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Clinical practice guidelines (CPGs) are intended to improve health and reduce disease burden, yet their global status and their association with disease burden remain unknown. This longitudinal ecological study analyzes the quantity and quality of CPGs and explores their associations with disease burden. From 1995 to 2023, 10,657 CPGs were published, with the number of CPGs in 2023 being 5.15 times the number of CPGs in 1995. Europe contributed 40.2% of all CPGs, while CPGs from Africa accounted for only 0.8% of and showed minimal growth. Treatment CPGs contributed the most (45.0%), while diagnosis CPGs showed the fastest growth. Prevention CPGs (8.4%) and nursing CPGs (5.3%) and rehabilitation CPGs were less common, with rehabilitation CPGs showing the slowest growth. The 3-year update rate of CPG is 23.8%. Quality assessment of 1633 CPGs using AGREE II revealed substantial variation across domains. Overall 37.8% CPGs achieved scores above 70% per domain, while the "applicability" domain had the lowest median score (36%). Cross-region CPGs and CPGs for neoplasms exhibited the highest AGREE II scores, with the median score exceeding 70% in 3/6 domains. In contrast, CPGs from Asia and CPGs for cardiovascular diseases did not exceed the 70% threshold in any domain. Neither the quantity nor the quality of CPGs was significantly associated with disease burden at the national level. In conclusion, this study reveals that the quantity of CPGs has increased over the past three decades, although geographic and thematic differences remain. Future opportunities for CPG development include improving the AGREE II scores and focusing on less represented areas including rehabilitation, nursing and prevention.

Fecal microbiota transplantation (FMT) has shown promise in overcoming resistance to immune checkpoint inhibitors (ICIs) in early-phase cancer trials. We investigated the safety, feasibility and efficacy of FMT from ICI responders to patients with advanced cancers progressing on ICIs. This was a single-arm phase IIa basket trial (MITRIC; NCT05286294) including patients with ICI-refractory cancer. Long-term ICI responders were used as FMT donors. Patients received FMTs in combination with ICIs; two FMT administrations (by colonoscopy) were scheduled before the first radiological evaluation after 6 weeks, and up to three later FMTs were allowed (by enema). Co-primary endpoints were the evaluation of FMT-related adverse events and objective response rate. Feasibility, clinical benefit rate, progression-free survival (PFS), overall survival (OS), implant engraftment, immune response and biomarkers were among the secondary objectives. The study enrolled 12 patients with melanoma (n=9), head and neck squamous cell carcinoma (HNSCC; n=1), renal cell carcinoma (n=1) or microsatellite instability-high pancreatic cancer (n=1). FMT was well tolerated, whereas immune-related toxicity occurred in 6/12 patients. All patients received the first FMT; 10/12 patients also underwent the second FMT. No objective responses were observed, while 5/12 patients recorded stable disease. Clinical benefit per-protocol (stable disease >6 months) was achieved in a patient with melanoma, who had regression of some lesions and remains alive after 33 months without further systemic treatment. Mixed responses with regression of some lesions were observed in another melanoma patient, and in a patient with HNSCC. The median PFS was 1.5 months, and median OS was 10.1 months. Sequencing of fecal samples indicated engraftment after the first FMT in most patients. Mass cytometry analysis of peripheral blood cells suggested that an activated and differentiated T-cell signature was associated with improved PFS and OS, while a naïve T-cell phenotype and a myeloid-dominant environment were unfavorable. CD14+ monocytes and serum interleukin-8 increased at group level over time. FMT in combination with ICIs was safe and feasible in patients with advanced cancers, but with limited clinical activity. Further studies are required to clarify the potential benefit of FMT, identify the appropriate patient population and define criteria for donor selection. NCT05286294.

Spatially offset Raman spectroscopy (SORS) and its surface-enhanced variant, SESORS, have emerged as transformative techniques in analytical chemistry and biomedical diagnostics, enabling non-invasive, depth-resolved molecular sensing. SORS facilitates the probing of endogenous signals from subsurface layers, while SESORS enhances sensitivity through plasmonic nanostructures, allowing sensitive targeted detection of exogenous diagnostic agents. Spatially offset Raman spectroscopy is now making significant strides in clinical diagnostics. This review focuses on the rapid advancements in the field over the past five years, including applications in cancer diagnosis, monitoring of therapeutic response, bone health assessment, and tracking of neurotransmitters and SERS-labelled nanoparticles through tissue depths exceeding 7 cm in a tissue wrapped mouse model and 14 cm from within a porcine tissue stack. The integration of tailored nanostructures, resonant Raman labels, and advanced optical instrumentation has expanded the versatility of SESORS, with modalities such as transmission, backscattered, and resonance-enhanced configurations offering flexibility across diverse clinical scenarios. Despite challenges in standardisation, biocompatibility, and instrumentation, interdisciplinary progress in optics, nanotechnology, and spectroscopy continues to drive innovation. This review critically examines the recent evolution, classification, and biomedical applications of SORS and SESORS, highlighting their limitations and future potential in translational medical research.

Somatic mutations rewire the ubiquitin-proteasome system (UPS) to support tumor growth, but the proteome-wide consequences of cancer-driver alterations on UPS composition remain incompletely understood. Using harmonized proteogenomic data from up to 11 CPTAC cohorts, we performed an integrated pan-cancer analysis of UPS protein dysregulation, prognostic associations, and mutation-driven remodeling. We show that mRNA poorly predicts UPS protein abundance, that a defined set of E3 ligases is recurrently dysregulated across cancers, and that somatic mutations (most strikingly TP53 loss) produce coherent UPS protein-quantitative trait locus (pQTL) signatures. Two case studies (UBR5 and TRIM28) illustrate orthogonal modes of UPS rewiring: a mutation-driven axis in which TP53-mutant tumors elevate UBR5 to support replication stress tolerance, and a lineage-driven axis in which TRIM28 engages tissue-restricted regulatory networks with opposing prognostic effects in glioblastoma versus head and neck cancer. Each axis exposes context-specific therapeutic vulnerabilities, including sensitivity to DNA damage response inhibitors (UBR5-high) and lineage-specific drug responses (TRIM28-high). Together, these analyses define a mechanistic framework for how cancer-driver mutations reshape proteostasis through the UPS and nominate mutation- and lineage-defined dependencies for precision degrader therapy. The harmonized pan-tissue atlas and the UbiDash interactive resource that underpin parts of this analysis are reported in our companion paper [1].

Mortality disparity in cancer between Black individuals and White individuals in the US has been declining since its peak in the early 1990s, but a comprehensive analysis of cancer type-specific contributions to this reduction is lacking. To examine cancer type-specific contributions to the reduction in the all-cancer excess mortality rate among Black individuals compared with White individuals between the peak period (1991-1995) and a contemporary period (2019-2023) in the US. A cross-sectional temporal analysis was conducted between March 2025 and October 2025. Non-Hispanic Black individuals and White individuals who died of cancer between 1991 and 2023 were included in the analysis. The data were compiled by the National Center for Health Statistics. Race as recorded on death certificates. All-cancer and cancer type-specific excess mortality rates per 100 000 for Black individuals compared with White individuals were calculated by sex. Changes in excess mortality rates were estimated by subtracting the current excess mortality rate for 2019 to 2023 from the peak excess mortality rate for 1991 to 1995. Cancer type-specific contributions to all-cancer excess mortality rate reduction were expressed as percentages. The analysis included 2 461 715 cancer deaths in 1991 to 1995 (11.8% non-Hispanic Black individuals; 47.4% were female) and 2 674 385 cancer deaths in 2019 to 2023 (13.2% non-Hispanic Black individuals; 47.2% were female). Between 1991 to 1995 and 2019 to 2023, the all-cancer excess mortality rate decreased by 98.4 (95% CI, 96.1-100.7) per 100 000 in Black males and by 21.7 (95% CI, 20.3-23.1) per 100 000 in Black females. Among males, lung (31.1% [95% CI, 29.9%-32.4%]), prostate (25.8% [95% CI, 24.7%-27.0%]), and esophageal (12.1% [95% CI, 11.7%-12.5%]) cancer accounted for the largest share of reductions in the all-cancer excess mortality rate. Among females, colorectal (18.8% [95% CI, 16.6%-20.9%]), cervical (16.9% [95% CI, 15.8%-18.0%]), and lung (15.1% [95% CI, 12.3%-17.9%]) cancer accounted for the largest share of reductions in the all-cancer excess mortality rate. Despite these gains in excess mortality rate reduction, prostate, lung, and stomach cancer remained among the top 5 cancer types with the largest contemporary excess mortality rate among males, whereas colorectal cancer increased in rank (from fifth to second) as well as liver cancer (from tenth to fifth), reflecting comparatively smaller or stalled progress. Breast cancer and uterine corpus cancer ranked highest during the contemporary period in excess mortality rate among Black females compared with White females (7.2 per 100 000 and 4.9 per 100 000, respectively), followed by myeloma, colorectal, and pancreatic cancer. In this cross-sectional temporal analysis, the marked reductions in excess mortality indicate meaningful progress achieved for several major cancer types through population-wide cancer control efforts. Persistent or worsening disparities in breast cancer and uterine corpus cancer and substantial residual disparities in prostate cancer highlight priorities for equitable interventions. These findings may guide more targeted and efficient cancer prevention and control strategies to further accelerate progress.

Electronic health records (EHRs) provide rich longitudinal disease histories, but existing methods for analysing these data typically treat diseases in isolation1 and rarely integrate germline genetics. Here we present ALADYNOULLI, a Bayesian generative framework that jointly models longitudinal EHR diagnoses, age and polygenic risk to recover latent time-varying disease signatures and patient-specific signature loadings; the model is formulated as a mixture of probabilities rather than a probability of a mixture2, correctly accommodating simultaneous and chronic conditions. Applied to three independent biobanks (UK Biobank3, Mass General Brigham4 and All of Us; total n > 683,000) spanning up to 52 years of follow-up and 348 diseases, the model recovers 21 replicable signatures with high cross-cohort composition preservation (median of 80%) and reveals biological subtypes within diagnostic categories (Cohen's d up to 4.25; P ≤ 1 × 10-8 for 95% of comparisons). Signatures are concordant with established disease biology: carriers of familial hypercholesterolaemia5 enrich in the cardiovascular signature; carriers of clonal haematopoiesis of indeterminate potential6 in the inflammation signature; and a rare variant burden in LDLR, TTN and BRCA2 (refs. 7,8) aligns with disease specificities. A signature-based genome-wide association study identifies 151 genome-wide significant loci including cardiovascular associations missed by single-trait analyses. An explicit likelihood enables inverse probability weighting for selection bias9 while preserving biological signal. For disease prediction, ALADYNOULLI outperforms Pooled Cohort Equation (PCE), PREVENT and Gail at 1-year and 10-year horizons; disease-level (PheCode) predictions complement code-level foundation models such as Delphi-2M (ref. 10).

