued both the cardiovascular abnormalities and transcriptomic profile observed in Vhl conditional knockout mice, implicating Hif2 (hypoxia-inducible factor 2) as a key mediator. Human KD cardiac samples showed expression of HIF2 in coronary lesions and surrounding inflammatory infiltrates, confirming hypoxia pathway activation in severe KD. Our findings establish HIF2 as a central driver of coronary inflammation, vascular remodeling, and thrombotic complications resembling those observed in severe KD. The Vhl/Wt1 conditional knockout mouse model recapitulates key cardiovascular features of KD and offers a valuable platform for mechanistic studies and therapeutic exploration.

Aging-associated loss of chromatin compaction is linked to derepression of retrotransposable elements (RTEs) in mouse and human tissues. Whether such RTE transcription contributes to the microglia activation that is common in aged brains is unknown. Here, we show that DAXX, a histone chaperone and RTE repressor, is downregulated during aging, preserves microglia homeostasis and inhibits cellular senescence. Loss of Daxx in young-adult microglia drives a reactive phenotype marked by chromatin decompaction at RTEs, loss of homeostatic markers, cell cycle re-entry and behavioral changes. This state leads to DNA damage and microglial depletion, followed by replacement with DAXX-deficient/Apoehigh microglia displaying features of senescence. Sustained induction of senescence relies on promyelocytic leukemia protein, a DAXX-interacting factor and interferon target. Together, these findings highlight the importance of heterochromatin maintenance in preserving adult microglial identity and plasticity, with broader implications for brain homeostasis, healthy aging and behavior.

Here we present the pathology workflow of the Mutographs project, a 7-year, multicancer genomics initiative led by the Sanger Institute and funded by Cancer Research UK. Mutographs aims to elucidate global differences in cancer incidence through whole-genome sequencing and mutational signature analysis of selected cancers with unexplained variation of incidence rate worldwide. The Protocol was developed by the International Agency for Research on Cancer, where pathology assessment and paired tumor-blood DNA extraction were performed. We provide details for handling frozen cancer tissues for multistep pathology evaluation and quality control, tumor-enrichment methods (such as macrodissecion and laser-capture microdissection) for certain tumor types, and manual and automated DNA extraction. This Protocol has achieved an overall success rate of 88% in providing high-quality DNA for whole-genome sequencing, aiding in the decision to exclude low-quality or noneligible tissues. This Protocol provides a reproducible, fully integrated pathology-molecular pipeline in challenging conditions to ensure high-quality DNA and reliable data interpretation at an unprecedented multinational scale and could be considered a model for high-quality tissue processing in international genomics initiatives. It is based on routine practice of pathology and requires practical experience of processing frozen tissues and morphological assessment of frozen tissues by digital pathology.

As primary WNT receptors, Frizzled (FZD) receptors behave as nonclassical GPCRs; however, their engagement with downstream transducers is largely unknown. Previous studies have suggested that β-arrestin recruitment to FZD receptors depends on its interaction with Dishevelled (DVL) and that this process regulates both canonical and non-canonical pathways. Here, we reveal that FZD6, which mainly mediates non-canonical WNT signalling, directly binds to β-arrestin 1 (βarr1) and report the cryo-EM structure of the FZD6-βarr1 complex, which revealed a unique shallow pocket in FZD6 for βarr1 engagement and identified arrestin-specific motifs that are distinct from those observed in previously reported FZD receptor-transducer complexes. Collectively, our findings establish a direct arrestin recruitment mechanism in FZD receptors that shares key features with arrestin engagement in classical GPCRs, suggesting that the engagement of core GPCR transducers may modulate WNT signalling specificity. These insights position FZD receptors as druggable targets akin to classical GPCRs, opening new avenues for targeting FZD receptors for a wide range of diseases, including cancer.

Endoplasmic reticulum (ER)-phagy receptors have elusive physiological functions beyond ER remodeling. To gain insight into these, we screen for cytoplasmic interactors of well-known receptors in mouse pancreatic ductal adenocarcinoma cells. Our data subsequently reveal a non-canonical action of PRKAR1A, a subunit of protein kinase A (PKA), in binding and activating FAM134B/C. At a molecular level, an amphipathic helix in the otherwise-disordered C terminus of FAM134B directly binds PRKAR1A dimers. Multimodal imaging reveals that this interaction occurs at interorganellar contact sites between the ER and liquid-like condensates of PRKAR1A. Mechanistically, ER-condensate contacts promote clustering of FAM134B/C with LC3B and recruit lysosomes, forming degradation hubs. Unexpectedly, these structures are found to control cytoplasmic signaling through FAM134B/C-mediated lysosomal sequestration of RhoA. Underscoring the physiologic relevance of this, these cytoplasmic-facing actions of FAM134B/C result in RhoA-dependent cell morphologic and migratory control in response to cyclic AMP (cAMP) stimuli. Thus, interorganellar contacts expand the roles of FAM134B/C ER-phagy receptors beyond ER degradation.

Tumor cells rely on sustained protein synthesis despite fluctuating metabolic stress. To examine how metabolic state directly influences translational output, we investigated lactate utilization. Intracellular accumulation of lactate, a central glycolytic product, acutely represses mRNA translation. Mechanistically, alanyl-tRNA synthetase 1 (AARS1) charges tRNAs with lactate instead of amino acids. Unlike the rapid and selective transfer of alanine to cognate tRNAAla, slower lactyl transfer permits lactate modification of non-cognate tRNAs, broadly compromising elongation fidelity. Functionally, this direct metabolic control over a fundamental process of the central dogma reshapes the translatome, operating as an intrinsic metabolic brake that aligns biosynthetic capacity with energy state. Notably, aggressive tumors elevate lactate transporters, limiting intracellular lactate accumulation and evading translational repression. Pharmacological blockade of monocarboxylate transporters restores intracellular lactate accumulation, re-establishes translational repression, and impairs tumor progression in mice. These findings uncover a metabolite-tRNA charging event directly rewiring translational output and reveal a metabolic vulnerability with therapeutic potential.

Generative AI (Gen-AI) has shown a remarkable impact in several biological research areas, from protein folding and de novo design to pathogenic mutation prediction. However, it remains unclear whether these molecular-level successes can translate to cellular and multicellular insights relevant to fields ranging from immunology to cancer and neurodegeneration. This arises from the intricate nature of the molecular mechanisms that determine cellular and organismal behavior, the lack of sufficient training data, and the multicellular nature of most pathophysiologic phenotypes. Novel Gen-AI frameworks are likely needed to integrate prior biological knowledge, such as molecular interaction networks, as well as guiding principles focusing the community's attention on solving biologically and translationally relevant problems. Drawing inspiration from Hilbert's list of 23 mathematical problems that have focused the mathematical community's attention for more than a century, we propose fifteen grand AI challenges to focus the biomedical community's attention on critically relevant questions, most of which still lack effective predictive methodologies.

Cellular senescence is a stress-response program that coordinates cell-autonomous growth arrest and non-cell autonomous remodeling of the tissue environment. Initially defined through its role in aging and tumor suppression, senescence is now recognized as a highly complex program shaped by cell-intrinsic and environmental factors. This complexity places senescence as a broad stress response mechanism operating at the intersection of metabolic dysfunction, chronic inflammation and tissue repair. Given its constant exposure to metabolic, inflammatory and toxic stress alongside its high regenerative demands, the liver is particularly prone to engage senescence as a protective mechanism to maintain tissue integrity. In this context, senescence exerts dual and often opposing effects. This review discusses the multifaceted roles of senescence along the liver disease trajectory, from its early protective functions in tissue repair to its later pathologic contributions to chronic liver disease and tumor progression, underscoring how cell type, stage of senescence program, disease stage and microenvironmental cues shape these divergent outcomes. We highlight the potential of emerging therapeutic interventions that modulate senescence, including pro-senescence, senolytic, senomorphic, and immune-modulating strategies. Advancing our understanding of the multifaceted roles of cellular senescence will be essential to translate this knowledge into therapies that selectively modulate senescent cell states, harnessing their beneficial effects while limiting their detrimental roles in chronic liver disease and cancer.

Direct targeting of the oncoprotein MYC has not yet been successful. We here report a novel dual protein degrader, GT19630, which binds directly to MYC and G1 to S phase transition protein 1 (GSPT1). GT19630 disrupts a novel feedforward loop of MYC and GSPT1, where MYC promotes transcription of GSPT1, and GSPT1 senses the stop codon of MYC to properly terminate its translation. The agent induces integrated stress response and abrogates oxidative phosphorylation through inhibition of the TCA cycle, resulting in apoptosis. GT19630 has superior activity compared to GSPT1- targeting molecular glues. GT19630 induces profound anti-proliferative effects and apoptosis at low nanomolar concentrations in a multitude of leukemia and lymphoma cell lines and primary samples, including those with TP53 mutations. GT19630 is highly active in vivo in models of therapy-resistant hematologic malignancies, including Burkitt's lymphoma, acute myeloid leukemia (AML) and multiple myeloma. CD34+ AML blasts overexpress MYC protein compared to normal hematopoietic stem/progenitor cells (HSPCs) and GT19630 induces greater cytotoxicity in AML cells compared to normal HSPCs. Further, GT19630 restores sensitivity to venetoclax and profoundly prolongs survival in vivo in venetoclax-resistant AML. GT19630 was well tolerated in humanized Crbn mice. In conclusion, our data support the development of the MYC/GSPT1 degrader GT19630 as a therapeutic strategy of MYC-driven hematologic malignancies.

Thalidomide, initially marketed as a sedative and anti-emetic, was withdrawn in the 1960s due to catastrophic teratogenic effects. Decades later, thalidomide was reconsidered as an anticancer agent, resulting in FDA approval against multiple myeloma. This success spurred the development of structural analogs lenalidomide and then pomalidomide as cancer therapies (immunomodulatory imide drugs, IMiDs). Subsequent landmark studies have revealed that IMiDs act as molecular glue degraders, co-opting the E3 ubiquitin ligase cereblon (CRBN) to induce the polyubiquitination and proteasomal degradation of several proteins (e.g., the transcription factors IKZF1/3). From 2015 onwards, IMiDs have also been widely incorporated into the design of PROteolysis-TArgeting Chimeras (PROTACs) to recruit additional target proteins to CRBN and trigger target depletion. IMiDs have proven particularly suited for PROTAC development due to their efficient co-opting of CRBN and structural simplicity. Intense medicinal chemistry and chemical biology efforts have expanded CRBN-binding chemotypes far beyond classical thalidomide analogues, yielding a diverse array of molecular glues and PROTAC handles with tailored neosubstrate profiles and properties. In this review, we systematically organize the synthetic landscape of CRBN binders from a chemotype-centric perspective. After providing a historical overview of thalidomide, we classify CRBN binders into four major families: phthalimido glutarimides (thalidomide derivatives), isoindolinone-2-yl glutarimides (lenalidomide-type scaffolds), glutarimides linked to alternative heterocyclic or aromatic motifs, and emerging glutarimide-free CRBN modulators. For each class, representative examples are discussed alongside modular routes and recurrent derivatization vectors that have enabled both clinically used IMiDs and next-generation molecular glue degraders and PROTACs. By integrating structural insights with synthetic strategies, we provide a structure-guided roadmap for chemists and chemical biologists seeking to navigate, rationally expand, and functionally tailor the growing chemical space of small-molecule CRBN binders.

The clinical efficacy of photodynamic therapy (PDT) is fundamentally limited by the scarcity of efficient photosensitizers (PSs) and the oxygen dependence of singlet-oxygen-mediated cytotoxicity. Here we report pentaperylene decaimide selenide (PPD-Se), a nanographene-derived photoelectronic material that functions as a high-performance Type-II photosensitizer. PPD-Se exhibits broadband absorption (300-650 nm), enhanced intersystem crossing enabled by a selenium-induced heavy-atom effect, a small ΔEST (0.50 eV), and a high 1O2 quantum yield (ΦΔ = 0.40). To address hypoxia-limited PDT, PPD-Se nanoparticles were covalently integrated with microalgae to construct an algae@PPD-Se biohybrid, in which PPD-Se is shielded from premature activation yet undergoes glutathione (GSH)-triggered release in the tumor microenvironment. Cleavage of disulfide linkages restores the photosynthetic activity of algae, enabling light-driven O2 production that alleviates local hypoxia and simultaneously boosts PPD-Se-mediated ROS generation. The biohybrid exhibits enhanced intracellular uptake, amplified ROS production, and potent apoptosis induction under white light-emitting diode (LED) irradiation (400-700 nm, 1 mW·cm-2). In vivo, algae@PPD-Se significantly downregulates HIF-1α, restores intra-tumoral oxygenation, and achieves marked tumor growth inhibition without observable systemic toxicity. This study introduces a dual-functional optoelectronic-biological PDT platform that couples a newly designed nanographene photosensitizer with photosynthetic oxygenation, offering a mechanistically driven strategy to overcome the oxygen dependency of PDT.

Accurate capture and molecular analysis of circulating tumor cells (CTCs) from whole blood are crucial for early cancer diagnosis, prognosis and personalized therapy. Conventional enrichment methods tend to induce cellular damage, compromising cell viability and limiting downstream functional analysis. To address this, we present an Acousto-SERS Integrated Trapping Chip (ASSIT Chip) which combines acoustic sorting with surface enhanced Raman scattering (SERS) fingerprinting for label-free, non-invasive CTC identification and recovery. In this platform, interdigital transducers generate a finely tuned acoustic field that gently sorts CTCs from whole blood via contactless manipulation. The sorted cells are then transported to an integrated SERS region for label-free spectral fingerprinting. From acoustic sorting to optical identification, the fully non-invasive workflow minimizes cellular damage. Additionally, the dual-mode approach integrates physical size selection with chemical biomolecular identification, thus significantly improving the enrichment purity. Through precise control of acoustic and hydrodynamic fields, the ASSIT Chip achieves high sorting efficiency (86.7%) and recovery rate (96.9%). By employing a microsphere-templated plasmonic substrate, label-free SERS detection accurately distinguishes CTCs from blood cells with an accuracy of 96.6%. More importantly, the retrieved CTCs retain high viability and undiminished proliferative capacity, enabling subsequent cell culture and downstream biological studies. The validation with whole blood samples confirms that the ASSIT Chip provides a robust platform for cancer diagnostics and functional CTC analysis, holding considerable promise for clinical translation and personalized medicine.

The advancement of nanoplasmonics and meta-optics has positioned localized surface plasmon resonance (LSPR) biosensing as an attractive technology for point-of-care testing (POCT), overcoming key limitations of traditional propagating surface plasmon resonance (PSPR) in terms of miniaturization and cost. However, the multistep and complex surface chemistry required for conventional nanostructure-based LSPR biosensors remains a major obstacle to their reproducible and efficient fabrication. To address this issue, we developed a more streamlined surface modification strategy based on magnetron sputtering of a uniform graphene-like carbon (GLC) nanofilm onto ultrasmooth gold nanogroove arrays (UGNA), followed by electrochemical oxidation to enable direct biomolecular coupling. This graphene-like oxide-modified UGNA (GO-UGNA) eliminates the need for intermediate linking layers, significantly simplifying the fabrication process while maintaining efficient probe immobilization. Utilizing a miscellaneous-protein strategy combined with initial rate analysis, the biosensor detected tumor markers carbohydrate antigen 19-9 (CA19-9), carbohydrate antigen 15-3 (CA15-3), and carbohydrate antigen 72-4 (CA72-4), achieving broad linear ranges (1-1000 U/mL) in both wavelength-based and intensity-based detection modes. The limits of detection were 0.410 U/mL, 0.486 U/mL, and 0.690 U/mL for CA19-9, CA15-3, and CA72-4 in wavelength mode, whereas those in intensity mode were 0.119 U/mL, 0.103 U/mL, and 0.458 U/mL, respectively. Clinical sample analysis demonstrated excellent concordance with a commercial electrochemiluminescence system (Roche Cobas 8000 e801). With its rapid modification process (∼10 min) and reproducible fabrication, this work establishes a promising platform for developing practical, low-cost POCT biosensors with clinical-grade accuracy.

Accurate evaluation and reliable quantification of telomerase activity in cancer cells are essential for early cancer diagnosis and treatment. In this work, we developed a sensitive and reliable electrochemical method for telomerase detection, based on the electrocatalytic signal amplification of acetaminophen (AP) by monodispersed gold nanorods (Au NRs) and ordered mesoporous carbon (OMC). Signal switching is achieved via the incorporation of dideoxycytidine, while cobalt ions regulate sequence extension, enabling the efficient immobilization of Au NR-labeled signal probes on the electrode surface. The proposed biosensor exhibits excellent stability, specificity, and repeatability, highlighting its strong potential for sensitive and accurate in vitro cancer diagnostics. This strategy may contribute to the advancement of early cancer screening technologies. However, challenges remain for clinical translation, including the complexity of real biological samples and the need for standardized large-scale fabrication. Addressing these limitations will be critical for the development of next-generation biosensing platforms.

Cancer remains a major global health challenge, highlighting the need for effective biomarker monitoring. Carcinoembryonic antigen (CEA), a widely used tumor-associated marker, is crucial for cancer diagnosis. Surface-enhanced infrared absorption (SEIRA) spectroscopy, which detects unique molecular vibrations, offers a promising approach for trace cancer biomarker detection. However, conventional nanoantenna-based SEIRA is limited by small high-field regions and weak molecule-metasurface coupling, restricting signal enhancement. To address these challenges, this work presents a multi-hotspot coupled metasurface with high degrees of freedom, optimized via a particle swarm algorithm. By expanding the effective hotspot area and optimizing the loss ratio of the metasurface, we significantly enhanced the molecule-metasurface normalized coupling strength (μeff) and amplified the CEA molecular enhancement signal (ΔA). This ΔA enhancement results from the combined effects of μeff and the optimized loss ratio (ξ). Experimental results demonstrate excellent linearity for CEA detection across a wide concentration range from 0.1 ng/mL to 10 μg/mL, with a minimum distinguishable concentration as low as 0.1 ng/mL. This performance represents an order-of-magnitude improvement over conventional nanoantenna-based SEIRA sensors. This strategy offers promising potential for sensitive detection of other disease biomarkers, advancing human health monitoring.

Noninvasive detection of bladder cancer remains challenging due to the limited analytical performance of current urine-based assays under realistic detection conditions. Here, we report a linear programmable DNA nanostructure that integrates multivalent aptamer recognition with CRISPR/Cas12a signal transduction for detecting tumor-derived urinary exfoliated cells. Assembled via hybridization chain reaction, the multivalent scaffold was shown to enhance ligand-cell binding stability under mechanically perturbed detection processes, thereby supporting mechanically stable biological recognition and yielding an approximately 14-fold increase in cellular binding affinity compared with monovalent aptamers. The optimized architecture (MAP12) enables detection with a LOD of 1.1 cells/mL in model systems and achieves high diagnostic performance in clinical urine samples (92% sensitivity, 88% specificity; AUC = 0.9424), supporting dual signal readouts via fluorescence and lateral flow devices (LFD). This work establishes a DNA nanostructural strategy for reliable, rapid, and noninvasive cancer cell detection under realistic operational conditions.

Resistance to tyrosine kinase inhibitors (TKIs) severely limits their long-term efficacy in renal cell carcinoma (RCC). Here, we identify prolyl 4-hydroxylase subunit alpha 3 (P4HA3) as a critical driver of TKI resistance and elucidate its underlying mechanism. P4HA3 was markedly upregulated in TKI-resistant RCC cells, and its overexpression promoted proliferation and resistance to sunitinib and axitinib, whereas P4HA3 depletion restored drug sensitivity and suppressed tumor growth in vitro and in vivo. Mechanistically, P4HA3 inhibited ferroptosis by stabilizing the cystine transporter SLC7A11, thereby maintaining redox homeostasis and suppressing lipid peroxidation. P4HA3 directly interacted with SLC7A11 and induced hydroxyproline modification at residues P153 and P161, which attenuated NEDD4-mediated ubiquitination and proteasomal degradation of SLC7A11. Moreover, a hydroxylase-inactive P4HA3 mutant failed to stabilize SLC7A11, suppress ferroptosis, or promote TKI resistance, demonstrating the essential role of P4HA3 enzymatic activity. Importantly, pharmacological inhibition of P4HA3 using Tubuloside A or 1,4-DPCA disrupted the P4HA3-SLC7A11 axis and significantly enhanced TKI efficacy across RCC cell lines, patient-derived organoids, and xenograft models. Together, these findings establish a hydroxylation-dependent P4HA3-SLC7A11-NEDD4 axis that drives ferroptosis evasion and TKI resistance in RCC and highlight P4HA3 as a promising therapeutic target.

The proliferation of hepatocytes is essential for liver regeneration. Hepatocytes harbor supernumerary centrosomes, which can lead to multipolar spindles, chromosome misalignment and prolonged mitosis. How hepatocytes overcome these mitotic challenges to efficiently regenerate is unclear. To understand how hepatocytes cope with mitotic challenges, we established a high-quality in vivo CRISPRi screen in regenerating mouse liver. By cross-referencing with gene essentiality data in cell lines, we identified genes required specifically by hepatocytes for proliferation. Knockout mice, RNA sequencing and proteomic approaches were used to understand the function of the candidate gene and the mechanism in regulating hepatocyte proliferation. NEURL1B, a rarely studied E3 ligase, is specifically required for hepatocyte cell division in vivo. Inhibiting NEURL1B results in enlarged, polyploid hepatocytes, impaired liver repopulation, and resistance to liver cancer. This dependence on NEURL1B arises from a unique spindle pole organization mechanism used by hepatocytes. At the start of mitosis, hepatocytes exhibit multiple microtubule-organizing centers (MTOCs) organized by NUMA1. These MTOCs undergo extensive remodeling, including clustering and inactivation to form bipolar spindles. NEURL1B localizes to and destabilizes MTOCs, with NUMA1 identified as a substrate. By destabilizing MTOCs, NEURL1B facilitates inactivation of extra MTOCs and enables mitosis progression. Human single nucleus RNA-seq and cell culture data support a conserved role for NEURL1B in human hepatocytes. Our approach to studying mitosis in vivo reveals a unique mechanism used by hepatocytes to manage supernumerary centrosomes and identifies NEURL1B as a critical regulator of this pathway, paving the way to new treatments for liver regeneration and cancer.

Host-derived lipids undergoing enzymatic or nonenzymatic oxidation play critical roles in regulating inflammation. Polyunsaturated fatty acids, cholesterol, and cholesterol intermediates can be enzymatically oxidized and serve as signaling mediators controlling tissue homeostasis and immunity. Spontaneously generated oxidized lipids, including nonenzymatically oxidized phospholipids (oxPLs), result from oxidative stress and accumulate during inflammation, affecting cellular metabolism, immune cell functions, and cell fate. These distinct classes of oxidized lipids not only share overlapping inflammatory roles but also exhibit divergent effects depending on their molecular structures and cellular targets. This Review highlights the double-edged nature of oxPLs: Although their transient production triggers protective responses, their accumulation sustains inflammation, contributing to tissue damage. We also discuss the emerging roles of oxPLs in cell death programs, immune cell activation, and stromal cell functions, which are critical processes favoring tumor growth. Overall, we highlight how oxidized lipids orchestrate immune responses and explore their contribution to infectious diseases and cancer.

Immunoglobulin G (IgG)-based monoclonal antibodies are effective therapies for cancer, autoimmune diseases, and migraine. However, they are actively transported across the placenta by the neonatal Fc receptor (FcRn), limiting their use during pregnancy. Using mouse models and an ex vivo human placental perfusion system, we show that although FcRn binds albumin independently of IgG, albumin is not transported to the fetus in mice or across human placental tissue. Fusion of IgG to albumin markedly reduced transplacental transport in both models while preserving the prolonged plasma half-life conferred by FcRn. Similarly, fragment antigen-binding fragments fused to engineered albumin with enhanced FcRn binding showed minimal fetal exposure. In a mouse model of fetal and neonatal alloimmune thrombocytopenia, albumin fusion of an anti-human platelet antigen IgG reduced fetal antibody transfer and attenuated thrombocytopenia in the offspring. These findings identify albumin as an attractive fusion partner for biologics intended to minimize fetal exposure during pregnancy.

The cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway promotes tumor immunogenicity, but intratumoral STING agonists, despite strong preclinical efficacy, have shown limited clinical efficacy. The mechanisms restricting therapeutic STING activation remain unclear. In mice, intratumoral delivery of the endogenous STING ligand cyclic guanosine monophosphate-adenosine monophosphate (GMP-AMP) (cGAMP) using viruslike particles (VLPs) preferentially activates STING in dendritic cells and primes circulating tumor-specific T cells. Using this system, we investigated mechanisms limiting effective STING-based immunotherapy. STING-induced type I interferon signaling was dispensable for cGAMP-VLP-mediated tumor control. In contrast, dendritic cell autophagy was required for generating circulating antitumor CD8 T cells and for regulating baseline neutrophil levels in lymph nodes. cGAMP-VLP overrode this regulation, inducing neutrophil accumulation in tumors and draining lymph nodes that limited efficacy. Neutrophil depletion enhanced tumor control through mechanisms involving neutrophil elastase and programmed cell death 1 ligand 1. These findings reveal that defective dendritic cell autophagy and neutrophil-mediated immunosuppression, rather than insufficient interferon signaling, hinder the effectiveness of intratumoral STING immunotherapy in preclinical mouse models.

Chimeric antigen receptor (CAR) technology has revolutionized B-cell malignancy treatment by enabling T cells to effectively recognize and target lineage-specific surface antigens. However, CAR T cells show limited efficacy against myeloid neoplasms and solid tumors due to challenges in identifying suitable surface targets. In this study, we present a CAR targeting the intracellular WT1 oncoprotein, cross-presented by surface HLA class II (HLA-II) alleles. WT1-CAR T cells, derived from an antibody raised solely against a WT1 peptide, recognized the WT1330-348 peptide promiscuously presented by 18 out of 20 tested HLA-II alleles, overcoming traditional HLA restrictions. WT1-CAR T cells specifically recognized leukemic cells in a WT1- and HLA-II-dependent manner and mediated an antitumor response in vitro and in vivo. This approach broadens CAR-targetable antigens beyond traditional HLA restrictions and offers a promising therapeutic option to a wide and genetically diverse patient population. Leveraging the promiscuous binding of HLA-II-peptide complexes, we developed a CAR T-cell approach targeting an intracellular oncoprotein WT1 presented across diverse HLA-II families. Our study establishes a framework for CAR therapies against intracellular antigens, extending potential CAR T-cell applications to new cancer types and patient populations.

MALT1 (mucosa-associated lymphoid tissue lymphoma/leukemia protein 1)-TRAF6 [tumor necrosis factor receptor (TNFR)-associated factor 6] interaction drives lymphocyte activation and adaptive immunity, but it also contributes to maintaining immune homeostasis. MALT1 exists in two isoforms that differ only by either encoding two (MALT1A) or one (MALT1B) TRAF6 binding motif (T6BM). The human mutation MALT1 E806D in T6BM2, expressed in both MALT1A and MALT1B, has been associated with an immune disorder combining symptoms of immune deficiency and autoimmunity. Here, we report that the orthologous germline mutation MALT1 E814D is sufficient to induce a fatal autoimmune syndrome in mice. We demonstrate that species-specific differences in the effects of T6BM2 disruptions can be attributed to alterations in MALT1 splicing and that immune homeostasis is restored by genetically enforcing expression of MALT1A in MALT1 E814D mice. Thus, alternative MALT1 splicing allows tuning of TRAF6 association, thereby functioning as a molecular rheostat to balance between optimal immune activation and maintenance of peripheral tolerance.

ETS variant transcription factor 2 (ETV2) serves as a foundational transcription factor for endothelial lineage specification. However, the lineage-specific cofactors that orchestrate with ETV2 during endothelial fate commitment remain elusive. Here, we demonstrate that ETV2 drives the rapid forward programming of human pluripotent stem cells (hPSCs) into endothelial cells (ECs) by direct remodeling of endothelial-specific enhancers. Crucially, we identify T cell acute lymphocytic leukemia protein 1 (TAL1), which is traditionally characterized as a hematopoietic regulator, as an indispensable cofactor for ETV2-mediated endothelial commitment. Distinct from its role in murine development, TAL1 deficiency in hPSCs not only aborts the endothelial program by impairing H3K27ac deposition at key enhancers but also triggers a profound lineage redirection toward a mesenchymal fate. Mechanistically, TAL1 physically interacts with ETV2 to recruit the p300, thereby facilitating a permissive chromatin environment for endothelial identity. By leveraging an hPSC-based differentiation model, our findings establish TAL1 as a master gatekeeper of human EC specification and provide a molecular blueprint for how ETV2-centric complexes synergistically govern human cell fate.

Acute myeloid leukemia (AML) is an aggressive hematological malignancy arising from hematopoietic stem and progenitor cells (HSPCs). Current treatments often fail to eradicate AML; therefore, new therapeutic strategies are essential. Here, we reveal that RNA terminal uridylyl transferase enzymes 4 and 7 (TUT4/7) are druggable therapeutic targets, whose genetic deletion suppresses AML growth, induces apoptosis, and improves the survival in leukemic mouse models. Notably, a preclinical TUT4/7 inhibitor promotes cell death in samples from patients with AML and synergizes with venetoclax. Mechanistically, TUT4/7 inactivation suppresses mevalonate pathway gene expression, compromising the cholesterol synthesis pathway. Current AML therapies often cause severe hematopoietic toxicity. Although Tut4/7 deletion results in inflammatory activation throughout the hematopoietic system, this is permissive to a normal life span and Tut4/7 deficiency does not compromise HSPC function. Together, these findings identify TUT4/7 as druggable targets, whose inactivation suppresses AML while sparing normal hematopoiesis. In combination with venetoclax, this represents a promising therapeutic strategy.

Regulatory T cells (Treg) infiltrate most tumors, and increased Treg infiltration is correlated with reduced survival in patients with cancer. Although Tregs suppress antitumor immunity, they are also integral for preventing harmful inflammation in nontumor tissues, hindering the development of cancer therapeutic strategies targeting all Tregs systemically. In this study, we used intratumoral (IT) delivery of diphtheria toxin (DT) in Foxp3DTR mice to deplete Tregs within tumor tissues while leaving peripheral Tregs intact. IT delivery of DT reduced Treg frequencies in the tumor, which promoted potent tumor control without autoimmunity. Interestingly, tumor control was principally mediated by CD4+ T cells, whereas CD8+ T cells only contributed when CD4+ T cells were absent. Although conventional dendritic cells (cDC) were required to clear tumors, either type I DCs or type II DCs (cDC2) alone were sufficient to promote tumor control. Distant secondary tumors, mimicking metastases, were also controlled by IT Treg ablation in the primary tumor. Mechanistically, IT Tregs suppressed antitumor T-cell responses by blocking the acquisition and presentation of tumor antigen by cDC2s. Importantly, similar mechanisms of control were observed using a clinically translatable IT Treg-depleting anti-CCR8 antibody. Collectively, these findings reveal a distinct therapeutic strategy that leverages CD4+ T cells upon ablation of Tregs within tumors. Localized ablation of intratumoral Tregs activates an alternative arm of the immune system by acting on type II conventional dendritic cells and CD4+ T cells, inducing prolonged suppression of tumor growth.

The influence of genetic ancestry on cancer development and treatment response is underappreciated within pediatric neuro-oncology. Pediatric brain tumors (PBT) comprise the deadliest malignancies among childhood cancers. Poorer prognoses are accentuated in minority populations and remain skewed despite attempts to address social determinants of health and clinical trial underrepresentation. Ancestral differences in tumor biology and pharmacogenomics may contribute to outcome discrepancies. As such, we sought to review the role of genetic ancestry in PBT biology and treatment response and investigate whether foundational preclinical models adequately reflect ancestral heterogeneity. Population-linked genetic events are correlated with biological tumor differences but remain nascently explored. Likewise, there are indications that ancestral variations may influence treatment outcomes. Yet, despite data supporting response differences between ancestries in non-brain tumor populations, few therapeutics have been pharmacogenetically tested. Additionally, assessment of ancestral identity of brain cancer cell lines demonstrated lack of genetic ancestral heterogeneity, with European and Asian overrepresentation. Hence, therapeutic insights are being constructed with a preselected population lacking genetic diversity, decreasing the breadth of clinical translation and further exacerbating health disparities. This review proposes steps to address this gap and promote ancestral heterogeneity in treatment development within pediatric neuro-oncology.

Transcription factors (TFs) are aspirational therapeutic targets, as their dysregulation drives altered cell states. Yet many disease-relevant TFs are disordered and lack canonical binding pockets, frustrating direct small-molecule inhibition. Indirectly targeting the effector molecules that modulate TF function is a promising, underexplored alternative. Here we report a strategy for capturing cancer-specific protein-protein interactions using context-dependent µMap photoproximity labeling. With an intein-based method for catalyst conjugation in biochemically intact nuclei, we capture unique c-Myc interactomes in healthy and cancerous prostate cells and mine them for druggable vulnerabilities. We identify STE20-like kinase (SLK), a cancer-specific interactor that stabilizes c-Myc, drives epithelial morphology and is essential for tumorigenesis. Mechanistically, SLK phosphorylates c-Myc at serine 329, antagonizing GSK3β-dependent phosphodegron phosphorylation. This interaction is associated with a splicing change promoting nuclear localization of the long SLK isoform. Patient data link this isoform to c-Myc target expression across tumor types; the interaction validates across diverse tissues.

Multidisciplinary tumour boards (MDTs) are the standard for gastrointestinal oncological decision-making but remain resource-intensive. Whether specialty-specific role prompting induces genuinely distinct clinical reasoning in large language models (LLMs)-or merely role-appropriate language around an invariant output-has not been systematically tested. We applied five zero-shot prompting frameworks and a majority-vote ensemble to GPT-5 across 100 gastrointestinal oncology cases with MDT-validated decisions: a simulated MDT, multi-expert deliberation, three specialist personas, and a majority-vote ensemble. Concordance with MDT recommendations ranged from 78% to 87%, with no significant inter-framework differences (Cochran's Q = 8.46, p = 0.133). Specialty-characteristic language was near-universal (97-100%) but uncorrelated with accuracy. Embedding analysis revealed high semantic similarity across personas (cosine similarity 0.805-0.836; η² = 0.049), contrasting with substantially greater output separation under multi-expert deliberation (η² = 0.554-0.581). GPT-5 reliably adapts linguistic style to clinical personas but produces limited specialty-specific output diversity, supporting its role as a decision-support adjunct rather than an autonomous specialist simulator.

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

TIE2 is an endothelial receptor tyrosine kinase (RTK) essential for vascular integrity, and constitutively active TIE2 mutants are involved in venous malformations (VMs). VMs are currently treated by surgery or sclerotherapy, but effective pharmacologic options remain limited, especially for surgically challenging Blue Rubber Bleb Nevus Syndrome (BRBNS). TIE2 activation has typically been assessed by immunoblotting of phosphoproteins, yet scalable assays applicable to pharmacological evaluation are still lacking. Here, we present a bioluminescence resonance energy transfer (BRET) biosensor that quantifies receptor-proximal TIE2 activation by monitoring recruitment of GRB2, a major adaptor in RTK signaling. This BRET sensor detects angiopoietin-1 (ANG-1) at physiological plasma concentrations and is compatible with a high-throughput format. Furthermore, it captures constitutive activity of TIE2 mutants associated with VMs and enables pharmacological evaluation of TIE2 variants. Notably, the T1105N-T1106P variant, frequently observed in BRBNS, exhibited relatively higher sensitivity to the clinical oncology drugs regorafenib and lenvatinib. To validate selected BRET-defined pharmacological profiles in an endothelial context, we used HUVEC-derived HUEhT-2 cells and established endothelial TIE2-GRB2 BRET measurements, followed by downstream AKT/ERK signaling analysis and tube formation assays. These endothelial assays largely supported the selected HEK293T BRET findings, particularly the pharmacological profile of T1105N-T1106P mutant, at receptor-proximal, downstream signaling, and functional levels. Together, this BRET-based biosensor establishes a two-step framework: HEK293T BRET enables efficient primary profiling, whereas endothelial assays provide orthogonal validation to support biological interpretation. This framework offers a practical strategy for prioritizing pharmacological candidates for further endothelial and in vivo validation toward therapeutic development.

IDH1 and IDH2 are frequently mutated in various cancers, including acute leukemias. However, the distinct mechanisms by which mutant IDH1 or IDH2 drive hematopoietic neoplasms remain poorly understood. Here, we analyzed DNA methylation in IDH1- and IDH2-mutant acute myeloid leukemia and found neutrophil lineage-specific epigenetic alterations in IDH1-mutant patients that went along with severely impaired neutrophil differentiation. Transcriptional analysis of normal hematopoiesis in humans and mice revealed a strong physiological upregulation of IDH1/Idh1 in myeloid progenitors. To study the functional effects of Idh1 mutations on hematopoiesis in a preleukemic setting, we used a genetically engineered inducible mouse model expressing a heterozygous Idh1 mutation under control of the endogenous promotor. Our study revealed a cell-intrinsic block in neutrophil differentiation caused by repression of myeloid transcription programs in neutrophil progenitors. This included impaired expression of Cebpe, which encodes a key transcription factor regulating neutrophil differentiation. Reactivation of Cebpe expression, by overexpression of its upstream regulator Cebpa or following treatment with hypomethylating agents, restored differentiation, indicating that the differentiation block is reversible. In summary, we found a reversible, preleukemic impairment of neutrophil differentiation in IDH1-mutant hematopoiesis that correlates with elevated IDH1 expression in myeloid progenitors and likely explains the strong association of IDH1 mutations with myeloid neoplasms.

Lenalidomide, a maintenance treatment in multiple myeloma first-line therapy, increases the risk of secondary malignancies, including B-cell precursor acute lymphoblastic leukemia (B-ALL). We present a comprehensive molecular characterization of 57 patients with lenalidomide-associated B-ALL (LenB-ALL), revealing 3 mutational subgroups: (1) TP53mt (30%); (2) IDH2mt (p.R140Q) (23%); and (3) other, including NRAS/KRASmt. Remarkably, IDH2 R140Q mutations were highly enriched in LenB-ALL compared with those in primary B-ALL (P< .001). Furthermore, IKZF1 intragenic deletions, often subclonal and likely RAG recombinase-mediated, were observed in 54% (7/13) of IDH2mt patients with LenB-ALL. IDH2 mutations were not restricted to the leukemic clone: they persisted during measurable residual disease-negative remission and were identified in lymphoid as well as myeloid cell populations using fluorescence-activated cell sorting and single-cell RNA sequencing. This indicates a preleukemic origin of the IDH2 mutation within the context of clonal hematopoiesis. Transcriptomic and DNA methylation analyses revealed a distinct gene expression profile and a DNA hypermethylation phenotype in IDH2mt LenB-ALL, including IDH2mt-specific as well as lenalidomide-associated features. We propose that lenalidomide promotes the expansion of IDH2-mutated clonal hematopoiesis and, via IKAROS downregulation, induces a maturation arrest at the B-cell precursor stage. Subsequent genetic or epigenetic alterations render leukemogenesis independent of ongoing lenalidomide exposure. All these data define IDH2mt B-ALL as a distinct molecular subtype that is markedly overrepresented after lenalidomide treatment and highlight clonal hematopoiesis as a key contributing factor in the development of LenB-ALL.

Nucleotide metabolism is a fundamental biochemical process, providing the building blocks for DNA and RNA synthesis while driving critical cellular functions like energy transfer, signal transduction, and coenzyme synthesis. The precise regulation of nucleotide de novo synthesis, salvage, and degradation pathways is imperative for maintaining genomic stability and tissue homeostasis. Aberrations in these networks are increasingly recognized as primary drivers in various pathologies; notably, cancer cells exploit metabolic-functional reprogramming to enable rapid proliferation and therapeutic resistance. Despite the growing recognition of these metabolic vulnerabilities, an integrated understanding of how specific enzymatic dysregulations translate into diverse disease phenotypes remains fragmented. This review systematically synthesizes the physiological regulatory mechanisms governing nucleotide metabolism and describes the molecular underpinnings of its dysregulation across malignant, immune, and neurodegenerative disorders. We critically examine how specific metabolic enzymes are exploited to drive pathogenesis and highlight the complex interplay between nucleotide metabolites and inflammatory or oncogenic signaling cascades. Furthermore, we evaluate recent clinical innovations in precision medicine, focusing on novel inhibitors targeting key metabolic nodes. By mapping these intricate metabolic networks, this article provides a crucial basis for overcoming current therapeutic challenges, offering profound insights to accelerate the development of next-generation targeted therapies.

Recent phase I/II clinical trials have demonstrated that chimeric antigen receptor (CAR) T cells targeting the disialogangliosade GD2 represent a promising therapeutic option for pediatric patients with relapsed or refractory high-risk neuroblastoma (NB). However, incomplete and heterogeneous clinical responses highlight the need to improve CAR T-cell efficacy and persistence. We previously demonstrated the therapeutic benefit of combining the dual insulin-like growth factor 1 receptor/insulin receptor (IGF1R/IR) inhibitor linsitinib (LIN) with third-generation GD2.CAR T cells in diffuse intrinsic pontine glioma, where LIN induced tumor cell death and modulated the CAR T-cell phenotype. Here, we extended these findings to NB and explored the mechanisms of LIN-mediated CAR T-cell modulation. LIN, in combination with CAR T cells, significantly enhanced antitumor activity in LIN-sensitive NB cell lines. Mechanistically, we investigated ferrochelatase (FECH), a mitochondrial enzyme involved in heme biosynthesis, and a known off-target of LIN. LIN treatment or selective FECH inhibition with N-methyl protoporphyrin IX reduced intracellular heme, attenuated activation and exhaustion marker expression and promoted central memory characteristics associated with improved in vivo CAR T-cell persistence and functionality. Both treatments similarly decreased ATP production by reducing glycolysis and mitochondrial respiration in chronical antigen-activated CAR T cells. Collectively, these data reveal a dual mechanism of action for LIN, combining direct tumor cell cytotoxicity with metabolic reprogramming of CAR T cells linked to heme biosynthesis. These findings identify heme metabolism as regulator of CAR T-cell phenotype and function and support further investigation of FECH to enhance therapeutic efficacy in NB and beyond.

Acute myeloid leukemia (AML) remains challenging, especially for older or unfit individuals who develop resistance to venetoclax (VEN)-based regimens. Although TIM3 represents a promising AML target, clinical blockade has yielded suboptimal outcomes. Here, we develop VINCENT (VEN-integrated natural killer cell engager targeting TIM3), a therapeutic platform combining anti-CD16 and anti-TIM3 antibodies with nanoformulated VEN. In VEN-resistant AML cell lines and patient-derived xenograft models, VINCENT overcomes resistance by enhancing VEN delivery, eliminating TIM3+ blasts and dysfunctional T cells and activating natural killer (NK) cells. In primary AML samples, VINCENT selectively kills drug-resistant blasts, with efficacy correlating with the NK cell-to-TIM3+ blast ratio. Single-cell transcriptomics reveals VINCENT depletes high-TIM3 blasts and remodels the immunosuppressive microenvironment toward immune competence. Collectively, VINCENT simultaneously addresses VEN resistance, NK cell dysfunction and immune evasion, offering a personalized option for elderly or unfit individuals with AML failing standard VEN regimens.

The RAS family of oncogenes (KRAS, HRAS, NRAS) is among the most frequently mutated genes in human cancer. Therapeutic development has largely focused on inhibitors for KRAS codon 12 mutations, while mutant-selective inhibitors for Q61 variants remain elusive. A common mechanistic feature of G12 and Q61 mutants is the reduced efficiency of GTP hydrolysis, which enriches RAS in its active, signaling-competent state. Here we report small molecules that accelerate GTP hydrolysis in KRAS-Q61 mutants as an alternative therapeutic strategy. These compounds compensate for the loss of the catalytic residue Gln61 by introducing a general base into the active site, selectively enhancing hydrolysis of KRAS-Q61X (X = H, L, K, R) mutants by up to 20-fold. In mutant cancer cell lines, these compounds reduce GTP-bound RAS levels and suppress downstream signaling. This work establishes a mechanistic foundation for small-molecule 'GTPase activators' and offers a new paradigm for targeting RAS-driven cancers.

The clinical success of chimeric antigen receptor (CAR) T cell therapy requires scalable, non-invasive strategies for in vivo T cell engineering. Although mRNA delivery offers a promising alternative, lipid-nanoparticle-based carriers show limited efficiency for in vivo T cell transfection and typically require antibody conjugation. Here we report an inherent T cell-activating polymer-lipid nanoparticle that enables ligand-free, efficient mRNA transfection and activation of T cells in vivo. This mRNA delivery vehicle, composed of p-toluenesulfonyl arginine (RT)-modified oligoethylenimine-based lipid nanoparticles (ERTLNPs), preferentially mediated mRNA transfection in the spleen following systemic administration. Without exogenous stimulation, ERTLNPs intrinsically activated T cells, triggering robust mRNA expression and proliferation. Mechanistically, ERTLNPs engaged the PI3K/AKT/mTOR signalling axis to reprogram T cell metabolism, promoting expansion and restraining exhaustion. The systemic delivery of mRNA encoding fibroblast activation protein CAR via ERTLNPs contributed to the in situ generation of functional CAR T cells, which efficiently eliminated pathological fibroblasts in models of cancer and fibrosis, with minimal off-target effects. This ligand-free, metabolically reprogramming mRNA delivery system provides a clinically translatable approach for in vivo CAR T cell generation.

Ubiquitination regulation is essential for maintaining protein homeostasis. Dysregulated ubiquitination is increasingly recognized to participate in malignant progression. Fbxw11, a substrate-recognition subunit of SCF-type E3 ligases, is involved in tumorigenesis and progression. However, its role and molecular mechanism in acute myeloid leukemia (AML) have not been elucidated. Here we demonstrat that lower levels of Fbxw11 were detected in AML patients and correlated with worse prognosis. Furthermore, overexpression of Fbxw11 delayed AML progression by reducing leukemia stem cell (LSC) frequency and inhibiting leukemia cell migration. Mechanistically‌, quantitative proteomics combined with co-immunoprecipitation and ubiquitination assays identified dihydropyrimidinase-like 2 (Dpysl2) as a direct substrate of Fbxw11, leading to its poly-ubiquitylation and proteasomal degradation. Moreover, overexpression of Dpysl2 reversed the effects of Fbxw11 on LSC levels and migration of AML cells as well as accelerated AML progression in vivo. In addition, inhibition of Dpysl2 with lacosamide, an FDA-approved antiepileptic drug, phenocopied Fbxw11 restoration and delayed AML progression. Lacosamide combined with cytarabine further prolonged the survival of AML mice. Our results suggest Fbxw11 as an intrinsic suppressor in AML progression by regulating the ubiquitination and degradation of Dpysl2, providing new insights into ubiquitination regulation and highlighting the potential of targeting the Fbxw11-Dpysl2 axis as a therapeutic strategy for AML.

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

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

Mutations in the transcription factor gene Wilms Tumor 1 (WT1) are one of the leading causes of congenital glomerular disease, characterized by severe urinary protein loss and glomerular scarring. No disease-modifying therapies exist for WT1 glomerulopathies, and affected children rely on dialysis or kidney transplantation. We evaluated a previously uncharacterized treatment in a mouse model with an orthologous human mutation in Wt1 (Wt1+/R394W) that replicates the pathology of WT1 glomerulopathy. Lipid nanocomplexes were engineered to target integrin αvβ3 for efficient delivery of mRNA to primary podocytes and glomerular endothelial cells in vitro. A minimally invasive ultrasound-guided renal artery injection enabled precise and specific kidney localization in vivo, a finding not replicated by systemic administration. Nanocomplex-derived protein localized in glomeruli for up to 7 days in healthy and diseased mice. This platform was then used to perform an interventional preclinical trial in Wt1+/R394W mice, delivering angiopoietin-1 (Angpt1) mRNA, a vascular growth factor critical for glomerular health that is reduced in Wt1+/R394W podocytes. Angpt1 nanocomplex therapy reduced albuminuria, preserved glomerular endothelial integrity, prevented podocyte loss, and alleviated glomerulosclerosis in Wt1+/R394W mice. These findings demonstrate the therapeutic potential of this targeted approach for WT1 glomerulopathy and provide a foundation for clinical translation to improve outcomes in children with glomerular disease.

Clip reactions featuring highly selective and efficient bond cleavage are powerful synthetic tools. Herein, a radiation-induced clip reaction of sulfilimine is established. Sulfilimines are quantitatively decomposed to phenothiazines and amines by radiation with a record-high radiolytic yield of 270.9 nM/Gy. This high efficiency and selectivity are attributed to the near-unity utilization of hydrated electrons and hydroxyl radicals, the two primary radiation-generated reactive species. The sulfilimine bond undergoes reductive cleavage by hydrated electrons to generate a phenothiazine radical cation and an amine. Then, phenothiazine radical cation is reduced to phenothiazine by hydroxyl radicals and water. The clip reaction exhibits a broad substrate scope, rapid reaction kinetics, and robust performance under typical interferences, particularly under biological conditions. By pairing with sulfilimine-based click reactions, it enables the radiation-triggered activation of bioactive molecules in living systems. Notably, the cytotoxicity of the sulfilimine-caged monomethyl auristatin E (MMAE) is dramatically reduced to be about 1/800 of MMAE. Upon 1 Gy γ-irradiation in cancer cells, the anticancer bioactivity can be restored. Moreover, the sulfilimine-caged MMAE shows a remarkable in vivo anticancer effect with γ-ray irradiation. This work establishes a new modality of bioorthogonal chemistry, expanding the chemical toolkit for spatiotemporally controllable bond cleavage in living systems. It also provides a molecular framework for radiotherapy-enhanced, spatiotemporally targeted chemotherapy.

Light-activated control over drug activity offers a powerful strategy to improve therapeutic selectivity, yet most photoresponsive systems rely on ultraviolet or short-wavelength visible light with limited tissue penetration. Here we report two ruthenium(II) polypyridyl complexes bearing extended donor-π-acceptor ligands and a photocaged kinase inhibitor, imatinib, that enable efficient drug release under deep-red light (660 nm) irradiation. Systematic photophysical and photochemical studies demonstrate that π-conjugation engineering red-shifts metal-to-ligand charge-transfer absorption while preserving clean photoinduced release of coordinated imatinib from the ruthenium center. In BCR-ABL-positive leukemia cells, these complexes exhibit minimal dark toxicity but pronounced light-dependent cytotoxicity, accompanied by apoptosis and suppression of oncogenic BCR-ABL phosphorylation. Effective activity in 3D tumor spheroids was also observed, confirming efficient penetration and photoactivation in a physiologically relevant model. Together, these results establish a molecular design strategy for developing deep-red-light-activated Ru(II) photocages and highlight their potential for spatiotemporally controlled kinase inhibition in cancer therapy.

Innate immunity constitutes the body's first line of defense, deploying evolutionarily conserved mechanisms to detect and respond to pathogenic threats and tissue injury. In contrast to the adaptive immune system's specificity and memory, innate immunity relies on germline-encoded pattern recognition receptors (PRRs) to sense pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs). This review outlines the major PRR families and their downstream signaling pathways, culminating in the production of cytokines and inflammasome activation. Additionally, this review explores the functional integration of cellular responses and examines the sophisticated regulatory circuits, including transcriptional, posttranscriptional, epigenetic, and metabolic mechanisms, that calibrate innate immune responses. Importantly, we highlight how dysregulated innate immunity underlies a wide spectrum of diseases, from autoinflammatory and autoimmune disorders to chronic inflammatory states and cancer. Recent insights into trained immunity and innate immune memory further redefine the scope of innate defenses, revealing that metabolic and epigenetic remodeling can prime innate cells for enhanced secondary responses. Finally, we discuss the therapeutic potential of modulating innate pathways, including PRR agonists as vaccine adjuvants or anticancer agents, and PRR inhibitors in autoimmune and inflammatory diseases. By integrating current mechanistic knowledge with emerging therapeutic strategies, this review underscores the central role of innate immunity in health and disease while pointing toward promising avenues for future research and clinical translation.

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

The Tn antigen, a truncated O-glycan, is frequently elevated in pancreatic ductal adenocarcinoma (PDAC). Multiple therapeutic approaches targeting Tn have been developed, but they have not demonstrated clear efficacy signals in early phase clinical studies. Improving Tn-targeted strategies in PDAC will require both overcoming the immunosuppressive tumor microenvironment and defining pathways by which truncated O-glycans promote growth and immune evasion. Here, we showed that Tn reshapes the tumor immune landscape of PDAC. Expression of Tn antigen on PDAC cells enhanced proliferation in vitro and tumor growth in vivo. Tn expression remodeled the immune microenvironment, skewing tumor-associated macrophages toward M2-like phenotypes, reducing cross-presenting dendritic cells, and expanding myeloid-derived suppressor cells (MDSCs). Single-cell RNA sequencing confirmed expansion of MDSCs and downregulation of antigen processing and presentation in the immune cell infiltrate of Tn+ tumors. Tumor-intrinsic transcriptomic analyses revealed activation of TNF-α/NF-κB signaling and induction of IL-34, a cytokine linked to monocyte survival and differentiation in Tn antigen expressing tumors. Additionally, high Tn expression in both organoids derived from pancreatic cancer patients and in PDAC mouse models was associated with increased IL-34 expression. Genetic deletion of Il34 in PDAC cells attenuated Tn-driven tumorigenesis and reduced MDSC infiltration, while recombinant IL-34 promoted myeloid cell differentiation and proliferation in vitro. Together, these findings establish a glyco-immune-cytokine axis in which truncated O-glycans contribute to IL-34-mediated immunosuppression, providing mechanistic insight and potential therapeutic targets in PDAC.

Recent advances have demonstrated the application of microcavity-containing hydrogel microparticles, known as nanovials, for massively parallel and high-throughput screening of therapeutic T cell populations for adoptive cell therapies. Nanovial cavities coated with peptide-MHC (pMHC) or antigen tetramers selectively bind to their cognate T cell receptor (TCR) or chimeric antigen receptor (CAR) to activate T cells and capture secreted cytokines. However, binding of tetramers or recombinantly expressed antigen by T cells may not reflect physiological T cell activation or cytotoxicity, as the binding interface is not fully representative of the natural immunological synapse formed between T cells and professional antigen-presenting cells (APCs). Here, we leverage the recent discovery of an ESCRT- and ALIX-binding region (EABR) sequence to generate antigen-presenting vesicles and cancer-mimicking exosomes from standard HEK293T and Expi293F cell cultures. EABR-mediated vesicles present natural, full-length oncologically relevant membrane proteins embedded in lipid bilayers to functionalize the nanovial cavity with cell-like membranes. Hydrogel nanovials functionalized with the EABR-mediated vesicles show improved T cell capture of 1G4 T cells and enhanced induction of secretion in HER2 CAR-T cells compared to hydrogel surfaces functionalized with recombinantly expressed soluble proteins.

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.

Metastatic microenvironments vary widely not only in their biochemical composition but also in their mechanical properties. Here, we examined how the mechanical rigidity of the metastatic niche affects metastases seeding and the local efficacy of antitumor immunosurveillance. Cancer cells stiffened in response to increasing environmental rigidity, a biophysical change that mechanically sensitized them to killing by cytotoxic lymphocytes. In immunodeficient mice, rigidity sensing by cancer cells yielded robust bone colonization, accompanied by marked stiffening of the cancer cells themselves. Conversely, in immunocompetent hosts, stiffer cancer cells were selectively eliminated, and bone metastasis was suppressed. In patients, metastatic cell stiffness was associated directly with environmental rigidity and inversely with immune infiltration. Expression of Spp1, encoding the secreted glycoprotein osteopontin, defined a subset of cancer cells that expanded in the bone, and deletion of Spp1 limited environmentally induced cancer cell stiffening, bone colonization, and immune vulnerability. Thus, environmental mechanosensing regulates both metastases seeding and antitumor immunity, providing an immunological basis for metastatic site selection.

Clinical translation of nanomedicines remains constrained by a persistent gap between static preclinical assays and the dynamic shear environment of the human vasculature. This review argues that fluid shear stress is an underappreciated contributor to clinical attrition, acting alongside well-established factors including biomarker-target mismatch, enhanced permeability and retention heterogeneity, preclinical-clinical discordance in tumor biology, and manufacturing variability. Circulating nanocarriers traverse a mechanical environment in which wall shear stress spans from 0.1 dyn/cm2 in hepatic sinusoids to over 1000 dyn/cm2 in stenotic regions. Emerging quantitative studies indicate that these forces can modulate carrier stability, protein-corona composition, and drug release in ways that static in vitro assays cannot resolve, with the cholesterol-rigidified bilayer of Doxil and the shear-activated platforms of Korin and colleagues providing representative positive control and mechanistic evidence. We synthesize recent progress at the interface of computational fluid dynamics and machine learning. Physics-informed neural networks, deep operator networks, and graph neural networks feature prominently in this synthesis and underpin an integrated framework for mechanistically informed pharmacokinetic prediction. Reported benchmarks show surrogate models reconstructing hemodynamic fields with under 5% error relative to high-fidelity CFD while reducing data requirements 5- to 10-fold, and convolutional surrogates trained on approximately 1800 patient-specific simulations predicting time-averaged wall shear stress with 2.5% mean absolute error. These surrogate models integrate hemodynamic descriptors (TAWSS, OSI, and RRT) with distributional particle features (D10, D50, and D90). We outline the evidentiary and methodological path toward biomechanically informed nanomedicine development aligned with emerging regulatory frameworks, while acknowledging that routine regulatory acceptance of computational shear modeling remains a future milestone rather than current practice.

Clinical translation of nanomedicines is greatly hindered by insufficient understanding of their in vivo process, yet a key challenge lies in quantifying the encapsulated versus free drug forms in tissues and cells. Herein, we present a facile, versatile anti-PEG single-chain variable-fragment antibody (PEG-scFv)-based method enabling quantitative measurement of both forms in various biofluids (e.g., interstitial fluid, cytoplasm). By this method, we map the in vivo process of PEGylated liposomal doxorubicin (sLip/Dox) at unprecedented resolution. In the bloodstream, doxorubicin remains largely encapsulated in liposomes (>99%). In liver as the main organ for drug elimination, less drug was distributed in the interstitium (>80% encapsulated) but more in liver cells (mainly in Kupffer cells) released in a time-dependent manner, accompanying doxorubicin transferred to hepatocytes most in free form by 12 h postinjection. After extravasation into tumors, there was a limited access of sLip/Dox to tumor cells, confining most of the drug in the interstitium mainly being encapsulated (more than 75%), and the internalized fraction underwent a gradual release process in both tumor-associated macrophages and tumor cells. These findings revealed that for sLip/Dox, which primarily underwent drug release intracellularly, cellular internalization rates could be the key factor in determining its in vivo performance. Given widespread PEGylation on developing nanomedicines and the cost-effectiveness of scFv production, PEG-scFv offers a broadly applicable tool for dissecting in vivo processes of nanomedicines to establish dose-effect relationships like small-molecule drugs, further to guide rational nanotherapeutic design.

Treatment with immune checkpoint inhibitors (ICIs) has radically improved outcomes for patients with microsatellite instability-high (MSI-H) metastatic colorectal cancer (mCRC). Intrinsic and acquired resistance, however, remains an important concern. Pyrimidine pathway regulates the innate immune response and may offer an opportunity to enhance ICI efficacy via modulation of pyrimidine metabolism. Identification of primary, adaptive and acquired resistance mechanisms and development of actionable strategies to overcome resistance and expand the benefit of ICI is paramount. Our group is the first to show that the dihydropyrimidine dehydrogenase (DPD; gene DPYD) metabolic pathway predicts the efficacy of ICI in MSI-H tumors. Retrospective analysis of germline single nucleotide polymorphisms was done in patients with MSI-H CRC cohort (Veneto Institute of Oncology; OIOV-IRCCS, Padua, Italy) treated with immunotherapy. Dpyd knockout or overexpressed MC38 MSI-H syngeneic mice models were treated with anti-programmed cell death protein 1 (PD-1) and anti-cytotoxic T-lymphocyte associated protein 4 (CTLA-4) alone or in combination and their effect on tumor growth and its microenvironment was studied. In our real-world data analysis, genetic variants in DPYD were associated with progression-free survival and tumor response in patients with MSI-H mCRC treated with ICI. In preclinical studies, complete tumor elimination was observed in 100% of mice bearing Dpyd knockout tumors treated with a combination of anti-PD-1 and anti-CTLA-4, further validated by pharmacologically inhibiting DPD using eniluracil. Our results showed that the pyrimidine pathway has a strong association with immunotherapy treatment response in CRC. These findings may support the development of novel treatment strategies exploiting DPD inhibition in MSI-H tumors and their quick integration into clinic.

Controversies persist regarding the optimal regimens of neoadjuvant and adjuvant therapies for locally advanced esophageal squamous cell carcinoma (ESCC). This real-world cohort study aimed to compare the efficacy of neoadjuvant chemotherapy (NCT), chemoradiotherapy (NCRT), and chemoimmunotherapy (NCIT), and to clarify the survival contributions of adjuvant strategies. We retrospectively enrolled 1818 ESCC patients across four high-volume Chinese medical centers. After propensity score matching, pathological tumor response and tumor downstaging followed the hierarchy NCRT > NCIT > NCT, yet no significant differences were detected in overall survival (OS) or progression-free survival (PFS). Subgroup analyses revealed superior OS benefits of NCIT in never‑smokers, nondrinkers, and patients with clinical N2-3 disease compared with NCT. While NCRT showed greater PFS benefit than NCIT as clinical T stage advanced, its OS advantage was restricted to obese individuals. Adjuvant immunotherapy extended PFS for patients receiving prior NCIT, whereas adjuvant chemotherapy delivered survival benefits to those with pathological lymph node metastasis. We constructed optimal treatment rule (OTR) decision trees with moderate-to-good predictive capacity to guide selection of neoadjuvant and adjuvant therapies based on survival benefit. These results highlight the necessity of precision perioperative therapy for ESCC. The established OTR decision trees can support individualized clinical decision-making.

Triple-negative breast cancer is an aggressive subtype comprising 10-20% of all breast cancer cases and has a worse prognosis than other subtypes. This open-label, multicenter, single-arm, phase 2 clinical trial evaluates the safety and efficacy of ivonescimab combined with chemotherapy as first-line treatment in female patients with locally advanced unresectable or metastatic triple-negative breast cancer who have not received previous systemic therapy. Eligible patients received ivonescimab 20 mg kg-1 intravenously every 2 weeks and paclitaxel 90 mg m-2 or nab-paclitaxel 100 mg m-2 intravenously on days 1, 8 and 15 of each 4-week treatment cycle. The primary endpoints were safety and the investigator-assessed objective response rate per RECIST v.1.1. A total of 36 patients were enrolled. As of July 15, 2025, the median follow-up duration was 22.1 months. The primary endpoints were met. Treatment-related adverse events occurred in 36 (100.0%) patients. Grade ≥3 treatment-related adverse events were reported in 21 (58.3%) patients. No treatment-related deaths occurred. Immune-related adverse events occurred in 15 (41.7%) patients, with grade ≥3 events in 4 (11.1%) patients. Of the 35 patients evaluable for treatment efficacy, the objective response rate was 80.0% (95% confidence interval: 63.1-91.6), including 2 (5.7%) complete responses and 26 (74.3%) partial responses. Ivonescimab plus chemotherapy demonstrated encouraging antitumor activity as first-line therapy for patients with previously untreated advanced triple-negative breast cancer and merits further investigation. ClinicalTrials.gov identifier: NCT05227664 .

Orally administered small-molecule programmed death ligand 1 (PD-L1) inhibitors may have the potential to improve patient outcomes in the treatment of a range of cancers compared with their antibody-based counterparts. A small molecule might achieve better tumor tissue penetration, and oral administration could significantly improve convenience and access for patients. Three phase 1 open-label, non-randomized, dose escalation, and expansion studies evaluated the safety, preliminary efficacy, pharmacokinetics (PK), and pharmacodynamics (PD) of three agents in patients with advanced solid tumors: INCB086550 (NCT03762447), INCB099280 (NCT04242199), and INCB099318 (NCT04272034). Overall, 138, 182, and 104 patients received INCB086550, INCB099280, and INCB099318, respectively. Most had previously received ≥2 lines of cancer therapy for advanced or metastatic disease; 9.6%-16.5% had received prior immunotherapy. All three agents were rapidly absorbed and showed stable dose-dependent PK. With INCB086550, 88 patients (63.8%) had ≥1 treatment-related treatment-emergent adverse event (TEAE), and 19 (13.8%) had ≥1 treatment-related grade ≥3 TEAE. In total, 14 patients (10.1%) had a nervous system-associated TEAE for which an immune-mediated etiology could not be ruled out; events were predominantly peripheral sensory and motor neuropathies. With INCB099280 and INCB099318, 144 (79.1%) and 69 (66.3%) of patients had ≥1 treatment-related TEAE, and 25 (13.7%) and 12 (11.5%) had ≥1 treatment-related grade ≥3 TEAE, respectively. The most frequent immune-related adverse events were skin reactions (INCB099280 and INCB099318) and hepatitis (INCB099280). No dose-limiting toxicities (DLTs) occurred during dose escalation with INCB086550 or INCB099318; two DLTs occurred in two patients with INCB099280 (grade 2 vomiting with 600 mg once daily and grade 2 maculopapular rash with 800 mg two times per day). Overall objective response rates for INCB086550, INCB099280, and INCB099318 were 10.9% (95% CI 6.2% to 17.3%; n=15), 8.8% (95% CI 5.1% to 13.9%; n=16), and 8.7% (95% CI 4.0% to 15.8%; n=9), respectively. Target engagement and PD activity were demonstrated, including PD-L1 binding, and increases in cytokine and chemokine production, as well as T-cell activation and proliferation. Both INCB099280 and INCB099318 had an acceptable safety profile, with preliminary evidence of antitumor activity. The risk of immune-mediated neuropathy led to discontinuation of the clinical program for INCB086550.

Psoriasis progresses to psoriatic arthritis in up to 30% of patients, causing irreversible joint damage and substantial healthcare burden. Whether biologic therapy reduces psoriatic arthritis risk remains unclear. The aim of this paper was to evaluate the association between biologic therapy and psoriatic arthritis risk in psoriasis patients. We conducted a systematic review and meta-analysis of cohort studies (PROSPERO: CRD420251240466), searching PubMed, EMBASE, and Cochrane Library from inception to December 2025. We included cohort studies of adults with psoriasis without prior psoriatic arthritis that reported adjusted risk estimates for incident psoriatic arthritis and used an 'on-drug' analytical framework to account for changes in treatment status over time. Pooled hazard ratio (HRs) with 95% confidence interval (CI) were calculated using fixed- or random-effects models. In total, 15 cohort studies including 124,138 psoriasis patients with over 778,690 person-years of follow-up were included. Biologic use was associated with a 46% lower psoriatic arthritis risk versus non-biologic therapy (pooled HR 0.54, 95% CI 0.43-0.68), consistent versus non-systemic therapies (HR 0.57, 95% CI 0.33-0.97) and methotrexate (HR 0.48, 95% CI 0.45-0.51). Interleukin (IL)-17 inhibitors (HR 0.65, 95% CI 0.49-0.87; 3 studies) and IL-12/23 or IL-23 inhibitors (HR 0.46, 95% CI 0.36-0.60; 4 studies) were associated with lower psoriatic arthritis risk than tumor necrosis factor inhibitors. IL-23 inhibitors showed greater protection than IL-17 inhibitors (HR 0.67, 95% CI 0.58-0.77), while IL-23 inhibitors and IL-12/23 inhibitors did not differ (HR 0.88, 95% CI 0.69-1.13). Biologic therapy for psoriasis, particularly IL-23/IL-17-targeted agents, is associated with reduced incident psoriatic arthritis risk. These findings suggest early pathway suppression may delay clinical psoriatic arthritis diagnosis, though causal inference is limited by the observational data.

Prostate cancer (PCa) frequently metastasizes to bone, marking incurable disease. This progression is driven by an immunologically cold bone tumor microenvironment that fosters resistance to therapy. To define the mechanisms underlying this uniquely immunosuppressive niche, we applied spatial single cell analyses across primary tumors and metastatic sites. Bone metastases showed marked suppression of tumor-intrinsic type I interferon (IFN-I) signaling and loss of antigen presentation, features that were strongly associated with reduced bone metastasis-free survival. Tumor-intrinsic IFN-I expression correlated with memory T cell infiltration, whereas the bone myeloid compartment was enriched for protumor macrophages and showed reduced dendritic cell (DC) activation and antigen presentation. Digital spatial profiling of matched tumors revealed a broad loss of IFN-I-regulated immunostimulatory and checkpoint molecules. Notably, B7-H3, a putative negative regulator of IFN-I, was highly expressed in bone metastases and inversely associated with antigen presentation. These findings define bone-specific mechanisms of immune resistance and highlight therapeutic vulnerabilities that could inform precision therapeutic strategies for PCa.

Cancer remains a major global health burden and the second leading cause of mortality worldwide. Recent advances in cancer immunotherapy have emphasized the critical role of the tumor microenvironment (TME) in determining therapeutic outcomes, leading to the classification of tumors into immunologically "hot" and "cold" phenotypes. Cold tumors are characterized by low immunogenicity, limited immune cell infiltration, and a highly immunosuppressive microenvironment, resulting in poor prognosis and resistance to immune checkpoint inhibitors. Despite the development of multiple immunotherapeutic strategies, effective activation of antitumor immunity in cold tumors remains a major clinical challenge. Current approaches aim to initiate immune responses through priming strategies such as cancer vaccines and adoptive T-cell transfer, while simultaneously overcoming immunosuppressive signaling via immune checkpoint blockade. Additional strategies include depletion of myeloid-derived suppressor cells and enhancement of co-stimulatory pathways. However, these approaches are often limited by inefficient delivery, poor tumor penetration, and systemic toxicity. Nanotechnology has emerged as a promising platform for tumor microenvironment reprogramming. Nanocarriers enable targeted delivery of immunomodulatory agents, enhance antigen presentation, and improve immune activation while overcoming biological barriers such as dense stroma and abnormal vasculature. By integrating nanotechnology with immunotherapy, new opportunities arise to convert cold tumors into hot, immune-responsive phenotypes, thereby improving therapeutic efficacy and clinical outcomes.