Patients with cancer usually report limitations of their functional capacity, which may range from subclinical impairment of cardiopulmonary exercise reserve to poor quality of life with physical, cognitive, or psychosocial consequences. Exercise training is a potent multi-targeted approach to control pre-existing and new risk factors. Preliminary evidence shows that it is associated with a lower risk of cancer therapy-related cardiotoxicity and increased self-reported well-being in patients with cancer. Current evidence demonstrates that supervised exercise therapy, including high-intensity interval training, is safe and well-tolerated and reduces risk in subjects with cancer in the pre-, active-, and post-treatment settings. The present consensus document will discuss the role of exercise training in cardio-oncology, focusing on patients with cardiovascular diseases induced by cancer treatment and on those who received cardiotoxic therapies.

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

Although there are several ongoing clinical trials using neoantigen peptide-based cancer vaccines, challenges still exist to implement in clinical approval such as poor antigen stability, inefficient delivery, and inadequate immune activation. To address these limitations, we developed a polymer-based polyvalent peptide and adjuvant (SPPA) that co-delivers lipid-conjugated tumor-specific peptides with Toll-like receptor 7/8 (3M-052) and a STING (2'3'-cGAMP) agonist. This nanoplatform enables efficient peptide encapsulation, sustained release, and targeted delivery to antigen-presenting cells (APCs), thereby enhancing both innate and adaptive immune responses. We synthesized and characterized a library of lipid-conjugated tumor-associated and neoantigenic peptides. In vitro, SPPA significantly upregulated pro-inflammatory genes and cytokine secretion, confirming robust innate immune activation and demonstrated effective cellular uptake and lymphatic trafficking. In vivo, SPPA alone or in combination with anti-PD-1 antibody (αPD-1) elicited strong cytotoxic T lymphocyte (CTL) responses and inhibited tumor growth in four aggressive syngeneic mouse models: Triple-negative breast cancer (4T1), HER2+ breast cancer (TUBO), lung carcinoma (LLC1), and renal cell carcinoma (RENCA). The combination therapy led to pronounced tumor growth inhibition, survival benefit, and immune cell infiltration, including elevated CD8+IFN-γ+ T cells and M1 macrophages, and reduced regulatory T cells and MDSCs. Spatial transcriptomics revealed localized transcriptional reprogramming, with downregulation of extracellular matrix genes and activation of inflammatory pathways. Collectively, these findings establish SPPA as a potent and versatile nanovaccine platform capable of inducing durable antitumor immunity, especially when combined with immune checkpoint blockade. This approach offers strong translational potential for personalized immunotherapy across diverse solid tumor types.

Molecular machines rely on dynamic, low-affinity interactions to perform their functional roles. We developed PhIX-MS (photo-induced in situ crosslinking-mass spectrometry), a structural proteomics workflow to capture topological information for such transient interactions in cells by UV-activated crosslinking. Applying PhIX-MS with cryo-electron microscopy (cryo-EM) to proteasomes, we mapped the redox sensor TXNL1 at the proteasome regulatory particle (RP), including its dynamic thioredoxin-like domain near RPN2/PSMD1 and RPN13/ADRM1, where it is ideal for reducing substrates prior to proteolysis. RPs without the proteolytic core particle (CP) were structurally resolved while bound to TXNL1 and/or the chaperone PSMD5/S5b, which inserts its C terminus into the ATPase pore, causing extensive structural rearrangements. Additionally, PhIX-MS and AlphaFold identified the ubiquitin ligase UBE3C/Hul5 at RPN2, RPN3, and a dynamic RPN10 region, tethering UBE3C above the substrate entry channel. Our integrative approach enables the localization of native, low-affinity protein interactions and is broadly applicable to dynamic macromolecular assemblies.

DNA methylation is a crucial epigenetic modification whose abnormal alterations are closely associated with various tumors and are considered potential biomarkers for cancer diagnosis. However, achieving highly sensitive and selective detection of low-abundance methylated DNA in complex biological samples remains a significant challenge. This study developed a functionalized glass micropipette electrochemical sensing strategy based on the enzyme-responsive signal switching effect for highly sensitive detection of methylated DNA. This system utilizes glass micropipettes modified with polydopamine nanotubes (PDA-NTs) to construct an ion-transport interface, where the capture probe (cpDNA) is immobilized to achieve specific recognition of target sequences. The detection mechanism relies on sequence differences introduced by sodium bisulfite conversion and the selective cleavage action of the nucleic acid exonuclease Exo III: upon forming stable double-stranded structures with the cpDNA, methylated DNA triggers Exo III-mediated structural changes at the interface. This modulates ion transport behavior within the channels, generating a current response that switches from "Off" to "On", enabling effective differentiation between methylated and unmethylated DNA. This sensing system exhibits excellent linear response to methylated DNA within the 1 aM-100 fM range, with a detection limit as low as 8.5 aM, and enables reliable detection in 10% human serum samples. This strategy achieves sensitive detection through an enzyme-responsive signal switch without requiring PCR amplification or complex labeling procedures, providing a simple and effective method for rapid analysis of DNA epigenetic modifications.

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

RNA modifications, such as N6-methyladenosine (m6A), N1-methyladenosine (m1A), 5-methylcytosine (m5C), 7-methylguanosine (m7G), pseudouridine (Ψ), and adenosine-to-inosine (A-to-I) editing, constitute a dynamic epitranscriptomic network that profoundly regulates RNA metabolism and gene expression. Their dysregulation is increasingly recognized as a hallmark of cancer. This review critically synthesizes the multifaceted roles of RNA modifications to bridge the gap between descriptive epitranscriptomic mapping and functional tumor biology. We systematically evaluate how writers, readers, and erasers dictate transcript stability and translation efficiency, driving tissue-specific tumor evolution across diverse malignancies. Crucially, we explore the intersection of RNA modifications and the tumor immune microenvironment, detailing their mechanisms in orchestrating immune evasion, altering antigen presentation, and regulating immune checkpoints. Furthermore, we examine how epitranscriptomic reprogramming dictates cellular responses to chemotherapy, radiotherapy, targeted treatments, and immunotherapy. By comprehensively analyzing these mechanisms, this review aims to facilitate the translation of epitranscriptomic findings into clinical applications, laying a theoretical foundation for targeted anti-tumor strategies.

Cancer is one of the leading causes of global morbidity and mortality and is characterized by its high heterogeneity, genomic instability and adaptive plasticity. Applying Darwin's theory of evolution, the concept of tumor evolution has improved our understanding of the biological behavior of advanced cancer, while technological limitations have long left how to dynamically characterize the evolutionary process unsolved. In recent years, the development of high-throughput sequencing, single-cell and spatial omics, lineage tracing, computational modeling, and noninvasive biopsy technology has helped explain tumor heterogeneity and track the evolutionary trajectory of tumors, which has attracted widespread attention in tumor evolution. Here, we discuss the models and drivers of tumor evolution, focusing on how cancer cells adapt and evolve under multidimensional selective pressure, including the intracellular, extracellular, and exogenous levels. We further summarize key signaling pathways and molecular mechanisms involved in genomic instability, epigenetic regulation, metabolic reprogramming, tumor microenvironment remodeling, immune escape and therapy-induced selection. In addition, emerging therapeutic strategies guided by tumor evolution are also discussed, emphasizing that the identification of key genetic and epigenetic targets, together with dynamic monitoring of clonal changes, is crucial for overcoming treatment resistance and improving patient outcomes. By systematically summarizing the signaling pathways and molecular mechanisms underlying tumor evolution, this review aims to provide new targets and conceptual frameworks for promoting precision treatment strategies in oncology in the future.

Circulating tumor cells (CTCs) are emerging as pivotal biomarkers for early cancer metastasis detection, prognosis assessment, and real-time therapeutic monitoring. However, their extreme rarity and phenotypic heterogeneity pose significant challenges for efficient capture and viable recovery. Inspired by the dynamic recognition and adhesion mechanisms of leukocytes under physiological conditions, we developed a red light-responsive leukocyte-inspired multiscale hierarchical interface (g@SoS-PACF) for the specific capture and noninvasive release of CTCs. The platform features a microsphere-nanosphere-polymer brush architecture that mimics key structures and functions of leukocytes, thereby enabling topography-assisted anchoring, enhanced interfacial complementarity, and antifouling performance. In addition, this hierarchical interface increases the local density and mobility of targeting ligands, significantly improving recognition efficiency across heterogeneous CTC subpopulations. A photoresponsive azobenzene-cyclodextrin host-guest system is embedded to enable red light-triggered, reversible trans-cis isomerization, allowing gentle release of intact CTCs. In vitro studies revealed capture efficiencies above 83% across diverse EpCAM-expressing tumor cells, with release efficiencies exceeding 94% and post-release viability greater than 89%. Importantly, clinical validation using blood samples from 15 cancer patients demonstrated reliable isolation of phenotypically diverse CTCs. This work presents a smart bioinspired system with broad implications for rare cell sorting, liquid biopsy technologies, and next-generation precision oncology.

In recent years, heavy-atom-free photosensitizers have been recognized as an important class of agents for photodynamic therapy (PDT). In particular, redox-activated heavy-atom-free photosensitizers inspired by the redox imbalance of the tumor microenvironment are emerging as a promising strategy to improve the specificity and therapeutic efficiency of PDT. This review highlights design strategies of redox-activated heavy-atom-free photosensitizers (PSs), including donor-acceptor-based frameworks, thionation, aggregation-induced emission (AIE) driven intersystem crossing enhancement, and the integration of responsive moieties for reactive oxygen species (ROS), glutathione (GSH), cysteine (Cys), and hydrogen sulfide (H₂S). In addition, recent advances in the development of redox-activated heavy-atom-free PSs over the past three years are summarized. Finally, main challenges, including hypoxic tumors, limited tumor-targeting efficiency, and limited light penetration into deep-seated tumors, as well as future prospects for this field, are discussed.