Macrophages have emerged as promising candidates for cell-based immunotherapies due to their intrinsic plasticity, tissue-infiltrating capabilities, and central roles in orchestrating immune responses. Their phenotypic and functional diversity within tissues and the tumor microenvironment has driven interest in harnessing these cells for therapeutic purposes. Integrating recent knowledge in signaling cascades balancing the activity of macrophages in combination with genetic engineering have enabled the development of macrophages with improved functions, including the introduction of synthetic receptors such as chimeric antigen receptors (CARs). These superior macrophages orchestrate innate immune activity with antigen-specific targeting, offering distinct advantages over conventional CAR-T and CAR-NK cell therapies, especially in solid malignancies. Emerging preclinical and early clinical data support the feasibility, safety, and therapeutic potential of macrophage-based strategies. However, successful clinical translation requires overcoming key challenges in standardization, scalable manufacturing and regulatory compliance of cell products. This review integrates current knowledge in the diversity of macrophage signaling which feeds into engineering techniques, therapeutic applications, and manufacturing innovations. Leveraging concepts of macrophage tissue plasticity highlights the potential of macrophages as next-generation cell therapeutics with broad potential in oncology and beyond, bridging fundamental immunology with translational medicine. Continued interdisciplinary research will accelerate clinical adoption and expand indications across diseases.

Tumor-associated macrophages (TAMs) critically modulate solid tumor progression and immunotherapy resistance, yet their profound context-dependent heterogeneity defies the traditional M1/M2 dichotomy. In this review, we propose the 'niche-metabolism axis' framework, deconstructing the tumor microenvironment into dynamically interacting functional niches (e.g., hypoxic cores and tertiary lymphoid structures). Within these niches, specific physicochemical stresses substantially influence localized TAM metabolic reprogramming. Metabolites such as lactate and lipids act not merely as substrates but as signaling mediators and epigenetic donors, shaping TAM phenotypes. Consequently, TAMs acquire spatial metabolic imprints and evolve into 'ecosystem engineers' that actively remodel the immune microenvironment. This framework highlights that therapeutic paradigms should shift from nonselective TAM depletion to niche-specific metabolic-epigenetic reconstruction, providing a precise roadmap for designing next-generation engineered macrophage therapies.

Cancer immunotherapy has revolutionized oncologic treatment, yet its efficacy remains constrained by tumor immune evasion and the immunosuppressive tumor microenvironment. Addressing these multifaceted barriers calls for strategies that can both eliminate cancer cells and simultaneously amplify antitumor immunity. Pyroptosis, a lytic and highly immunogenic form of programmed cell death mediated by gasdermin proteins, has recently emerged as a compelling approach to reprogram the tumor immune landscape. By releasing abundant damage-associated molecular patterns and pro-inflammatory cytokines, pyroptosis not only destroys tumor cells but also promotes antigen presentation and robust local-to-systemic immune responses. With the rapid advancement of nanotechnology, pyroptosis-engineered nanomedicine has gained increasing interest for synergizing cell killing and immune activation to enhance cancer immunotherapy. This review summarizes recent progress in the molecular mechanisms of pyroptosis and highlights emerging nanomedicine strategies designed to precisely induce pyroptosis within tumors. The discussion further extends to design principles, activation modalities, and therapeutic outcomes in preclinical models. Finally, current challenges and future directions toward clinical translation are outlined. Collectively, this review integrates pyroptosis biology with nanoengineering concepts to inspire next-generation immunotherapeutic interventions against cancer.

Dietary interventions can influence cancer progression, yet the role of low-protein diets (LPDs) in pancreatic ductal adenocarcinoma (PDAC) immunotherapy is unclear. Here we show that an LPD suppresses PDAC progression in male mice by remodeling the gut microbiota and activating antitumor immunity. LPD promoted immune activation and drove an immunostimulatory tumor-associated macrophage phenotype. Microbiota depletion abolished these effects and fecal microbiota transplantation from LPD-fed donors transferred the protective phenotype to recipients. Mechanistically, LPD enriched Blautia coccoides, which produced uridine diphosphate (UDP)-galactose to activate the macrophage P2Y14R-STAT1 axis, inducing an immunostimulatory phenotype. Combining LPD, B. coccoides or UDP-galactose with anti-PD1 improved survival over anti-PD1 alone. In persons with advanced PDAC, reduced fecal B. coccoides and serum UDP-galactose correlated with poor outcomes. These findings establish that LPD reshapes the gut microbiota and metabolites to enhance antitumor immunity through the UDP-galactose-P2Y14R-STAT1 axis, offering a dietary strategy to improve PDAC immunotherapy.

Efficient delivery of large, negatively charged self-amplifying RNA (saRNA) into dendritic cells (DCs) is critical for next-generation cancer vaccines. However, this remains challenging due to the high sensitivity of DCs to chemical carriers and high-voltage electroporation. In this study, an integrated nanopore-electroporation (NEP) microdevice was developed by combining 200 nm track-etched polycarbonate membrane, bidirectional PDMS microfluidic channels, and Pt/ITO electrodes to localize the electric field and induce membrane permeabilization at low voltage (≤30 V). Multiphysics simulations revealed that 200 nm nanopores concentrated the electric field at the cell-membrane interface, generating transmembrane potentials exceeding 3 V. Using DC2.4, the NEP system achieved 75% propidium iodide (PI) uptake at 25 V with 90% viability, confirming controllable nanoscale perforation. Direct delivery of GFP-encoding saRNA achieved approximately 50% transfection efficiency with sustained protein expression for more than 96 h, significantly outperforming mRNA at an equal dose. Long-term viability (>85% at 96 h) and negligible cytotoxicity demonstrated the excellent biocompatibility of the device. This reagent-minimal, modular NEP platform thus provided a high-efficiency, low-toxicity route for saRNA delivery into hard-to-transfect immune cells, offering a versatile engineering framework for DC-based cancer immunotherapy, RNA vaccine development, and broader cell gene-modification applications.

Quantitative assessment of chimeric antigen receptor T (CAR-T) activity in solid tumors remains challenging, as immune engagement can induce early mechanical softening of tumor spheroids before overt cell death becomes detectable. To address this limitation, we developed Cytotongue, a three-dimensional (3D)-printed microfluidic aspiration system for real-time, treatment-integrated aspiration-response phenotyping of live tumor spheroids. Using human epidermal growth factor receptor 2 (HER2)-positive BT-474 breast cancer spheroids exposed to CAR-T cells, the Cytotongue system quantified aspiration-induced elongation dynamics and assay-specific deformation indices derived from an empirical biphasic deformation framework. This approach enabled detection of CAR-T-induced mechanical softening at low effector-to-target ratios (1:1-2:1) within 24 h, conditions under which conventional propidium iodide (PI) staining showed minimal response. System-level validation demonstrated robust performance, with high Z' factors (0.74-0.93), large effect sizes, and low coefficients of variation. Moreover, Cytotongue distinguished CAR-T-associated mechanical softening accompanied by apoptosis from doxorubicin-induced deformation responses despite comparable PI readouts. Collectively, this work introduces a treatment-integrated microfluidic aspiration system that enables real-time mechanical phenotyping of tumor spheroids, providing a new physical dimension for evaluating immunotherapeutic and drug responses.

Cellular senescence and immune dysfunction intersect as central biological processes shaping disease progression and therapeutic responses across ageing-related pathologies. Rather than acting independently, senescent cells (SnCs) and immune cells engage in a dynamic and bidirectional crosstalk that influences immune surveillance, inflammatory signaling, and tissue remodeling. This review focuses on the molecular and cellular mechanisms through which SnCs modulate immune composition and function, emphasizing immune checkpoint ligands, stress-associated surface molecules, and senescence-associated transcriptional programs governing senescence-associated secretory phenotype (SASP) expression. We further examine how immune-targeted therapies can, in turn, remodel senescent niches by promoting immune-mediated clearance or altering senescence-associated phenotypes. Integrating evidence from cancer to metabolic and chronic inflammatory diseases, we discuss how senotherapies reshape immune landscapes and how immunotherapies impact senescent cell burden and function. Building on these mechanistic insights, we propose that the senescence-immune axis constitutes a tractable therapeutic vulnerability that can be exploited through rational combination strategies. Finally, we outline five conceptual frameworks and biomarker-guided trial archetypes for integrating senotherapies and immunotherapies in a bidirectional crosstalk, aiming to enhance therapeutic efficacy while mitigating ageing- and treatment-associated toxicities. By reframing senescence and immunity as interconnected therapeutic targets, this review provides a mechanistic foundation for next-generation combinatorial interventions across ageing-associated diseases.

Tertiary lymphoid structures (TLSs) drive antitumor immunity, but their functional heterogeneity translates into inconsistent immunotherapy responses in hepatocellular carcinoma (HCC), raising the question of which immune niches actually predict efficacy. Using spatial multi-omics analysis, we uncover a distinct niche, the stem-like lymphoid niches (SLNs), that correlates with stronger responses to PD-1/PD-L1 blockade. Unlike conventional TLS, SLNs feature a specialized niche of CD70+ type 2 innate lymphoid cells (ILC2s) and progenitor exhausted T (Tpex) cells. Intriguingly, intratumoral Bacillus cereus promotes SLN accumulation via triggering endothelial IL-33 release, which rewires ILC2s metabolism and enhances H3K4me3-mediated epigenetic activation of CD70, Lta, and Ltb genes, reinforcing the niche's lymphoid-organizing function. We identify the bacterial phosphoinositide-specific phospholipase C (PI-PLC) as the effector behind this endothelial IL-33 release. Combining PI-PLC with αPD-L1 synergistically amplifies antitumor immunity, highlighting SLNs as a promising target for boosting immunotherapy in HCC.

Effective cancer immunotherapy is hindered by immunosuppressive crosstalk within the tumor microenvironment. We engineered a tumor immune microenvironment-on-a-chip (TIMoC) that recapitulates the vascularized, hypoxic, and spatially organized niche of human solid liver tumors. We employed TIMoC to dissect the reciprocal interaction between macrophages and natural killer (NK) cells. Macrophages induced NK cell dysfunction, while dysfunctional NK cells promoted M2 macrophage polarization. This bidirectional impairment created a self-perpetuating immunosuppressive loop. TIMoC served as an in vitro screening tool, confirming the limited efficacy of TIGIT blockade in a multicellular context and revealing synergistic anti-tumor activity for combinations of a macrophage-reprogramming agent (resiquimod) with NK cell-targeting antibodies. By incorporating patient-derived organotypic tumor spheroids and autologous immune cells, the personalized TIMoC platform modeled patient-specific responses and evaluated effective drug combinations, demonstrating its potential to guide precision immunotherapy. This work elucidates a key immunosuppressive axis and introduces a versatile platform for rationally designing combination immunotherapies.

Nonclassical major histocompatibility complex class I (MHC-I) molecules, including human leukocyte antigen E (HLA-E), HLA-F, HLA-G, MHC-I-related protein 1 (MR1), and the CD1 family, constitute a conserved antigen-presenting system that regulates immune surveillance, tissue homeostasis, and tolerance through specialized interactions with innate and unconventional T cells. Although these molecules have long been implicated in cancer, infection, autoimmunity, and transplantation, their distinct immunobiology and therapeutic potential have largely been considered in isolation. Recent advances in structural immunology, single-cell and spatial profiling, engineered immune cell technologies, and early clinical studies have established nonclassical MHC-I pathways as tractable targets for immunotherapy. In this review, we synthesize the biology, disease-associated functions, and therapeutic targeting of these molecules, integrating immune checkpoint blockade, antibody-based therapeutics, and MR1- and CD1-restricted cellular immunotherapies into a unified framework. We further highlight shared immunological principles, emerging clinical translation, and opportunities for universal, off-the-shelf immune interventions.

Effector regulatory T (eTreg) cells are indispensable for maintaining peripheral immune tolerance and can also interfere with antitumor immunity. However, how epigenetic modifications regulate eTreg differentiation and peripheral maintenance remains largely unknown. Here, we identify the histone deubiquitinating enzyme Bap1 as a pivotal regulator of eTreg cell fate via its H2A deubiquitination activity. Treg-specific deletion of Bap1 markedly impairs eTreg differentiation and promotes ferroptosis through two phases, culminating in a fatal systemic autoimmune syndrome. In the differentiation phase, Bap1 drives the resting Treg (rTreg)-to-eTreg transition by orchestrating metabolic reprogramming through H2AK119 deubiquitination-mediated activation of Stat5, mTORC1, and OXPHOS signaling pathways. In the maintenance phase, Bap1 protects eTreg from ferroptosis through the GSH-Gpx4 axis and MUFA-mediated lipid metabolism. Yy1 interacts with Bap1 at specific genomic loci to regulates the pathways responsible for eTreg differentiation and resistance to ferroptosis. Finally, acute ablation of Bap1 in Treg cells enhances antitumor immunity by disrupting intratumoral eTreg survival and remodeling the tumor microenvironment. Collectively, our findings identify Bap1 as a key epigenetic-metabolic integrator that governs eTreg cell fate to maintain immune tolerance and restrains antitumor immunity. Schematic overview of the mechanism by which Bap1-mediated deubiquitination determines immune tolerance and anti-tumor immunity by modulating eTreg cell differentiation and ferroptosis. Created with BioRender.com. Created in BioRender. Liu, H. (2026) https://BioRender.com/5f76yo3 .

Multidrug resistance (MDR) in colorectal cancer (CRC) arises from interacting tumour-cell, pharmacological and microenvironmental programmes that undermine both drug activity and delivery. Chemotherapy-associated MDR is the principal focus of this review, while resistance to targeted therapy and immunotherapy is considered where it directly affects delivery design or patient selection. Bioactive compounds can modulate several resistance pathways, but their translational value is constrained by poor solubility, instability, rapid metabolism and inadequate exposure at resistant lesions. This review critically examines when nanomedicine can make such mechanistic activity pharmacologically and translationally meaningful. We link resistance biology to delivery functions, evaluate bioactive chemosensitiser candidates against human exposure, compare major nanomedicine design strategies, and assess nano-bio interactions, repeat-dose safety, manufacturing and clinical positioning. Across the evidence, three limitations recur. First, concentrations associated with chemosensitisation for curcumin, resveratrol, epigallocatechin gallate and quercetin generally exceed measured human parent-analyte exposure. Second, many resistance claims rely on parental or poorly characterised models, whereas studies combining defined resistance provenance with functional mechanism and exposure confirmation remain uncommon. Third, human studies demonstrate feasibility, tissue exposure or treatment response, but not mechanism-specific reversal of CRC MDR by a bioactive compound or bioactive nanomedicine. Progress therefore depends less on adding new particle classes than on matching a necessary formulation function to a defined resistance or spatial barrier, quantifying active exposure in the relevant compartment, validating mechanism in appropriate models, and integrating repeat-dose safety, scalable manufacturing and biomarker-guided clinical development.

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

Systemic inhibition of interleukin-1b (IL-1b) has been shown to reduce the incidence of lung cancer in patients in years after treatment, but knowledge gaps surrounding its activation and role in the tumor microenvironment hinder cancer interception. We developed activity-based technologies to probe inflammation in early lung cancer and identified a translational target candidate. Probes sensitive to IL-1b-activating proteases were designed and applied to a murine model of inflammatory lung cancer, Kras/Trp53-mutant mice with SIINFEKL expression (KPS). Our nanosensors revealed elevated caspase-1 expression and activity in tumors, highlighting the importance of caspase-1 processing IL-1b during cancer development. We conducted a preclinical combination therapy trial by administering IL-1b blockade and caspase-1 inhibition. We observed significant reduction in lung cancer, including complete ablation of tumor incidence in nearly 20% of KPS mice. Our approach to understanding the interplay of protease activity and cytokine activation supports new strategies to mitigate inflammation and intercept lung cancer progression.

The development and validation of prognostic and predictive biomarkers in breast cancer is limited by the availability of well-annotated datasets linking tumor molecular features to treatment response and survival outcomes. To address this need, we generated an extensive mouse models dataset comprised of 26 immunocompetent mammary tumor models spanning diverse genetic backgrounds, epithelial-mesenchymal states, the basal-luminal axis, and distinct immune microenvironments. For each model, survival was measured under no treatment, immune checkpoint inhibition (ICI), and carboplatin/paclitaxel chemotherapy, and RNA-sequencing was performed on baseline tumors and on 7-day on-treatment samples for both regimens. Baseline murine tumor gene expression features were used to train a machine learning Elastic Net model that predicted survival outcomes on multiple human breast cancer datasets with performance comparable to that of existing prognostic assays. Models trained for ICI benefit, using either the untreated or 7-day ICI treated samples, predicted ICI benefit on human ICI treated datasets, with the 7-day treated tumor model showing better performance. A predictor of carboplatin/paclitaxel response developed from the murine mammary tumor data performed well in mice but did not generalize to human chemotherapy cohorts. Finally, comparison of multiple computational approaches, including XGBoost, random forests, and support vector regression, showed that all methods successfully predicted survival outcomes, with Elastic Net offering the best performance and interpretability. These results indicate conserved cancer biology between mouse and human tumors for prognosis and ICI response and establish a large preclinical dataset with linked phenotypic and genomic data as a resource for biomarker discovery.

Mutant KRAS inhibition has revolutionized the treatment of lung adenocarcinoma. Unfortunately, responses to this form of targeted therapy are often of limited duration because of the development of resistance. Targeted protein degradation, including using PROTACs (PROteolysis-TArgeting Chimeras), presents an alternative approach to targeting oncogenic drivers in cancer. In this issue of Cancer Research, Martín and colleagues developed a dTAG-KRASG12V syngeneic mouse model that allows for the study of the effects of degrading the KRASG12V oncoprotein in vivo. The authors discovered that degrading the KRASG12V oncoprotein leads to regression of the resulting lung adenocarcinoma tumors. Most of the regression was based on cancer cell-intrinsic responses, although the tumor microenvironment also underwent substantial remodeling. Despite the initial efficacy of the treatment, the authors found that prolonged PROTAC KRAS degrader treatment eventually resulted in relapse. Resistance to PROTAC treatment seemed to be driven by dysregulation of the ubiquitin-proteasome system that is required for the activity of the PROTAC degraders. Despite developing resistance to the PROTAC degraders, the resulting tumors were still dependent on the KRAS oncoprotein, meaning that they were still sensitive to conventional KRAS inhibitors. Thus, PROTACs that degrade the KRAS oncoprotein are a promising modality for the treatment of lung adenocarcinomas. Resistance to PROTACs may differ from conventional KRAS inhibitors, suggesting potential strategies for overcoming such resistance. See related article by Martín et al., p. 4115.

Immune cell microbots with self-propelling and navigating capabilities have become an exciting field of research. Isolation, activation, and adoptive transfer of cytotoxic immune cells in vitro are some of the most common technical methods. However, the ex situ activation and adoptive transfer of cytotoxic immune cells carry the risk of systemic inflammatory responses and functional exhaustion. To address this bottleneck, we report a CD5+ dendritic cell microbot (CD5+DC robot) constructed by engineering natural CD5+ dendritic cells to phagocytose magnetic nanoparticles coated with tumor cell membranes. The PD-L1 on the surface of CD5+ cells is preblocked to enhance immune activation capacity. Under switched exogenous rotating/conical magnetic fields, CD5+DC robots form chain-like clusters for upstream motion or ribbon-like clusters for downstream motion within the vasculature, enabling precise aggregation at intestinal targets. Then, they navigate along tumor chemokine gradients, penetrating the extracellular matrix barrier via positive chemotaxis to infiltrate deep into tumor tissues and trigger cascading activation of resident immune cells. Distinct from traditional cellular microbot therapies that rely on self-mediated cytotoxicity, this CD5+DC robot system activates durable antitumor immunity by reprogramming innate immune cells within the tumor microenvironment. This strategy offers a novel pathway toward precision-targeted therapy with excellent biocompatibility and functional stability.

The limited efficacy of immunotherapies in advanced prostate cancer stems from a tumor microenvironment (TME) in which myeloid-driven immune suppression, stromal remodeling, and metabolic barriers converge to limit antitumor immunity. In this study, we characterized the immunometabolic properties of an ultrasmall prostate-specific membrane antigen-targeting silica particle therapy as a first-in-class strategy to reprogram the Toll-like receptor (TLR)-ferroptosis axis in MYC-driven prostate cancer. As single agents, these particles suppressed lipid and steroid biosynthesis, disrupted lipid peroxidation control, and impaired nutrient flux, sensitizing tumors to ferroptosis. Coordinated redox remodeling, stromal reprogramming, and innate immune activation reversed myeloid suppression and promoted CD8+ T-cell infiltration. When combined with CSF-1R inhibition and immune checkpoint blockade, the particles suppressed tumor growth, extended survival beyond 100 days, and achieved up to 50% complete remission in MYC-overexpressing models. These findings position TLR-ferroptosis axis remodeling as a mechanistic blueprint for rational, particle-driven immunotherapies with broad translational potential in prostate cancer and other immunologically refractory malignancies. Clinically validated, PSMA-targeted ultrasmall core-shell silica particles reprogram immunometabolic pathways via a TLR-ferroptosis axis, enabling tumor microenvironment remodeling and potentiating checkpoint blockade in prostate cancer, with translational implications for treatment-resistant disease.

Pancreatic ductal adenocarcinoma (PDAC) relies on elevated autophagy to support metabolism, proliferation, and immune evasion. Inhibiting autophagy has been reported to improve response rates in patients with PDAC. In this work, we identified a mechanism to explain how the loss of autophagy in PDAC triggers reprogramming of the tumor microenvironment (TME) to ultimately stimulate an antitumor response. Autophagy inhibition in PDAC recruited macrophages via the CXCL1/2-CXCR2 axis. Simultaneously, the loss of autophagy resulted in a decrease in the canonical "don't eat me" ligand CD47 on tumor cells, thereby inducing their susceptibility to macrophage phagocytosis. Although CD8+ T cells were critical to the antitumor immune response to autophagy inhibition in PDAC, they were not directly involved in cytotoxicity but played a critical role in stimulating macrophage phagocytosis of tumor cells. Taken together, this study strongly supports the implementation of autophagy inhibition in pancreatic cancer and highlights a crucial link between PDAC biology and the TME-macrophage cross-talk that effectively promotes tumor cell killing. Inhibition of autophagy in pancreatic cancer promotes antitumor immunity through a stepwise process of recruiting macrophages and enhancing phagocytosis of cancer cells, providing a potential therapeutic strategy for this deadly tumor type.

Chronic alcohol consumption is an established risk factor for esophageal squamous cell carcinoma (ESCC). Elucidating the underlying mechanisms linking metabolic stress to esophageal carcinogenesis could help identify prevention and treatment strategies. Through a transancestry meta-analysis encompassing more than 200,000 individuals from East Asia, we identified a germline variant (rs1051921 C>T) at the MLXIPL locus [encoding carbohydrate-responsive element-binding protein (ChREBP)] that markedly elevated ESCC risk among alcohol drinkers. Functionally, the risk allele facilitated N6-methyladenosine (m6A) modification of ChREBP transcripts, enhancing their stability through recognition by the m6A reader YTHDF1. Alcohol further potentiated this regulatory axis by promoting ACSS2-mediated conversion of acetate into acetyl-CoA and increasing H3K27ac enrichment at the promoters of key m6A regulators. Increased ChREBP expression activated the transcription factor ATF3, which triggered endoplasmic reticulum stress and epithelial-mesenchymal transition, thereby conferring anoikis resistance. In parallel, ATF3 promoted an immunosuppressive tumor microenvironment through the upregulation of PD-L1 and VEGFA, leading to the exclusion of CD8+ T cells and the expansion of granulocytic myeloid-derived suppressor cells. In vivo intervention via ATF3 knockdown, anti-PD-L1 treatment, or pharmacologic perturbation of ChREBP-related metabolism with metformin or an ACSS2 inhibitor significantly suppressed ESCC progression and restored intratumoral CD8+ T-cell infiltration. These findings establish ChREBP as an alcohol-sensitive metabolic-epitranscriptomic switch that integrates genetic susceptibility with immune evasion to drive esophageal carcinogenesis. Alcohol-responsive m6A-ChREBP signaling links genetic susceptibility to metabolic stress and immune evasion in esophageal cancer, revealing actionable targets for risk stratification, prevention, and immunometabolic intervention in high-risk drinkers.

Fusobacterium nucleatum is a key microbial driver of colorectal cancer progression. In this study, we identified an intercellular signaling axis through which F. nucleatum remodels the immune microenvironment. Analysis of 54 clinical colorectal cancer specimens revealed that high intratumoral F. nucleatum load correlated significantly with PD-L1 upregulation in tumor-associated macrophages, diminished CD8+ T-cell cytotoxicity, and poor patient prognosis. Mechanistically, F. nucleatum infection activated NF-κB signaling in colorectal cancer cells to induce the transcription of the long noncoding RNA mitotically associated long noncoding RNA (MANCR), which was selectively packaged into exosomes via the ESCRT-III/ALIX complex and transferred to macrophages. In recipient macrophages, exosomal MANCR interacted with HNRNP U to increase PD-L1 mRNA stability, leading to sustained PD-L1 surface expression. In humanized mouse models, F. nucleatum exposure inhibited CD8+ T-cell infiltration and suppressed granzyme B activity, thereby compromising antitumor immunity and facilitating tumor proliferation and metastasis. These findings demonstrate that F. nucleatum exploits a tumor-derived exosomal long noncoding RNA network to remotely manipulate macrophage plasticity. Therefore, targeting the F. nucleatum/MANCR/PD-L1 axis may represent a viable strategy to overcome immune resistance in the colorectal cancer microenvironment. Fusobacterium nucleatum exploits tumor-derived exosomal MANCR to stabilize PD-L1 in macrophages, providing a mechanism of bacterial-driven immune evasion and a potential therapeutic target to restore antitumor immunity in colorectal cancer.

Recent drug discovery breakthroughs have led to the approval of KRASG12C inhibitors in lung adenocarcinoma. Unfortunately, clinical responses are often hampered by the rapid onset of resistance. Proteolysis-targeting chimeras (PROTAC) have emerged as promising alternatives to traditional inhibition. However, there is limited mechanistic understanding of KRAS degradation in vivo. In this study, we developed a preclinical lung adenocarcinoma mouse model and demonstrated that targeted oncogenic KRAS degradation induces rapid tumor regression primarily due to cancer cell-autonomous mechanisms. However, transcriptional, histologic, and immunophenotypic analyses revealed a substantial remodeling of the tumor microenvironment. Notably, disease relapse observed during prolonged PROTAC treatment stemmed mostly from proteolysis machinery dysregulation, indicating resistance mechanisms distinct from those reported upon KRAS inhibition. Collectively, these findings highlight the therapeutic potential of KRAS degradation in lung adenocarcinoma, providing insights into both cell-intrinsic and cell-extrinsic mechanisms that accompany antitumor responses and support the ongoing clinical exploration of this approach. KRAS degradation induces rapid regression of lung adenocarcinoma tumors mainly via cancer cell-intrinsic mechanisms, offering a complementary strategy to target KRAS given the short duration of clinical responses to inhibitors. See related commentary by Garcia Borrego and Misale, p. 3903.

Transcriptional intratumoral heterogeneity (ITH) is a hallmark of aggressive cancers. Investigation into the ITH programs that drive tumor metastasis and immune evasion could help identify potential treatment and prevention approaches. Through single-cell RNA sequencing analysis of upper aerodigestive squamous cell carcinoma (UASCC) cells and patient tumors, we uncovered a hybrid epithelial-mesenchymal transition (hEMT) ITH program linked to metastatic dissemination. The transcription factor ETS1 was identified as a master regulator of the hEMT program, directly activating prometastatic genes and promoting distant spread in vivo. Unexpectedly, ETS1 also orchestrated an immune-cold tumor microenvironment by transcriptionally activating STAT1 and CD274 (PD-L1), suppressing T lymphocyte infiltration, and elevating immune checkpoint molecules. Clinically, high ETS1 expression in tumors strongly correlated with poor survival and resistance to immune checkpoint blockade across multiple cohorts. Drug screening demonstrated that ETS1-high cancers were vulnerable to HSP90 inhibitors (e.g., alvespimycin), which suppress ETS1 by disrupting hypoxia-inducible factor 1 alpha-mediated transcriptional activation. Together, this work reveals ETS1 as a dual driver of tumor distal metastasis and immune evasion in UASCC, while nominating HSP90 inhibition as a tailored treatment strategy for ETS1-driven tumors. These findings provide a roadmap for targeting aggressive ITH subsets and overcoming immunotherapy resistance. Transcriptional intratumoral heterogeneity drives metastasis and immune evasion via an ETS1-regulated hybrid EMT program, which can be targeted with HSP90 inhibitors to treat aggressive, immunotherapy-resistant tumors.

The cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway is a central sensor of innate immunity that plays critical roles in recognizing cytosolic DNA and initiating antitumor immune responses. However, this pathway exhibits extensive spatiotemporal duality and context dependency in tumor regulation. In recent years, modulating this pathway to convert immunologically "cold" tumors into immunologically "hot," inflamed tumors has emerged as a cutting-edge strategy to reverse resistance to immune checkpoint inhibitors (ICIs). This review outlines the molecular mechanisms underlying the activation and regulation of the cGAS-STING pathway, with emphasis on its complex role in orchestrating the tumor immune phenotypic switch. A nuanced analysis of the pathway's duality distinguishes between acute immunostimulatory activation and chronic, pro-tumorigenic inflammation driven by chromosomal instability (CIN). Furthermore, current evidence regarding direct and indirect T-cell modulation, as well as pathway-mediated remodeling of the tumor microenvironment (TME) across diverse malignancies, is discussed in detail. Crucially, the current bottlenecks in clinical translation are described, including evaluation of the failure of first-generation agonists and the promise of next-generation delivery platforms such as antibody-drug conjugates (ADCs) and nanoparticle systems. Finally, a novel strategic framework is proposed involving mapping of specific STING-targeted modalities to distinct TME phenotypes, such as immune-desert, immune-excluded, and exhausted-inflamed states. A detailed understanding of the cGAS-STING axis has substantial value in providing theoretical guidance and supporting clinical translation in the development of next-generation combination immunotherapies, as well as broadening the patient population benefiting from clinical interventions.

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

Immunotherapeutic approaches for cutaneous squamous cell carcinoma (cSCC) remain limited to programmed cell death protein 1 (PD-1) blockade. Although genomics studies have characterized key driver mutations in cSCC, preclinical models that faithfully recapitulate both the genetic landscape and immune microenvironment of the human disease, that could drive the development of novel, effective therapies, are lacking. To address this need, we generated genetically engineered mouse models harboring inducible p53wmR172H expression and Cdkn2a deletion in stratified epithelia. These mice spontaneously developed well-differentiated and spindle cell cSCCs, whose histopathological and immunological features mimicked those observed in patients. We established syngeneic cell lines and characterized their genomic and transcriptomic profiles through whole exome sequencing and RNA sequencing. We used these syngeneic tumor models to test the effect of STING agonist, PD-1, VISTA and CTLA-4 blockade applied at different schedules and combinations. Finally, immune checkpoint molecules detected in the syngeneic models were validated in human cSCC tissue arrays. Whole exome sequencing confirmed alterations overlapping with human cSCC, including mutations in Notch, interferon signaling, and cytokine pathways. Transcriptomic analysis revealed the upregulation of immunoregulatory genes, including Cd274, Lgals9, and Il33, suggesting a suppressive tumor immune microenvironment (TIME). Preclinical therapeutic evaluation in this model demonstrated that PD-1 blockade and the treatment with a STING agonist elicited partial responses. The combination of STING agonist with checkpoint inhibition targeting either PD-1/CTLA-4 or PD-1/VISTA significantly enhanced antitumor immunity in tumors resistant to anti-PD-1 monotherapy. These results establish a robust, genetically defined, immunocompetent platform for modeling cSCC and evaluating novel immunotherapeutic strategies for treating patients with this disease. Our findings also identify VISTA and CTLA-4 as promising immune checkpoints to target in cSCC and support clinical evaluation of this combination of immune checkpoint blockade to overcome immune checkpoint inhibitor resistance and improve clinical outcomes.

Systemic administration of bispecific T‑cell engagers (BiTEs) targeting CEACAM6 has shown therapeutic potential but is limited by on‑target/off‑tumor toxicity and short serum half‑life. To overcome these challenges, we engineered an oncolytic herpes simplex virus (oHSV) that delivers and locally produces anti‑CEACAM6/CD3 BiTE directly within the tumor microenvironment (oHSV‑anti‑CEA6/CD3). In immunocompetent mouse models of colon (CT26) and breast (4T1) cancer, a single intratumoral injection of oHSV‑anti‑CEA6/CD3 promoted tumor regression, increased infiltration of CD8⁺CD44⁺ T cells both locally and systemically, and induced durable antitumor immunity. When combined with PD‑1 blockade, this localized BiTE‑arming strategy synergistically enhanced therapeutic efficacy, leading to superior tumor control and long‑term immunological memory without additional toxicity. Mechanistically, the virus exerts a dual action: direct oncolysis and sustained in situ BiTE production, enabling targeted T‑cell activation and tumor elimination while avoiding systemic BiTE exposure. Unlike previous reports of BiTE‑armed oncolytic viruses or simple combinations with immune checkpoint inhibitors, our approach leverages the oHSV platform to achieve spatiotemporally controlled T‑cell redirection coupled with virus‑mediated innate immune stimulation. This work provides a safe, effective, and readily translatable strategy for solid tumor immunotherapy.

Cancer progression is closely associated with dysregulated redox homeostasis within the tumour microenvironment (TME), where reactive oxygen and nitrogen species influence tumour survival, immune evasion, and therapeutic resistance. While physiological redox signalling supports antitumour immunity, persistent oxidative stress promotes immunosuppression and limits the efficacy of immunotherapies, making redox modulation an attractive therapeutic strategy. Naturally occurring polyphenols have emerged as promising redox-regulating agents due to their antioxidant, anti-inflammatory, and immunomodulatory properties. Beyond scavenging reactive species, they regulate pathways involved in immune activation, metabolic reprogramming, ferroptosis, and tumour-immune interactions. However, their clinical application is hindered by poor bioavailability, rapid metabolism, and limited tumour accumulation. Nanotechnology-based delivery systems, including liposomes, polymeric nanoparticles, metal-based nanoplatforms, biomimetic vesicles, and stimuli-responsive carriers, have been developed to overcome these limitations. These platforms enhance polyphenol stability, targeted delivery, and controlled release while enabling synergistic interactions with immunotherapy. Importantly, they can function as programmable redox-immunomodulatory systems capable of remodelling the TME, inducing immunogenic cell death, promoting ferroptosis, and activating innate and adaptive immune responses. This review discusses the interplay between redox regulation and tumour immunity, examines the immunomodulatory mechanisms of polyphenols, and highlights recent advances in nanocarrier-based delivery strategies. Challenges related to biological heterogeneity, biomarker development, manufacturing scalability, regulatory approval, and clinical translation are also considered, together with future perspectives for precision redox medicine based on polyphenol-enabled nanotechnologies.

Inflammatory cancer-associated fibroblasts (iCAFs) are a highly plastic stromal population that critically shapes tumor progression, immunosuppression, and therapeutic response in esophageal squamous-cell carcinoma (ESCC). Epithelial-intrinsic programs are increasingly recognized as key determinants of fibroblast reprogramming within the tumor microenvironment, yet the underlying mechanisms remain incompletely understood. Here, we identified hepatoma-derived growth factor (HDGF) as a pivotal epithelial-intrinsic regulator that drives iCAF formation in ESCC. Mechanistically, nuclear HDGF functioned as a transcriptional activator by directly binding the ENO1 promoter, thereby upregulating the expression of the glycolytic enzyme enolase 1, enhancing aerobic glycolysis, and promoting lactate secretion from tumor cells. Tumor-derived lactate was subsequently taken up by CAFs and induced histone H4 lysine 12 lactylation (H4K12la), which epigenetically activated NF-κB signaling and promoted iCAF formation. Functionally, HDGF-induced iCAFs promoted tumor progression through activation of the IL-6/JAK1/STAT3 axis and established an immunosuppressive microenvironment characterized by increased recruitment of regulatory T cells and reduced infiltration of CD8+ T cells, thereby facilitating immune evasion. Therapeutically, blockade of ENO1 effectively disrupted the glycolysis-lactylation cascade, markedly suppressing tumor growth and iCAF formation in vivo. Moreover, ENO1 inhibition reprogrammed the immunosuppressive tumor microenvironment and significantly enhanced the efficacy of anti-PD-1 therapy. Collectively, our findings reveal an HDGF/ENO1/H4K12la/iCAF axis that links tumor metabolic reprogramming, stromal inflammatory activation, and immunosuppression in ESCC, identifying this axis as a promising therapeutic target for overcoming immunotherapy resistance.

As the most prevalent subtype of colorectal cancer, microsatellite-stable colorectal cancer (MSS CRC) is resistant to T cell-focused immune checkpoint blockade due to its low mutational burden and immunosuppressive tumor microenvironment. Despite reprogramming tumor-associated macrophages toward a tumoricidal M1-like state being a promising alternative, its efficacy is limited by paradoxical upregulation of programmed cell death-ligand 1 (PD-L1) on M1-like macrophages and their inherently poor antigen cross-presentation capacity. Here, we report that poly-metformin (PMet) can mimic the intracellular domain of PD-L1, competitively inhibit the membrane anchoring of PD-L1, and thereby effectively downregulate cell-surface PD-L1. To evaluate the therapeutic efficacy of PMet in immunosuppressive MSS CRC, we further developed an oral probiotic outer membrane vesicle (OMV) gene delivery system, siYthdf2/PMet@Akk-OMV, and demonstrated its ability to reduce PD-L1 levels on both macrophages and tumor cells while synergistically enhancing macrophage cross-presentation of tumor antigens, leading to potent activation of specific antitumor immunity and significant tumor growth inhibition. This work thus provides both theoretical and experimental foundations for treating cancers resistant to traditional immune checkpoint blockade therapy.

Radiofrequency ablation (RFA) is the primary curative therapy for early hepatocellular carcinoma (HCC). However, incomplete ablation inevitably occurs in clinical practice, leaving residual tumors that are highly immunosuppressive, refractory to immune checkpoint inhibitors, and prone to recurrence and metastasis. To address this, we developed a novel nanoformulation, OMVbsAb-MAL, by modifying bacterial outer membrane vesicles (OMVs) with DEC205/PD-L1 bispecific antibodies and maleimide (MAL) groups. Following local administration after RFA, OMVbsAb-MAL forms thioether bonds with tumor proteins, thereby capturing abundant tumor-associated antigens (TAAs) released in situ. The nano-sized OMVbsAb-MAL penetrates the tumor stroma and traffics efficiently to tumor-draining lymph nodes. Significantly, the bispecific antibodies on the nanoplatform enable specific DC recognition, mediating targeted delivery of TAAs to DCs within tumors and lymph nodes. Concurrently, OMVbsAb-MAL blocks PD-L1, thereby restoring DC function and alleviating PD-L1-mediated immune suppression. In combination with TAAs, the immunogenic vesicles markedly promote DC maturation, resulting in robust antigen presentation and potent T cell activation. Consequently, OMVbsAb-MAL synergizes with RFA to elicit both local and systemic antitumor immune responses, effectively suppressing residual and distant tumors, and thus provides a promising strategy for integrating RFA with immunotherapy in HCC.

Tumor immunotherapy has emerged as a transformative strategy for cancer treatment. However, its clinical efficacy remains limited by the immunosuppressive tumor microenvironment (TME), which restricts immune cells infiltration, suppresses effector cells activity, and promotes immune escape. Recent advances in catalytic nanomedicine have highlighted single-atom nanozymes (SAzymes) as a promising platform for overcoming these barriers. Owing to their atomic precision and tunable electronic structure, SAzymes exhibit highly efficient enzyme-like catalytic activities and enable precise regulation of the TME. Through catalytic therapy, SAzymes can alleviate hypoxia, regulate lactate accumulation, disrupt redox homeostasis, and reprogram immunosuppressive immune cells populations. Importantly, SAzyme-mediated catalytic therapy can induce immunogenic cell death (ICD), thereby enhancing antitumor immune responses. In addition, the well-defined atomic architecture of SAzymes provides a unique opportunity to establish quantitative structure-activity relationships (QSAR), enabling rational optimization of catalytic performance through modulation of their composition and coordination environment. This review systematically summarizes the mechanisms by which SAzymes remodel the TME and enhance cancer immunotherapy, discussing recent advances in atomic level design principles and multimodal synergistic therapy. Finally, current challenges and future perspectives, including machine learning-guided SAzymes design, biosafety and clinical translation, are discussed to guide the next generation of SAzyme-based catalytic immunotherapies.

Antibody-mediated blockade of the PD-1/PD-L1 immune checkpoint has revolutionized cancer treatment. However, their efficacy is often limited by the compensatory upregulation of PD-L1 synthesis, which sustains surface expression upon blockade. FOXM1, a pivotal oncogenic transcription factor overexpressed in diverse cancers, directly transactivates PD-L1 expression, presenting a strategic upstream therapeutic target. To concurrently suppress both membrane-bound and newly synthesized intracellular PD-L1, we modularly designed a dual-functional nanoassembly via the co-assembly of a fluorinated FOXM1-inhibitory peptide and a PD-L1-targeting aptamer. This nanoassembly leverages fluorination to enhance nanoassembly stability and cytosolic delivery efficiency to achieve a programmable two-stage PD-L1 suppression. The aptamer module mediates tumor-targeted binding and blocks surface PD-L1, after which the internalized nanoassembly releases the peptide module to transcriptionally suppress PD-L1 via FOXM1 inhibition. Consequently, this nanoassembly achieves potent PD-L1 downregulation, addressing the limitations of conventional antibody blockade that primarily targets surface proteins. In murine models, this dual-inhibition strategy robustly reinvigorates antitumor immunity, significantly suppressing tumor growth and metastasis. Our work establishes a dual-functional nanoassembly that programmably controls PD-L1 expression, presenting a promising approach to circumvent adaptive resistance in cancer immunotherapy by durably targeting the source of PD-L1 expression.

Chimeric antigen receptor (CAR) T-cell therapy has shown limited efficacy in solid tumors, largely due to immunosuppressive mechanisms within the tumor microenvironment (TME). While tumor-associated glycans are known to protect malignant cells from immune attack, the contribution of N-glycans expressed by non-malignant TME populations to CAR-T cell dysfunction remains poorly defined. We investigated the role of N-glycans in non-malignant TME populations, focusing on M2-like macrophages and hepatic stellate cells in liver metastasis of colorectal (CRC) and pancreatic cancer (PDAC). Using in vitro co-culture systems, transcriptomic analysis, and tumor-bearing humanized mouse models, we assessed how pharmacologic or genetic disruption of key nodes of the N-glycosylation pathway (MGAT5, MAN2A1 and ST6GAL1) in immune and stromal compartments shapes T-cell function. In patient samples, a branched N-glycan signature was associated with transcriptional programs characteristic of tumor-promoting macrophages and stromal cells, linking N-glycosylation to an immunosuppressive TME. Disruption of N-glycan synthesis in non-malignant TME cells reduced their immunosuppressive and tumor-supporting functions. Single-cell RNA sequencing of tumor-bearing humanized mice showed depletion of protumor IL1β+ macrophages and diminished inhibitory macrophage-T cell interactions following N-glycosylation blockade. Selective MGAT5 disruption in immune and stromal compartments suppressed immunosuppressive programs and enhanced CAR-T cell antitumor activity independently of tumor cell glycosylation. These findings show that N-glycans expressed by non-malignant TME cells restrain CAR-T cell responses in CRC and PDAC, highlighting MGAT5-dependent branching as a potentially actionable axis and supporting a broader role for multiple nodes of the N-glycosylation pathway.

Acquired resistance limits the durability of programmed cell death protein-1 (PD-1) blockade in lung adenocarcinoma, yet the tumor-intrinsic programs and immune circuits that drive acquired resistance relapse remain poorly defined. The purpose of this study was to identify tumor-intrinsic mediators of acquired resistance and determine how they remodel antitumor immunity. An orthotopic bioluminescence-tracked Lewis lung carcinoma (LLC1) lung adenocarcinoma model was established in immunocompetent mice treated with anti-PD-1. Tumor-intrinsic regulators were identified by an in vivo genome-wide CRISPR loss-of-function screen and validated using inducible tetracycline-off knockdown. Prostaglandin E2 (PGE2) signaling was interrogated through tumor-cell Ptgs2 knockdown/deletion, 16,16-dimethyl PGE2 administration, selective EP2/EP4 antagonists, and celecoxib treatment. Natural killer (NK)-cell function was analyzed by flow cytometry, immunofluorescence, RNA sequencing, cAMP measurement, calcium flux assays, mouse and human NK-cell co-culture cytotoxicity assays, and NK-cell adoptive transfer. Celecoxib was used to evaluate the therapeutic potential of pharmacologic PGE2 blockade in vivo. Public immunotherapy datasets were analyzed to assess the clinical relevance of PTGS2. The orthotopic LLC1 model captured key features of heterogeneous anti-PD-1 responses, including relapse after initial regression. The CRISPR screen identified Ptgs2 as a key driver of acquired resistance. Tumor-derived PGE2 progressively increased in resistant tumors, and its genetic silencing overcame resistance by restoring NK-cell infiltration and function. Mechanistically, PGE2 signaled through EP2/EP4 receptors to elevate cAMP and induce CREM, thereby suppressing NK-cell cytotoxicity and cytokine production. This axis was validated in human NK cells. Pharmacologic inhibition of cyclooxygenase-2 with celecoxib reversed acquired resistance, an effect abrogated by NK-cell depletion. Tumor-derived PGE2 is an important contributor to acquired resistance to PD-1 blockade in lung adenocarcinoma. Therapeutic disruption of the EP2/EP4-cAMP-CREM axis restores NK-cell function and overcomes acquired resistance.

Soluble programmed death-ligand 1 (sPD-L1) is a promising non-invasive biomarker for early cancer diagnosis and immunotherapy monitoring. However, electrochemical detection of sPD-L1 in serum remains challenging due to severe biofouling that compromises sensitivity and specificity. We developed a one-step, dual-antifouling electrochemical interface by co-assembling phase-transited bovine serum albumin (PTB) and a cyclic multifunctional peptide (CP). PTB forms a dense, amyloid-like nanofilm that acts as a robust protein shield, while CP introduces a zwitterionic antifouling loop and a PD-L1-binding motif. This synergistic architecture significantly suppresses non-specific protein adsorption while enabling oriented target recognition. The sensor achieves a detection limit of 37.4 pg/mL and accurately distinguishes cancer patients from healthy individuals using neat serum without dilution or pretreatment. The PTB/CP dual-antifouling platform offers a simple, sensitive, and clinically translatable strategy for sPD-L1 detection, with broad potential for point-of-care cancer diagnostics and immune monitoring.

Metastatic colorectal cancer (mCRC) is one of the deadliest cancers with very poor response to immune checkpoint blockade (ICB). Standard therapies employ chemotherapy combined with epidermal growth factor receptor (EGFR) blocking antibodies, which are only effective in a fraction of patients with RAS/RAF wild-type tumours. We have previously shown that EGFR deletion in myeloid cells of CRC, rather than in the cancer cells themselves, reduces tumour growth. Here, we investigate to which extent EGFR blockade in myeloid cells increases anti-tumour immunity, thus sensitising CRC to ICB. Using a syngeneic preclinical CRC liver metastasis model based on the transplantation of murine RAS mutant CRC organoids into mice lacking EGFR in myeloid cells, we observe a reduction in metastasis development accompanied by increased intratumoural T-cell infiltration. Importantly, we demonstrate that EGFR deletion reduces the capacity of granulocytic myeloid-derived suppressor cells (G-MDSCs) to suppress CD4+ T-cell proliferation. RNA-seq analysis of sorted MDSCs and T-cells uncovered an EGFR-dependent signature involved in immunosuppression, which in proficient-mismatch-repair (pMMR) CRC patients is associated with worse overall survival. Therapeutically, lifting immunosuppression by EGFR deletion in myeloid cells sensitised tumours to anti-PD-L1 treatment, thus preventing liver metastasis development. These results imply that anti-EGFR therapies combined with ICB might be successful in preventing metastasis of RAS mutated CRC with high infiltration of suppressive EGFR+ myeloid cells.

Prostate cancer (PCa) remains one of the most prevalent and lethal cancers in men, particularly in its metastatic form. Despite therapeutic advancements, resistance to therapy often leads to a castration-resistant state with genetic and molecular alterations that promote immune evasion and disease progression. Chronic inflammation and the immunosuppressive tumor microenvironment (TME) further exacerbate immune dysfunction. The suboptimal clinical success of current immunotherapies in advanced PCa underscores the urgent need to better understand the mechanisms underlying immune escape during disease progression and in response to therapeutic interventions. This review explores the key immune-evasive features of PCa, the contribution of the TME, and the immunotherapeutic strategies that have been clinically evaluated to improve outcomes in patients with advanced disease.

Gastrointestinal (GI) adenocarcinomas pose a significant therapeutic challenge due to highly immunosuppressive tumor microenvironments that limit effective antitumor immune responses. Dense stromal fibrosis, immune exclusion and poor responses to immune checkpoint inhibitors are hallmarks of treatment-resistant malignancies, most notably pancreatic ductal adenocarcinoma (PDAC) and subsets of gastric cancer. In this regard, IL-15 superagonists such as N-803, NIZ985, RLI, and NKTR-255 have emerged as promising immunotherapeutic candidates, as they selectively expand natural killer (NK) cells and CD8+ T-cells without the systemic toxicity and regulatory T-cell activation observed with IL-2 therapy. IL-15 superagonists have emerged as more effective therapeutic agents when used in conjunction with local inhibitors of TGF-β and IL-10, stromal remodeling, and vascular normalization. They are supported by increasingly conclusive mechanistic, preclinical and early clinical data. A combination of these interventions circumvents extracellular matrix-provoked immune barriers, reverses VEGF-induced endothelial dysfunction, and enhances the trafficking of lymphocytes into tumor cores. Moreover, several localized therapeutic delivery strategies, such as endoscopic delivery, implantable depots, biomaterial scaffolds, and nanocarriers, can improve intratumoral cytokine retention and reduce systemic inflammatory toxicity. Together, these advances define a new paradigm for cytokine-immunotherapy: IL-15 superagonists serving as key mediators of reprogramming of the tumor microenvironment. Therapeutic strategies targeting IL-15 have the potential to restore cytotoxic immunity, improve access to immune cells, and promote responsiveness to the checkpoint blockade, making them a promising yet investigational approach to converting immune-cold GI malignancies into more treatment-responsive disease states, though further clinical validation is required.

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

Fasting-mimicking diets (FMD) have been reported to increase the anti-tumor efficacy in preclinical studies in various cancers. FMD can impact the tumor microenvironment (TME), and elucidating the mechanisms and cells mediating the effects of FMD may uncover combination treatment strategies. Here, we performed single-cell transcriptomic sequencing to characterize the TME changes induced by FMD intervention in a mouse model of spontaneous breast cancer. The sequencing data indicated that FMD suppressed tumor cell stemness, promoted apoptosis, and increased the infiltration of innate and adaptive immune cells, especially natural killer (NK) cells and effector CD8+T cells. Importantly, FMD induced phenotypic reprogramming of cancer-associated fibroblasts (CAFs), leading to a decrease in the immunosuppressive inflammatory CAF (iCAF) subset. Mechanistically, FMD decreased platelet-derived growth factor C (PDGFC) secretion in tumor cells by reducing glucose and inhibiting glycolysis to reprogram CAFs. PDGFC increased activation of the JAK/STAT3 pathway, which induced iCAF differentiation. Combining FMD with PDGFR inhibitors increased the efficacy of anti-PD-L1 immunotherapy in vivo. Overall, this study reveals a mechanism of metabolic-immune rewiring through which FMD suppresses tumor progression, providing preliminary evidence for the potential of combination FMD-based strategies in breast cancer treatment.

The clinical efficacy of cancer immunotherapy is fundamentally constrained by the synergistic evasion mechanisms of antigen concealment and effector immune cell suppression. To overcome these barriers, we proposed a therapeutic paradigm that sequentially dismantles the tumor's "immune camouflage" and relieves "metabolic coercion," thereby enabling systematic remodeling of the tumor immune microenvironment. A cascade-acting nano-Proteolysis Targeting Chimera (PROTAC) engine was fabricated via phosphatidylcholine-driven self-assembly, enabling tumor-targeted delivery and pH-responsive drug release within the tumor microenvironment. The system initially disrupted the tumor's immune evasion barrier by inducing cellular senescence and enhancing MHC-I-mediated antigen presentation. It subsequently reprogrammed tumor metabolism through targeted degradation of the BRD4/c-Myc axis, which alleviated nutrient competition and reversed the metabolic suppression of tumor-infiltrating T cells. This sequential intervention potently enhanced dendritic cell activation and antigen-presenting capacity, promoted the infiltration and reactivation of cytotoxic T lymphocytes, and synergistically induced PD-L1 degradation. Collectively, these coordinated effects established a durable antitumor immune response that potently suppressed both primary tumor growth and distant metastasis in a triple-negative breast cancer model. Overall, this work established a cascaded immune remodeling paradigm, conceptualized as unmasking and unblocking, offering a broadly applicable therapeutic strategy to overcome immunosuppression in solid tumors.

Innovations in therapeutic modalities and targeted drug delivery are two key approaches to improving the effectiveness of cancer treatment. With the development of therapeutic concepts and nanotechnology, recent research has shifted from tissue- or cell-level drug delivery to organelle-specific delivery to amplify therapeutic effects. Among these organelles, the endoplasmic reticulum (ER) has emerged as a particularly promising target because of its extensive membrane network, which serves as a major site for protein synthesis, lipid metabolism, calcium homeostasis, and intracellular signaling. The disruption of ER function can trigger severe cellular stress and apoptosis, providing a rationale for ER-targeted cancer therapy. However, traditional small-molecule drugs often exhibit poor ER specificity, rapid degradation, and limited intracellular accumulation, significantly restricting their therapeutic potential. In contrast, ER-targeting multifunctional nanoplatforms have demonstrated superior advantages, including enhanced stability, precise localization, controlled drug release, and multimodal therapeutic capabilities. In this Review, we systematically summarize the recent advances in ER-targeting nanotherapeutics and their applications in chemotherapy, phototherapy, immunotherapy, biotherapy, and combination therapy. Furthermore, we discuss the advantages, challenges, and mechanistic insights of these strategies, aiming to provide perspectives on accelerating the clinical translation of ER-targeting precision drugs.

Three-dimensional (3D) cell models, such as spheroids and organoids, are powerful tools to enhance the predictive power of preclinical in vitro studies and support clinical translation of anti-cancer therapies. However, their integration into drug discovery pipelines remains limited, particularly for high-throughput screening, where rapid and reliable viability assessment is essential. Conventional viability assays, often dye-based or destructive, remain labor-intensive and poorly suited for high-throughput applications and downstream re-use of the models. In this study, we demonstrate, for the first time, that electrical impedance spectroscopy (EIS) enables label-free viability profiling of individual spheroids. For this purpose, a differential EIS sensor was integrated into a plastic microfluidic chip in flow-through configuration with self-aligned facing electrodes. Using six probing frequencies, we measured the EIS responses of liver cancer spheroids exposed to varying concentrations of dimethyl sulfoxide and doxorubicin, a chemotherapeutic compound. We identified an optimized frequency pair that enabled us to monitor spheroid viability, independent of spheroid size. Comparison with conventional viability assays, namely fluorescence imaging and ATP quantification, confirmed that the EIS method reliably reflected viability changes and revealed inter- and intra-treatment group differences owing to the method's single-spheroid resolution. The same frequency pair could be used to detect viability changes for both compounds, suggesting transferability of the EIS method across treatments. Furthermore, we showed that utilizing additional EIS features improved classification of spheroids with subtle viability differences. Together, these results demonstrate that EIS is a promising approach for noninvasive, single-spheroid viability profiling and a valuable tool for high-throughput drug screening with spheroid models.

High levels of immune suppression are a common intrinsic mechanism of resistance in metastatic breast cancer that calls for developing immunotherapeutic combinations to broaden treatment responses. Histone deacetylase (HDAC) inhibitors can sensitize tumors to dual checkpoint inhibition in patients. Here, we investigated the tumor microenvironment (TME) of breast metastases by combining experimental and clinical data with theory to elucidate the mechanism of response to treatment with the HDAC inhibitor entinostat combined with anti-PD-1 and anti-CTLA-4. Knowledge-guided subclustering of single-cell RNA-sequencing (scRNA-seq) data and cell circuit analyses from murine breast-to-lung metastases identified 39 cell states and salient interactions, of which myeloid, T cell, and B cell subpopulations were most affected. Analyses of patient biopsies and blood via spatial proteomics and flow cytometry corroborated the preclinical findings, showing increased T cell and B cell activation, mature tertiary lymphoid structures, and increased CD8+ T cell-macrophage distances in responders to entinostat + nivolumab + ipilimumab. Combination treatment increased immunoglobulin production in patients and mice, and murine studies demonstrated increased tumor-targeting IgG and implicated B cells as necessary for treatment response. Inhibition of the ICAM1 and IFNγ pathways in myeloid cells partially recapitulated treatment effects on CD8+ T cells observed via scRNA-seq. Mathematical modeling of tumor-immune dynamics implicated simultaneous modulation of multiple TME interactions as required for response to the combination treatment. Overall, this study identifies lymphoid and myeloid cell contributions to response to treatment with HDAC inhibitors and immune checkpoint blockade, providing a framework for discovering interactions driving responses in complex TMEs.

The high incidence and mortality rates of tumors have resulted in significant social and economic burdens, posing a major global threat to human life and societal development. In recent years, molecular targeted therapy for tumors has become a research hotspot. C-MET, the receptor for hepatocyte growth factor (HGF), plays a crucial role in the HGF/C-MET signaling pathway, which is involved in various processes such as tumor cell growth, invasion, migration, angiogenesis, epithelial-mesenchymal transition, tumor microenvironment remodeling and therapeutic resistance. Several C-MET-targeting strategies have been developed, including small-molecule tyrosine kinase inhibitors (TKIs), monoclonal antibodies (mAbs) against C-MET or HGF, antibody-drug conjugates (ADCs), nucleic acid aptamers, soluble receptors, natural compounds, and proteolysis targeting chimeras (PROTACs) targeting MET. These inhibitors have demonstrated encouraging anti-tumor effects in both preclinical and clinical studies, with several already available on the market. However, further research is still needed on the activation mechanisms of the HGF/C-MET signaling pathway and its interactions with other receptor tyrosine kinases, which will aid in identifying suitable patients for these treatments. This review provides a comprehensive overview of the structure, regulation, signaling pathways, and functions of C-MET, along with recent advances in C-MET inhibitors, offering valuable insights for cancer therapy.

Chimeric antigen receptor (CAR) T-cell therapy has revolutionized oncology, and its foundational logic-precise antigen recognition coupled with durable effector activity-extends naturally to chronic non-malignant diseases sustained by long-lived pathological cells. These include viral reservoirs, autoreactive B and plasma cells, activated fibroblasts, alloimmune clones, and senescent cells that remodel tissue niches and evade clearance by conventional therapies. This review highlights how CAR-based strategies can be adapted across diverse disease settings by redirecting engineered immune responses toward disease-sustaining cellular compartments. Co-stimulatory domains such as CD28, 4-1BB, and OX40 enhance persistence and effector function; programmed cell death protein 1 (PD-1)-CD28 switch receptors reverse inhibitory signaling; and cytokine-resistant CARs incorporating dominant-negative transforming growth factor-β (TGF-β) receptors maintain activity within suppressive microenvironments. We discuss these approaches across infections, including human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV); autoimmunity involving CD19- and B-cell maturation antigen (BCMA)-directed depletion strategies and CAR-engineered regulatory T cells (CAR-Tregs); fibrosis targeting fibroblast activation protein (FAP); hemophilia using B-cell antibody receptor (BAR)-CARs against factor VIII and factor IX inhibitors; transplantation employing human leukocyte antigen (HLA)-specific CAR-Tregs; and senescence-associated pathologies targeting urokinase plasminogen activator receptor (uPAR) and natural killer group 2D ligands (NKG2DLs). Early clinical experiences in systemic lupus erythematosus, systemic sclerosis, myositis, and multiple sclerosis, together with preclinical successes in chronic infections and fibrotic disease, demonstrate both feasibility and durable disease modification. By extending CAR-T therapy beyond oncology, these applications position programmable cellular immunotherapy as a broadly adaptable platform for eliminating persistent pathological cells, remodeling diseased tissue environments, and restoring long-term immune homeostasis.

γδ T cells represent a promising avenue for cancer immunotherapy. The Vγ9Vδ2 T-cell receptor (TCR), which is expressed by the predominant subset of γδ T cells, responds to phosphoantigen (pAg)-engaged butyrophilins (BTNs) on various cancer cells. However, the molecular mechanism underlying the pAg-mediated activation of Vγ9Vδ2 TCRs remains a subject of debate. Here, we employed an integrative approach to elucidate the mechanism of pAg reactivity in Vγ9Vδ2 T cells. Our results demonstrate that BTNs form higher-order oligomers in the absence of pAg. Upon pAg binding, these higher-order oligomers dissociate into separate tetramers, enabling Vγ9Vδ2 TCR engagement. This pAg-induced dissociation of higher-order BTN oligomers is critical for pAg-mediated activation of γδ T cells. Our findings reveal a mechanism of BTN higher-order oligomer dissociation-driven pAg sensing, providing valuable insight for future immunotherapeutic strategies.

Hypoxia, a hallmark of solid tumors, poses a significant challenge in cancer therapy due to its association with poor prognosis and resistance to conventional treatments. Oncolytic viruses represent a promising treatment strategy, as they selectively replicate within cancer cells and lyse them, potentially including those in hypoxic tumor regions. Here, we examined how hypoxic conditions influence the activity of enadenotucirev (EnAd), a clinically relevant group B oncolytic adenovirus previously detected in hypoxic areas of xenograft tumors. We demonstrated that hypoxia enhances virus production by boosting transcription and translation of immediate-early, early, and late adenoviral genes. The immediate-early gene E1A was upregulated within 2 h (17-fold) after virus entry under hypoxia, driven by a conserved hypoxia-response element (HRE) in its promoter. Mechanistic studies revealed that the hypoxia-inducible factor (HIF)-2α and HIF-1β heterodimers bind to this HRE, transactivating E1A. By inducing E1A expression, hypoxia also elevated viral genome synthesis, structural protein production, and therapeutic transgene expression, underscoring the potential of EnAd to target the hypoxic tumor microenvironment. This is the first report of a functional HRE in a human adenovirus, conserved across 59 adenovirus genotypes, and identifies hypoxia as a driver of enhanced oncolytic activity with implications for adenovirus-based therapies in solid tumors.

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.

Bile acids have emerged as compartmentalized immunometabolic signals that link host metabolism, microbial ecology, and tumor immunity. Altered bile acid profiles are common across malignancies and are associated with tumor progression, immune tone, and responsiveness to immunotherapy. Bile acids shape the gut microbiota, and in turn, microbial enzymes diversify the bile acid pool, generating distinct bile acid species that can remodel the tumor immune landscape. Depending on species identity, concentration, and context, bile acids can support immune surveillance or enforce immune escape by reshaping antigen priming, lymphocyte fitness, myeloid suppression, and immune cell trafficking. Here, we synthesize emerging concepts defining a microbiome-bile acid-immune axis in cancer and highlight therapeutic opportunities to harness bile acid signaling as next-generation strategies in oncology.

Cancer vaccines have underperformed clinically, largely because antigen-specific CTL priming and expansion are constrained by insufficient costimulatory activation and increased DC-mediated coinhibition. Here, we present a personalized cancer vaccine platform, STAR, which engineers stimulator of interferon gene (STING)-activating autologous tumor-derived microvesicles (TMVs) to concurrently amplify costimulatory signaling and relieve DC coinhibition during antigen cross-presentation. TMVs provide broad tumor antigen cargo and intrinsic STING activity; loading manganese phosphate nanoparticles (MnPs) potentiates cGAS-STING signaling, drives DC maturation, and enhances antigen cross-presentation. Importantly, concurrent delivery of PD-L1-targeting siRNA (siPD-L1) effectively inhibits DC-associated PD-L1, freeing CD80 to engage CD28 and limiting PD-1/PD-L1 signaling and thereby significantly enhancing antigen-specific CD8+ T cells. In mouse models, STAR vaccination elicited durable prophylactic immunity and achieved substantial therapeutic efficacy against melanoma and colorectal tumors without additional immunotherapies. Notably, patient-specific STAR vaccines derived from surgically resected tumors effectively prevented postoperative recurrence and metastasis in aggressive melanoma and triple-negative breast cancer models, thereby highlighting their translational potential. Overall, the STAR vaccine platform offers a versatile and clinically applicable approach to enhancing personalized cancer immunotherapy across diverse cancer types.

Toll-like receptors (TLRs) are considered general sensors of bacterial encounters. Here, we examined whether other pattern recognition receptors are commonly activated during bacterial infection. TLR-independent interferon (IFN) responses were induced in macrophages in response to diverse bacterial encounters. Of the cytoplasmic receptor families examined, the cyclic dinucleotide (CDN) sensor STING was required for IFN responses to evolutionarily diverse bacteria. Various bacterial CDNs were present in murine tissues; these activated stimulator of interferon genes (STING) after bacteriolysis in phagolysosomes in a manner requiring two CDN transporters. Importantly, bacterial CDNs were increased in colonic biopsies from patients with inflammatory bowel disease. Systemic delivery of dead, CDN-laden bacteria promoted anti-tumor immunity in mice. Detection of diverse CDNs, including pyrimidine-based CDNs, was an evolutionarily conserved feature of STING, with distinct binding modes for purine- and pyrimidine-based CDNs. Thus, a phagocytosis-CDN-STING connection places cytoplasmic sensing as a common outcome of host-bacteria interactions that set the immune tone of a tissue, with implications for host defense.

Gastrointestinal stromal tumor (GIST) is the most common gastrointestinal mesenchymal tumor, driven by tyrosine-protein kinase (KIT) and platelet-derived growth factor receptor A (PDGFRA) mutations. Specific variants, such as KIT exon 11 deletions, carry prognostic and therapeutic implications, whereas wild-type variants derive limited benefit from tyrosine kinase inhibitors. Given the limited reproducibility of established clinicopathologic risk models, deep learning (DL) applied to whole-slide images (WSI) emerged as a promising tool for molecular classification and prognostic assessment. We analyzed 8398 GIST cases from 21 centers in seven countries, including 7,238 with molecular data and 2,638 with clinical follow-up. DL models were trained on WSIs to predict mutations, treatment sensitivity, and recurrence-free survival (RFS). DL predicted mutational status in GIST from WSIs, with area under the curve of 0.87 for KIT and 0.96 for PDGFRA, and high performance was observed for subtypes, including KIT exon 11 del-inss 557 to 558 (0.67) and PDGFRA exon 18 D842V (0.93). For therapeutic categories, performance reached 0.84 for avapritinib sensitivity and 0.81 for imatinib sensitivity. DL models predicted RFS, with hazard ratios of 8.44 in the overall cohort and 4.74 in patients receiving adjuvant therapy. Prognostic performance was comparable with pathology-based scores, with highest discrimination in the overall cohort and in patients without adjuvant therapy. DL applied to WSIs enables prediction of molecular alterations, treatment sensitivity, and RFS in GIST, performing comparably with established risk scores across international cohorts, providing a baseline for future multimodal predictors. Deep learning on histology predicts KIT and PDGFRA mutations and stratifies recurrence-free survival in a large international cohort of gastrointestinal stromal tumors from multiple centers.

In fecal immunochemical test (FIT)-based colorectal cancer (CRC) screening, colonoscopy after a positive FIT often identifies no CRC or advanced neoplasia, leaving management poorly standardized. We mapped evidence on definitions, extracolonic evaluation, colonoscopy quality, residual CRC risk, and screening re-entry after a positive FIT and nonexplanatory colonoscopy. This registered scoping review followed PRISMA-ScR guidance. PubMed/MEDLINE, Scopus, and Web of Science were searched until April 2026, along with reference-list screening and targeted searching of official screening program/policy documents. Sources were charted by one reviewer; included sources, uncertain eligibility decisions, and key findings underwent second-reviewer verification. We included 10 studies on definitions, 11 empirical studies plus three contextual evidence syntheses on evaluation beyond the colon, 11 core empirical studies on colonoscopy quality/residual CRC risk, and nine empirical studies on management and screening re-entry, supplemented by policy mapping. Definitions varied substantially, supporting "nonexplanatory colonoscopy" as a pragmatic umbrella term. Routine upper gastrointestinal or small-bowel evaluation was not supported in otherwise asymptomatic individuals. Reassurance was quality dependent; residual CRC risk was low in the short term, but not zero. Screening re-entry pathways varied and were not evidence based. Management should first be to establish whether the index colonoscopy is reliable enough to justify reassurance. After a trusted negative examination, the evidence does not support routine upper gastrointestinal or small-bowel evaluation, or routine early repeat colonoscopy in otherwise asymptomatic individuals. Residual CRC risk remains low but not zero, and optimal screening re-entry remains uncertain.

As cancer care becomes increasingly complex, patients may experience prolonged delays between diagnosis and definitive surgical management. While consolidation and expansion of health systems, along with centralization of care, may further amplify these delays, a comprehensive assessment of contemporary waiting times within cancer surgery care pathways in the US is lacking. To evaluate national trends and predictors of delays from diagnosis to initiation of first-course therapy among patients ultimately undergoing cancer surgery. This retrospective cohort study included patients with nonmetastatic, clinical stage I through III breast, colon, lung, pancreatic, gastric, or esophageal cancer in the National Cancer Database from January 2012 to December 2023 who underwent definitive surgical resection. Waiting time from diagnosis to first-course therapy, defined as up-front surgery or neoadjuvant therapy preceding surgery, was assessed. Data were analyzed from August 2025 to June 2026. The primary outcome was the waiting time from diagnosis to first-course therapy. Secondary outcomes included factors associated with prolonged (≥30 days) or extreme (≥60 days) delays. A total of 2 731 059 patients were identified. Mean (SD) patient age was 63.5 (12.8) years, and 2 325 110 patients (85%) were female. From 2012 to 2023, median (IQR) time from diagnosis to first-course therapy increased across all malignancies (breast: 34 days [22-50] in 2012-2015 to 45 days [32-64] in 2022-2023; colon: 20 days [7-34] to 31 days [15-49]; lung: 41 days [27-60] to 53 days [35-77]; pancreas: 23 days [14-35] to 32 days [22-44]; gastric: 35 days [21-51] to 49 days [33-70]; and esophageal: 38 days [27-54] to 48 days [35-66]; P value for trend <.001 for all) and among both patients undergoing up-front surgery and those receiving neoadjuvant therapy prior to surgery. Delays were more pronounced at academic compared with community hospitals and among patients referred for care. Predictors of longer waiting time included Medicaid insurance (5 of 6 cancers), lowest-quartile income (6 of 6 cancers), Black race (5 of 6 cancers), increased travel distance (4 of 6 cancers), care in the West region (6 of 6 cancers), and treatment at academic institutions (6 of 6 cancers). Receipt of robotic operations was linked with longer waiting times for nonbreast malignancies (5 of 5 cancers). Among patients undergoing cancer surgery, waiting times to initiation of first-course therapy have steadily increased since 2012, particularly at high-volume academic centers and among patients referred for definitive care. With continued consolidation and expansion of health systems, system-level strategies are urgently needed to monitor and mitigate delays in the delivery of surgical care for cancer.