Copper, an essential trace element with dual functions in cancer progression, drives tumor growth via oncogenic signaling, metabolic plasticity, and extracellular matrix remodeling. By contrast, copper overload triggers cuproptosis, a form of mitochondrial proteotoxic cell death mediated by the FDX1/LIPT1/DLAT/Fe-S regulatory axis. To date, a unified theoretical framework integrating copper metabolism, tumor microenvironment (TME) remodeling, and antitumor immunity remains lacking. In this review, we reframe the TME as a structured copper ecosystem in which both cellular components and the extracellular matrix are modulated by copper, and propose a contextual copper signaling network, in which copper-mediated tumor cell fate is determined by the labile copper pool, the metabolic state, and tumor cellular heterogeneity. We further delineate a unified causal chain linking cuproptosis-driven immunogenicity and cGAS-STING activation to immune cell activation and PD-L1 modulation. Therapeutically, copper chelation and cuproptosis induction strategies have demonstrated promising efficacy, and combining cuproptosis induction with existing antitumor therapies may reverse therapeutic resistance and enhance treatment efficacy. Future studies need to validate cuproptosis-related signatures as predictive biomarkers for precision oncology and refine copper-targeted therapies to minimize systemic toxicities.

Lactylation is a recently identified metabolism-associated post-translational modification that provides a mechanistic link between tumor metabolic reprogramming and epigenetic regulation. Although aberrant lactate accumulation in tumor tissues has long been recognized, a comprehensive and unified understanding of how lactate-derived modifications contribute to tumor initiation and progression remains lacking. Here, we use lactate metabolism as a conceptual framework to systematically review the biogenesis and regulatory networks of lactylation, with a particular focus on its functions and underlying molecular mechanisms in key malignant processes, including tumor proliferation, invasion and metastasis, angiogenesis, immune evasion, radiotherapy resistance and systemic therapy resistance. Furthermore, we review emerging anti-tumor therapeutic strategies targeting lactate metabolism and tumor lactylation, highlighting that glucose metabolism-driven lactate accumulation and lactylation play a pivotal role in sustaining tumor cell survival and immunosuppressive phenotypes. Inhibition of lactate metabolism markedly attenuates tumor-associated lactylation and exhibits substantial synergistic efficacy when combined with other anti-tumor therapies. Overall, this work advances a systematic understanding of the roles of lactylation in tumor malignant progression and provides an important conceptual and theoretical foundation for the development of novel cancer therapeutic strategies targeting protein lactylation.

Plasma cell-free DNA (cfDNA) fragmentomics offer promising cancer biomarkers, but their molecular regulation remains elusive. Here, we investigate the role of epigenomic modifications in cfDNA fragmentation. We identify strong correlations between cfDNA fragmentomic features and various epigenetic marks measured in cfDNA. We further segment the genome into different chromatin states using histone modification signals, revealing consistent associations with cfDNA fragmentomics. The association is further validated by histone modifier perturbation experiments, confirming chromatin organization as a key regulator of cfDNA fragmentation. CfDNA fragmentomic features associated with Transposon Elements (TEs) outperform genome-wide metrics in cancer diagnosis, reflecting cancer type-specific patterns. Leveraging these insights, we develop TEANA (Transposon Element Analysis in cfDNA), an AI-empowered model using a small set of TE fragmentomic features for pan-cancer detection and tumor-origin prediction, achieving robust performance across independent cohorts. Hence, chromatin states drive cfDNA fragmentation, and dysregulated TEs provide highly informative biomarkers for cancer diagnosis.

Neutrophils are dynamic regulators of cancer progression rather than short-lived bystanders. Within the tumor microenvironment (TME), tumor-associated neutrophils (TANs) adopt heterogeneous, context-dependent states that extend far beyond the conventional N1/N2 dichotomy. Recent single-cell and spatial transcriptomic insights reveal that cytokine, stromal, metabolic, and epigenetic cues orchestrate this remarkable TAN plasticity. Once activated, TAN-derived proteases, reactive oxygen species, and neutrophil extracellular traps (NETs) drive angiogenesis, immune evasion, metastasis, and therapy resistance. Conversely, specific TAN subpopulations can directly kill tumor cells, present antigens, and synergize with adaptive immunity. This review synthesizes recent breakthroughs in TAN recruitment, heterogeneity, metabolic and epigenetic reprogramming, and cellular crosstalk. Furthermore, we evaluate therapeutic strategies targeting molecular signaling and NET formation, alongside emerging approaches to reprogram or engineer neutrophils, such as chimeric antigen receptor-engineered neutrophils (CAR-Ns). Finally, we address current barriers to clinical translation and outline how to achieve context-dependent modulation of neutrophil states to preserve essential host defenses during TAN-targeted oncology therapies.

This study reports a robust DNA polymer spherical nucleic acid (PSNA) nanomachine based on domain-crosslinked self-assembly for highly selective and ultrasensitive imaging of microRNAs (miRNAs) in living cells. The nanomachine is constructed from DNA micelles formed via self-assembly of amphiphilic monomers followed by radical-initiated polymerization. This design effectively overcomes key limitations of conventional spherical nucleic acids, including poor biostability, inefficient cellular uptake, and nonspecific signal leakage, by providing a covalently stabilized architecture with high DNA loading capacity. A target-recognizing lock-walking strand and a fluorophore-labeled track strand containing the substrate sequence are immobilized on two distinct PSNA particles, respectively. Upon specific activation by endogenous miRNA-21 within tumor cells, the DNA walker is released and initiates a cyclic walking process that binds to the substrate strands and triggers a fuel-driven strand displacement cycle, leading to significant signal amplification. This walking mode is multi-legged and spatially confined. It enables high-speed reaction kinetics, substantially enhances fluorescence signals, and improves imaging clarity. The resulting PSNA nanomachine exhibits excellent stability and amplification capability, permitting ultrasensitive miRNA detection under low-background conditions. Moreover, this platform has been successfully applied for in situ imaging of miRNA in various cell lines, demonstrating strong potential for early clinical diagnostics and real-time monitoring of therapeutic responses.

Recent advances in large language models have extended to genomic applications, yet model robustness relative to context is unclear. Here, we demonstrate two intrinsic biases (input sequence length and nucleotide position) affecting SegmentNT results, a model included with the Nucleotide Transformer that provides nucleotide-level predictions of biological features. We demonstrate that nucleotide position within the input sequence (beginning, middle, or end) alters the nature of SegmentNT's raw prediction probabilities, which can be standardized to improve prediction consistency. While longer input sequence length improves model performance, diminishing returns suggest a surprisingly small input length of ∼3072 nucleotides might be sufficient for many applications. We further identify a 24-nucleotide periodic oscillation in SegmentNT's prediction probabilities, revealing an intrinsic bias potentially linked to the model's training tokenization (6-mers) and architecture. We identify potential approaches to account for these biases and provide generalizable insights for utilizing nucleotide-resolution functional prediction models.

FDA-approved oncolytic herpes simplex virus-1 (oHSV) therapy has emerged as a promising viro-immunotherapy for solid tumors. However, tumor- and tumor microenvironment (TME)-associated adaptations following viral treatment, such as feedback immune suppression, neoangiogenesis, and enhanced tumor aggressiveness, often hinder complete tumor eradication. A deeper understanding of the molecular mechanisms underlying resistance to oHSV is crucial to enhancing its clinical impact. We recently discovered that oHSV induces Insulin-like growth factor 2 (IGF2) secretion, shaping an immunosuppressive TME. Similarly, radiotherapy (RTx) activates the IGF1/IGF1R and YAP1 signaling pathways, further promoting therapeutic resistance. In this study, we investigated how oHSV-induced Insulin-like growth factor 1 receptor (IGF1R) signaling drives feedback pro-survival and proliferative pathways in tumor cells and evaluated the therapeutic potential of combining IGF1R blockade with oHSV and RTx. We first demonstrated that oHSV activates IGF1R signaling in vitro and in vivo, promoting tumor proliferation. While IGF1R-targeted monotherapies have shown limited cytotoxicity, its combination with oHSV led to a modest but significant increase in cytotoxicity across tested breast cancer (BC) and primary glioblastoma (GBM) cells in vitro and in vivo xenograft models. Furthermore, we observed that co-treatment with oHSV and RTx robustly activated both IGF1R and YAP1 in resistant cells, revealing the IGF1R/YAP1 axis as a key mediator of resistance to dual oHSV and RTx therapy. Notably, the triple combination of oHSV, RTx, and IGF1R blockade yielded synergistic anti-tumor effects, abolished YAP1 expression and nuclear localization, and significantly enhanced survival in orthotopic BC and GBM models. Collectively, these findings identify the IGF1R/YAP1 axis as a critical driver of resistance to oHSV and RTx and provide a strong rationale for the clinical evaluation of this triple-combination strategy to enhance therapeutic efficacy in patients with BC and GBM.

Ferroptosis is a unique form of programmed cell death that involves multiple organelles. Although traditionally viewed as a 'degradation workshop', accumulating evidence reveals that the lysosome serves as a central hub for iron metabolism and signal transduction, orchestrating the overall fate of cellular ferroptosis across spatiotemporal dimensions. In this review, we propose the concept of the 'lysosome-ferroptosis axis' and outline its roles in metabolic signaling, autophagy, and lysosomal membrane permeabilization. We further discuss the involvement of this axis in neurodegenerative, tumor, and cardiometabolic diseases, with the aim of providing new insights for targeted therapeutic strategies.

Long noncoding RNAs (lncRNAs) regulate the tumor microenvironment (TME), yet their cell-intrinsic roles within immune populations of the TME remain underappreciated. In this review, we shift focus from the cancer cell to the immune compartment, systematically reviewing how immune cell-intrinsic lncRNAs govern CD8+ T cell exhaustion, CD4+ T cell polarization, NK cell cytotoxicity, dendritic cell antigen presentation, and macrophage inflammatory programming. We highlight lncRNAs that function as molecular switches-tipping immune cells between antitumor effector and immunosuppressive states-and examine how exosomal lncRNAs extend these regulatory circuits across cellular boundaries within the TME. Finally, we evaluate opportunities for lncRNA-based biomarkers and therapies designed to target the immune landscape, outlining a framework for integrating immune-intrinsic lncRNA biology into precision immuno-oncology.