Identifying patients at high risk for recurrence after endoscopic resection of T1 colorectal cancer (CRC) remains challenging. This study aimed to identify recurrence risk subtypes and develop an interpretable risk stratification framework. This retrospective study analyzed 1123 patients with T1 CRC treated with endoscopic resection alone across 26 Japanese institutions (July 2009-December 2016). Patients were divided into development (68%) and evaluation (32%) cohorts based on institutional stratification. K-means clustering was applied to clinicopathologic variables to identify recurrence risk subtypes. A decision tree classifier was subsequently developed to generate transparent risk stratification rules. Three distinct subtypes were identified in the development cohort. Subtype 1 exhibited a numerically higher recurrence rate (5.4%) than Subtype 2 (0.9%) and Subtype 3 (1.4%). Although subtypes 2 and 3 showed comparable recurrence rates, they were clearly differentiated by morphology (flat vs. polypoid). In the evaluation cohort, Subtype 1 continued to show a numerically higher recurrence (4.8%) compared with subtypes 2 (1.6%) and 3 (1.1%). The decision tree model stratified recurrence risk hierarchically: submucosal invasion <1000 μm indicated low risk, whereas invasion ≥1000 μm required morphologic assessment, with polypoid lesions classified as high risk and flat lesions further stratified using a 2000-μm threshold. Three clinically distinct recurrence risk subtypes were identified in T1 CRC following endoscopic resection, suggesting that morphologic subclassification of T1b lesions may refine stratification beyond conventional depth-based criteria. The decision framework offers a preliminary exploratory basis for recurrence risk assessment in this population.

Endoscopic and radiological staging of early rectal cancer (ERC) remains suboptimal, contributing to inaccurate treatment allocation and the risk of under- and overtreatment. This study aimed to evaluate the feasibility and safety of the Pocket-detection method with Real-time Intraprocedural Muscle-retracting sign Evaluation (PRIME) resection algorithm for ERC. We retrospectively analyzed a prospective registry of rectal lesions managed according to the PRIME algorithm between August 2023 and October 2025. The muscle-retracting sign (MRS) status guided adaptive dissection plane selection: endoscopic submucosal dissection (ESD) if MRS negative; endoscopic intermuscular dissection (EID) or full-thickness resection (knife-assisted [kFTR] or device-assisted [FTRD]) for MRS-positive lesions. Primary outcomes were technical success and the rate of R0 endoscopic resection. The secondary outcomes were the technical success of deep-margin optical diagnosis and adverse events. 47 lesions were included. MRS prevalence was 34.0% (16/47). Technical success of endoscopic resection was 97.9% (46/47; 95%CI 88.7%-99.9%). ESD was performed in 31 lesions, EID in nine, kFTR in five, and FTRD in one. R0 resection was achieved in 89.1% (41/46; 95%CI 76.4%-96.4%) and R0 vertical margin in 95.7% (44/46; 95%CI 85.2%-99.5%). Technical success of deep-margin optical diagnosis was 100% (47/47; 95%CI 92.5%-100%). Nonsevere (AGREE classification grade I-IIIa) adverse events occurred in 10.6% of patients (5/47; 95%CI 3.5%-23.1%). The MRS-guided adaptive endoscopic resection algorithm (PRIME) was feasible and safe for ERC achieving high R0 rates. These findings support its validation in larger studies with longer follow-up.

Colonoscopy is important for colorectal cancer screening, but real-world data on complications and their associated costs are limited. We assessed direct and indirect complications within 7 days after screening colonoscopy and estimated excess costs by complication type. We examined 365 685 individuals, aged 60-69, invited to screening in Region Stockholm, Sweden, from 2008 to 2021, to assess excess healthcare utilization after screening colonoscopy following a positive fecal occult blood test (FOBT). Healthcare events within 7 days postcolonoscopy were compared with 1 : 5 propensity score-matched controls, calculating between-group differences in healthcare utilization and costs. 11 177 screening colonoscopies were included. Bleeding occurred most frequently (47/10 000, relative risk [RR] 52.6, 95%CI 21.0-131.6), among the 86/10 000 individuals who had at least one healthcare event within 7 days postcolonoscopy. Compared with the propensity score-matched controls, the colonoscopy group had at least a threefold higher proportion of unplanned outpatient visits (RR 3.0, 95%CI 2.2-4.2) and hospital admissions (RR 4.2, 95%CI 3.0-5.8), driven by direct complications. No increase in indirect complications (e.g. cardiovascular or cerebrovascular) was observed. The excess cost was €32.88 per colonoscopy. Four patients died within 30 days of colonoscopy, with no increase in all-cause mortality compared with propensity score-matched controls (3.6 vs. 10.0 per 10 000; RR 0.4, 95%CI 0.1-1.0). Healthcare utilization and costs increased after screening colonoscopies compared with propensity score-matched controls, driven by direct complications. However, because few screened individuals undergo colonoscopy, and complications occur in fewer than 1% of procedures, excess costs are marginal relative to program expenditure.

Transarterial chemoembolization (TACE) remains a cornerstone therapy for intermediate-to-advanced hepatocellular carcinoma (HCC); however, the optimal embolic platform remains uncertain. This multicenter, retrospective, real-world study (Clinical trial registration number: ChiCTR2500113198) conducted across China compared a novel temperature-sensitive liquid embolic agent, TempSLE-TACE (T-TACE), with conventional drug-eluting bead TACE (D-TACE) in 328 patients with Barcelona Clinic Liver Cancer (BCLC) stage B/C HCC. Following inverse probability of treatment weighting (IPTW), T-TACE achieved significantly superior objective response rates (ORRs) compared with D-TACE according to both RECIST 1.1 criteria (54.11% vs. 27.78%, P < 0.001) and mRECIST criteria (73.56% vs. 54.93%, P = 0.002). T-TACE was additionally associated with significantly prolonged progression-free survival (median PFS: 12.0 vs. 9.0 months; HR = 0.67, P = 0.001) and overall survival (median OS, 24.0 vs. 15.0 months; HR = 0.49, P < 0.001). Moreover, T-TACE demonstrated a favorable safety profile, with lower incidences of hepatic and gastrointestinal toxicities, including any-grade alanine aminotransferase elevation and hyperbilirubinemia. Subgroup analyses further demonstrated consistent OS, PFS, and ORR benefits across major clinical subgroups, with effect sizes remaining significantly favorable in high-risk populations, including advanced portal vein tumor thrombosis type Vp4, baseline AFP > 1000 ng/mL, and PIVKA-II > 2000 mAU/mL. Exploratory histo-molecular and spatial transcriptomic analyses suggested that T-TACE may promote immune microenvironment remodeling through enhanced Th17-cell infiltration and CD8⁺ T-cell activation, whereas incomplete embolization after D-TACE was more frequently associated with residual intermediate-state tumor cells and an immunosuppressive microenvironment. Collectively, these findings provide preliminary evidence supporting T-TACE as a promising real-world therapeutic strategy for intermediate-to-advanced HCC.

Desmoplastic stroma defines pancreatic ductal adenocarcinoma (PDAC), a highly lethal malignancy. This stroma drives tumor progression, immune evasion, drug resistance, and poor drug delivery. Although targeting tumor stroma has shown preclinical promise, most clinical trials have failed due to limited translational success. A major hurdle is the stroma's extreme heterogeneity and dynamic nature, meaning uniform treatments fail. Emerging evidence supports the existence of distinct 'stromal states' with differential biological functions and therapeutic vulnerabilities. Characterizing these states provides a framework to stratify patients and guide precise, stroma-directed therapies. This review summarizes the clinical landscape of PDAC stroma-targeting strategies, discusses distinct stromal states, and outlines emerging opportunities to exploit the stroma as a therapeutic axis in PDAC to improve clinical trial outcomes.

As renal tumor classification continues to evolve and new hereditary entities emerge, accurate recognition of hereditary renal neoplasms is increasingly important. Advances in precision genomics, genetic testing, and international registries will improve early detection, risk stratification, and precision management.

Background Artificial intelligence (AI)-assisted colonoscopy reduces costs in colorectal cancer screening, but its incremental value relative to non-AI resect-and-discard (RD) strategies for diminutive polyps (≤ 5 mm) remain unclear. We aimed to compare the clinical, economic, and environmental outcomes of AI-assisted and non-AI RD strategies with standard of care (SoC). Methods In our Markov model simulating 6 million individuals aged ≥45 years undergoing screening colonoscopy, 4 strategies were evaluated: SoC, non-AI RD, CADe-assisted RD (CADe-RD), and CADe+CADx-assisted RD (CADe+CADx-RD). The primary outcome was total quality-adjusted life years (QALYs), with secondary outcomes including cost and carbon dioxide (CO₂) emissions. Results Non-AI RD achieved the lowest total cost ($36.6B) and carbon emissions (0.90 million kg CO₂), while yielding the highest QALYs (51.37 million), although differences in QALYs across strategies were small (<0.01%). CADe-RD and CADe+CADx-RD remained cost-saving relative to SoC ($36.9B and $37.4B vs $37.8B, respectively) but were associated with higher costs than non-AI RD. CADe modestly increased adenoma detection without improving QALYs, while the addition of CADx increased costs and emissions and resulted in slightly lower QALYs. Sensitivity analysis showed that CADe+CADx-RD matched or exceeded CADe-RD QALYs only within a limited region of high diagnostic performance. At currently reported CADx performance levels, CADe+CADx-RD yielded lower QALYs and higher costs than CADe-RD. CADe+CADx-RD achieved non-inferiority in QALYs relative to CADe-RD only within a limited region of high diagnostic performance. Conclusions In our non-deterministic model, non-AI RD provided the optimal balance of clinical benefit, cost savings, and lowest environmental impact.

Wilson's disease (WD) is an inherited and treatable disorder of copper metabolism with hepatic or neurological manifestations. It continues to pose challenges in clinical management, and long-term outcome data remain limited. This study investigated hepatic involvement and clinical outcomes in cohorts from Taiwan. Longitudinal data from 146 patients with WD, retrieved from the medical database of a tertiary referral center, were analyzed (mean age at diagnosis 20.7 ± 12.4 years; mean follow-up 16.8 ± 10.8 years; maximum 51.8 years). All patients received standard treatment and care. Nine patients (6.2%) underwent liver transplantation, 6 patients (4.1%) died, and 3 patients (2.1%) developed hepatocellular carcinoma (HCC). Thrombocytopenia <100 K/μL at diagnosis was associated with reduced survival (P=0.0001). Univariate Cox regression identified thrombocytopenia at diagnosis as a strong predictor of liver transplantation or death (P = 0.0001; hazard ratio (HR): 8.41; 95% CI: 2.97-23.78). Consistently, thrombocytopenia <100 K/μL was associated with reduced survival (P < 0.0001) in an external cohort of 155 patients. In the original cohort, the estimated annual HCC risk was 0.12% (95% CI: 0.03-0.36), increasing to 0.86% (95% CI: 0.18-2.5) in those with thrombocytopenia at diagnosis (P=0.029). An exploratory Firth penalized logistic regression showed that older age at diagnosis was associated with higher odds of developing HCC (odds ratio: 1.096; 95% CI: 1.009-1.191; P = 0.031). WD patients have a high rate of liver fibrosis, regardless of whether they present with neurological and hepatic symptoms, and are at risk of developing HCC, warranting lifelong hepatic surveillance.

Hepatocellular carcinoma (HCC) exhibits substantial molecular and morphological heterogeneity. The recently updated 6th edition of the World Health Organization Classification of Digestive Tumours recognizes nine specific HCC subtypes, each characterized by distinct morphomolecular and clinical profiles. Although these subtypes account for approximately 35% of all HCC cases, the remaining 65% are classified as conventional HCCs. These subtypes are categorized based on their prognostic implications: the unfavorable prognosis group comprises macrotrabecular-massive, sarcomatoid, and neutrophil-rich HCCs; the comparable prognosis group includes steatohepatitic, fibrolamellar, and chromophobe HCCs; and the favorable prognosis group consists of lymphocyte-rich and clear cell HCCs. By contrast, scirrhous HCC is associated with various clinical outcomes. Furthermore, this classification includes two newly recognized patterns: vessels encapsulating tumor clusters and CTNNB1-mutated HCC. The imaging features of HCC are significantly correlated with its molecular-pathological characteristics. In particular, aggressive subtypes frequently present with specific imaging hallmarks such as rim arterial phase hyperenhancement, diffuse arterial phase hypo-enhancement, and intratumoral necrosis on T2-weighted imaging, which serve as critical noninvasive predictors of clinical outcomes. However, a definitive diagnosis of specific subtypes based solely on imaging remains challenging. This review provides a comprehensive overview of the molecular, pathological, and radiological characteristics of HCC subtypes and patterns, emphasizing that a detailed understanding of these features is essential for informed therapeutic decision-making and accurate prognostic assessment.

Gut microbiome dysbiosis causes various intestinal diseases. However, an undefined composition and potential biosafety risks limit the applicability of traditional fecal microbiota transplantation (FMT). Synthetic microbial communities (SynComs), which are compositionally defined and rationally designed emerging live biotherapeutics, offer a novel alternative to FMT. This review establishes strict boundaries between SynComs and traditional donor-derived preparations, comparatively evaluating "top-down" and "bottom-up" construction strategies. We explored the mechanisms underlying the SynComs-mediated synergistic restoration of intestinal homeostasis via direct targeted antagonism and modulation of the host immune network. Moreover, we systematically evaluated the current research landscape of SynComs in Clostridioides difficile infection, inflammatory bowel disease, and colorectal cancer. This review examines fundamental challenges, including host colonization resistance, chemistry, manufacturing, and control barriers, biosafety risks, and microbiokinetic regulatory frameworks, thereby addressing the translational gap. Our analysis of current literature provides a theoretical basis for the clinical translation of SynComs as emerging live biotherapeutics.

Dense stromal barriers and lysosomal sequestration severely restrict the extracellular transport and intracellular trafficking of nanomedicines in pancreatic cancer, while the robust glutathione (GSH)-dependent antioxidant defense further compromises therapeutic efficacy. Here, we developed a proton sponge-enabled dual-driven nanomotor, HA/PEI-DOX@MMSN@Pt (HPDMP), for MRI-guided pancreatic cancer therapy. The asymmetric Pt domain generated near-infrared (NIR)-induced self-thermophoretic motion as the principal externally controllable driving mechanism, whereas endogenous H2O2 provided auxiliary catalytic propulsion, together facilitating nanomotor transport through the dense tumor stroma and enhancing intratumoral penetration. Following cellular uptake, PEI-mediated proton sponge activity facilitated lysosomal escape and increased cytosolic DOX availability. Meanwhile, tumor-microenvironment-responsive degradation of the Mn-doped framework consumed GSH and released Mn2+, thereby enhancing reactive oxygen species (ROS) generation, ferroptosis-associated oxidative damage, and T1-weighted MRI contrast. HA modification further promoted CD44-mediated tumor targeting. Under NIR irradiation, HPDMP achieved pronounced antitumor efficacy with favorable biosafety. This work provides a barrier-oriented nanomotor strategy for pancreatic cancer therapy.

Ferroptosis provides promising method for tumor therapy but is limited by the low efficiency and safety concerns of conventional iron delivery systems. Herein, we report efficient ferritin-mimetic nanominerals engineered via a small molecule-mediated iron phosphate mineralization strategy, which emulates the natural iron storage and release processes to selectively trigger tumor ferroptosis. Using two ferroptosis inducers, palbociclib and Fin56, as model small molecules, the resulting nanominerals exhibit uniform size distribution, robust stability, and 72% iron incorporation efficiency, enabling precisely controlled ferrous ion release and dual-drug co-delivery within tumor tissues to achieve a triple amplification of ferroptosis. The amplified ferroptosis response further elicits a potent antitumor immune effect through immunogenic cell death, ultimately resulting in remarkable therapeutic efficacy with minimal side effects both in vitro and in vivo. This small molecule-mediated iron phosphate mineralization strategy offers a superior alternative to traditional coordination self-assembly for nanomedicine construction through enhanced programmability, improved stability and higher iron loading, thereby advancing ferroptosis-related combination therapy and synergistic therapy.

The coexistence of atherosclerosis and cancer is increasingly common, yet the influence of tumors on atherogenesis remains poorly understood. In this study, we investigated the impact of colorectal cancer (CRC) on atherosclerosis and elucidated the underlying mechanisms. We analyzed clinical data from patients with concurrent atherosclerosis and CRC and established three murine models of atherosclerosis comorbidity: ApoE-/- and Ldlr-/- mice bearing subcutaneous MC38 tumors, and APCmin/+ApoE-/- mice with spontaneous intestinal adenomas. We found that both patients and mice with concurrent tumors exhibited reduced plaque burden and enhanced plaque stability. Integrative multi-omics analysis comprising single-cell RNA sequencing, spatial transcriptomics, metabolomics, and bulk RNA sequencing revealed that tumors systemically deplete arginine, leading to adaptive metabolic reprogramming of intraplaque T cells toward an oxidative phosphorylation (OXPHOS)-dominant state. This metabolic shift was associated with reduced cytotoxic T-cell infiltration, suppressed pro-inflammatory effector programs, and weakened T cell-macrophage interactions, collectively establishing a low-inflammatory plaque microenvironment. Importantly, dietary arginine supplementation restored T-cell activation and reversed the CRC-mediated atheroprotective effects. These findings reveal a critical role of T-cell immunometabolic reprogramming in cancer-atherosclerosis comorbidity, demonstrate the pronounced context-dependent effects of arginine in advanced atherosclerosis, and highlight plaque-targeted immunometabolic interventions as a promising strategy to modulate plaque progression and stability.

Liver sinusoidal endothelial cells (LSECs) form a specialized discontinuous microvascular interface between sinusoidal blood and the hepatic parenchyma. Functioning as a central regulator of hepatic homeostasis, LSECs integrate metabolic, inflammatory, and hemodynamic signals to maintain sinusoidal permeability, immune tolerance, vascular tone, and lipid exchange. During acute injury or sustained hepatic stress, LSECs may lose their differentiated sinusoidal phenotype and acquire features of capillarization, including fenestrae loss, subendothelial matrix accumulation, impaired scavenging activity, and attenuation of KLF2-eNOS-NO signaling. Capillarized LSECs are not merely a byproduct of hepatic damage; instead, they may function as early endothelial sentinels and important contributors to the amplification of parenchymal injury, inflammation, fibrogenesis, and tumor immune evasion. This review summarizes the physiological functions of LSECs and examines mechanisms through which LSEC dysfunction contributes to acute liver injury, metabolic dysfunction-associated steatotic liver disease, viral hepatitis, cirrhosis, and hepatocellular carcinoma. By comparing conserved mechanisms with disease-specific spatial and molecular triggers, this review highlights how LSEC phenotypic switching disrupts the angiocrine-immune-fibrotic axis and contributes to disease progression. Recent advances in single-cell and spatial omics, LSEC subpopulation heterogeneity, and capillarization-aware therapeutic delivery are further discussed. Finally, therapeutic reprogramming of dysfunctional LSECs toward a quiescent, fenestrated, and tolerogenic phenotype is proposed as a strategy to restore sinusoidal homeostasis and improve treatment of liver diseases.

Pancreatic tumors are characterized by a prominent desmoplastic stroma that can account for up to 90% of the tumor. Given the marked upregulation of CD44, a family of transmembrane glycoproteins, in pancreatic cancer-associated fibroblasts (CAFs), we investigated its role in regulating myofibroblastic and inflammatory CAF phenotypes. Conditional deletion of Cd44 in fibroblastic cells in Cd44fl/fl;PdgfrβCreERT2 mice significantly reduced tumor growth and was associated with a significant reduction in intratumoral regulatory T cells (Treg). Consistently, in human CAFs CRISPR/Cas9-edited to delete CD44, the fibroblasts' morphology changed drastically: CAFs lost their elongated phenotype and adopted a round shape, reflecting their inactivation. This was accompanied by a significant downregulation of activation markers, unresponsiveness to exogenous stimuli, and reduced contractile activity. Furthermore, CD44 ablation decreased extracellular matrix production and altered the immunomodulatory cytokine secretion, highlighting its role in both fibrosis and immune regulation. Finally, CD44-deficient CAFs exhibited a reduced capacity to suppress anti-tumor immune responses by failing to induce immunosuppressive programs in dendritic cells (DCs) and by increasing cytotoxic T cell (CTL)-mediated tumor cell killing. Collectively, we identify CD44 as a regulator of CAF activation and immunosuppressive function, linking stromal remodeling to immune evasion in pancreatic cancer and supporting CD44 as a potential therapeutic strategy to reprogram the tumor stroma and enhance anti-tumor immunity.

The nanocomposite hydrogels fail to achieve clinical transformation due to their unsatisfactory nanomedicine escapement from hydrogel and suboptimal tumor selectivity. Herein, this work highlights a successful development of a polysaccharide-based dynamic organic nanocomposite hydrogel, in which crosslinked nano-prodrugs (NPs) are dispersed within liquid hydrophobic orthoester (OE) and subsequently encapsulated in polysaccharide-based hydrogels crosslinked via dynamic imine bonds. These dynamic imine linkages endow the hydrogel with injectability, self-adaptability, and tumor extracellular pH-responsive biodegradability. OE enables efficient NPs release from the hydrogel, enhances their deep penetration into tumor tissues, and undergoes further degradation to restore NPs to their original particle size. The NPs enhance selective drug accumulation through tumoral intracellular pH-triggered aggregation and enable targeted drug release in response to specific intracellular pH/glutathione conditions, ultimately promoting apoptosis and cytotoxicity through a synergistic interaction between cisplatin and demethylcantharidin. These superior attributes allow for complete tumor suppression without adverse effects. Consequently, this polysaccharide-based dynamic organic nanocomposite hydrogel demonstrates substantial potential for localized tumor-targeted therapy, greatly improving its clinical applicability.

Activation of antigen-presenting cells (APCs) via the stimulator of interferon genes (STING) signaling pathway is a promising strategy for cancer therapy. However, the rational engineering of a targeted synergistic immunomodulator and its morphological impact for STING activation remains elusive. Herein, we developed an APC-targeting lentinan-STING agonist immunomodulator with flexible and assembled morphological forms for cancer therapy. Lentinan (LN) exhibited intrinsic APC targeting and immune activation characteristics, and the conjugation between lentinan and a STING agonist (DMXAA) not only decreased the toxicity of lentinan toward APC cells, but also realized synergistic STING activation via lentinan and DMXAA. The flexible and assembled morphological forms were readily achieved by tuning the DMXAA loadings of the immunomodulators, denoted as LN-DMXAA(L) and LN-DMXAA(H). The flexible immunomodulator LN-DMXAA(L) with lower DMXAA loading potently induced macrophage reprogramming and dendritic cell maturation as compared with the assembled immunomodulator LN-DMXAA(H) and free drug DMXAA. In a 4 T1 tumor-bearing mice model, LN-DMXAA(L) significantly boosted tumor growth inhibition rate to 78.3% as compared with 46.2% for LN-DMXAA(H) and 27.8% for DMXAA by activating APCs and CD8+ T cells via the STING signaling pathway. Thus, this work highlights the conception of a flexible immune-activating polysaccharide-STING agonist immunomodulator for cancer therapy.

Accurate detection and nondestructive collection of circulating tumor cells (CTCs) remain an enormous challenge due to their extremely low concentration. Herein, an electrochemical cytosensor was developed based on living cell-based bionic micro-nano robots (LCMNRs) and necklace-like AuPt porous hollow nanocomposites (nlAuPt phNCs). The LCMNRs can efficiently cleave D-RNA by the synergistic effect of Mg2+-dependent DNAzyme activity and autonomous motility of living cells, and can simultaneously recognize the triple membrane markers of CTCs based on DNA proximity effect. The nlAuPt phNCs, as signal labels, exhibited high specific surface area and multiple excellent enzyme-mimetic activities, particularly prominent electrocatalysis toward H2O2. The developed cytosensor demonstrated a superior analytical performance, with a wide linear range (3 to 3 × 105 cells mL-1) and an exceptionally low detection limit of 1 cell mL-1. Additionally, it exhibited the capability to distinguish among multiple types of cancer cells and enable the isolation of CTCs while largely maintaining cell viability. The analysis of clinical blood samples confirmed the cytosensor had outstanding anti-interference ability, along with a specificity of 100% and a sensitivity of 86.7%. Collectively, this developed cytosensor holds great promise as a powerful tool for the precise detection of CTCs and the early monitoring of tumor metastasis.

Surface stress, as an indicator of cellular state, responds to morphological changes, signaling, and disease processes, and is therefore highly significant for early diagnosis and precise disease monitoring. However, cell surface stress can become very subtle when the external stimulus is weak. Detecting these weak stress signals with high sensitivity to enable reliable and precise biosensing is thus an essential challenge. Here, we report a surface-stress biosensor based on multiphysics coupling that demonstrates label-free electrical detection of trace cell populations (as few as 20 cells·mL-1) and is applicable across diverse adherent cell types. The differing surface stresses generated by various cells on the biosensor surface were amplified by incorporating ferromagnetic materials and applying an external magnetic field. The chip employs interdigitated electrodes and a non-floating thin-film design to ensure that weak electrical signals from the densely packed palladium-nanoparticle film can be detected while minimizing the influence of the film's weight on biosensor performance. The biosensor provides a wide detection range from 200 to 2 × 104 cells·mL-1, with high sensitivity and a detection limit as low as 20 cells·mL-1 for normal human hepatocyte (L02) and human hepatocellular carcinoma cells (HepG2) after treatment with sorafenib. Moreover, we further elucidate the principle of magnetic sensitization through theoretical calculations. This work represents a significant advancement in high-sensitivity biosensing for the personalized treatment and management of hepatocellular carcinoma and provides insights into novel in-vitro diagnostic strategies.

Designing nanozymes with enhanced catalytic performance remains a fundamental challenge in functional nanomaterials; concurrently, achieving synergistic multifunctionality constitutes an unmet need in nanozyme engineering. This study introduces an ultrasmall heterostructure nanozyme via interface electronic modulation. The heterostructure nanozyme is constructed through the facile growth of ultrasmall Cu4SnS4 crystals on SnS nanosheets, named as SCS. A spherical aberration-corrected transmission electron microscope is used to unveil the atomic arrangement characteristics of SCS heterostructures for the first time. The spontaneous transfer of electrons from SnS to Cu4SnS4 is revealed by density functional theory (DFT) calculations, demonstrating that the interface creates an ideal environment for accelerated mass and electron transfer. The enhanced peroxidase-like activity (129-fold) and photothermal efficiency are mediated through heterostructure-induced electron enrichment. The SCS nanozyme enables triple-modal lateral flow immunoassay signals (colorimetric, enhanced colorimetric, and photothermal) for carcinoembryonic antigen detection, achieving a 15-min rapid assay with an ultralow limit of 0.209 ng mL-1. It also facilitates visual tumor-normal tissue differentiation based on its superior catalytic performance. This work provides a paradigm for activity-oriented nanozyme design via interfacial electronic modulation.

Drug delivery strategies with excellent spatiotemporal controllability and biocompatibility hold exciting prospects in personalized medicine and biopharmaceuticals. Among them, all-optical delivery strategies have attracted significant attention due to their low invasiveness and high positioning accuracy. However, current all-optical schemes predominantly rely on conservative optical gradient force trapping or transient scattering forces propulsion, which cannot sustain stable driving and limit delivery speed. Here, we present an all-optical delivery strategy based on Airy beam, enabling high-speed and subcellular-precision delivery through the synergistic regulation of gradient-scattering forces. The dominant scattering force of Airy beams propels drug carriers at speeds exceeding 400 µm/s, while the gradient force ensures a subcellular delivery precision of ~1 µm during high-speed propulsion. Quantitative measurements using a custom axial imaging module reveal at least a 7-fold speed enhancement over conventional all-optical methods. When applied to anticancer drug delivery, the localized mechanical forces generated by high-speed propulsion enhance stable adhesion of carriers to the cell membrane, thereby promoting drug uptake and accelerating tumor cells apoptosis. This work establishes Airy beams as a powerful tool for drug delivery, opening a new avenue for high-performance all-optical delivery platforms.

Tumor-derived lactate has long been regarded as a metabolic waste product. However, accumulating evidence indicates that lactate also functions as a signaling molecule that actively remodels the tumor immune microenvironment. How lactate-driven post-translational modifications in immune cells contribute to immune evasion in hepatocellular carcinoma (HCC) remains incompletely understood. This study aimed to identify the immune cell population responsible for lactylation-driven immunosuppression in HCC and to elucidate the molecular mechanism by which lactylation rewires macrophage metabolism to impair CD8+ T cell-mediated antitumor immunity. Selective in vivo immune cell depletion models were employed to define the key immune mediators of lactate-induced immunosuppression. Proteomic screening, site-directed mutagenesis, and lipidomic profiling were used to characterize lactylation targets and lipid metabolic alterations. Functional assays, including signaling pathway analyses, cytokine measurements, and tumor immune profiling, were performed in both in vitro systems and mouse HCC models. Macrophages were identified as the principal immune cell type mediating lactylation-dependent immunosuppression in HCC. Alanyl-tRNA synthetase 1 (AARS1) functioned as a non-canonical lactyltransferase, catalyzing lactylation of carnitine palmitoyltransferase 1A at lysine 675. This modification impaired long-chain fatty acid transport into mitochondria, leading to cytosolic accumulation of oleic acid (OA). OA directly disrupted cGAS binding to cytosolic DNA, thereby suppressing STING activation and type I interferon (IFN-I) production. Attenuated IFN-I signaling resulted in reduced major histocompatibility complex-I expression on tumor cells and impaired CD8+ T cell-mediated recognition and cytotoxicity. These findings uncover a lactate-lipid metabolism axis that links tumor-derived lactate to innate immune suppression in HCC. Targeting AARS1-mediated lactylation represents a potential therapeutic strategy to restore macrophage immunostimulatory function and enhance antitumor immunity.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent liver disease that can progress to metabolic dysfunction-associated steatohepatitis (MASH), a condition marked by inflammation and fibrosis that increases the risk of cirrhosis, liver failure, and hepatocellular carcinoma. Loss-of-function (LoF) variants of hydroxysteroid 17-β dehydrogenase 13 (HSD17B13), a lipid droplet-associated enzyme, protect against progression from MASLD to MASH and more severe types of liver disease. We generated a knock-in mouse model harbouring the human orthologue of the HSD17B13 P260S patient variant and investigated the molecular mechanisms underlying its protective phenotype upon dietary challenge, by applying comprehensive spatial transcriptomics, histological analysis, matrix-assisted laser desorption/ionisation (MALDI) imaging, and in vitro cellular characterisation. Male Hsd17b13P260S mice showed reduced liver fibrosis (p < 0.0001) upon dietary challenge, reflecting the protective effect seen in patients and supporting the model's translatability. Despite similar transcript levels, HSD17B13 P260S protein in the liver was significantly lower compared to wild-type (WT). Interestingly, HSD17B13 WT protein lost its periportal zonation when switching mice from a chow to a high trans-fat diet (HTF), whereas the HSD17B13 P260S protein retained periportal expression. Male Hsd17b13P260S mice showed increased steatosis (p < 0.05), while spatial transcriptomics revealed the suppression of pro-fibrotic and immune response-related genes. MALDI imaging identified significant differences in phospholipids across genotypes and diets. Both HSD17B13 WT and P260S variant protein localised to lipid droplets, but the latter showed reduced enzymatic activity. Lastly, we observed reduced expression of CCND1 (p < 0.05) and an increase in multinucleated hepatocytes (p < 0.0001) in livers from male Hsd17b13P260S compared to WT upon HTF feeding. Our study suggests that the Hsd17b13P260S mouse model is translatable towards human physiology and useful for understanding the protective mechanisms of the human HSD17B13 P260S LoF variant. Lower HSD17B13 P260S protein abundance, reduced enzymatic activity and retained periportal localisation may contribute to protection from liver disease progression by maintaining zonated transcriptomic and lipidomic profiles. These findings provide key insights into HSD17B13 biology and its role in MASLD to MASH progression. Novartis Biomedical Research Postdoctoral program, Sigrid Jusélius Foundation, Forendo Pharma.

Remodeling of fatty acid metabolism is increasingly recognized as a critical feature of tumor progression, yet the contribution of very long-chain fatty acid (VLCFA) remains incompletely understood. The elongation of VLCFAs, which is mediated by four consecutive enzymes in the endoplasmic reticulum (ER), has been implicated in tumor progression. However, the molecular mechanisms underlying VLCFA elongation enzymes and their specific contributions to tumorigenesis remain largely elusive. Here, we demonstrate that trans-2-enoyl-CoA reductase (TECR) is upregulated in colorectal cancer (CRC). Structural and biochemical analyses revealed a conserved catalytic mechanism for TECR-mediated trans-2-enoyl-CoA reduction. Moreover, we show that TECR forms a stable complex with 3-hydroxyacyl-CoA dehydratase (HACD), to cooperatively drive VLCFA elongation. A unique U-shaped loop in HACD is critical for recognizing TECR. Disruption of the HACD-TECR interaction interface significantly suppresses CRC cell growth. Collectively, these findings elucidate the molecular mechanism of HACD-TECR-mediated VLCFA elongation and suggest a potential therapeutic strategy for CRC treatment by modulating VLCFA metabolism.

Pancreatic cancer remains one of the deadliest malignancies, with gemcitabine-based chemotherapy as the mainstay treatment for most patients, yet resistance emerges almost universally. A defining feature of pancreatic cancer is its dense, fibroblast-rich stroma, in which heterogeneous cancer-associated fibroblasts (CAF) actively shape tumor biology and therapeutic response. Here, we elucidated a stromal-metabolic mechanism through which chemoresistant CAFs confer gemcitabine resistance. We identified a subset of mitophagy-competent CAFs that enhanced pancreatic cancer gemcitabine resistance. The epithelial-mesenchymal transition transcription factor ZEB1 acts as a master regulator of this CAF-driven chemoresistance program, being upregulated and epigenetically activated via SETD1A-mediated H3K4 methylation in gemcitabine-resistant CAFs. ZEB1 promotes BNIP3-mediated mitophagy in CAFs, leading to increased nucleotides secretion, which competitively inhibited gemcitabine incorporation into cancer cells while simultaneously supplying pyrimidine metabolism substrates for pyrimidine metabolism. Concurrently, ZEB1 transcriptionally activated CXCL8, engaging the CXCR1/2-MEK/ERK pathway in tumor cells and further augmenting pyrimidine metabolism via the RRM1/E2F1/G6PD axis, collectively diminishing gemcitabine cytotoxicity. Notably, combined inhibition of CXCR1/2 or G6PD with gemcitabine robustly suppressed tumor growth and restored chemosensitivity both in vitro and in vivo. These findings uncover a key stromal-metabolic axis in pancreatic cancer, linking mitophagy CAF activity to metabolic remodeling in tumor cells and identifying ZEB1 and its downstream network as actionable targets to overcome chemoresistance. ZEB1 reprograms mitophagy-competent CAFs to promote gemcitabine resistance by enhancing nucleotide supply and activating tumor pyrimidine metabolism, supporting therapeutic targeting of the ZEB1-CXCL8-pyrimidine axis in pancreatic cancer.