RAS mutations are among the most prevalent oncogenic drivers in solid tumors and are consistently associated with suboptimal responses to radiation therapy (RT). Within this family, KRAS is the dominant isoform and a central regulator of tumor stress adaptation. Increasing evidence indicates that oncogenic KRAS orchestrates radioresistance through coordinated tumor-intrinsic and microenvironmental mechanisms. Cell-intrinsically, KRAS enhances DNA damage repair, replication stress tolerance, redox buffering, and ferroptosis defense. The KRAS-NRF2-53BP1 axis exemplifies this program by accelerating non-homologous end joining and enabling rapid repair of radiation-induced DNA double-strand breaks. Concurrently, KRAS reshapes the tumor microenvironment by promoting myeloid recruitment, metabolic rewiring, impaired antigen presentation, and immune checkpoint upregulation, thereby constraining the immunogenic effects of RT. The rapid evolution of RAS-directed therapeutics, including allele-specific, ON-state, dual-state, and pan-RAS inhibitors, as well as emerging degraders and molecular reprogramming strategies, has transformed a historically "undruggable" target into a clinically actionable vulnerability. Preclinical evidence indicates that KRAS inhibition can restore radiosensitivity and partially recondition antitumor immunity. However, adaptive resistance frequently converges on MAPK pathway reactivation and persistent immune suppression. Integrating next-generation RAS inhibitors with RT and immune-directed therapies may therefore represent a critical strategy for achieving durable tumor control in KRAS-mutant cancers.

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

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.

A critical challenge in the bench-to-bedside translation of controlled drug release strategies is the sharp decline in reaction efficiency as biological complexity increases. A platform capable of maintaining bioorthogonal-like drug release─remaining minimally perturbed by physiological environments─would address an unmet clinical need. This is particularly relevant for radiotherapy-mediated drug release, where the oxidative activation of prodrugs is often compromised by the rapid quenching of reactive intermediates in vivo. Herein, we engineer a hafnium-based nanoscale metal-organic layer platform that leverages a unique "surface-confinement effect" to overcome this challenge. By covalently tethering prodrugs to the Hf-nMOLs surface, we constructed two-dimensional nanoreactors that spatially localize the activation process within an interface enriched with reactive species. This design effectively insulates the activation step from biological scavengers, ensuring efficient payload release efficiency across increasing biological complexity. When loaded with the topoisomerase I inhibitor Exatecan, the Hf-nMOLs system achieved an intratumoral drug-release G-value of 568 nM·Gy-1, resulting in potent radiosensitization and significant tumor-growth suppression under low-dose X-ray irradiation. This work presents a versatile strategy for robust radio-chemotherapeutic combinations, achieving the simultaneous release of diverse payloads activated by radiotherapy. Our findings also suggest that engineering nanoscale surface confinement may provide a generalizable materials strategy to help confer bioorthogonality to otherwise labile activation reactions.

Targeted protein degradation (TPD) has emerged as a powerful strategy to eliminate disease-relevant proteins, yet current approaches remain largely constrained to hijacking ubiquitin ligases. We previously introduced ByeTACs, bifunctional molecules that directly recruit proteins to the proteasome for E-ligase independent degradation. Here, we report "Truly" degraders, a new class of dual-mechanism molecules that combine a ligand for the proteasomal receptor Rpn13 with a ligand for cereblon (CRBN) to simultaneously engage both ubiquitin-independent and ubiquitin-dependent degradation pathways. Structure-guided design identified an optimal linker length that supports efficient substrate processing, with the PEG4 derivative (Truly-4) inducing robust depletion of both Rpn13 and CRBN in several cancer cell types. Remarkably, Truly-4 is the first noncovalent small molecule shown to degrade full-length Rpn13, a target previously approached using covalent or domain-restricted strategies. Mechanistic studies confirmed that degradation of Rpn13 proceeds via CRBN-dependent E3 ligase activity, whereas CRBN degradation occurs through an E-ligase independent process, consistent with a ByeTAC mechanism. Importantly, Truly-4 induces selective cytotoxicity in hematologic and solid cancer cell lines but not in healthy cells, despite comparable Rpn13 depletion, indicating that dual degradation can uncouple target engagement from toxicity. These findings establish a generalizable framework for engineering bifunctional degraders that program the proteasome to execute parallel degradation mechanisms and highlight proteasome receptors as druggable nodes for selective destruction of disease-relevant proteins.

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

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

Lipid nanoparticle-encapsulated mRNA (mRNA-LNP) vaccines trigger the potent differentiation of antigen-specific cytotoxic CD8 T cells in addition to antibody production. Despite its high immunogenicity, the cellular mechanisms by which mRNA-LNP induces such unusual immune responses remain largely unclear. Here, we show that mRNA-LNP induces potent and long-lasting CD8 T cell expansion through an antigen presentation mechanism that differs from that of conventional adjuvants. In mice immunized with mRNA-LNP, the number of antigen-specific CD8 T cells was one order of magnitude higher than that induced by combining antigen proteins with immunostimulants such as lipopolysaccharide or polyinosinic:polycytidinic acid. Intramuscularly administered mRNA-LNPs were mainly taken up by migratory type 2 conventional dendritic cells in draining lymph nodes, resulting in notably strong and persistent antigen presentation through major histocompatibility complex class I. Furthermore, CD8 T cell induction by mRNA-LNP required migratory dendritic cells but not the traditional cross-presentation pathway that is otherwise essential for antiviral and antitumor immunity. Thus, the mRNA-LNP formulation exerts unconventional immune responses through a different antigen-presentation pathway from conventional component vaccines.

Cancer therapy has advanced substantially through targeted therapies and immunotherapy; however, durable clinical responses remain limited by the development of drug resistance. Increasing evidence identifies cancer stem cells (CSCs) as central drivers of therapeutic failure, tumor recurrence, metastasis, and minimal residual disease. CSCs possess self-renewal and differentiation capacities together with remarkable adaptability under therapeutic stress, enabling long-term tumor maintenance and regeneration. CSC-mediated resistance arises through coordinated intrinsic and extrinsic mechanisms. Intrinsically, CSCs employ multiple survival programs, including cellular quiescence, enhanced DNA damage response and repair, ATP-binding cassette transporter-mediated drug efflux, apoptosis evasion, and metabolic reprogramming. Extrinsically, these mechanisms are reinforced through dynamic interactions with the tumor microenvironment (TME), particularly hypoxic and perivascular niches that support stemness and therapeutic tolerance. Importantly, CSCs are increasingly recognized as dynamic cellular states rather than fixed populations and exhibit marked plasticity through reversible transitions between stem-like and non-stem states, frequently mediated by epithelial-mesenchymal transition (EMT). This plasticity promotes intratumoral heterogeneity and replenishes resistant cell populations. In this review, we provide a comprehensive synthesis of the molecular and microenvironmental mechanisms underlying CSC-driven drug resistance and critically discuss emerging therapeutic strategies targeting CSC plasticity, niche interactions, metabolic adaptation, and immune evasion. Collectively, these insights support the development of integrated multi-target therapeutic approaches to improve long-term clinical outcomes.

Tumor-associated macrophages (TAMs) primarily arise from infiltrating monocytes, yet the mechanisms guiding their differentiation remain unclear. Here, we show that human macrophages rely on autocrine macrophage migration inhibitory factor signaling to suppress p53 during M2-like transition. Disruption of this pathway led to activation of p53 and, unexpectedly, the nuclear receptor NR4A1, inducing a senescence-like state resembling interleukin (IL)-1β⁺ and IL-4 Induced 1 (IL4I1)⁺ TAM subsets observed across multiple cancers. These TAM-like macrophages exhibited a transcriptional program driven by NR4A1, similar to that induced in IL-1β⁺ TAMs by the combined action of tumor necrosis factor α (TNF) and the cyclooxygenase-2-dependent arachidonic acid (AA) metabolite prostaglandin E2. They also upregulated the AA-selective acyl-CoA synthase ACSL4, which promoted cell survival and restrained IL-1β release despite elevated IL1B expression. This effect was mediated through induction of the IL4I1⁺ TAM marker CD38, which drove the production of IL-10. Mechanistically, ACSL4 preserved the homeostatic function of stimulator of interferon genes (STING). Loss of ACSL4 resulted in extensive macrophage death and converted STING from a homeostatic regulator into a driver of IL-1 cytokine release. Finally, we show that the CDK4/6 inhibitor abemaciclib repolarized TAM-like macrophages toward a more inflammatory phenotype through off-target inhibition of ACSL4. Abemaciclib enhanced inflammatory signaling by modulating ectodomain shedding, increasing TNF while reducing the release of its natural antagonist, TNF receptor II. Together, these findings clarify mechanisms underlying scRNA-seq-defined TAM phenotypes, identify ACSL4 as a potential therapeutic target, and reveal how abemaciclib can promote inflammatory responses in cancer patients.

It has been highly challenging to reliably detect various cancers at an early stage when tumors are just millimeters in size. To address this, we developed a tumor microenvironment (TME)-responsive nanoprobe, PR-KAd@CD-AuNC, enabling cross-validated cancer detection through in vivo second near-infrared (NIR-II) fluorescence imaging and in vitro colorimetric urinalysis. The nanoprobe consists of three integrated components: a renal-clearable signal-output segment (cyclodextrin-functionalized gold nanocluster, CD-AuNC), a tumor-targeting and size-controlling component (PR), and a matrix metalloproteinase-2 (MMP2)-cleavable linker (KAd). PR, composed of 8-arm poly(ethylene glycol) for prolonged circulation and c(RGDfK) peptides for αvβ3 integrin targeting, directed selective tumor accumulation after intravenous injection of PR-KAd@CD-AuNC. The intrinsic emission of CD-AuNC above 1100 nm enabled high-resolution, real-time NIR-II fluorescence imaging with attenuated photon scattering, allowing precise tumor delineation. Within the TME, specifically overexpressed MMP2 cleaved the KAd linker, releasing ∼2 nm CD-AuNC fragments from the ∼10 nm parent nanoprobe. Being smaller than the ∼5.5 nm renal filtration threshold, these fragments were renally excreted. Concurrently, the peroxidase-like activity of CD-AuNC catalyzed tetramethylbenzidine oxidation to produce a visible blue signal, providing a simple and low-cost urinalysis method suitable for broad cancer screening applications. This dual-modality strategy effectively distinguished cancers from inflammation and other diseases, detecting small tumors for multiple cancer types with a sensitivity surpassing that of computed tomography (CT) imaging, which makes it a promising early detection approach. Furthermore, it was also employed to dynamically assess cancer therapeutic efficacy (i.e., immunotherapy, chemotherapy, and surgical resection), suggesting clinical value for guiding treatment decisions.

Although checkpoint inhibitors have revolutionized cancer treatment, responses are largely restricted to targeting CTLA-4 and PD-(L)1. TNFR2 has been identified as a promising target, but further therapeutic development requires a better understanding of whether agonists or blockers should be used and whether FcγR interactions promote efficacy. Here, we engineered two distinct α-TNFR2 antibody types: ligand-blocking FcγR-engaging versus non-ligand-blocking agonist antibodies. Both showed potent anti-tumor efficacy across multiple syngeneic mouse tumor models and combined effectively with α-PD-1. The ligand-blocking antibody depleted intratumoral Tregs and reprogrammed the myeloid compartment, directing monocytes towards a pro-inflammatory macrophage and dendritic cell trajectory. Interestingly, this effect depended on both inhibitory and activating FcγRs, reflecting the simultaneous action on Tregs and myeloid cells. In contrast, the agonist directly co-stimulated T and NK cells, partially through FcγR-independent mechanisms. Both antibodies converged on activating tumor-specific CD8+ T cells mediating tumor rejection. Clinical assessment of both ligand-blocking (BI-1808) and agonist (BI-1910) α-TNFR2 antibodies is currently ongoing.

Antibody-drug conjugates (ADCs), a revolutionary tumor therapeutic modality integrating the targeting specificity of monoclonal antibodies and the cytotoxicity of payloads, have undergone four generations of development, with over 20 approved drugs and more than 300 candidates in clinical trials globally. Despite their remarkable success in treating both hematological malignancies and solid tumors, traditional ADCs still face intractable challenges, including heterogeneity in target antigen expression, insufficient internalization, payload-mediated drug resistance, and severe treatment-related adverse events. This review elaborates on the latest design optimization strategies and research advances of next-generation function-enhanced ADCs, focusing on four core directions: bispecific ADC construction, internalization efficiency improvement, binding affinity enhancement, and payload combination innovation.

Therapeutic resistance remains a prevalent and intractable clinical challenge across a broad spectrum of human malignancies. Despite extensive investigations, the intricate molecular networks by which the tumor microenvironment (TME) mediates such resistance are not fully understood. In this study, we identified nucleotide-binding oligomerization domain-containing proteins 1 and 2 (NOD1/2) as pivotal regulators of adaptive resistance to diverse antitumor therapies, including immune checkpoint blockade (ICB), adoptive T-cell therapy, and cytotoxic chemotherapy. In murine tumor models, genetic ablation of NOD1/2 or receptor-interacting protein kinase 2 (RIPK2), as well as pharmacological inhibition of RIPK2, remodeled the TME by decreasing immunosuppressive macrophages and boosting CD8⁺ T cell infiltration and cytotoxicity. Mechanistically, NOD1/2 activation in macrophages upregulated programmed death-ligand 1 (PD-L1) expression via the RIPK2/NF-κB signaling axis, establishing an immunosuppressive TME that impaired CD8⁺ T cell-mediated antitumor immunity. Notably, in the clinically relevant setting of immunotherapy resistance, targeted suppression of NOD1/2 signaling in patient-derived peripheral blood mononuclear cells (PBMCs) restored and potentiated ICB responsiveness in patient-derived tumor organoids. Bioinformatic analyses further demonstrated that NOD1/2-associated gene signatures were significantly enriched in tumor-associated macrophages post-therapy. Our findings define NOD1/2 as a novel innate immune checkpoint that orchestrates therapy-induced adaptive resistance and highlight this pathway as a promising target to overcome treatment resistance in refractory cancers.

Zn2+ offers significant advantages for cancer therapy via mitochondrial disruption, immune activation, and metal homeostasis perturbation, but faces challenges of rapid clearance and systemic toxicity. Existing zinc‑based (e.g., zinc oxide (ZnO), and zinc peroxide) nanoparticles often lack internal voids and sufficient surface area, restricting drug loading and functionalization. In this study, we developed a crystallinity‑gradient selective etching method to tailor biodegradable hollow mesoporous zinc nanoparticle (HMZN). Typically, zinc peroxide nanoparticles with low‑crystallinity core and high‑crystallinity shell were synthesized, and ammonia water was used to selectively etch the inner low‑crystallinity core, producing HMZN. Hemin (HEM) and Gd‑poly(acrylic acid) macrochelates (GP) were then loaded to construct HEM@HMZN@GP for dual-source H2O2 surge-driven tumor ferroptosis therapy. In acidic tumor microenvironment, Zn2+, H2O2, HEM, and GP can be released from HEM@HMZN@GP. The released Zn2+ damages mitochondrial electron transport chain (ETC) and generates superoxide anions (•O2 -), which can be rapidly catalyzed by superoxide dismutase into H2O2. The dual-source of H2O2 markedly elevates local oxidative stress. HEM can be enzymatically converted by heme oxygenase‑1 to Fe2+, which reacts with the abundant H2O2 via Fenton reaction to produce hydroxyl radicals, initiating lipid peroxidation and ferroptosis. GP enhances T1‑weighted magnetic resonance imaging signals, facilitating drug delivery visualization in tumors.

Herbal self-assembled nanoplatforms (H-SANs) represent an emerging strategy in herbal nanomedicine by integrating the intrinsic bioactivity of phytochemicals with supramolecular nanoscale organization. Unlike conventional nanocarriers that may be limited by low drug loading, excipient burden, premature release, and scale-up complexity, H-SANs are carrier-free or predominantly bioactive systems formed through noncovalent interactions, including π-π stacking, hydrogen bonding, hydrophobic forces, electrostatic interactions, and metal coordination, among polyphenols, flavonoids, alkaloids, terpenoids, saponins, and related phytochemical components. Reported assemblies can achieve high drug loading, exhibit stimuli-responsive structural changes under disease-associated cues such as acidic pH, redox imbalance, or enzyme overexpression, and can be further engineered with targeting motifs. Preclinical studies have explored their applications in oncology, inflammation, neurodegeneration, metabolic disorders, cardiovascular disease, antimicrobial therapy, wound repair, kidney injury, and bone disorders, where H-SANs may act as bioactive scaffolds as well as delivery systems. However, their translational development remains constrained by dilution-dependent instability, incomplete understanding of in vivo disassembly and parent-drug regeneration, protein corona formation, tumor transport heterogeneity, raw-material variability, manufacturing reproducibility, regulatory classification, and limited human-relevant pharmacokinetic and safety evidence. Emerging strategies, including AI-assisted modeling, rational multicomponent co-assembly, continuous manufacturing, and improved biological validation, may help address these challenges. Overall, H-SANs provide a promising but still largely preclinical framework for high-payload herbal nanomedicine, and their clinical value will require rigorous physicochemical characterization, reproducible manufacturing, and cautious translational validation.

The ability to selectively edit specific RNA modifications is needed to understand their roles in cellular function and disease. However, existing approaches, particularly catalytically inactive Cas (dCas)-based systems, have limited applicability owing to their reliance on eraser proteins, and their performance can vary depending on the modification type and target context. Here we introduce the RNA Modification-Blocking (RModBlock) strategy, which uses chemically modified antisense oligonucleotides (ASOs) with locked nucleic acid to precisely inhibit modifications at targeted sites. Most RNA modification writers, including those for m5C and pseudouridine, require specific structural contexts. Using representative modifications with and without eraser proteins, m5C and pseudouridine, respectively, we demonstrated that RModBlock ASOs blocked the formation of their structural context, inhibiting modified bases by up to 97% in human cells. RModBlock ASOs also inhibited m6A, despite its writer protein not requiring a strict structural context, suggesting broad applicability. Moreover, this strategy achieves comparable or superior performance when dCas13-eraser-based systems are applicable. Finally, by inhibiting cancer-relevant modifications and through in vivo delivery to the mouse liver, we highlight its therapeutic potential. This showcases the RModBlock strategy as a precise, efficient and versatile approach for manipulating RNA modifications, with broad applicability in basic and translational research.

Deubiquitinases regulate key oncogenic and immune pathways but have proven challenging to exploit therapeutically. Among them, ubiquitin-specific protease 22 (USP22) has emerged as a molecule of interest due to its involvement in both tumor-intrinsic programs and tumor-immune interactions. In this review, we synthesize current evidence describing how USP22 modulates oncogenic transcriptional states and immune evasion. Tumor-intrinsically, USP22 has been implicated in stabilizing select oncogenic factors and shaping chromatin accessibility in ways that can reinforce proliferation, survival, stem-like properties, and metastatic potential. In parallel, USP22 influences antitumor immunity by modulating MHC-I-mediated antigen presentation, immune checkpoint expression, and the fitness of intratumoral regulatory T cells. We discuss emerging pharmacological approaches to target USP22, the limitations of current inhibitor strategies, and the importance of distinguishing direct enzymatic functions from indirect transcriptional consequences. Together, these insights suggest that USP22 inhibition may offer a therapeutic strategy with dual effects on tumor biology and antitumor immunity.

In the 19th century, Rudolf Virchow observed lymphocyte infiltration in tumors and suggested a potential link between cancer and inflammation. Current explanations for the origin of tumor-infiltrating lymphocytes are based on the interaction between cancer cells and the immune system. This model was proposed before it was recognized that tumors are frequently infected or colonized by microbial elements, suggesting the presence of bacteria-specific and virus-specific T cells within tumors. While several aspects of this microbial experience remain under investigation, their impact on the tumor microenvironment and antitumor immune response may be significant, as the pathogens may elicit the direct or indirect recruitment of tumor-infiltrating lymphocytes. This "Infection Hypothesis" regarding the origin of tumor-infiltrating lymphocytes complements the classical tumor immunoediting hypothesis, offers new avenues for a comprehensive understanding of the tumor ecosystem, and highlights the need for more nuanced cancer treatment approaches that account for the potential interconnected roles of lymphocytes and pathogens in the tumor milieu.