Colorectal cancer (CRC) is a highly prevalent malignancy worldwide. Early diagnosis remains challenging, and its strong propensity for metastasis is a major contributor to poor patient prognosis. Current approaches for surgical guidance and photodynamic therapy (PDT) in CRC are hindered by key limitations. Imaging techniques used for intraoperative navigation often lack sufficient sensitivity and specificity, while conventional PDT often employs "always-on" photosensitizers that lack tumor selectivity, posing a risk of collateral damage to healthy tissues. To address these limitations, this study leveraged the overexpression of β-N-acetylhexosaminidase (Hex) in CRC to design and develop an activatable near-infrared phototheranostic probe (INA) for tumor diagnosis and therapy. In vitro experiments demonstrated that INA is effectively activated by endogenously overexpressed Hex in CRC cells. Under light irradiation, it exhibits potent phototoxicity against CRC cells and three-dimensional tumor spheroids, with minimal dark toxicity. In tumor-bearing mouse models, INA enabled tumor-specific fluorescence imaging, real-time detection of microtumors for surgical navigation, and effective PDT, while demonstrating favorable biocompatibility. With its simple molecular architecture, INA is positioned as a highly competitive and readily translatable candidate for integrated diagnosis and therapy in CRC.

Photodynamic therapy (PDT) has been shown to improve survival and quality of life in patients with unresectable extrahepatic cholangiocarcinoma. However, its therapeutic efficacy is frequently limited by the survival of residual tumor cells that can re-enter the proliferative cycle. Our previous studies showed that residual cholangiocarcinoma cells can re-enter the proliferative cycle following PDT accompanied by hypoxia-induced activation of the HIF-1α survival pathway and intracellular antioxidant programs. To address this residual viability, we developed a glutathione-responsive targeted nanosystem (TSH NPs), in which the aggregation-induced emission (AIE) photosensitizer TPA-Ph-RDN is conjugated to the chemotherapeutic agent hydroxycamptothecin (HCPT) via a disulfide linkage. This design enables HCPT to exert direct cytotoxic effects while simultaneously suppressing the HIF-1α-mediated hypoxic adaptation pathway, thereby reducing residual tumor viability and enhancing PDT efficacy. This strategy achieves a dual-mechanism, two-pronged therapeutic effect. Both in vitro and in vivo studies demonstrated that TSH NPs elicited markedly synergistic antitumor activity, highlighting their potential as a promising therapeutic approach for improving clinical outcomes in patients with eCCA.

The clinical translation of AI in medical imaging faces critical challenges including scare annotated data, long-tailed pathological distributions, and privacy constraints in virtual imaging trials. To address these limitations, we propose a Multi-conditional Diffusion framework with Texture Constraints (MDTC) for synthesizing clinically reliable lesions in CT images. The key innovation lies in the joint integration of anatomical mask guidance and a Gray-Level Co-occurence Matrix-based texture classifier into the diffusion network. This dual-constraint mechanism uniquely enforces structural fidelity and pathologically heterogeneous characteristics in synthetic lesions, effectively preserves pathological heterogeneity and simultaneously enhances the authenticity of the generated lesionst in existing methods. The experiments on hepatocellular carcinoma and pulmonary nodule datasets demonstrate that the proposed MDTC method achieves favorable performance in terms of texture fidelity, a significant improvement in PSNR, and notable optimization in FID compared to classic generation models. Furthermore, downstream classifiers trained on synthetic data remain capable of maintaining the vast majority of baseline classification performance, while data augmentation for rare pulmonary nodules significantly improved the classification efficacy. This work establishes a new paradigm for generating diagnostically meaningful synthetic data, effectively alleviating critical bottlenecks in virtual imaging trial. The experimental results demonstrate that MDTC-generated lesions preserve structural authenticity and have potential to mitigate texture homogenization, thereby enabling robust downstream diagnostic model training and offering a practical, privacy preserving solution to expand virtual imaging trials toward underrepresented disease groups and real world clinical workflows.

Standard slide-based two-dimensional (2D) histopathology severely undersamples spatially heterogeneous tissue, with each thin 2D section representing <1% of the entire biopsy volume. Recent advances in non-destructive three-dimensional (3D) pathology, such as open-top light-sheet microscopy, enable comprehensive high-resolution imaging of large clinical specimens. Since manual review of these massive and complex 3D datasets is infeasible in clinical practice, we present TRICARE, a deep-learning triage framework that identifies high-risk 2D cross sections within 3D pathology datasets to enable time-efficient pathologist evaluation, which offers a lower-risk route for accelerated adoption by retaining pathologists for final diagnoses. TRICARE assigns risk scores to all 2D levels within a tissue volume by leveraging context from a subset of neighbouring depth levels, outperforming models in which predictions are based on isolated 2D levels. In two use cases-risk stratification based on prostate cancer biopsies and screening for dysplasia/cancer in endoscopic biopsies of Barrett's esophagus-AI-triaged 3D pathology, enabled by TRICARE, demonstrates the potential to improve the detection of high-risk diseases compared with slide-based 2D histopathology while optimizing pathologist workloads.

Interindividual variability in response to cancer therapy is a major challenge in the management of colorectal cancer (CRC). The gut microbiome contributes to differential therapeutic efficacy and toxicity by modifying the pharmacokinetics and pharmacodynamics of anticancer drugs. Furthermore, bacterial products interact with tumor and immune cells, altering therapeutic outcomes. This review focuses on the impact of pharmacomicrobiomics on CRC therapy. We describe how gut microbiota affects drug metabolism on a mechanistic level and outline the interactions of specific microbes and their products with chemo-, targeted, and immunotherapies employed in CRC. Finally, we provide an overview of current strategies, including probiotics, engineered bacteria, and fecal microbiota transplantation, that exploit the gut microbiome to improve therapeutic efficacy and reduce toxicity.

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

Despite advances in combined photothermal (PTT) and photodynamic (PDT) therapy for melanoma, therapeutic efficacy remains limited by tumor hypoxia, intracellular glutathione (GSH), insufficient tumor targeting, and photobleaching of photosensitizers. Herein, tumor-targeting and GSH self-depleting hyaluronic acid (HA)/tannic acid (TA)-engineered Prussian blue (PB) oxygen nanogenerators (ONs) loaded with IR780 (IHTPB ONs) are developed to enhance synergistic PTT/PDT. The IHTPB ONs exhibit uniform morphology, excellent colloidal stability, acidity/GSH-responsive IR780 release, high photothermal conversion efficiency (60%), and outstanding photothermal stability. Importantly, PB-mediated GSH oxidation and TA-mediated GSH conjugation enable dual-mode GSH depletion, while PB catalyzes oxygen generation to alleviate tumor hypoxia and promote IR780-mediated singlet oxygen production. Following CD44-mediated cellular uptake, IHTPB ONs effectively deplete intracellular GSH and, under near-infrared irradiation, induce robust reactive oxygen species generation and hyperthermia, leading to mitochondrial dysfunction, lipid peroxidation, apoptosis, and ferroptosis. In vivo, IHTPB ONs exhibit superior tumor accumulation and significantly enhanced antitumor efficacy compared with free IR780 and non-targeted nanoparticles, resulting in prolonged survival of melanoma-bearing mice. This work provides an effective nanoplatform integrating tumor targeting, oxygen self-supply, and dual-mode GSH depletion to potentiate synergistic PTT/PDT for melanoma therapy.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal human malignancies, characterized by persistently poor survival rates and limited effective therapeutic options. Ferroptosis, an iron-dependent form of regulated cell death driven by uncontrolled lipid peroxidation, has recently emerged as a potentially targetable vulnerability in PDAC. In this Review, we synthesize current understanding of the core molecular mechanisms governing ferroptosis, including dysregulated iron metabolism, lipid peroxidation pathways, and failure of antioxidant defense systems. We further discuss the complex regulatory networks that shape ferroptosis sensitivity in PDAC, highlighting the contributions of endoplasmic reticulum stress, mitochondrial dysfunction, autophagy, and DNA damage-associated signaling pathways. Within the broader landscape of PDAC biology, ferroptosis exerts context-dependent and sometimes opposing effects, influencing both tumor initiation and the development of therapeutic resistance. Accordingly, PDAC cells engage diverse epigenetic, metabolic, and microenvironmental adaptations to evade ferroptotic cell death, thereby sustaining resistance to therapy. Finally, we review the rapidly expanding spectrum of ferroptosis-based therapeutic strategies, encompassing small-molecule inducers, natural products, and, in particular, advanced nanomedicine platforms designed to enhance intratumoral drug delivery, overcome stromal barriers, and enable coordinated induction of ferroptosis while remodeling the tumor microenvironment.

Macroautophagy (MA) and chaperone-mediated autophagy (CMA) are lysosomal degradation pathways with context-dependent roles in cancer. However, how MA- and CMA-associated transcriptional states jointly relate to cancer molecular features and drug responses remains unclear. Here, we developed a transcriptome-based framework integrating MA- and CMA-associated gene signatures to define relative MA and CMA states across cancer types. These states were associated with distinct patterns of genomic instability, oncogenic signaling, immune features, and pharmacogenomic profiles. In pooled cancer cell-line analyses, MAhigh CMAhigh states were associated with relative resistance to EGFR tyrosine kinase inhibitors (EGFR-TKIs), whereas MAhigh CMAlow states showed greater sensitivity, nominating an autophagy-associated pharmacological pattern for mechanistic investigation. Using KRAS-mutant pancreatic cancer cells as an autophagy-dependent, therapy-resistant model, we found that LAMP2A depletion enhanced sensitivity to EGFR-TKIs in an MA-dependent manner. All-trans retinoic acid (ATRA), a pleiotropic retinoid that modulated CMA-related activity in this system, phenocopied key effects of genetic CMA suppression and potentiated sensitivity to EGFR-TKIs. CMA suppression was associated with increased autophagic flux, TFEB nuclear localization, and ULK1 phosphorylation changes consistent with MA activation. Moreover, transcriptomic analysis reveals that CMA suppression sensitizes cancer cells to EGFR-TKIs at least partially through downregulation of SEMA6D. SEMA6D depletion enhanced autophagic flux, increased lysosomal capacity, and partially contributed to the response to combined EGFR-TKI and ATRA treatment. In PANC-1 xenografts, ATRA potentiated EGFR-TKI-mediated tumor suppression without significant toxicity. Together, these findings establish a transcriptome-based MA-CMA framework for prioritizing context-dependent autophagy-associated vulnerabilities and provide focused mechanistic support for MA-CMA crosstalk in KRAS-mutant pancreatic cancer models.

The poor prognosis of head and neck squamous cell carcinoma (HNSCC) is primarily attributed to lymphatic or systemic metastasis. The cellular identity plasticity provides carcinocytes with the motility phenotype fluctuations, which indicates the intensified metastasis. Aiming at elucidating this lineage evolution, this study integrated single-cell RNA sequencing data from 12 primary lesions, 12 lymphatic metastases, and 12 distant metastases to portray the pan-metastatic cellular landscape. With the escalation of metastasis, there was a dramatic increase in the proportion of cancer-associated fibroblasts (CAFs). The spatiotemporal RNA alternative splicing and trajectory tracing methods with/without histopathological constraint observed that the SCGB3A1+ malignant epithelial cells, which become conspicuous in metastasis, undergo partial epithelial-mesenchymal transition (pEMT) to exhibit a CXCL14+ CAF phenotype, and B2M+ CAFs serve as a transdifferentiation terminal. Moreover, multimodal genetic detection identified WD repeat domain 54 (WDR54) as a potential initiator of cellular identity reshaping. Functional experiments demonstrated that WDR54-launched dedifferentiation preserves lineage plasticity via epithelial signature erosion and markedly enhances cellular invasion, migration, and MDSC-mediated immune evasion. Homologous protein structural congruence alignment and CUT&Tag profiling revealed that WDR54 initiates the TGF-β-driven pEMT program by selective H3K4me3 and H4K16ac modifications. Clinically, the practical utility of WDR54 as a diagnostic, prognostic and therapeutic target has received validation from a real-world perspective. In summary, the research establishes WDR54 as an epigenetic regulator that initializes cell identity and triggers metastasis in HNSCC.

Enhancer of zeste homolog 2 (EZH2) is frequently overexpressed in cancer, correlating with adverse clinical outcomes. However, how EZH2 overexpression supports tumorigenicity is not fully elucidated. Here we identify a previously unexplored tumour-promoting axis involving EZH2-SREBP2 association. EZH2 and SREBP2, a master regulator of lipid metabolism, cooperate to drive high expression of mevalonate pathway genes, enhancing cholesterol biosynthesis and sustaining tumour growth. Transcriptional activation domains of EZH2 and SREBP2 bind p300 directly, mediating proto-oncogene activation. Furthermore, we employed proteolysis targeting chimeras (PROTACs) to target this non-canonical EZH2 function. Independent EZH2-targeting PROTACs degrade both EZH2 and SREBP2, downregulating SREBP2-associated gene-expression programmes and inhibiting tumour growth. Collectively, this study unveils an EZH2-SREBP2 regulatory axis that promotes cholesterol biosynthesis and fuels tumorigenesis, shifting the current paradigm of EZH2's oncogenic functions.

Lineage-determining transcription factors (TFs), such as p63 in epithelial tissues, establish normal tissue identity but can also drive carcinogenesis. The epigenetic basis for these divergent effects, however, is unclear. Here, we show that p63 reorganizes three-dimensional (3D) chromatin topology in squamous cell carcinoma (SCC) through cooperation with the tumor-selective partner TF, FOXK1. Multi-omic profiling across normal and malignant states reveals that p63 enhances connectivity between cis-regulatory elements near oncogenes and promotes a secondary enhancer looping architecture, in which p63 and FOXK1-enriched enhancers indirectly contact target promoters via FOXK1-associated intermediate anchors lacking p63 binding. FOXK1 mediates p63-dependent chromatin looping, as FOXK1 loss selectively weakens p63-dependent loops and reduces transcription of associated target genes. The p63-FOXK1 interaction is observed in human tumor samples but not in normal tissues. Together, our findings define an epigenetic mechanism by which a lineage master TF and tumor-restricted partner TF reshape 3D chromatin structure to drive oncogenic transcription, with implications for tumor cell identity and therapy.

KRAS mutations are a major driver of pancreatic ductal adenocarcinoma (PDAC). RASA2, a RAS GTPase-activating protein, modulates KRAS protein levels in wild-type contexts, suggesting it could play a potential role in PDAC. Here, we systematically investigated the biological function and molecular mechanisms of RASA2 in PDAC. Integrative analyses of multiple datasets and clinical samples demonstrated that RASA2 was consistently upregulated in KRAS-mutant PDAC and significantly associated with poor prognosis and metastatic progression. Gain- and loss-of-function studies revealed that RASA2 markedly enhanced PDAC cell migration and invasion in both KRAS-mutant and KRAS-wild-type models, suggesting that its pro-metastatic activity is largely independent of KRAS mutational status. Transcriptomic and mechanistic analyses revealed that RASA2 activated GLI1 through a TGFβ2-dependent, non-canonical Hedgehog pathway. Mechanistically, RASA2 interacted with RTF1 to promote H2BK120 ubiquitination at the TGFB2 promoter, thereby enhancing TGFβ2 transcription and activating downstream GLI1 signaling. Pharmacological inhibition of TGFβ signaling or genetic silencing of GLI1 effectively suppressed RASA2-driven migratory, invasive, and metastatic phenotypes in vitro and in vivo. Collectively, these findings reveal a mechanism by which RASA2-dependent epigenetic and transcriptional reprogramming promotes metastatic progression and nominate the RASA2- TGFβ2-GLI1 axis as a potential therapeutic target in PDAC.

The lethality of pancreatic ductal adenocarcinoma (PDAC) is driven, in part, by cellular plasticity that facilitates dedifferentiation and dissemination. Although transcriptional programs underlying these processes are well characterized, the contribution of translational control to PDAC cell-state regulation in vivo needs to be further understood to develop strategies to restrain malignant plasticity. Using a genome-wide CRISPR/Cas9 screen in immunocompetent hosts, we identified the noncanonical initiation factor eIF4G2 (DAP5/NAT1) as a translational checkpoint that restrains PDAC progression. Loss of eIF4G2 accelerated tumor growth, promoted poorly differentiated, basal-like histology, and triggered widespread metastasis. Ribosome profiling revealed that eIF4G2 supports the translation of a discrete cohort of mRNAs with long, guanine-cytosine (GC)-rich, structured 5' untranslated regions, including tumor suppressors such as Pten and transcriptional regulators such as Crebbp. Accordingly, loss of eIF4G2 was accompanied by secondary transcriptional enrichment of migration and wound-healing programs and induction of basal-like markers. In human PDAC, eIF4G2 expression was reduced in poorly differentiated lesions, and functional eIF4G2 perturbation in patient-derived PDAC cells increased clonogenic growth, whereas enforced eIF4G2 expression suppressed colony formation. Computational inference from human PDAC datasets revealed that reduced eIF4G2 activity correlated with increased metastasis, enhanced basal-like features, and poorer patient survival. Together, these findings establish noncanonical translation initiation as a determinant of PDAC cell-state control and identify eIF4G2 as a barrier to malignant plasticity and metastatic dissemination. The translational checkpoint regulator eIF4G2 preserves epithelial identity and suppresses metastasis in pancreatic cancer, revealing selective translation as a determinant of subtype, prognosis, and therapeutic stratification.

Enhanced glycolysis leads to pathological lactate accumulation and promotes malignant tumor progression through protein lactylation. However, the mechanisms by which lactylation enhances RNA processing to drive prostate cancer metastasis remain unclear. Specifically, we found that pathologically accumulated lactate induces the synthesis of a novel circRNA, circPTK2 (hsa_circ_0135904), which is highly expressed in PCa and predicts poor patient prognosis. Functionally, lactylated EIF4A3-induced circPTK2 promotes PCa metastasis in vitro and in vivo. Mechanistically, we revealed that lactylated EIF4A3 interacts with the flanking introns of exons 16-20 on PTK2 pre-mRNA to trigger sustained circPTK2 biogenesis. Subsequently, circPTK2 upregulates the expression of SLC4A4 by directly sponging miR-582-5p, which in turn activates the PI3K/AKT signaling pathway to drive the epithelial-mesenchymal transition (EMT) and metastasis of PCa. Crucially, SLC4A4-mediated intracellular pH (pHi) homeostasis promotes intracellular lactate accumulation, thereby enhancing EIF4A3 lactylation and establishing a positive regulatory axis. Importantly, circPTK2 overexpression significantly accelerated subcutaneous tumor growth and facilitated lung metastasis in PCa mouse models. Our findings identify a novel mechanism by which lactate metabolism drives circRNA biogenesis, where the lactylation of EIF4A3 promotes the aberrant synthesis of circPTK2. Furthermore, we characterized a unique mechanism involving a lactylation-driven EIF4A3/circPTK2/SLC4A4 positive regulatory axis which promotes distant metastasis in PCa. Targeting this lactylation-mediated axis represents a promising therapeutic strategy for metastatic disease.

Cancer stem cells (CSCs) are tumor cell subsets with self-renewal, multilineage differentiation, and tumor-initiating capacity that sustain cancer initiation, progression, metastasis, and relapse. Targeting CSCs therefore represents a promising route to improve the durability of cancer treatment. However, translation of this approach into routine care has been slow because of the biological complexity and clinical constraints. This review discusses current concepts of CSC origin and plasticity, the criteria used to define CSCs across different tumor types, and the marker systems as well as high-resolution technologies that are used to track CSC states. Developmental pathways, growth factor and cytokine cascades, as well as microenvironmental and stress responses that control CSC maintenance and therapy resistance are explored with a focus on their tractability as drug targets. We then discuss mechanisms through which CSCs escape chemotherapy, radiotherapy, and targeted agents. We review current efforts to use these pathways in designing small molecules, antibodies, cellular therapies, and vaccines aimed at CSC compartments. Heterogeneity within and between tumors, dynamic interconversion between CSC and non-CSC states, and support from specialized niches are considered as major barriers for clinical trial design, biomarker development, and response assessment. Emerging single-cell, spatial, and lineage tracing approaches, together with organoid and ex vivo platforms, are reviewed as tools that can bridge preclinical models and patient samples and guide the development of CSC-directed combination regimens. The goal is to outline translational principles that can guide future strategies for integrating CSC-focused interventions with established therapies to improve long-term disease control.

Transcription factors often exhibit a striking paradox: they function as tumor suppressors in one context and promote oncogenesis in another. The underlying mechanisms of this context dependence have remained elusive. We propose a novel conceptual framework, systemic redox switching, to resolve this paradox. Based on our research on upstream stimulatory factor 2 (USF2) and convergent observations on other factors, our model suggests that redox regulation is network-embedded rather than driven by discrete cysteine switches. We propose that transcription factors occupy distinct regulatory regimes (homeostatic, adaptive, and survival states) which are connected by threshold-like, hysteretic transitions. Beyond classical graded input-output views, this framework explicitly posits discrete, hysteretic regime transitions at the transcriptional-network level and links them to a minimal dynamical model of the USF2-TFEB-NRF2-redox motif. These transitions convert continuous redox inputs into distinct changes in promoter occupancy and transcriptional programs. USF2 exemplifies a kinase-integrated, non-canonical switch that decodes mitochondrial and autophagy signals via phosphorylation (e.g., Ser155) and context-dependent cooperation with NRF2 and HIFs. Extending this logic to diverse archetypes (NRF2, HIFs, FOXOs, c-MYC, and AHR) demonstrates the framework's generalizability. The model is experimentally tractable; targeted perturbations of primary sensing modules (e.g., KEAP1 mutation, PHD inhibition, or USF2 phosphorylation disruption) should predictably alter regime transitions. By reframing paradoxical behaviors as controlled state transitions, this framework provides a unifying, network-level understanding with direct implications for targeting therapies in cancer, metabolic diseases, and age-related pathologies.

Histamine is a key signaling molecule in pathophysiology that can exhibit significant regulatory roles in diverse health and disease states. Besides the well-studied noncovalent interactions between histamine and its receptors, protein histaminylation is a recently discovered mechanism of action through which histamine regulates cellular signaling pathways in a covalent modification manner. Histaminylation is an emerging protein post-translational modification (PTM), where an isopeptide bond is formed between the histamine primary amine and the γ-carboxyl group of glutamine through a transamidation reaction catalyzed by transglutaminase 2 (TGM2). However, due to the lack of efficient pan-specific antibodies targeting histaminylated glutamine, the histaminylation proteome in cells remains poorly explored. Here, we report the design and development of a novel Nτ-propargylated histamine (Nτ-PH) probe as well as its successful application in chemical proteomic profiling of the histaminylation proteome in cancer cells. Notably, new TGM2-catalyzed epigenetic marks on core histones, e.g., H2AX-Q84 and Q104 histaminylation, have been identified from cancer cells and verified. Lastly, the crosstalk between H2AX histaminylation and γH2AX formation was discovered in this study, suggesting that TGM2-mediated histaminylation plays a critical role in DNA damage responses.

Metabolic reprogramming and the evasion of regulated cell death are two canonical hallmarks of cancer, yet the mechanistic threads connecting these distinct biological phenomena remain incompletely understood. The "Warburg effect," characterized by robust aerobic glycolysis, results in the massive accumulation of lactate. Once dismissed as a metabolic waste product, lactate has recently emerged as a pivotal signaling molecule and an epigenetic precursor for lysine lactylation (Kla), a novel post-translational modification that fundamentally reshapes the chromatin landscape. Concurrently, ferroptosis-an iron-dependent form of non-apoptotic cell death driven by unrestrained lipid peroxidation-has garnered intense interest as a therapeutic vulnerability in therapy-resistant tumors. However, a paradoxical observation persists highly glycolytic tumors, despite generating abundant ROS (reactive oxygen species), often display intrinsic resistance to ferroptosis. In this review, we propose that histone and non-histone lactylation serves as the critical "epigenetic bridge" coupling metabolic flux to ferroptosis evasion. We dissect the molecular machinery by which the "Lactate-Kla axis" transcriptionally activates antioxidant defenses (e.g., GPX4, SLC7A11) and remodels the tumor microenvironment to suppress ferroptotic triggers. Furthermore, we delineate how targeting the writers and erasers of lactylation can dismantle this metabolic shield, offering a rational strategy to re-sensitize refractory tumors to ferroptosis-inducing therapies. This synthesis highlights a new frontier in cancer biology where metabolism, epigenetics, and cell fate decisions converge.

Ferroptosis is an iron-dependent form of regulated cell death that has emerged as one of the mechanisms underlying cisplatin-induced nephrotoxicity. Here, we report an iron-chelating polymeric micelle for controlled cisplatin activation and ferroptosis suppression to address the above issue. The block copolymer, methoxyl polyethylene glycol-poly(glutamic acid) (mPEG-PGlu) was coupled with an iron chelator, deferiprone (Dfp) to generate the tailored amphiphilic conjugate, mPEG-P(Glu-Dfp) that could coordinate with activated cisplatin and self-assemble into micelles (Pt@MDfp). The cisplatin-loaded mPEG-PGlu micelles (Pt@MGlu) and free cisplatin were employed as controls. Upon endocytosis, the intracellular labile iron could replace cisplatin due to the higher affinity between Dfp and Fe2+. This coordination exchange resulted in controlled cargo release and suppressed lipid peroxidation and inflammation in renal tubular cells (HK-2). The proof-of-concept was also validated in the subcutaneous 4T1 mouse breast tumor model using plasma and urine biomarkers of ferroptosis, inflammation, and renal injury as the indices, which concurred well with the histological staining of kidney tissue and fibrosis assessment. Moreover, due to the controlled cargo activation, Pt@MDfp significantly enhanced the in vivo antitumor efficacy compared to Pt@MGlu and free cisplatin. Collectively, the coordination exchange approach is promising in controlled delivery of cisplatin for efficacy enhancement.

Epithelial-mesenchymal transition (EMT) mediated metastasis remains the primary contributor to cancer-related mortalities worldwide, highlighting the critical need for therapeutic strategies that simultaneously eradicate primary tumors and suppress metastatic progression. However, conventional photodynamic therapy (PDT), particularly oxygen-dependent type-II photosensitizers, suffers from hypoxia-limited efficacy and may even induce EMT under suboptimal treatment conditions. Herein, we report enhanced type-I reactive oxygen species (ROS)-generating nanoparticles (NPs) based on the coassembly of two structurally similar small molecules (TQTT-NO and TQTT-NH), integrating light-controlled nitric oxide (NO) release to synergistically inhibit tumor growth and EMT. By leveraging precise molecular structure matching, the coassembled NPs (TQTT-NO/NH NPs) enable efficient intermolecular electron transfer, as revealed by the photocurrent results and Gibbs free energy calculations, thereby favoring type-I ROS generation under white-light irradiation while simultaneously triggering on-demand NO release. The developed TQTT-NO/NH NPs effectively suppress transforming growth factor-β (TGF-β)-induced EMT, inhibit cancer cell migration and invasion in vitro, and markedly reduce primary tumor growth and lung metastasis in a murine tumor model under light activation. Overall, this work establishes a generalizable molecular coassembly strategy for enhancing type-I PDT and EMT regulation, offering a promising paradigm for next-generation antimetastatic phototherapeutic platforms with translational potential.

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

Accurate clinical diagnosis of triple-negative breast cancer (TNBC) usually needed tedious, time consuming, high-cost tissue section analyzation procedures by using conventional clinical medical approaches. Thus, it is an urgent to develop low-cost, rapid, high-sensitivity TNBC diagnostic approaches. Herein, we have proposed a temperature-compensation short-period microfiber long-period grating (MFLPG) sensor coated with Au nanostars (Au NS)-Nb2CTX MXene coupled with localized surface plasmon resonance (LSPR) effect for the high-sensitivity tracking tumor-derived exosomal programmed cell death ligand-1 (PD-L1) in collected clinical human samples. For sensing exosomes particles in pure phosphate buffered saline (PBS) solution, the sensor device could exhibit an ultra-high detection sensitivity of 0.852 nm/log10 (particles/mL), and an ultra-low limit-of-detection (LOD) of 12.9 particles/mL within the exosomes concentration ranging from 101 particles/mL to 108 particles/mL. More significantly, the MFLPG sensor could differentiate breast cancer patients from healthy controls, and it could also differentiate TNBC patients from some other non-TNBC patients' samples. Therefore, this work could pave a new method for rapid, high-sensitivity and accurate diagnosis of TNBC in clinical samples.

Molecular residual disease (MRD) detection has potential to transform the selection of adjuvant therapy. Most evidence is from tumor-informed assays. Tissue-free assays not requiring sequencing of the primary tumor may simplify workflows if they offer accuracy similar to that of tumor-informed assays. To evaluate tissue-free circulating tumor DNA (ctDNA) analysis in patients with triple negative breast cancer (TNBC) and compare the tissue-free assay with tumor-informed assay results. This was a prognostic and exploratory analysis of patients with TNBC at moderate to high risk of recurrence who were participating in the ctDNA surveillance period of c-TRAK TN, a multicenter phase 2 clinical study. Plasma samples were collected from participants every 3 months for up to 2 years after the completion of adjuvant therapy and analyzed prospectively with digital polymerase chain reaction (dPCR). Data for the analysis were from a database lock on September 28, 2021, with subsequent follow-up through January 18, 2023. Data were analyzed from July 2025 to May 2026. Tissue-free assay leverages cancer differential methylation patterns to detect ctDNA. Recurrence-free survival by tissue-free ctDNA detection status. Comparison between tissue-free ctDNA detection and both dPCR and whole-exome sequencing-powered multivariant tumor-informed assays. The analysis included 1026 plasma samples from 159 patients (mean [SD; range] age, 51.4 [11.4; 25.0-78.0] years; 159 females [100%]). The tissue-free assay detected ctDNA in 54 patients (34.0%), with detection strongly associated with risk of recurrence (HR, 27.2; 95% CI, 13.7-54.2; P < .001). Among patients with ctDNA detected by both the tissue-free assay and dPCR (42 patients [26%]), tissue-free detection occurred at an earlier time point in 14 patients (33.3%); dPCR detection occurred before tissue-free detection in no patients. Median lead time to recurrence was 7.9 (95% CI, 6.1-10.5) months for tissue-free vs 5.8 (95% CI, 3.3-10.0) months for dPCR (HR, 0.57; 95% CI, 0.34-0.95; P = .03). Concordance between tissue-free and multivariant tumor-informed assays was good. Among patients detected by both assays (41 of 133), 12 of 41 (29.3%) had ctDNA detected earlier by multivariant tumor-informed assay with 1 (2.4%) earlier by the tissue-free assay. Median lead times to recurrence were 7.6 (95% CI, 4.6-10.5) months with tissue-free and 7.1 (95% CI, 5.7-10.0) months with multivariant tumor-informed assay (HR, 1.46; 95% CI, 0.87-2.44; P = .15). In this prognostic study, tissue-free ctDNA detection during surveillance was strongly prognostic for recurrence in patients with TNBC. Comparable lead times between the tissue-free and multivariant tumor-informed assays support tissue-free MRD detection in a clinical trial setting, permitting ctDNA testing when tissue is not available.

Triple-negative breast cancer (TNBC) exhibits profound resistance to immunotherapy, largely due to a tumor microenvironment constrained by dysregulated lipid metabolism and persistent hypoxia. Excessive fatty acid uptake mediated by the scavenger receptor CD36 (Cluster of differentiation 36) imposes metabolic stress on immune cells while simultaneously fueling tumor growth, whereas hypoxia independently enforces immune exhaustion and macrophage immunosuppression. Despite extensive efforts, targeting either pathway alone has yielded limited therapeutic benefit, underscoring the need for coordinated immunometabolic intervention. Here, we report a living biohybrid strategy that simultaneously regulates lipid metabolism and oxygen availability to reprogram the TNBC microenvironment. By engineering photosynthetic Synechococcus cyanobacteria with surface-displayed anti-CD36 antibodies, we construct a dual-functional system (Syne@aCD36) capable of blocking CD36-mediated fatty acid uptake while generating oxygen in situ under light stimulation. This coordinated regulation alleviates lipid-induced metabolic stress and hypoxia-driven immune suppression, thereby restoring CD8⁺ T cell cytotoxicity, reversing exhaustion phenotypes, and promoting pro-inflammatory macrophage polarization. In an aggressive 4T1 TNBC model, Syne@aCD36 induces robust tumor growth inhibition, enhances immune infiltration, and exhibits minimal systemic toxicity. This work establishes a previously unexplored paradigm for immunometabolic therapy by integrating metabolic checkpoint blockade with photosynthetic oxygenation, providing a promising strategy for immunometabolic remodeling in lipid-rich and hypoxic tumor microenvironments. STATEMENT OF SIGNIFICANCE: We focus on the fact that excessive fatty acid uptake and tumor hypoxia cooperatively drive immunosuppression and T cell exhaustion in the tumor microenvironment. Our Syne@aCD36 platform possesses two innovations: 1. A dual-function living therapeutic system. Syne@aCD36 blocks CD36-mediated fatty acid uptake on tumor and immune cells while simultaneously generating oxygen through photosynthesis under light stimulation, thereby alleviating lipid metabolic stress and tumor hypoxia in situ. 2. Synergistic immunometabolic reprogramming. By concurrently regulating lipid metabolism and oxygen availability, Syne@aCD36 restores CD8⁺ T cell cytotoxicity, reduces exhaustion phenotypes, and promotes pro-inflammatory macrophage polarization, leading to enhanced immune infiltration and tumor growth suppression. Our study provides a strategy for tumor immunotherapy through integrated regulation of metabolic checkpoints and the tumor microenvironment.

Lactylation, a lactate-dependent posttranslational modification, is involved in cancer development. Here, we showed that GCN5L2 physically interacts with nuclear SIX1 and acts as a key regulator of SIX1 lactylation. GCN5L2 depletion promotes APC/Cdh1 binding to SIX1, increases SIX1 ubiquitination, and reduces SIX1 protein stability. Functionally, the GCN5L2-SIX1 axis promotes glucose uptake, glycolysis process, and energy supply in TNBC cells. GCN5L2 knockdown attenuates the growth and migration of TNBC cells, and increases the sensitivity of TNBC cells to paclitaxel. Overexpression of SIX1 reverses the GCN5L2 knockdown-induced glycolysis and growth suppression in TNBC cells. In addition, GCN5L2 expression is enriched and positively correlated with SIX1 in clinical TNBC tissues. More importantly, we demonstrated 9‴-Methyl salvianolate B as a candidate modulator of the GCN5L2-SIX1 complex via an unbiased screening. 9‴-Methyl salvianolate B suppresses the GCN5L2-SIX1 axis and blocks the glycolysis, proliferation, metastasis, and paclitaxel resistance of TNBC cells. Overall, this study highlights the significance of the GCN5L2-SIX1 axis in sustaining glycolysis and TNBC development, and provides experimental evidence of 9‴-Methyl salvianolate B as a candidate modulator of the GCN5L2-SIX1 axis, presenting a promising strategy for TNBC management.