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

Beyond their canonical role in bioenergetics, mitochondria are now recognized as critical signaling platforms that orchestrate innate immune responses. Central to this function is mitochondrial dynamics-the controlled equilibrium between fission and fusion-which serves as a critical structural and thermodynamic checkpoint for cellular fate and immunological status. A substantial body of evidence indicates that pathological mitochondrial fission, frequently driven by Dynamin-related protein 1 (Drp1), is a hallmark of numerous inflammatory conditions. Mechanistically, fragmented mitochondria release damage-associated molecular patterns (DAMPs) and induce acute ATP suppression, metabolically "licensing" NLRP3 activation by collapsing the ATP hydrolysis potential (ΔGATP). Recent breakthroughs have redefined this axis, distinguishing between physical damage and metabolic triggers, such as pyrimidine imbalance via the YME1L-SLC25A33 axis. Furthermore, the immunogenicity of DAMPs is strictly context-dependent; oxidized or "fragile" mtDNA containing ribonucleotides act as hyper-immunogenic ligands for cytosolic sensors like cGAS-STING. Emerging evidence further highlights that endosomal-mitochondrial crosstalk, intercellular mitochondrial transfer, and lipid-driven metabolic rewiring profoundly govern macrophage polarization and tissue homeostasis. Conversely, promoting mitochondrial fusion and robust quality control preserves organellar integrity and attenuates inflammatory cascades. This review critically synthesizes current literature, deconstructing the molecular linkages between organelle structure and metabolic signaling. By exploring the consequences in sepsis, neuroinflammation, osteoarthritis, and cancer, this treatise evaluates the pharmacological potential of modulating mitochondrial dynamics-ranging from direct Drp1 inhibitors and unfractionated heparin to metabolic stabilizers (e.g., GLP-1 receptor agonists), multi-pronged disruptors (e.g., Antimycin A), targeted nanomedicine, and communication-driven mitochondrial transplantation-positioning this axis as a promising frontier for precision pharmacology.

In this issue, Shu et al.1 identify the RNA-binding protein PCBP1 as a PARP1 interactor and negative regulator, a function relieved by SIRT7 deacetylation of PCBP1 upon DNA damage. Tumor studies indicated clinical relevance of this pathway, especially in response to radiotherapy, in modulating PARP1 activity as a therapeutic strategy.

Regulatory T cells (Tregs) are essential for maintaining immune tolerance. We recently identified chaperone-mediated autophagy (CMA), a selective lysosomal degradation pathway, as a critical regulator of Treg function. Treg activation induces CMA, but this response is markedly diminished with aging. Mice lacking CMA specifically in Tregs develop systemic inflammation, impaired immune tolerance and reduced lifespan. We confirm that CMA is a fundamental mechanism supporting Treg suppressive function as CMA-deficient Tregs are unable to suppress intestinal inflammation in a model of inflammatory bowel disease and fail to block the anti-oncogenic immune response activated in a syngeneic tumor model. Mechanistically, CMA supports metabolic fitness, remodels immune-related protein networks, and promotes degradation of the m6A RNA demethylase FTO, linking lysosomal proteostasis to epitranscriptomic control of IL-2 responsiveness. Restoration of CMA in aged mice improves Treg function, highlighting CMA as a promising therapeutic target for both inflammatory diseases and cancer immunotherapy.

Chitinase-3-like-1 (CHI3L1, also known as YKL-40) has been recognized as a biomarker of inflammation and tissue remodeling and has now emerged as a pseudoenzymatic immune checkpoint. Recent structural, immunological, and translational studies redefine it as an active regulator of immune suppression rather than a passive disease marker. Despite lacking catalytic activity, CHI3L1's conserved chitinase fold and glycan-decorated surface acts as a modular scaffold linking cytokine, metabolic, and stress signals to immune suppression and fibrotic remodeling. Through its chitinase-like structure it engages with receptor triad complex, including IL-13Rα2, TMEM219, and Galectin-3, activating signaling pathways such as MAPK, PI3K/AKT, and TGF-β/Smad. Through these interactions, CHI3L1 has been reported to promote fibroblast activation, angiogenesis, and immune suppression, creating environments where immune cells are excluded or silenced. These checkpoint-like effects are not limited to cancer; they also appear in fibrosis, infection, and neuroinflammatory diseases, linking CHI3L1 to a broad spectrum of immune-resistant conditions. This review integrates structural glycobiology, receptor pharmacology, and immunometabolic mechanisms to reveal CHI3L1 as a unifying regulator of stromal-immune crosstalk. It further highlights preclinical and clinical evidence demonstrating that antibody, RNA-based, and small-molecule inhibitors of CHI3L1 restore immune surveillance and limit pathological remodeling. Targeting this pseudoenzymatic checkpoint alongside canonical PD-1 and CTLA-4 blockade represents a promising strategy to overcome immune resistance and normalize pathogenic tissue remodeling across diverse diseases.

How transcription factors (TFs) and their binding sites organize and engage nucleosomes at natural genomic locations remains poorly understood. Here, we develop Benzonase-seq to measure the rotational phasing of nucleosomes in human cells and enhance chromatin immunoprecipitation (ChIP)-exo (v6) to measure rotational phasing on the same DNA molecule bound by a TF. Unbound CTCF sites were found to be rotationally accessible on nucleosomes, and this rotational accessibility is encoded by classical dinucleotide periodicities. CTCF binding results in nucleosome displacement to adjacent DNA phasing sequences. Upon examining 40 TF classes, unbound sites were found to be phased either inward or outward or to lack phasing. In all examined cases, TF binding (e.g., NFIA and FoxA) results in adjacent rotational and translational phasing, which is not dinucleotide encoded. Benzonase-seq also more robustly maps nucleosome and subnucleosome positions in hard-to-map CpG islands. These findings provide a clearer view of how TFs engage and position nucleosomes to shape the natural chromatin landscape.

Poly (ADP-ribose) polymerase 1 (PARP1) is a DNA damage sensor and one of the initiating enzymes essential for DNA repair. Understanding how to control PARP1 activity is critical for developing strategies to maintain genome stability. Here, we report that the RNA-binding protein poly(rC)-binding protein 1 (PCBP1) interacts with and inhibits PARP1 activity under physiological conditions, and that this inhibition is relieved in the early phase of the DNA damage response. Mechanistically, histone deacetylase SIRT7 mediates the deacetylation of PCBP1 at K314 and K351, disrupting its interaction with the DNA-binding domain of PARP1 and thereby permitting PARP1 activation. Modulating PCBP1 expression or acetylation through gene editing or antagonistic peptides affects the efficiency of DNA damage repair in both cell and murine models. Reduced PCBP1 levels are associated with poor survival in cancer patients following radiotherapy. This regulatory role for PCBP1 in PARP1 activity may form the basis of a therapeutic strategy that combines the inhibition of PCBP1 deacetylation with DNA-damaging agents.

We report a fluorescence chip based on a membrane-anchored, bipedal DNAzyme walker assembled on small extracellular vesicles (sEVs) surfaces via catalytic hairpin assembly (CHA). Unlike previously reported single-arm or proximity-ligation-dependent walkers on nanoparticle supports, this CHA-triggered DNAzyme walker is designed with a bipedal architecture and operates directly on the sEVs membrane. In our design, CHA simultaneously generates two DNAzyme arms on the same sEVs, enabling faster substrate cleavage kinetics and superior signal amplification. The assay achieves a theoretical detection limit of 2.8 particles/μL, approximately 1.5-fold lower than a comparable single-arm architecture. Dual recognition of the sEVs lipid bilayer and surface EpCAM protein (using a cholesterol-modified substrate and an anti-EpCAM antibody) effectively eliminates false-positive signals from free EpCAM or soluble interferents. The entire reaction cascade is integrated onto a chip platform, improving reproducibility and point-of-care potential. The chip performs robustly in fetal bovine serum and clinical serum samples, with recovery rates of 99.1-103.8% and strong correlation with nanoparticle tracking analysis. By establishing a CHA-assisted, bipedal walking mechanism on biological membranes, this work opens a new paradigm for designing efficient DNA nanomachines on membrane-enveloped targets.

Photodynamic therapy (PDT) demonstrates remarkable versatility by activating diverse non-apoptotic cell death pathways, effectively circumventing apoptosis resistance and enhancing tumor eradication. Key mechanisms include autophagic cell death, regulated necrosis-driven immune activation, ferroptosis-mediated oxygen replenishment, pyroptosis-induced immunogenic cell death (ICD), and paraptosis. These pathways collectively highlight PDT's capacity to disrupt tumor survival mechanisms and stimulate systemic anti-tumor immunity. However, several challenges remain, including precise spatiotemporal control of ROS, hypoxia mitigation, and selective photosensitizer delivery. Advances in nanotechnology, hypoxia-responsive agents, and combination therapies (e.g., immune checkpoint inhibitors or chemotherapy) hold promise for overcoming these limitations. In this review, we discuss multiple types of non-apoptotic cell death pathways activated by PDT and the underlying mechanisms of each distinct cell death process. We also review the clinical applications and current challenges of leveraging these pathways to enhance tumor treatment in PDT. Future research should prioritize the development of subcellular-targeted photosensitizers, deeper light-penetration technologies, and biomarker-guided personalized regimens for more precise and effective PDT.

Chromosome mis-segregation events that remain unresolved during cytokinesis threaten genome stability. Persistent ultrafine DNA bridges engage the Aurora B-dependent abscission checkpoint (termed NoCut), which delays abscission by phosphorylating components of the ESCRT complex. Here we show that NoCut surveillance repurposes human ESCRT-III, the membrane-remodeling complex that seals the reforming nuclear envelope in anaphase. In response to persistent ultrafine DNA bridges, ESCRT-III transfers from the reforming nuclear envelope to the mis-segregated DNA bridge and ESCRT-III complexes protect the DNA from damage, as evidenced by increased DNA damage upon CHMP1B depletion. Complementary in vitro assembly reactions show that the human ESCRT-III proteins CHMP1B and IST1 can copolymerize into double-stranded filaments that encase double-stranded DNA and nucleosomes and prevent nuclease digestion and cGAS recognition, demonstrating that ESCRT-III complexes can directly bind and protect DNA. Lastly, cells expressing a DNA-binding mutant of CHMP1B exhibit cytokinesis failure and binucleation when ultrafine DNA bridges persist, revealing a mechanism of safeguarding genome stability.

The successful development of KRASG12C mutation-specific covalent inhibitors made direct targeting of KRAS a reality, yet the limitations in mutational coverage and susceptibility to resistance have spurred the pursuit of broader and more durable strategies. This review highlights the most recent developments on innovative strategies beyond traditional inhibitors, including molecular glues (MGs) and protein-protein interaction (PPI) modulators. Molecular glues, such as the RMC macrocyclic series (e.g., RMC-6291 as a G12C-selective MG, RMC-6236 as a pan-MG), function by recruiting endogenous chaperone proteins to form ternary complexes that sterically block KRAS-effector interactions or allosterically restore GTPase activity. This macrocyclic platform enables rational design of mutation-selective and broad-spectrum MGs. Another parallel advancement involves the design of small molecular MGs to strengthen KRAS engagement with E3 ligases (e.g., NEDD4-1 or LZTR1), thereby resulting in enhanced PPI or degradation of KRAS. Complementarily, PPI inhibitors (e.g., BBO-10203, VVD-699) disrupt critical oncogenic interfaces between KRAS and PI3Kα, offering a strategy to inhibit oncogenic signaling with fewer side effects. Collectively, balancing broad-spectrum versus selective targeting of KRAS, attaining optimal drug-like properties and establishing rational combination therapies with other modalities remain key challenges. The ongoing evolution of MGs and PPI modulators heralds a major expansion of the therapeutic frontier for KRAS-driven cancers and is redrawing the boundaries of precision oncology.

Antibody-drug conjugates (ADCs) have significantly advanced cancer therapy by enabling the selective delivery of cytotoxic agents to tumour cells. However, ADC efficacy remains constrained by its dependence on a single target antigen, which limits tumour targeting and promotes resistance in heterogeneous tumours with variable and low antigen expression1-6. Here we introduce an in vivo bioorthogonal ligation strategy that generates functional antibody-ADC click constructs following systemic administration. This platform provides a modular and translatable approach for enhanced targeted drug delivery in heterogeneous tumours. We conjugate therapeutic antibodies and ADCs with trans-cyclooctene and tetrazine moieties for sequential administration to enable in vivo ligation of an antibody with an ADC after systemic delivery. The antibody-ADC click approach demonstrated improved antitumour activity relative to standard ADC monotherapy or antibody plus ADC combinations in preclinical models of HER2 and EGFR co-expression. These included tumours with low, ultralow, negative or heterogeneous HER2 expression and resistant or ineligible for conventional HER2-directed ADCs. This modular strategy leverages receptor biology and bioorthogonal chemistry for optimal therapeutic efficacy and does not require extensive antibody re-engineering. Moreover, the antibody-ADC click approach can be extended to other receptor pairs, which makes it a flexible modular platform to address heterogeneity and resistance to targeted therapies across different tumour types.

Aberrant glycosylation is a nearly universal hallmark of cancer cells. Tumor-associated glycans can reshape cancer cell biology and act as crucial suppressors of anti-tumor immunity. By engaging immune inhibitory receptors or by modifying the function of immune checkpoints, glycans dampen natural immune responses and immunotherapy efficacy. Therefore, glycans serve both as direct targets for anticancer interventions and as adjuvants aimed at potentiating immunotherapy success. In this Review, we provide an overview of tumor glycosylation and its impact on cancer immune surveillance. Finally, we discuss recent therapeutic developments that highlight the potential of glycans as precision cancer medicines.

The stimulator of interferon genes (STING) pathway is a cornerstone of innate immunity and a promising therapeutic target for autoimmune diseases, inflammation, and cancer treatment. Lysine crotonylation, a recently discovered post-translational modification, regulates various cellular processes; however, its role in STING activation remains unclear. Here, we identified dehydrogenase/reductase (SDR family) member 9 (DHRS9) as a critical metabolic regulator of the STING signaling pathway. DHRS9 deficiency impaired activation of the cGAS-STING pathway, suppressed antiviral immunity against HSV-1, and exacerbated viral replication. Mechanistically, DHRS9 converts butyryl-CoA into crotonyl-CoA, thereby enhancing histone crotonylation (H3K14cr and H3K18cr) at the STING promoter to drive its transcription. AAV-mediated DHRS9 delivery significantly enhances antiviral and antitumor immunity, demonstrating its robust therapeutic potential. This study reveals a metabolic-epigenetic axis that regulates STING expression, offering new therapeutic strategies for immune-related diseases.

Sleep integrity has been recognized as a crucial contributor to maintain health and cellular homeostasis. Current evidence supports a bidirectional relationship between sleep disorders and different molecular mechanisms such as protein homeostasis (proteostasis). Nevertheless, few studies have been performed to understand the relation between proteostasis network (PN), sleep disorders and the development of age-related diseases. Hence, in this review we focus on describing the effect of sleep disorders over proteostasis defined as the dynamic balance of protein synthesis, folding, trafficking, and degradation essential for maintaining cellular functions. First, we synthesize emerging evidence about dysfunction of the PN and sleep disorders -including insomnia, sleep-disordered breathing and circadian misalignment. Then, we discuss a conceptual framework in which sleep disruption and the concomitantly PN imbalance leads to many chronic diseases including neurodegenerative, metabolic, cardiovascular, breathing disorders, cancer, cataracts, and sarcopenia. Finally, we suggest that sleep disturbances act as an active modifier of PN that may influence susceptibility to many age-related diseases, and emphasize the need of longitudinal human studies to define the temporal and mechanistic relationships within the sleep-proteostasis-aging axis.

Bifunctional glycosylases, OGG1 for purines and NTH1 for pyrimidines, repair oxidized DNA bases via consecutive glycosylase and AP-lyase reactions, yet their catalytic relationships and lyase activity's biological relevance remain unresolved. Here, we solved crystal structures of archaeal and human Ogg1 and Nth1 captured in key damage-recognition and catalysis-ready states, complemented by ab initio molecular dynamics simulations of their complete reaction trajectories. We thereby define a unified catalytic mechanism for OGG1 and NTH1 conserved over three billion years, distinct from the canonical oxocarbenium-ion mechanism of monofunctional glycosylases. While divergent in their oxidized substrate recognition, their chemistry converged on ribose protonation and ring opening that precede the deglycosylation step. Acid-base catalysis mediated by a conserved aspartate-lysine dyad lowers the C-N bond cleavage barrier, while the excised 8-oxoG base in OGG1 or a conserved aspartate in NTH1 facilitates the AP-lyase reaction. Moreover, structures of human OGG1 bound to product DNA and to product DNA plus a potent small-molecule agonist F51, within the catalytic pocket, reveal that agonists accelerate enzyme turnover by promoting product release. Together, these findings clarify the catalytic logic of bifunctional glycosylases, enabling the development of chemical tools to interrogate lyase activity and therapeutics for oxidative damage in cancer and aging.

The roles of circulating complement C3, produced in hepatocytes, in immune defense have been extensively studied. Interestingly, the C3 gene evolved before the circulatory system, suggesting an essential but yet undefined role of locally produced C3. Here, we investigate the effect of C3 on cancer immunotherapy and find that cancer-associated fibroblast (CAF)-derived C3, not systemic, hepatocyte-derived C3, influences the efficacy of immune checkpoint blockade (ICB). Colorectal and lung tumors developed in CAF-specific C3 knockout mice exhibit resistance to anti-PD-1 therapy, with increased immunosuppressive M2-like macrophage infiltration. Mechanistically, the C3 degradation product iC3b suppresses myeloid cell infiltration via complement receptor 3 signaling. Moreover, therapeutic targeting of this pathway restores sensitivity to PD-1 blockade therapy in immunotherapy-resistant mouse tumors. In human cancers, stromal C3 expression correlates with reduced M2-like macrophage infiltration and improved immunotherapeutic outcomes. Thus, our data demonstrate the role of locally produced C3 in the innate immune response, which may have been conserved across many organisms.

Mitochondrial tRNAs (mt-tRNAs) are central to energy production by translating essential oxidative phosphorylation subunits. Following transcription, mt-tRNAs undergo diverse processing steps, post-transcriptional modifications, and aminoacylation, which are critical for their functions. In this article, we review how human mt-tRNA-modifying enzymes deposit various post-transcriptional modifications onto mt-tRNAs, encompassing both well-characterized and less-understood marks. We also summarize the principles, peculiarities, and critical roles of mt-tRNA charging and proofreading, and highlight recently uncovered noncanonical functions of mitochondrial aminoacyl-tRNA synthetases (mt-aaRSs). Collectively, these recent findings demonstrate the dynamic regulatory mechanisms of mt-tRNA modification and aminoacylation, the extensive involvement of mt-aaRSs in cellular metabolic pathways, and the promising potential of targeting these enzymes in therapeutics.

Genes that enhance T-cell function represent promising targets for improving engineered T-cell therapies for cancer. While extensive CRISPR knockout screens have identified key genes enhancing T-cell persistence, employing Sleeping Beauty (SB) insertional mutagenesis, which induces both gain-of-function (GOF) and loss-of-function (LOF) mutations via the generation of fusion transcripts with endogenous genes, may uncover additional critical factors that previous approaches have overlooked. We developed transgenic mice carrying Doxycycline (Dox)-inducible SB mutagenesis system (DiSBey) in primary T cells. Using DiSBey, we conducted screens for genetic alterations enhancing T-cell persistence under chronic antigen exposure. Specifically, CD8+ T cells from Dox-fed DiSBey mice were subjected to repeated anti-CD3 stimulation over 18 days to mimic chronic antigenic stimulation. We then identified SB transposon genomic insertion sites and corresponding fusion transcripts from the persistent DiSBey CD8+ T cells using enhanced-specificity tagmentation sequencing and RNA sequencing, respectively. Under chronic stimulation, SB-mutagenized CD8+ T cells exhibited improved persistence and reduced terminal exhaustion phenotype. Across six independent screens, we identified 38 genes that were recurrently targeted by the SB transposon T2/Onc2 and differentially expressed under chronic anti-CD3 stimulation. T2/Onc2 insertions into Bach2 and Elmo1 were recurrently identified at the genomic level and were associated with altered nascent transcript expression. Bach2, known as a key regulator of T-cell memory formation and resistance to chronic viral infection-induced exhaustion but less characterized in engineered T cells for cancer therapy, was found to counteract exhaustion in vitro and enhance in vivo tumor persistence in the B16-Ova tumor model. Further, we showed that ectopic Bach2 expression levels influence engineered T-cell differentiation lineage, as low Bach2 overexpression retained more functional progenitor exhausted T cells and exhibited improved therapeutic efficacy. Finally, in human CART19-28ζ cells, BACH2 overexpression enhanced cytotoxicity and tumor control following chronic cancer stimulation. Controllable SB mutagenesis using DiSBey mice provides a novel platform for functional screening of genes that improve T-cell phenotypes important for their use as therapies. Our findings highlight a dose-dependent role of BACH2 in enhancing the function of engineered T cells under conditions of chronic antigenic stimulation.