Focal segmental glomerulosclerosis (FSGS) is a major cause of primary glomerular disease, characterized by progressive podocyte injury and loss. This study aimed to identify the RNA methylation regulator WTAP (Wilms' tumor 1-associated protein) as a novel pathogenic factor in FSGS. WTAP expression in podocytes was detected in kidney biopsies from patients with FSGS and in an adriamycin-induced FSGS mouse model. Two podocyte-specific Wtap-deficient mouse models were generated using the Cre-loxP system: a constitutive knockout (cKO) model and a tamoxifen-inducible knockout model. Loss- and gain-of-function strategies were used to investigate the functional role of WTAP in podocytes both in vivo and in vitro. WTAP expression was significantly decreased in podocytes from patients with FSGS and in the adriamycin-induced FSGS mouse model. Both cKO and tamoxifen-inducible knockout mice developed progressive glomerulosclerosis with characteristic features of FSGS. WTAP overexpression in podocytes attenuated podocyte injury both in vivo and in vitro. cKO mice exhibited alterations in the proportions of various renal cell types, including both renal intrinsic and inflammatory cells. Mechanistically, WTAP regulates SPP1 m6A methylation, thereby modulating its stability and expression. Notably, SPP1-mediated signaling emerged as a central hub coordinating pathological cell-cell interactions in cKO mouse kidneys, and neutralization of SPP1 significantly ameliorated renal injury in both cKO and adriamycin-induced FSGS mice. We successfully constructed 2 FSGS mouse models that are suitable for investigating the pathogenic mechanisms and therapeutic targets of FSGS. Our findings demonstrate that the WTAP/SPP1 (secreted phosphoprotein 1) signaling pathway contributes to the renal pathological changes in FSGS, suggesting that targeting this pathway may represent a promising therapeutic strategy.

Breast cancer is the most commonly diagnosed malignancy and a leading cause of cancer-related mortality. Hormone receptor-positive (HR+) tumors represent the most prevalent metastatic subtype, and early dissemination remains a major clinical challenge. Commensal dysbiosis, defined as an inflammatory gut microbiome with low biodiversity, promotes metastasis by inducing mammary gland inflammation. In this study, we investigated systemic mechanisms governing dysbiosis-induced metastasis. Metabolomic profiling revealed elevated primary bile acids (BA) in the dysbiotic fecal microbiome. Sequestration and supplementation approaches demonstrated that beyond driving metabolic disease and mammary gland inflammation, primary BAs orchestrated enhanced HR+ tumor dissemination via a prostaglandin E2 (PGE2)-dependent pathway. Analysis of The Cancer Genome Atlas showed that BA, insulin resistance, and PGE2 gene signatures are associated with reduced survival in patients with HR+ tumors. In complementary analyses using the Epic Cosmos electronic health record database, BA sequestrant use was associated with longer restricted mean survival time among patients with metastatic disease. Together, these findings reveal that commensal dysbiosis-associated loss of microbial BA metabolism elevates primary BAs and promotes HR+ metastatic progression through PGE2 signaling. Dysbiosis-induced bile acids drive systemic and mammary tissue-specific inflammation that promotes HR+ breast tumor metastasis, supporting the development of strategies targeting microbiome-derived metabolites to reduce metastatic risk in vulnerable populations.

Neutrophils are a prominent component of the tumor microenvironment (TME) that can have both pro- and antitumor functions. By analyzing neutrophils across different human cancers, we revealed an ICAM1high subset enriched in the TME, which was also observed in murine triple-negative breast cancer (TNBC) models. ICAM1high neutrophils exhibited an enhanced capacity for cell-cell adhesion specifically with tumor cells retaining epithelial features, and this adhesion conferred mutual advantages to both cell types. In contrast, cancer cells with mesenchymal-like phenotypes were vulnerable to neutrophil-mediated cytotoxicity due to decreased cell adhesion and elastase resistance. These opposite effects drove tumor evolution toward a dichotomy of neutrophil-enriched, epithelial-like ecosystems and macrophage-enriched, mesenchymal-like ecosystems. As ICAM1high neutrophils can reverse migrate from tissue into the circulation, the adhesive and reverse migratory properties together mediate metastatic intravasation. Spatial transcriptomic and tissue microarray analyses demonstrated interactions between tumor cells, neutrophils, and endothelial cells in human TNBC, particularly in non-Hispanic Europeans compared with African American patients. Together, this study demonstrated tumor-immune coevolution in which neutrophils instruct the phenotypes and metastatic behaviors of TNBC, which may preferentially occur in patients of certain ancestries. A distinct subset of ICAMhigh tumor-infiltrating neutrophils with increased adhesion and reverse transendothelial migration shapes tumor phenotypes and metastatic potential through mutually beneficial interactions with epithelial cancer cells.

Natural killer (NK) cells are effectors of innate antitumor immunity, yet their therapeutic potential in solid tumors remains largely unrealized. Breast cancer exemplifies this paradox: NK cells are present in circulation and detectable within tumors, but their cytotoxic activity is limited. Recent advances in single-cell and spatial profiling reveal that NK-cell failure in breast cancer does not result from simple immune absence but from multilayered constraints imposed by the tumor ecosystem. Soluble mediators, metabolic pressures, stromal architecture, and suppressive immune networks reprogram NK-cell identity and uncouple activation from cytotoxicity. Understanding how these constraints shape NK-cell states reframes breast cancer as a model of innate immune dysfunction and highlights new opportunities to reestablish NK-cell function through immunotherapies.

Metabolic reprogramming through enhanced glycolysis is a hallmark of cancer that supports tumor progression and promotes protumor immune responses. Methylglyoxal (MG), a reactive by-product of glycolysis, has recently emerged as an oncometabolite implicated in cancer progression and therapy resistance. Our previous work demonstrated that an imbalance between MG production and detoxification by the glyoxalase system, referred to as MG stress, contributes to progression and metastatic dissemination in triple-negative breast cancer (TNBC). However, the impact of MG stress on the tumor immune microenvironment remains poorly understood. Using two preclinical breast cancer models, we investigated the relationship between MG stress and immune modulation, with a focus on granulocytic myeloid-derived suppressor cells (g-MDSCs), major mediators of immune evasion. In silico analyses were performed to assess correlations between MG stress-related gene signatures and transcriptional markers of MDSC infiltration in patients with TNBC, as well as associations with anti-programmed cell death protein 1 (PD-1) immunotherapy response in melanoma cohorts. In vivo experiments combined the MG scavenger carnosine with PD-1 blockade in the immunotherapy-resistant 4T1 breast cancer model. MG stress was associated with the expansion of g-MDSCs in breast cancer models. Importantly, MG stress conferred metastatic potential to non-metastatic 67NR breast tumors, potentially through activation of the Nuclear Factor kappa B (NF-κB) pathway, increased granulocyte-macrophage colony-stimulating factor expression, and systemic expansion of g-MDSCs. In silico analyses revealed a positive correlation between MG stress-related gene signature and transcriptional markers of MDSC infiltration in patients with TNBC. Furthermore, this signature distinguished anti-PD-1 responder (low MG stress), from non-responders (high MG stress) in patients with melanoma. Therapeutically, combined targeting of MG stress with carnosine and PD-1 signaling significantly reduced g-MDSC accumulation in tumors, spleens, and lungs, and decreased lung metastatic burden in the 4T1 model. These findings identify MG stress as a driver of an immunosuppressive tumor microenvironment that may impair immunotherapy efficacy and promote metastatic progression in TNBC. Dual targeting of MG stress and PD-1 signaling represents a promising therapeutic strategy to overcome immune suppression and limit metastasis in immunotherapy-resistant breast cancer.

The retinoblastoma protein (Rb) is a tumour suppressor best known for repressing E2F transcription factors and halting cell cycle progression1. In hormone receptor-positive (HR+) breast cancer, CDK4/6 inhibitors activate Rb by preventing its phosphorylation, forming a key component of current endocrine therapy regimens2. How pharmacologically activated Rb remodels chromatin and influences transcription beyond cell cycle arrest remains poorly understood. Here we show that CDK4/6 inhibition induces redistribution of hypophosphorylated Rb to promoters and enhancers. Although Rb predictably binds to cell cycle gene promoters to repress transcription, at other sites, it unexpectedly promotes expression of oestrogen-responsive genes by integrating into oestrogen receptor (ER)-rich transcriptional hubs. CDK4/6 inhibition enhances ER target gene expression in breast cancer cells, patient-derived xenografts and clinical HR+ breast cancer samples in an Rb-dependent manner. This reprogramming is mediated in part by KDM5A, whose interaction with Rb contributes to gene regulation at these loci. Critically, components of this Rb-driven ER transcriptional program are pro-proliferative. In endocrine-sensitive tumours, this effect can be neutralized with anti-oestrogen therapy, explaining therapeutic synergy. In endocrine-resistant settings such as ESR1-mutant breast cancer, the program persists, limiting the therapeutic efficacy of CDK4/6 inhibition. These findings reframe Rb as a dual-function transcriptional regulator that, although enforcing cell cycle arrest, can also activate programs that counteract its tumour suppressor function.

Transcription factor SNAI1 guides plasticity and invasiveness in cancer. Using a complete SNAI1 knockout in mesenchymal, triple-negative breast cancer cells, unbiased genome-wide transcriptomic analysis revealed a marked under-expression of integrin-based adhesion and endocytic components. Utilizing this knockout cell model, complementary breast cancer cell models and functional screening of multiple differentially expressed genes, we found that the pioneering transcription factor FOXA1, whose expression is repressed by SNAI1, associates with several key mediators of the cellular phenotype. FOXA1 represses the small GTPase ARF6 and its exchange factor PSD4. In addition, some of the integrin and matrix metalloproteinase genes are regulated by the transcriptional FOXA1 signal. Accordingly, SNAI1 knockout cells presented poor adhesion to collagen type I or fibronectin, formed defective invadopodia and focal adhesions with weakened FAK/SRC signaling. SNAI1 knockout cells performed ineffective receptor-mediated internalization, including nanoparticle and extracellular vesicle (EV) uptake, exhibited reduced lysosomal content, lacked multivesicular bodies enriched in intraluminal vesicles and showed decreased EV secretion. Gain-of-function experiments demonstrated that SNAI1 has an impact on the PSD4/ARF6 signaling module, using FOXA1 as an intermediate factor to regulate EV release by tumor cells. We propose that the SNAI1-FOXA1 transcriptional mechanism operates at the level of membrane and vesicular trafficking control, which interlinks cell plasticity, adhesion and invasiveness through the extracellular environment, with the associated process of EV secretion.

Anthracycline-induced cardiotoxicity (AIC) limits life-saving chemotherapy and is driven by early metabolic remodeling. The nuclear receptor ERRα (estrogen-related receptor α) is a master regulator of cardiac energy metabolism, but the temporal dynamics of its downregulation, its causal role in AIC pathogenesis, and whether it can be pharmacologically activated to confer protection remain undefined. We performed temporal protein analysis in a porcine AIC model. Using cardiomyocyte-specificgain- and loss-of-function mouse models, we assessed the causal role of ERRα. Mechanistic studies included ChIP-qPCR, reporter assays, and microscale thermophoresis to investigate the natural compound formononetin. Human breast cancer patient-derived organoids were used to evaluate anticancer activity. ERRα expression was selectively downregulated in AIC pig hearts and cardiac tissue from chemotherapy-treated patients. Temporal analysis in pigs revealed that ERRα reduction occurred at the subclinical (6-week) stage, preceding overt cardiac dysfunction. Cardiomyocyte-specific ERRα overexpression activated mitochondrial gene programs, enhanced fatty acid oxidation, and preserved systolic function after doxorubicin challenge, whereas ERRα knockdown exacerbated bioenergetic failure and cardiac dysfunction. Through drug screening, we identified formononetin as a potent and selective ERRα agonist. Formononetin enhanced ERRα transcriptional activity, improved mitochondrial metabolism, and protected against AIC in both murine and porcine models. Mechanistically, ChIP-qPCR demonstrated increased ERRα occupancy at target gene promoters, and microscale thermophoresis confirmed direct binding of formononetin to the ERRα/PGC-1α complex, indicating allosteric stabilization. Finally, in human breast cancer patient-derived organoids, formononetin alone reduced viability and proliferation, and combined with doxorubicin further enhanced antitumor efficacy. ERRα downregulation is a causal early event in the pathogenesis of AIC. Formononetin acts as a first-in-class selective ERRα activator that improves cardiac metabolism and function while retaining anticancer activity, supporting its potential as a dual-action cardioprotective agent during anthracycline therapy.

In this study, superparamagnetic iron-carbon (Fe/C) nanoparticles (SPINs) engineered for alternating magnetic field (AMF)-based cancer therapy were synthesized using a co-precipitation approach with optimized precursor ratios. The resulting nanoparticles exhibited high magnetic performance, achieving saturation magnetizations of up to 149 emu/g, which are favorable for efficient AMF-mediated therapeutic activation. Consistent with this magnetic behavior, our previously published in vitro cytotoxicity studies demonstrated significant dose-dependent reductions in cell viability at relatively low nanoparticle concentrations, supporting their therapeutic potential. Based on these findings, in vivo AMF-mediated treatments were evaluated in DU145 prostate and MCF-7 breast cancer xenograft models using intratumoral SPIN injections at concentrations of 50 and 100 µg/mL. In the DU145 model, the 50 µg/mL dose produced gradual tumor regression, whereas the 100 µg/mL dose resulted in more rapid tumor reduction in responsive animals. Comparable dose-dependent therapeutic responses were observed in the MCF-7 model, with the higher SPIN concentration producing more pronounced tumor regression over the treatment period. Notably, thermal imaging revealed no detectable bulk heating during treatment, consistent with numerical modeling predictions, suggesting that the observed therapeutic efficacy occurred in the absence of conventional bulk hyperthermia. Collectively, these findings provide proof-of-concept evidence for achieving tumor regression without detectable bulk heating and support the potential of Fe/C SPINs as a promising platform for AMF-mediated cancer therapy. STATEMENT OF SIGNIFICANCE: • Alternating magnetic field (AMF)-driven superparamagnetic iron nanoparticles (SPINs) achieve significant tumor regression in two in vivo cancer models • Therapeutic effect arises from nanoscale magnetic dynamics, not bulk hyperthermia • Non-thermal mechanism challenges prevailing assumptions in magnetic hyperthermia • Findings establish a design paradigm for externally programmable nanotherapies.

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

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer with limited treatment options. Although PARP inhibitor (PARPi) offers great promise in treating TNBC with deficiency in homologous recombination (HR), most TNBC patients are HR-proficient. Furthermore, acquired resistance to PARPi remains as a challenge. Thus, there is an unmet need to identify new therapeutic target for developing advanced TNBC treatment strategy. I-SceI reporter assay and alkaline comet assay were used to analyze the role of Smyca in HR repair. Ingenuity pathway analysis was used to identify upstream regulators of Smyca-regulated transcriptome. RNA immunoprecipitation and RNA pull down were used to examine Smyca-FOXM1 interaction. Chromatin immunoprecipitation followed by sequencing was performed to identify FOXM1 target genes that are regulated by Smyca. Chromatin isolation by RNA purification was used to determine Smyca loading onto the promoters of FOXM1 target genes. Patient-derived organoid and xenograft mouse models were performed to evaluate the effect of Smyca on chemoresistance. Nanoparticle-assisted gapmer antisense oligonucleotides delivery was used to target Smyca in vivo. Co-culture of CD3 + T cells with TNBC cells and syngeneic mouse model were used to examine the effect of Smyca-FOXM1 targeting on anti-tumor immunity. The long non-coding RNA Smyca is highly expressed in TNBC. We show that Smyca is induced by genotoxic agents to enhance HR repair. Mechanistically, Smyca binds FOXM1 and promotes the recruitment of FOXM1 to the promoters of a set of HR and nucleotide metabolism genes, thereby promoting their expression. Smyca ablation induces BRCAness in HR-proficient TNBC, thereby sensitizing these tumors to platinum or PARPi. Furthermore, targeting Smyca-FOXM1 complex in combination with platinum or PARPi activates cGAS/STING pathway and tumor immunogenicity to enhance anti-tumor immune surveillance. Clinically, Smyca expression in breast cancer patients correlates positively with therapy resistance and negatively with HR deficiency, interferon signature, and infiltration of anti-tumor immune cells. Our study identifies an unprecedented role of Smyca in HR repair to promote TNBC survival and immune evasion in response to therapy and suggests Smyca as a potential target for sensitizing TNBC to chemotherapy, PARPi, or immunotherapy.

Long-term survival of extensive-stage small-cell lung cancer (ES-SCLC) remains rare, with most patients experiencing disease progression during maintenance therapy. Poly (ADP-ribose) polymerase (PARP) inhibitors have the potential to confer antitumor activity, modify tumor immunogenicity, and sensitize tumors to anti-programmed cell death protein 1/programmed death-ligand 1 therapy. We conducted this phase 2 trial to investigate the efficacy and safety of durvalumab plus olaparib as maintenance therapy in patients with ES-SCLC. This was a multicenter, single-arm, phase II trial that enrolled 60 patients with previously untreated ES-SCLC (NCT05245994). Patients received durvalumab (1,500 mg) combined with platinum-etoposide chemotherapy intravenously every 21 days for up to four cycles, followed by maintenance therapy with durvalumab (1,500 mg every 28 days) and oral olaparib (300 mg two times a day) until disease progression or unacceptable toxicity. Multi-omics analyses were performed to characterize molecular subtypes associated with clinical outcomes. The combination regimen demonstrated promising efficacy, with an alive and progression-free at 12 months rate of 25.0%, an objective response rate of 73.3%, a median progression-free survival of 6.8 months, and a median overall survival of 14.6 months. Multi-omics profiling identified a hypomethylation subgroup (cluster 1) that was associated with significantly improved survival outcomes. Further analysis revealed that this subtype exhibited enhanced antigen presentation machinery, a favorable cytokine profile, and suppression of DNA damage repair (DDR) pathways, potentially through elevated promoter methylation and transcriptional silencing of specific DDR genes, which together were associated with the favorable outcomes. This study presents the first prospective evidence supporting durvalumab plus olaparib as maintenance therapy in ES-SCLC. Multi-omics analysis identifies that DNA hypomethylation status may enrich for patients who benefit from PARP inhibition and immunotherapy. NCT05245994.

Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a rare yet highly aggressive subtype of non-small cell lung carcinoma (NSCLC) with limited therapeutic options. We conduct a comprehensive proteogenomic analysis of LCNEC using tumors and paired normal adjacent tissues from 107 patients (81 pure LCNEC and 26 combined LCNEC). APOBEC mutational signatures strongly correlate processes of tumor initiation and immune suppression, and KEAP1 mutations correlate with metabolic reprogramming in LCNEC combined with NSCLC. A conflicting relationship is observed between neuroendocrine and immune phenotypes. We identify three LCNEC subtypes with unique prognosis features, microenvironment dysregulation, genetic alterations, and potential therapeutic targets. Interleukin-33 (IL-33) emerges as a critical therapeutic biomarker associated with enhanced T cell infiltration and antitumor activity. We further optimize recombinant IL-33 (rIL33) with site-directed mutagenesis and develop PEGylated rIL33, demonstrating its prolonged circulation time in cynomolgus monkeys and superior immune agonist activity in mouse models. Overall, this study offers insights into LCNEC biology and provides promising innovative immunoagonist therapy strategies for lung cancer.

In this issue of Cancer Cell, Li et al. report a first-in-human phase 1 study of SYS6010, an epidermal growth factor receptor (EGFR)-targeting antibody-drug conjugate, in advanced solid tumors. The study highlights how EGFR-directed therapy moves beyond kinase inhibition toward antigen-guided cytotoxic delivery in non-small cell lung cancer.

SYS6010 is an antibody-drug conjugate targeting epidermal growth factor receptor (EGFR). We report the results of a phase 1 trial (ChiCTR2300072141) of SYS6010 in patients with non-small cell lung cancer (NSCLC). A total of 236 patients were treated. One dose-limiting toxicity occurred at 6.4 mg/kg; therefore, 4.2, 4.5, and 4.8 mg/kg were selected for cohort expansion. Treatment-related adverse events (TRAEs; any/grade ≥ 3) occurred in 99.6%/57.2% of patients. Common grade ≥3 TRAEs included neutropenia (30.9%), leukopenia (25.0%), and thrombocytopenia (17.4%). Objective response rate was 34.7% in EGFR-mutant NSCLC treated with EGFR tyrosine kinase inhibitors (TKIs) and platinum chemotherapy, 45.7% in EGFR-mutant NSCLC treated with EGFR TKIs, 20.0% in EGFR wild-type squamous NSCLC, and 35.7% in EGFR wild-type non-squamous NSCLC. Median progression-free survival and overall survival were 7.6 and 19.4 months, respectively, in EGFR-mutant NSCLC treated with EGFR TKIs and platinum chemotherapy. Overall, SYS6010 shows a manageable safety profile and encouraging antitumor activity in previously treated, advanced NSCLC.

Mefatinib, a novel second-generation epidermal growth factor (EGFR) tyrosine kinase inhibitor that has shown promising antitumor activity in targeting non-small cell lung cancer (NSCLC) with common and uncommon EGFR-activating mutations. In this phase III, randomized, double-blind trial in China, 336 eligible patients with advanced nonsquamous NSCLC harboring EGFR L858R or exon 19 deletion (ex19del) were assigned (2:1) to receive either mefatinib (60 mg daily, n = 223) or gefitinib (250 mg daily, n = 113). The primary endpoint was progression-free survival (PFS), assessed by an independent review committee (IRC). The trial is registered with chinadrugtrials.org.cn (CTR20192297). After a median follow-up of 15.9 months for mefatinib and 18.5 months for gefitinib, mefatinib demonstrated a significantly longer median IRC-assessed PFS compared to gefitinib (13.7 vs. 9.7 months; hazard ratio [HR] = 0.68; 95% confidence intervals [CI]: 0.53-0.87; p = 0.002). The 30-month overall survival rate was 60.2% for mefatinib and 54.3% for gefitinib. Patients with EGFR ex19del had comparable PFS for both treatment arms (p > 0.100), whereas patients with EGFR L858R had significantly longer median PFS when treated with mefatinib than gefitinib (13.7 vs 8.3 months HR = 0.55 [95% CI: 0.38-0.78]; p = 0.001). Patients with EGFR L858R had a 30-month overall survival rate of 56.6% with mefatinib and 43.7% with gefitinib. Treatment-related adverse events ≥grade 3 were reported in 45.7% of the mefatinib group and 24.8% of the gefitinib group. No new safety signals were observed for mefatinib. Mefatinib demonstrated superior efficacy to gefitinib with a similar tolerability profile in the first-line treatment of EGFR-mutated advanced NSCLC.

Bone metastasis (BM) frequently occurs in various types of cancer, particularly in prostate (PCa), breast (BCa), renal (RCC), and lung cancers (LCa), yet no reliable biomarker has been established. In this study, we evaluated the clinical utility of serum growth differentiation factor 15 propeptide (sGDPP), secreted by both osteoblasts and osteoclasts, as a diagnostic biomarker for BM across common solid tumors. A total of 799 participants were enrolled, including 107 healthy donors, 242 patients with PCa, 113 patients with BCa, 159 patients with RCC, and 178 patients with LCa. Among the cancer patients, 398 had no BM and 294 had BM. The diagnostic performance for BM was compared among conventional biomarkers: alkaline phosphatase (ALP), lactate dehydrogenase (LDH), estimated glomerular filtration rate, osteocalcin, bone-specific alkaline phosphatase, tartrate-resistant acid phosphatase 5b, procollagen type I N-terminal propeptide (PⅠNP), and sGDPP. Among all patients, ALP, LDH, PⅠNP, and sGDPP levels were significantly higher in patients with BM than in those without BM. Particularly, sGDPP showed the highest area under the curve values (0.91 in PCa, 0.80 in BCa, 0.81 in RCC, and 0.51 in LCa). Multivariate analysis showed sGDPP is an independent diagnostic biomarker for BM in PCa, BCa, and RCC (p < 0.01). Additionally, sGDPP in patients with osteoporosis did not significantly differ from that in healthy donors, suggesting that sGDPP increases only when tumor cells seed and proliferate in the bones. Overall, sGDPP demonstrated superior diagnostic performance compared to conventional biomarkers and may complement imaging tests in detecting BM.

Perioperative nivolumab significantly improved event-free survival (EFS) compared with placebo in patients with resectable non-small cell lung cancer (NSCLC) in the CheckMate 77T study (ClinicalTrials.gov NCT04025879 )1. Here, after randomization, 98 out of 229 patients who received nivolumab and 92 out of 232 patients who received placebo had evaluable biomarkers (41% of randomized patients). Of the 98 patients receiving nivolumab, 83 (85%) had detectable circulating tumour DNA (ctDNA) before initiating neoadjuvant treatment and 90 (92%) at neoadjuvant treatment completion. Of the 92 placebo-treated patients, 75 (82%) had detectable ctDNA at the treatment start and 78 (85%) at completion. Among the 98 nivolumab-treated patients, 76 (78%) had detectable and evaluable ctDNA before and after neoadjuvant treatment, and 50 of them (66%) had pre-surgical ctDNA clearance, and 25 out of 50 (50%) had pathologic complete response (pCR). For the placebo-treated group, these values were 64 out 92 (70%) for detectable and evaluable ctDNA before and after neoadjuvant treatment, and 24 out of 64 (38%) had pre-surgical ctDNA clearance, and 3 out of 24 (12%) had pCR. Furthermore, 4 out of 48 (8%) patients in the nivolumab group and 9 out of 44 (20%) in the placebo group who were negative for molecular residual disease (MRD) after surgery and before adjuvant treatment initiation became positive during the adjuvant treatment period; all had disease recurrence. EFS seemed to be prolonged with nivolumab (n = 60) versus placebo (n = 45) in patients with single or co-alterations in any of the KEAP1, STK11, CDKN2A and/or SMARCA4 driver genes (hazard ratio, 0.48; 95% confidence interval, 0.28-0.83). In a machine-learning model trained using biomarker-evaluable patients, top predictors of prolonged EFS included pre-surgical ctDNA clearance, non-N2 NSCLC, pCR, squamous tumour histology and nivolumab treatment. These findings provide insights into predictive markers for outcomes with perioperative nivolumab in resectable NSCLC.

Ultra-low-pass whole-genome sequencing (ULP-WGS) of cell-free DNA (cfDNA) offers a cost-efficient strategy for cancer detection, but its clinical application is limited by extreme data sparsity and poor model generalization. We developed Fragmentia-AI™ WGS, a mutation-calling-independent framework that uses a transformer-based multiple-instance learning architecture with sequential fine-tuning across tumor fraction (TF) strata to extract latent cancer-associated signals from ULP-WGS data. Model performance was evaluated in multiple independent cohorts, including a pan-cancer test set covering 17 cancer types, an external public dataset generated on a different sequencing platform, and a technical variability cohort with heterogeneous pre-analytical and experimental conditions. Clinical relevance was assessed by correlating model predictions with progression-free survival (PFS) in patients with advanced non-small cell lung cancer receiving chemoimmunotherapy. Sequential fine-tuning across TF strata significantly improved performance in low-TF samples, achieving a 35.6% relative increase in AUC compared with high-TF-only training (0.884 vs. 0.652). In the independent test cohort, the model achieved an overall AUC of 0.930, with consistent performance across TF strata and cancer types. External validation confirmed robust cross-platform generalizability (AUC: 0.929; sensitivity: 0.78; specificity: 0.92). The model maintained stable classification performance despite score fluctuations associated with pre-analytical and technical variables. Importantly, model-negative status, defined as a prediction score below the training-derived cutoff, remained significantly associated with improved PFS compared with model-positive status (HR = 0.49, 95% CI: 0.29-0.82) after multivariable adjustment. Collectively, this framework enables robust cancer detection and clinically meaningful risk stratification from highly sparse cfDNA sequencing data.

Air pollution is a leading environmental cause of lung cancer, yet the underlying biological pathways remain poorly understood. Identifying circulating biomarkers that capture early molecular responses may clarify how air pollutants contribute to carcinogenesis and help identify individuals at elevated risk. We conduct a prospective nested case-control study within two Cancer Prevention Study cohorts, profiling more than 1100 metabolites in pre-diagnostic plasma samples from 1357 participants. Residential concentrations of six major air pollutants are estimated at the time of blood draw. Here we show that eight circulating metabolites are associated with both air pollution exposure and subsequent lung cancer risk. Four metabolites, including γ-glutamylglutamine, phenylacetylglutamate, N-(2-furoyl)glycine, and 4-vinylguaiacol glucuronide, significantly mediate associations for particulate matter and ozone (adjusted q-value < 0.2). These findings suggest that air pollution may promote lung cancer partly through metabolic pathways related to inflammatory and oxidative processes, providing key insights into potential mechanisms and targets for prevention.

Cell targeting/permeabilization, organelle/biochemical pathway regulation, and drug resistance/metastasis/immunological expressions are considerations to advance cancer nanomedicine design. This study modulated mitochondria-targeting YKWYYRGAA (P1) peptide, into a multifunctional excipient via N-methylation and N-dimethylation, to synergise nano-chitosan conjugate in drug delivery and non-small cell lung cancer treatment. The spray-dried chitosan nanoparticles developed from P1, N-methylated YKWYYRGAA (P2) and N-dimethylated YKWYYRGAA (P3) were subjected to physicochemical testing, NRAS-mutated H1299 cell permeability/cytotoxicity/apoptosis and cell cycle arrest/drug resistance/metastasis/immunomodulation assessment, and in vivo pharmacokinetics/pharmacodynamics investigations. Methylated P2 increased cancer cell permeability/intracellular drug/nanoparticle uptake/drug targeting via sustained- and pH-stimuli responsive release and cytotoxicity unlike P1 and P3 which were ceased at membrane interface by excessive ionic/hydrophobic interactions. P2-grafted nanochitosan induced mitochondria-mediated apoptosis with minimal necrosis via ROS activation and FasL-linked death. It suppressed mTOR/MAPK signalling overcoming EGFR-resistance/EGFR mutation-independent pathways in tumorigenesis. It mitigated drug resistance via downregulating P-gp (efflux receptor) and GSTP1 (degrading enzyme) expressions, and epithelial-mesenchymal transition via interplay of E-cadherin against N-cadherin/snail/twist 1/vimentin/ezrin/MMP9 with MICA/ULBP1 suppression to reduce immunological lung tissue lysis. The inhaled P2-grafted nanochitosan provided a positive lung cancer recovery with reduced systemic exposure and hematological/biochemical toxicities. Single instead of dimethylation of YKWYYRGAA promoted the cascades of inter-dependent anti-cancer activities and efficacy of nanochitosan.

Developing novel strategies that can exploit a cell's endogenous machinery to selectively release toxic compounds in diseased cells, while leaving healthy cells unaffected, is a challenging task to accomplish. An established approach involves antigen recognition on tumor cells, enabling targeted delivery of prodrugs in which a cytotoxic compound is appended to an antibody by a cleavable linker. However, this strategy depends on the presence of highly expressed surface markers for which antibodies can be developed, limiting its applicability to cancers that display these markers. A new approach that could rely on disease-specific expression of an intracellular enzyme, which can be more broadly targeted, could expand the therapeutic potential of prodrugs clinically. Here, we demonstrate that the immunoproteasome, an isoform of the standard proteasome that is upregulated under conditions of inflammation, has the potential to be harnessed as a prodrug release enzyme. We conjugate the highly toxic and widely used therapeutic cargo MMAE onto an immunoproteasome-selective peptide and demonstrate its selective release in cancerous cells at low nM to pM concentrations, while healthy cells remain viable. We also establish the first translational validation that using immunoproteasome prodrugs in vivo leads to a significant reduction in tumor volume without significant toxicities in small cell lung cancer. We anticipate this approach to be broadly applicable, as the immunoproteasome is upregulated in a variety of cancers and does not require antibody recognition to be effective.

Early lung tumorigenesis depends not only on mutant clones but also on tumor-promoting niches that remain difficult to detect. In Cell, Pandya et al. identify a 14-protein plasma signature that predicts lung cancer years before diagnosis, report interleukin-1β-linked alveolar remodeling, and position tumor promotion as a target for precision prevention.

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

Pulmonary delivery of lipid nanoparticle (LNP)-based mRNA vaccines offers a promising strategy for localized lung cancer immunotherapy, yet how distinct pulmonary administration routes determine cellular targeting and therapeutic efficacy remains poorly understood. Here, we systematically evaluate intranasal and intratracheal delivery of mRNA-LNP vaccines and reveal a route-dependent immunological mechanism governing lung-targeted tumor vaccination. Although intratracheal administration yields only a 2.6-fold increase in total pulmonary protein expression compared to intranasal delivery, it produces a striking 26.8-fold enhancement in functional mRNA transfection efficiency within alveolar macrophages, the dominant antigen-presenting cell population in the alveolar space. This selective targeting reprograms alveolar macrophages toward an activated antigen-presenting phenotype, promoting efficient antigen presentation, robust CD8+ T-cell responses, and superior prophylactic and therapeutic efficacy in pulmonary tumor models. Notably, local depletion of alveolar macrophages completely abolishes the antitumor protection conferred by intratracheal vaccination, establishing their indispensable role in mediating pulmonary mRNA vaccine efficacy. Together, these findings provide mechanistic insights into lung-targeted mRNA cancer vaccination driven by alveolar macrophage engagement, providing critical insights for the design of next-generation LNP-based nanomedicines for lung cancer immunotherapy.

Cutaneous melanoma is rare in Patients of Color (POC), and the genomics are not well understood. Describe MAPK drivers and tumor mutational burden (TMB) of cutaneous and unknown primary (CUP) melanoma in POC and compare them to non-Hispanic White (NHW) patients. We analyzed a retrospective convenience cohort of 676 patients with advanced CUP melanoma undergoing clinically-warranted multigene sequencing with MSK-IMPACT. Self-identified (self-ID) POC were defined as non-White race and/or Hispanic ethnicity; this cohort was also analyzed by genetically inferred ancestry and skin tone using Monk scale. Self-ID POC (n=35) had frequent BRAF V600E mutations (43%) and, compared to NHW (N=641) patients, more "pan-wildtype" tumors of unknown driver (14% vs 4%, p=0.010). Among self-ID POC, skin tone and inferred ancestry showed moderate overlap, but did not consistently predict genomic features. Median TMB was higher for patients with light (N=185) vs medium/dark (N=8) skin tone (15.8 vs 2.6, p<0.0001). Skin tone assessments were only available for a subset of patients (N=193). BRAF mutations are underestimated given molecular testing preferentially sent in patients with BRAF-wildtype disease. CUP melanomas in self-ID POC had lower median TMB and were more likely to be pan-wildtype, reflecting differences in pathogenesis. Larger scale studies are needed to validate these findings.

Primary human myeloid cells hold promise for immunotherapies, yet efficient, scalable technologies for engineering and screening in these cells remain limited. Here we present a virus-like particle (VLP)-based toolkit that delivers diverse CRISPR editing modalities to human monocytes, macrophages and dendritic cells with high efficiency while preserving viability and innate immune responsiveness. VLP-mediated delivery of ribonucleoproteins supports gene knockout, base editing and epigenetic silencing. Combined with adeno-associated virus-mediated donor delivery, this approach enables site-specific integration of large DNA sequences by homology-directed repair. We developed SLICeVLP, which pairs sgRNA delivery by VPX-lentivirus with Cas9 protein delivery by engineered VLPs, and used it for pooled loss-of-function and Perturb-seq screens in human macrophages. We uncovered regulators of tumor necrosis factor (TNF) and CD80 expression, converging on TNFAIP3 as a central regulator of inflammatory polarization. TNFAIP3 ablation drove a proinflammatory state resistant to suppressive repolarization and enhanced cytotoxicity in chimeric antigen receptor macrophages. This system enables unbiased functional genomics in primary human myeloid cells, with implications for myeloid cell therapy design.

Canonical and noncanonical autophagic processes are integrated with innate and adaptive immunity and sterile or pathogen-induced inflammation. In canonical autophagy, double-membrane autophagosomes modified by ubiquitin-like ATG8 proteins in a process termed membrane atg8ylation sequester and eliminate intracellular targets such as invading microbes, defunct organelles, aggregates, and inflammatory molecules. Recently, a plethora of noncanonical processes that entail membrane atg8ylation of various intracellular organelles other than autophagosomes have been linked to immunity. This has led to confounding interpretations and conflation of diverse processes as autophagy. Here, we posit that these are divergent manifestations of a common ancestral homeostatic process of membrane atg8ylation and provide an overview of how they affect immunity and inflammation. These relationships are evident in model organisms and are reflected in human genetic predispositions to diseases with immune components. The membrane atg8ylation pathways affect acute and chronic inflammation, infections, autoimmunity, cancer, neurodegeneration, metabolic syndrome, diabetes, and other disorders.

Amomum tsaoko (AT) is mainly dried through natural drying (ND), open fire drying (OFD), air-source drying (ASD), or hot-air drying (HAD). To investigate the effects of drying methods on AT odorants and bioactivity, sensory science and network pharmacology methods were employed. Results indicated that the numbers of alcohols in HAD AT and alkenes in ND AT were the highest; the odorant concentration in ASD AT was the highest, whereas that in OFD AT was the lowest. (Z)-2-Decenol, germacrene D, 4-terpineol and neral were identified as differential markers. The bioactivity research outcomes on AT essential oil (ATEO) indicated that both HAD and ASD ATEO exhibited greater α-amylase and α-glucosidase activity inhibitory capabilities; ASD ATEO showed the highest inhibition rate (>97%) to A549 cancer cell and the strongest total reducing power; HAD ATEO had the highest inhibition rate (>85%) against SW872 cancer cells; ND ATEO exhibited the highest radical-scavenging efficiencies.

Precise imaging of intracellular biomarkers is crucial for accurate disease diagnosis and mechanistic insight. Although confined catalytic hairpin assembly (CCHA) can accelerate reaction kinetics, its performance is frequently compromised by nonspecific amplification, resulting in high background signals and limited imaging contrast. Here, we develop a tumor-upregulated endogenous apurinic/apyrimidinic endonuclease 1 (APE1)-triggered CCHA (AP-CCHA) system based on a DNA nanowire for selective microRNA (miRNA) imaging in living tumor cells. Using miR-21 as a proof-of-concept model, the mechanistic analyses demonstrated that conventional CCHA suffers from significant nonspecific signal amplification during both in vitro assembly and storage. For the proposed AP-CCHA system, we incorporated an APE1-activatable AP-H1 probe that improves molecular stability, suppresses nonspecific amplification, and consequently enhances the signal-to-noise ratio while preserving high reaction kinetics. As a result, the AP-CCHA system delivers a 3.5-fold improvement in sensitivity compared with CCHA and achieves an 18.2 pM detection limit for miR-21. Leveraging the elevated cytoplasmic APE1 levels in tumor cells, AP-CCHA remains quiescent in normal cells but is selectively activated in cancer cells, enabling high-contrast miRNA discrimination and precise in vivo tumor imaging, and effectively minimizing false positives. By integrating confinement-enhanced kinetics with endogenous enzyme activation, this strategy enables rapid, highly sensitive, and high-contrast biomarker imaging. The AP-CCHA platform thus provides a versatile analytical tool for nucleic acid-based diagnostics with strong potential for precision therapeutics and drug development applications.

Melanoma remains highly aggressive and refractory to conventional therapies, with current treatments often limited by toxicity and acquired resistance, underscoring the urgent need for targeted strategies with improved safety and durable efficacy. Here, we report a tyrosinase-guided cyclic peptide, c-RGDKYQ, that exploits the overexpression of tyrosinase in melanoma cells to trigger intracellular oxidation and in situ assembly of supramolecular nanostructures. These enzyme-instructed nanosystems selectively destabilize the actin cytoskeleton, thereby suppressing B16 melanoma cell migration, adhesion, and proliferation, and inducing apoptosis without requiring exogenous drug payloads or complex delivery carriers. This selective activity is strictly dependent on tyrosinase expression, as the linear control peptide lacking conformational constraint showed minimal cytotoxicity. In a murine B16 tumor model, c-RGDKYQ exhibits potent antitumor activity with minimal off-target effects and favorable tolerability. Transcriptomic profiling further confirms cytoskeletal collapse, evidenced by coordinated downregulation of actin/tubulin genes and compensatory upregulation of regulatory factors. This work establishes a proof-of-concept for a minimalist, enzyme-responsive peptide platform that achieves targeted melanoma therapy through physical cytoskeletal disruption. Beyond melanoma, this paradigm of enzyme-directed supramolecular assembly may offer a translatable framework for treating other malignancies characterized by dysregulated metabolic enzymes.

Head and neck squamous cell carcinoma (HNSCC) shows substantial intra- and inter-tumoral heterogeneity. We mapped tumor architecture across HPV-positive and HPV-negative HNSCC through spatial transcriptomics (n = 26). HPV-positive tumors display hypercellularity, higher lymphocyte presence, enriched hypoxia and reduced partial epithelial-to-mesenchymal transition (p-EMT) in malignant cells. We observed two distinct spatial architectures of p-EMT: p-EMT edge, where p-EMT is coupled to fibroblasts at the invasive front via TGFβ, and p-EMT core, in which tumor-infiltrating immunosuppressive macrophages and neutrophils induce p-EMT via oncostatin M in the core of tumor nests. These two p-EMT patterns were consistent across multiple samples from the same tumor, suggesting they are tumor-wide features. Together, these findings reveal that distinct interactions in the tumor microenvironment converge on a similar p-EMT cellular phenotype, but in a different spatial pattern that may have potential biological and clinical implications for our understanding of invasion, immune modulation and new targeted therapeutics for HNSCC.

Uveal melanoma is an aggressive tumor type that frequently results in fatal liver metastasis. A better understanding of the interactions between uveal melanoma and host cells in the tumor microenvironment that promote liver metastasis could lead to the development of therapeutic strategies. In this study, we used single-cell RNA-sequencing analysis of uveal melanoma-hepatic stellate cell (HSC) cocultures to explore the mechanisms by which uveal melanoma cells metastasize to the liver. HSCs enriched for uveal melanoma cell states that expressed genes implicated in cell survival, metabolic reprogramming, and growth differentiation factor 15 (GDF15)-mediated angiogenesis. The TGFβ family member GDF15 was associated with a metastatic uveal melanoma phenotype. Silencing of BAP1 in uveal melanoma cells led to increased GDF15 expression and accumulation of H3K27ac marks at the GDF15 promoter. Treatment of HSCs with GDF15 led to increased expression of extracellular matrix (ECM) proteins, inflammatory cytokines, and angiogenic factors, including IL8. Both exogenous GDF15 and IL8, as well as conditioned media from uveal melanoma-HSC cocultures, increased endothelial cell network formation in vitro, an effect that was blocked by anti-GDF15 antibodies. In multiple models of metastatic uveal melanoma, silencing of GDF15 inhibited the outgrowth of metastatic lesions, associated with reduced deposition of ECM and recruitment of endothelial cells. Together, these findings reveal that uveal melanoma liver metastasis development is dependent on GDF15-mediated remodeling of the liver microenvironment, which leads to an angiogenic response and matrix deposition that supports tumor growth. GDF15 functions as a growth factor that helps drive the development of liver metastases in uveal melanoma by contributing to angiogenesis and matrix deposition, allowing for metastatic outgrowth.

The unclear regulatory and transport mechanisms of Kirsten rat sarcoma viral oncogene homolog (KRAS) limit a comprehensive understanding of KRAS activity and the development of inhibitors. Wang et al. reported that farnesylation-induced KRAS phase separation drives colon cancer progression and resistance to G12C inhibitors. Statins can block this process, suppress tumor growth, and reverse drug resistance.

Gut microbial metabolites, particularly short-chain fatty acids (SCFAs), bile acid (BAs) derivatives, and tryptophan-derived metabolites (TDMs), act as signaling molecules, receptor ligands, and epigenetic modulators in regulating host metabolic and immune homeostasis. They exert local effects in the gut and reach distant tissues via hepatic, lymphatic, neural, and immune-hormonal routes, thereby regulating immunity, metabolism, and neural function. Disruptions in these pathways are associated with inflammatory diseases, neurodegeneration, and cancer. This review discusses the importance of SCFAs, BAs, and TDMs in different host metabolic and immunological contexts, dissects their molecular mechanisms of action on target cells, evaluates the therapeutic implications of targeting these microbial pathways, and outlines a roadmap for possible future clinical translations.

Male breast cancer is a rare but increasingly recognized disease with limited data on modifiable risk factors. Specifically, it is not known if the inverse association between cardiorespiratory fitness (CRF) and breast cancer risk reported in women is similar to the association between CRF and breast cancer in men. The study aimed to evaluate the association between CRF and the incidence of male breast cancer in a large, nationally representative cohort of U.S. Veterans. We analyzed data from 777,618 male Veterans who completed an exercise treadmill testing (ETT) between 1999 and 2024 within the Veterans Health Administration and had no evidence of breast cancer prior to the ETT. CRF was expressed in peak metabolic equivalents (METs). The cohort was categorized into 4 age-adjusted CRF categories based on the peak METs achieved. The primary outcome was incident male breast cancer, ascertained through the Veterans Affairs Computerized Patient Record System. Cox proportional hazards models were used to estimate hazard ratios (HRs) for breast cancer incidence across CRF categories, adjusted for age, race, body mass index, hypertension, diabetes (T2DM), chronic kidney disease, smoking, and alcohol use. Over a median follow-up of 10.7 years 518 men were diagnosed with breast cancer. After multivariable adjustment, age (HR = 1.24, 95% confidence interval (95%CI): 1.14-1.36) per decade, chronic kidney disease (HR = 2.13; 95%CI:1.41-3.23), hypertension (HR = 1.57; 95%CI: 1.28-1.92), T2DM (HR = 1.24; 95%CI: 1.02-1.51), and poor CRF (HR = 1.69; 95%CI: 1.24-2.30) were the strongest predictors of breast cancer. Compared to patients in the lowest CRF category, breast cancer risk was 41% lower for individuals in the highest CRF quartile (HR = 0.59; 95%CI: 0.44-0.81). Higher CRF was independently associated with a substantially lower risk of breast cancer in men. These findings suggest that CRF may represent a modifiable risk factor in the prevention of male breast cancer, underscoring the broader importance of physical fitness in reducing cancer risk across sexes.

Breast cancer (BC) mortality risk extends over decades; yet, the temporal prognostic patterns of histological grade across subtypes and stages remain unclear. Women with stage I-III BC diagnosed between 2000 and 2018 were identified from Surveillance, Epidemiology, and End Results registries (N = 767 218). Breast cancer-specific mortality (BCSM), as the primary endpoint, was analyzed using cumulative incidence functions including competing risk and annual hazard rates stratified by grade, estrogen receptor (ER) status, stage, and adjuvant chemotherapy. Restricted mean survival time differences were calculated to quantify absolute survival differences across follow-up intervals. Landmark survival analyses were carried out for 0 to <60, 60 to <120, and 120 to <180 months from diagnosis. Time-varying effects of grade were assessed through interval-specific modeling to evaluate nonproportional hazards. Overall, 23%, 44%, and 33% of cancers were grades 1, 2, and 3, respectively, accounting for 8%, 35%, and 57% of BC deaths. In each consecutive follow-up period, the contribution of grade 1 cancers to BCSM increased, and the contribution of grade 3 cancers decreased. Overall, 59% and 30% of deaths from grades 1 and 3 cancers, respectively, occurred after 5 years. In ER-positive disease, grade 3 tumors showed peak annual hazards between years 3 and 5, whereas grade 1 tumors had lower but sustained hazards extending beyond 10 years; the hazard curves converged between years 10 and 12. In ER-negative disease, early hazards were higher, and convergence also started earlier at years 5-8. Absolute risk was influenced by nodal status and tumor stage within each grade category and follow-up interval. Grade 1 cancers show sustained risk beyond 10 years, whereas grade 3 cancers exhibit front-loaded risk. These temporal risk patterns, together with other clinical and genomic data, could inform extended endocrine therapy decisions and surveillance strategies.

Inflammasome activation has emerged as a promising therapeutic target to reduce residual cardiovascular (CV) risk in patients on guideline lipid-lowering therapy. IL-1β and IL-6 have been intensively studied, whereas the role of the inflammasome-related cytokine IL-18 has gained less attention. Here we aimed to assess the associations of IL-18 with risk of coronary events and severity of atherosclerosis. The study included 4742 participants participating in a prospective Swedish Malmö Diet and Cancer (MDC) cohort study, 54 219 general population participants in the UK Biobank, and 1500 participants with Type 2 diabetes and/or known CV disease in the European multicentre SUMMIT VIP imaging study. Plasma levels of IL-18 were analysed using proximity extension assay and atherosclerosis by carotid ultrasound. The incidence of MI and CV death was registered during a 23-year follow-up for the MDC cohort and for a 15-year follow-up for the UK Biobank cohort. There were 617 cases of MI and 644 CV deaths in the MDC cohort and 2454 cases of MI and 1537 CV deaths in the UK Biobank cohort. Elevated IL-18 was associated with an increased risk of MI and CV death independent of hsCRP and IL-6 in both cohorts. In Cox proportional hazards regression models, this association was independent of major CV risk factors in the UK Biobank but not in the MDC cohort. IL-18 did not provide clinically meaningful risk reclassification in the UK Biobank cohort. The association of IL-18 with carotid atherosclerosis in the SUMMIT VIP cohort was weaker than that of IL-6. IL-18 is independently associated with risk of MI and CV death in the general population but does not improve clinical risk stratification. Targeting IL-18 directly or through inflammasome inhibition represents a promising approach for treatment of residual inflammatory risk in high-risk atherosclerosis patients that merits to be evaluated in future randomized clinical trials.

In ARANOTE (NCT04736199), darolutamide plus androgen-deprivation therapy (ADT) significantly reduced the risk of radiologic progression or death versus placebo plus ADT in patients with metastatic hormone-sensitive prostate cancer, with favorable safety. Here, we report on post hoc outcomes by disease volume/risk. Randomized patients received darolutamide plus ADT or placebo plus ADT (2:1). High-volume (HV) and high-risk (HR) disease were defined by the CHAARTED and LATITUDE criteria, respectively. End points included radiologic progression-free survival (primary; rPFS), time to initiation of subsequent systemic anticancer therapy, time to metastatic castration-resistant prostate cancer, time to prostate-specific antigen (PSA) progression, time to pain progression, PSA <0.2 ng/ml rates, overall survival, and safety. Of 669 patients, 472 had HV disease, 197 had low-volume (LV) disease, 400 had HR disease, and 269 had low-risk (LR) disease; baseline characteristics were generally balanced. Darolutamide consistently reduced the rPFS (HV: 40% [hazard ratio, 0.60; 95% confidence interval, 0.44 to 0.80]; LV: 70% [0.30; 0.15 to 0.60]; HR: 39% [0.61; 0.44 to 0.86]; LR: 60% [0.40; 0.25 to 0.62]), with similar trends for all secondary end points versus placebo. Treatment-emergent adverse events were similar between treatment arms across all subgroups. Darolutamide was well tolerated, and outcomes were improved versus placebo, regardless of disease volume/risk. Prior presentation: Presented at the American Society of Clinical Oncology Genitourinary (ASCO GU) Cancers Symposium, San Francisco, CA, February 13-15, 2025. Clinical trial registration: NCT04736199 (EudraCT 2020-003093-48).

The poor prognosis of cholangiocarcinoma (CCA) is largely driven by rapid, asymptomatic disease progression, which usually results in a late diagnosis in the absence of established screening strategies. An early, cost-effective, and universally applicable risk assessment strategy would therefore be valuable. We developed machine learning (ML) models on prospective, multimodal data from 487,495 UK Biobank (UKB) participants, of whom 649 developed CCA during follow-up. Data from England (80%) were utilised for ML development via five-fold cross-validation, and then all models were tested on withheld data from Scotland, Wales, and Newcastle (20%). Iterative ablation studies reduced inputs from >150 features across demographic data, lifestyle, health records, blood parameters, genomics, and metabolomics to models built on five and ten routinely available clinical parameters. These were externally validated in the Penn Medicine Biobank (PMBB; n = 2638; 28 CCA), All of Us Research Program (AOU; n = 330,433; 362 CCA), Japan Medical Data Centre Claims Database (JMDC; n = 8,425,522; 723 CCA) and TriNetX (n = 728,886; 1592 CCA). We show that ML models integrating biliary-disease associated health records and Gamma glutamyltransferase can stratify risk of future CCA. Evaluation on the UKB test set as well as three independent cohorts revealed robust performance and generalisability across ethnicities. We achieved AUROCs of 0.71 [95% CI: 0.703-0.711], 0.77 [95% CI: 0.764-0.778 ], 0.796 [95% CI: 0.795-0.798] and 0.8 [95% CI: 0.794-0.805] for UKB, PMBB, AOU, and JMDC respectively, with respective AUPRCs of 0.014 [95% CI: 0.009-0.018], 0.042 [95% CI: 0.037-0.048], 0.038 [95% CI: 0.033-0.042] and 0.001 [95% CI: 0.001-0.001]. In AOU, application of the Youden J-optimised threshold yielded a number needed to screen of 79. Separate models for intra- and extrahepatic CCA did not improve performance. In line with the pathophysiology, performance declined for longer intervals between assessment and event. A group-level analysis in the TriNetX cohort revealed hazard ratios of up to 82.5 [95% CI: 26.4-257.96]. We provide extensive interpretability results and release all source codes used to develop the presented models. We provide a comprehensive framework for early CCA risk stratification in the general population, identifying key predictors, and demonstrating the potential of data-driven models in personalised screening for hepatobiliary cancer. German Cancer Aid (grant #70115730), Junior Principal Investigator Fellowship programme of RWTH Aachen Excellence strategy.

The size of tumour-negative regional lymph nodes (LNnegs) may be associated with the host antitumour immune response in patients with oesophagogastric adenocarcinoma (OGAC). In the OE02 trial, larger LNnegs were independently associated with improved overall survival (OS); however, the prognostic relevance of LNneg size has not been validated in an independent OGAC population. Resection specimens from 1367 patients from two phase III trials (OE05, n = 652; ST03, n = 715) were analysed. The largest LNneg per patient was identified on digitised slides stained with haematoxylin-eosin using an automated segmentation pipeline. Lymph node ratio [LNratio; number of metastatic lymph nodes (LNs) divided by the total number of resected LNs] was calculated for patients with stage ypN1+ cancer. Anthracotic pigment burden was quantified using a deep learning model. Associations among LNneg size, primary tumour location, recurrence, and survival were evaluated. LNneg size and pigment burden differed by primary tumour location. LNnegs from oesophageal/junctional cancers were larger and had greater pigment burden than those from gastric cancers (all P < 0.0001). Larger LNnegs were associated with a lower recurrence rate (P = 0.033) and lower LNratio (P < 0.0001). Patients with LNnegs with a long axis diameter >10 mm had improved OS [hazard ratio (HR) 0.81, 95% confidence interval (CI) 0.70-0.93, P = 0.0147) and progression-free survival (HR 0.80, 95% CI 0.70-0.92, P = 0.0092). Presence of large LNnegs was independently prognostic after multivariable adjustment. This study validates LNneg size as a novel independent prognostic biomarker in patients with locally advanced OGAC treated with cytotoxic chemotherapy and surgery. Variation in LNneg size and pigment burden by primary tumour location may suggest anatomical or microenvironmental differences in LNneg biology that may affect LN assessment and immune-related characteristics. Further studies integrating quantitative microarchitectural and immune profiling of LNnegs are warranted to clarify the mechanisms driving LNneg enlargement and the potential functional relevance of pigment deposition.

Computer-aided detection (CADe) increases adenoma detection in colonoscopy, but its effectiveness in postpolypectomy surveillance remains uncertain. This study compared the adenoma detection rate (ADR) of CADe-assisted versus standard high definition colonoscopy during surveillance. We conducted a population-based, randomized controlled trial in the Galician colorectal cancer screening program. Colonoscopy days were randomized in a 1 : 1 ratio to high definition colonoscopy with or without CADe. The primary end point was the ADR. Prespecified subgroup analyses evaluated endoscopist baseline performance (median ADR in the standard arm). Effect estimates are reported as adjusted risk ratios (aRRs) with 95%CI. Of 5309 randomized surveillance colonoscopies, 4824 surveillance colonoscopies were included in the final analysis (2469 standard; 2355 CADe-assisted) after exclusion of incomplete examinations. The ADR did not differ between the groups (57.4% vs. 58.8%; aRR 1.02, 95%CI 0.95-1.10). CADe increased ADR among low performing (ADR <54.5%) endoscopists (45.5% vs. 52.1%; aRR 1.15, 95%CI 1.01-1.30), but not among high performing endoscopists (65.9% vs. 63.5%; aRR 0.96, 95%CI 0.88-1.05). In a high performing screening program, CADe did not increase ADR in surveillance colonoscopy. Its benefit may be limited to lower performing endoscopists.

Intestinal intraepithelial lymphocytes (IELs), including conventional CD8αβ T resident memory (Trm) cells and unconventional CD8αα T cells, promote tissue integrity. Here, we studied the G-protein coupled receptor signals regulating IEL positioning, homeostasis, and function. Deficiency in heterotrimeric G-protein subunit Gα13 or its effector Arhgef1 caused an intestine-specific loss of all types of CD8+ TCRαβ and TCRγδ IELs. Gα13-deficient IELs exhibited restricted intraepithelial movement and impaired maturation. Induction of intestinal CD8αβ+ Trm cells upon infection was intact in the absence of Gα13-signaling, but the cells had poor access to the villous niche and defective survival that could be rescued by increasing TGF-β or interleukin (IL)-15. In vivo CRISPR-Cas9 screening identified GPR132 as a Gα13-coupled receptor that regulates CD8αβ+ IEL homeostasis and migration to lysophosphatidylcholine. Mice bearing Gα13-deficient T cells suffered more severe colitis and increased colorectal tumor growth. The selective requirement for Gα13 signaling for IEL positioning and survival in the villous niche has implications for therapeutic intervention.

The immunosuppressive microenvironment is a hallmark of glioblastoma (GBM), limiting the efficacy of contemporary immunotherapies. While the central nervous system (CNS) relies on specialized cytokine networks to maintain immune homeostasis under physiological conditions, how these homeostatic signals are subverted during gliomagenesis remains poorly understood. Here, we report that interleukin-17D (IL-17D) is a CNS-intrinsic tumor suppressor whose expression is downregulated during gliomagenesis. Restoring IL-17D substantially extends survival in orthotopic GBM models by selectively reprogramming the myeloid compartment. Mechanistically, IL-17D engages its receptor CD93 to assemble a novel costimulatory receptor complex with SCARB1 in a lipid raft-dependent manner. This structural assembly engages Fyn/BCAP to trigger PI3K/AKT/NF-κB signaling, effectively inverting the function of SCARB1 from tolerogenic efferocytosis to active antigen presentation. Our study demonstrated that the restoration of IL-17D expression reshaped the GBM immune landscape, highlighting its potential as a therapeutic target to enhance the efficacy of immunotherapy.

Malignant gliomas are lethal brain tumors characterized by profound local immunosuppression and a radically remodeled myeloid landscape. Although these tumors mobilize resident microglia and infiltrating monocyte-derived macrophages, the mechanisms governing their phenotypic convergence and diversification remain elusive. Here, we integrated single-cell profiling and spatial transcriptomics of glioma-associated microglia in the GL261 model. We identified distinct microglial states that aligned with tumor architecture, most notably Cst7-expressing disease-associated microglia (DAMs) that aggregated at the tumor invasive margin and exhibited a conserved transcriptional signature shared across various central nervous system pathologies. Interferon-γ and toll-like receptor signaling sequentially tuned stage-specific DAM features, including transient MHC-II expression and sustained PD-L1 upregulation, thereby recalibrating the local immune equilibrium by reshaping bidirectional DAM-T cell interactions during glioma progression. Our findings highlight microglial state transitions as a stage-specific layer of immune regulation in glioma that shapes T cell fate and support targeting microglial plasticity to rebalance anti-tumor immunity.

Pre-metastatic niches composed of mainly myeloid cells are recognized as critical for tumor metastasis. However, whether adaptive immune cells also play an important role in pre-metastatic niche formation remains to be explored. CD4+ T cell accumulation in tumor-free lung tissues from mice bearing subcutaneous mouse tumors was detected by immunofluorescence/confocal microscopy. Tumor-conditioned media (TCM) from MB49-S1pr1 high mouse bladder tumor cells or ID8 ovarian tumor cells were administered to tumor-free mice to induce pre-metastatic niche formation. We used mice lacking functional Signal Transducer and Activator of Transcription 3 (STAT3) in T cells and Il17a‒/‒ mice to investigate the roles of STAT3 and interleukin (IL)-17. In vivo time-course experiments were performed to assess whether CD4+ T cell clusters contribute to CD11b+ pre-metastatic clusters. CD4+ T cell migration and chemokine receptor expression assays were employed to identify tumor factors driving CD4+ T cell recruitment. A co-culture system with human MRC-5 lung fibroblasts, healthy donor-derived CD4+ T cells, myeloid cells, and TCM derived from human cancer cells was used to evaluate CD4+ T cell-driven fibroblast activation and IL-17A dependency for myeloid cell migration. Microscopic analyses were performed to confirm CD4+ T cell clusters in tumor-free lymph node tissues from patients with prostate cancer and postmortem lung and liver specimens from patients with ovarian cancer. We demonstrate that CD4+ T cells accumulate in tumor-free lungs and promote tumor metastasis in mouse models. CD4+ T-cell pre-metastatic niche formation requires STAT3, which regulates Th17 CD4+ T cells. TCM drives IL-6-dependent CCR4/CCR6 upregulation on naive CD4+ T cells. CD4+ T cell clusters contribute to myeloid cell accumulation, and ablating STAT3 in T cells abrogates both T cell and myeloid cluster formation. IL-17 inhibition reduces myeloid lung infiltration. In human co-cultures, CD4+ T cells amplify TCM-induced fibroblast pre-metastatic niche-like activation and myeloid recruitment in an IL-17A-dependent manner. CD4+ and IL-17+ or p-STAT3+ clusters were also detected in non-metastatic tissues from patients with several cancers. CD4+ T cells form pre-metastatic niches through the STAT3-IL-17 axis, contributing to myeloid cell cluster formation, in part through amplifying fibroblast pre-metastatic niche-like activation. STAT3 and IL-17 in CD4+ T cells therefore are important for pre-metastatic niche formation and metastasis.

Suboptimal human-artificial intelligence (AI) interaction is a potential roadblock for implementation of AI in clinical practice. We aimed to evaluate the interaction between AI and endoscopists in the optical diagnosis of colorectal carcinoma (CRC). International endoscopists were invited to diagnose colorectal lesions online. After a pretest of 15 videos, the diagnosis could be adjusted in the subsequent 35 videos after receiving a simulated AI diagnosis. Histology was the gold standard. Endoscopists' diagnoses before and after AI were compared using the signal detection theory. Inappropriate response to AI was defined as decreased diagnostic performance by the endoscopist after AI. In 2730 diagnoses, the 78 participating endoscopists reached diagnostic accuracies of 71.4 % before and 73.7 % after AI (P < 0.001). Before AI, endoscopists missed only a few true CRCs (mean hit rate 0.86), at the cost of a mean false alarm rate of 0.35. Inappropriate response to AI was observed in 28.2 % of endoscopists. After AI, most endoscopists (41.0 %) shifted toward a more conservative attitude, tending to diagnose a lesion as non-CRC. Inappropriate response to AI is an issue in human-AI interaction for the optical diagnosis of CRC. Before using AI, endoscopists should become familiar with their own optical diagnostic performance, and that of the AI system, to possibly improve interaction.

Correction for 'Planar rotor-enabled quenching-resistant NIR-II fluorophores for high-contrast bioimaging and efficient cancer phototheranostics' by Weilong Chen et al., Mater. Horiz., 2026, 13, 6665-6678, https://doi.org/10.1039/d6mh00067c.

This study presents an application of delocalized hydrocarbon neutral monoradical, an olympicenyl radical (OR6), as a high-performance organic photothermal agent (PTA) for targeted photothermal therapy (PTT). By encapsulating OR6 into functionalized nanoparticles (dOR6-NPs), a remarkable near-infrared-II (NIR-II) absorption at 1056 nm and photothermal conversion efficiency (PCE) of 84.66% are achieved. To overcome the limitations of passive tumor targeting, a bioorthogonal "tag-and-target" strategy is developed using metabolic glycan labeling and copper-free click reaction. This approach significantly enhanced photothermal therapeutic efficacy both in vitro and in vivo. The dOR6-NPs induce significant cytotoxicity of cancer cells under 1064 nm laser irradiation, leading to nearly complete tumor suppression in a xenograft model while maintaining excellent biocompatibility and systemic safety. This work not only highlights olympicenyl radicals as a new class of PTAs but also establishes a robust click-reaction-based platform for precision cancer therapy.

Polysaccharides are important polymeric molecules widely recognized for their diverse pharmacological activities and long-standing relevance in traditional medicine. Levan, a fructose-based natural polysaccharide derived from microbial and plant sources is among the notable polysaccharides and it holds noteworthy potentials owing to its high yield and rate of production, chemical diversity and diverse biological effects. Levan exhibits an extensive range of physicochemical and biological properties, including, self-assembly, biodegradability, high biocompatibility, controlled release capability. Pharmacological studies have also demonstrated its antioxidative, immunomodulatory, and proapoptotic activities, in addition to its tumour growth suppression ability in both in vitro and in vivo cancer models. However, despite its various promising properties, levan remains understudied compared to other well-known polysaccharides such as β-glucans or fucoidans that are more widely studied. This review therefore seeks to bridge this gap by comprehensively discussing the anticancer mechanisms of levan, its recent research findings across various cancer cell types, its potential as an adjunct or alternative cancer therapy and its emergent applications of Levan-based nanoparticulate Systems. This serves as a link between traditional medicine and ethnopharmacological evidence that present levan as a potential agent with anticancer benefits.

Buckwheat polysaccharides (BWPs), key bioactive constituents broadly distributed across tissues of Fagopyrum Mill, have received growing attention for their nutritional value and health-promoting functions in functional food research. This review consolidates recent progress in BWP extraction and purification strategies, structural elucidation, bioactivities, and prospective applications. Current preparation approaches include hot-water extraction, enzyme-assisted extraction, ultrasound and microwave-assisted techniques, as well as emerging green processes. BWPs are mainly complex heteropolysaccharides composed of monosaccharides, including glucose, arabinose, xylose, galactose, mannose, and galacturonic acid, with arabinoxylan and pectin-like polysaccharides as representative types. BWPs exert diverse bioactivities, notably antioxidant effects, modulation of glucose and lipid metabolism, immunomodulatory and anti-inflammatory actions, anti-tumor potential, and improvements in gut microbiota composition. BWPs hold strong promise for applications in functional foods, nutritional interventions, and biomedical materials. Future studies should prioritize elucidating their mechanism of action, establishing clearer structure-activity relationships, and standardizing extraction processes to support scalable and high-value applications.

Cancer cachexia has long been attributed to circulating factors. Cross et al. now show that, in liver kinase B1 (Lkb1)-mutant lung tumors, local prostaglandin E2 acts on vagal neurons to drive anorexia and wasting, accelerated by fat. Defining metaboception as neural sensing of local signals-a new therapeutic concept.

The conformational switching of a protein from its water-soluble conformation to its membrane-inserted conformation is critical for many cellular processes: cellular entry of bacterial toxins via escape from the endosome, apoptotic regulation by the Bcl-2 protein family to preserve or disrupt the outer mitochondrial membrane, and tumor targeting by pH-Low-Insertion-P eptide pHLIP. Here, several empirical mechanisms of conformational switching are described, illustrating how the formation of membrane-competent conformation and subsequent membrane insertion can be triggered and modulated by pH and divalent cations, abided by changes in lipid composition of the target membranes. Despite structural similarities of the water-soluble conformations of the diphtheria toxin translocation (T) domain with that of proapoptotic BAX and antiapoptotic Bcl-xL, their corresponding insertion pathways and modes of conformational switching are markedly different. The conformational switching of the T-domain is initiated by protonation of key histidine residues in solution, while in the Bcl-2 proteins BAX and Bcl-xL, it occurs in the membrane proximity. Consequently, changes in the composition of the lipid bilayer (e.g., local enrichment in cardiolipin) can affect insertion and functioning of Bcl-2 proteins. Conformational switching of pHLIP occurs on membrane interfaces via pH-dependent and Ca2+-dependent mechanisms, which is strongly influenced by lipid composition.

Regulators are accelerating new approach methodologies in cancer drug development. Their capacity to replace animal studies should depend on the context of use. Organoids, chips, and computational models can answer local, measurable questions, whereas whole-body biodistribution, immune handling, organ clearance, chronic toxicity, and therapeutic index still require integrated physiological evidence.

Cellular senescence is a stress-response program characterized by durable proliferative arrest, but senescence entry does not determine a single biological outcome. Confusion between the fate of an individual cell and the behavior of a senescent-cell population has obscured interpretation across development, ageing and disease. This Review develops a fate-resolved framework that distinguishes continued arrest, cell death, physical shedding and, under restricted conditions, sustained proliferative recovery from population-level resolution or persistence. We reserve senescence escape for sustained proliferation by the same cell or lineage after validated senescence entry; failed resolution is instead one process that can produce persistence. We examine how checkpoint integrity, chromatin organization, metabolism, secretory signaling and immune surveillance shape these outcomes in development, tissue repair, ageing-associated disorders and cancer. The strength of evidence differs sharply by context. Longitudinal cancer models provide the clearest support for strict escape, alongside a restricted developmental lineage-tracing example, whereas fibrosis, neurodegeneration and metabolic or vascular disease more often support population persistence or impaired resolution without demonstrated cell-cycle re-entry. Accordingly, static markers, single-cell and spatial profiles, and computational trajectories can identify senescence-associated states but cannot establish subsequent fate without lineage tracing, live imaging or equivalent temporal evidence. Therapeutic strategies should therefore be matched to the dominant biological problem: preserving beneficial physiological senescence, resolving or modulating harmful persistent populations, or preventing recovery of residual senescent tumour cells. Because most approaches remain preclinical or early-stage, translation will require appropriate timing, cell-type specificity and protection of beneficial senescent populations.

Gene expression profiling (GEP) provides critical insights into the cellular states, phenotypes, and biological behaviors of biological systems. However, the traditional GEP acquisition methods have significant time limitations, which directly hinder the real-time monitoring of dynamic processes, such as cell development and disease progression. In this study, we propose the biological omics sparse sensing (BoSS) framework, which effectively resolves the compatibility issue between dimensionality-reduced sampling theory and biological reaction by introducing sparse sampling strategies and noise regularization constraints. Furthermore, we have first presented rapid capture and accurate reconstruction of GEP using multiplex RT-qPCR, increasing the efficiency of transcriptome data acquisition by 60-fold. The experimental results demonstrate that when evaluating transcriptional heterogeneity in cancer tissue blocks, BoSS exhibits excellent specificity and sensitivity in clustering analysis, while fully preserving key information related to biological states, thus ensuring that the accuracy of downstream data analysis is comparable to that of ribonucleic acid (RNA)-seq. This achievement indicates that the BoSS framework is expected to play an important role in scenarios, such as intraoperative assessment of cancer progression, personalized therapy, and predictive control of biological systems.