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

Pseudouridines (ψ) in mRNA are linked to alternative splicing, but their regulatory mechanisms remain unclear due to the lack of known reader proteins. Here, we identify SMD2, a core spliceosomal component, as a direct ψ reader. Through in vitro and ex vivo assays, we show that SMD2 preferentially binds to ψ-modified RNA over unmodified uridines in human cells. Specifically, SMD2 collaborates with ψ synthase (PUS) family enzymes to regulate alternative splicing by recognizing ψ residues near exon-intron junctions. Notably, SNRPD2, the gene encoding SMD2, is overexpressed in multiple cancers and is essential for tumor cell proliferation through modulating mRNA maturation. These findings establish a direct mechanistic link between ψ and spliceosomal function, which positions SMD2 as a key regulator of ψ-mediated splicing and a promising therapeutic target in cancer.

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.

Creatine supplementation is widely used in sports and increasingly popular among exercising individuals. Although the physiological role of creatine has been extensively studied, the creatine biology in pathological conditions remains poorly understood. Here we report that exogenous creatine supplementation promotes tumor metastasis via platelet activation mechanism in various mouse models and humans. Mechanistically, creatine supplementation increases megakaryocyte creatine levels and upregulates creatine kinase B (CKB). Unbiased phosphoproteomics reveals that a CKB-downstream, non-canonical STAT5B phosphorylation instigates various platelet functional genes, leading to hyperactive, metastasis-promoting platelets. Megakaryocyte-specific knockout of the creatine transporter Slc6a8 or Stat5b, as well as pharmacological inhibition of STAT5, ablates the creatine-augmented platelet hyperactivity and prevents consequent metastasis in mice. Importantly, creatine supplementation in healthy volunteers results in hyperactive peripheral platelets that increase metastasis risks. Together, our study sheds mechanistic insights into the creatine-induced metastasis and provides an anti-metastatic therapeutic paradigm by targeting megakaryocyte creatine metabolism.

Aryl C-glycosides represent a valuable class of carbohydrates with broad biological activity in drug discovery, yet their rapid and stereoselective synthesis from unprotected native saccharides remains a significant challenge. Here we report a nickel-catalyzed, ligand-promoted stereoselective C1-arylation of unprotected saccharides with aryl bromides, enabled by glycosyl sulfonylhydrazide donors that are accessible in one step from native sugars. Notably, a series of N,N'-disulfonylpyridinedicarboxamidine tridentate ligands was developed and found to be critical for achieving both high reactivity and anomeric selectivity. The present method accommodates a broad range of aryl and heteroaryl bromides and is applicable to diverse mono- and oligosaccharides. To showcase the synthetic utility of this approach, we constructed a Taxol-sugar conjugate library. In vitro anticancer activity studies revealed that most conjugates retained significant antiproliferative activity against the tested cancer cell lines, while compound 6n was identified as selectively targeting glucose-dependent cancer cells with markedly improved water solubility compared with Taxol, emerging as a promising lead compound for cancer therapy.

The development of far-red light-activatable photolabile protecting groups (PPGs) is crucial for precision medicine applications, yet remains challenging due to the low energy of far-red light photons and the frequent compromise of aqueous solubility. While conditional PPGs activated by specific biochemical conditions offer enhanced spatial control, their scope is limited by the need for external triggers or heterogeneous enzyme expression. Here, we report a new class of dual-responsive PPGs based on a silicon-xanthenium scaffold that are activated by both acidic pH and far-red light. Through systematic structural modification, we developed pcSiR718-OH, a photocage with a low molecular weight (<500 Da) and a red-shifted absorption maximum at 718 nm. pcSiR718CO2H exhibits efficient uncaging (ε × Φuncagingrel = 184 M-1cm-1) under mildly acidic conditions (pH 5.5-6.7) while remaining stable at physiological pH. Mechanistic studies reveal that photolysis can proceed via both homolytic and heterolytic pathways. We demonstrate the utility of this platform through the precise release of bioactive molecules─pomalidomide and gambogic acid─achieving spatiotemporal control over protein degradation in cellular models and tumor growth inhibition in a murine model. This work establishes a versatile strategy for designing environmentally responsive photocages for targeted therapy.

Intracellular proton-coupled electron transfer (PCET) governs cellular metabolism and fate, and its dysregulation is closely associated with disease progression. In normal cells, PCET into the electron transport chain supports ATP production and cellular growth. In cancer cells, however, overexpressed lactate dehydrogenase (LDH) redirects PCET to pyruvate, producing lactate and thereby sustaining tumor growth, proliferation, and metastasis. Reprogramming LDH-driven PCET in living tumors therefore represents an attractive antitumor strategy, yet no artificial catalyst has been shown to outcompete LDH and reprogram PCET in living cells. Here, we report an organoiridium catalyst, IrIII(Cp*)-CN, that captures proton-coupled electrons from NADH and transfers them to protons to generate H2 under ultrasound irradiation at physiological pH. In living cancer cells, IrIII(Cp*)-CN outcompetes LDH and redirects LDH-driven pyruvate reduction and lactate production toward proton reduction and H2 generation. This PCET-reprogramming process converts a tumor-promoting, lactate-producing reaction into a tumor-suppressive, H2-generating reaction, suppressing lactate and ATP production and markedly inhibiting tumor growth with minimal toxicity to normal cells and tissues. These findings establish a framework for reprogramming intracellular PCET via artificial catalysts that outcompete endogenous enzymes, offering a strategy for treating diseases associated with dysregulated cellular redox metabolism.

Compelling evidence shows that amino acids can influence tumor-immune cell communication. Their transfer across the plasma membrane involves multiple amino acid transporters within the solute carrier (SLC) family. Here, we present a new strategy to modulate SLC function by targeting the SLC G-quadruplexes (G4s). We first screen eight SLC genes in the promoter region and identify G4s, including one in the promoter region of glutamine transporter SLC38A2. Then, we design a CRISPR-chiral metallohelix system that selectively targets and stabilizes the SLC38A2 G4 in tumor cells, demonstrating evident enantioselectivity with the Λ-enantiomer being more effective. This leads to SLC38A2 downregulation, which shifts glutamine uptake from tumor cells to dendritic cells (DCs), thereby redirecting glutamine metabolism to activate DCs. Furthermore, we construct a CRISPR-DHX36 system to unwind the SLC38A2 G4 in DCs and upregulate SLC38A2 expression, further augmenting glutamine uptake to compete with tumor cells. The dual CRISPR system with the ability to "fold" and "unfold" the G4 motif in different cell types leads to a marked enhancement of CD8+ T cell activation and killing ability. Therefore, our work sheds new light on cancer immune regulation by modulating the metabolic communication between cancer cells and immune cells.

Cellular softness acts as a mechanical immune checkpoint, suppressing T-cell mechanosensing and cytotoxicity to drive tumor immune evasion. However, existing approaches to modulate membrane stiffness lack specificity, limiting targeted therapeutic potential. Here, we report a dual-spatially confined DNA nanowall assembly strategy to enhance tumor cell stiffness, thereby augmenting the efficacy of adoptive T-cell immunotherapy. In acidic tumor microenvironments, low-pH-triggered membrane-anchored probes (tissue-level confinement) selectively guide the construction of a DNA nanowall on the inner leaflet of the tumor cell membrane (cellular-level confinement). This biomechanical reprogramming markedly increases tumor rigidity. When combined with adoptive T-cell transfer (ACT) immunotherapy, this strategy restores T-cell mechanical force generation capacity, significantly improving tumor killing outcomes in vitro and in murine solid tumor models. Our findings underscore the critical role of biophysical signals in cancer-immune interactions, demonstrating that targeting mechanical immune checkpoints can potentiate antitumor immunity. Furthermore, these discoveries bridge biophysics and oncology, offering a paradigm for precision cancer therapy through the spatiotemporal control of cellular mechanics.

Chemically induced proximity (CIP) and chemically disrupted proximity (CDP) technologies have transformed biological research by enabling precise temporal control of protein-protein interactions and cellular functions. However, despite the extensive development of CIP systems, CDP tools with strong translational potential remain comparatively underdeveloped, calling for the need to expand the CDP toolkit. Here, we present a caffeine-operated dissociation system (CODS), a CDP platform activated by caffeine-an inexpensive and widely available small molecule from food or beverages with a well-established safety profile. Using an AI-guided de novo protein design framework, we reprogrammed an existing caffeine-responsive CIP module into a ligand-dependent dissociation system. CODS exhibits high sensitivity and rapid functional switching, with an EC50 below 90 nM and minute-scale dissociation and reassociation kinetics. We further demonstrate the utility of CODS across diverse cellular contexts, including caffeine-dependent suppression of gene expression, induction of programmed cell death via pyroptosis, and conditional deactivation of chimeric antigen receptor (CAR) T-cell activity. CODS establishes a broadly applicable CDP platform for tunable control of cellular functions and provides a potentially generalizable strategy for engineering chemically controlled dissociation systems from existing CIP architectures.

Arginine biosynthesis is frequently suppressed in cancer because of the loss of argininosuccinate synthase 1 (ASS1) expression, rendering cancer cells reliant on extracellular arginine. This feature has driven the development of systemic arginine-depleting strategies, which are clinically safe but offer limited clinical benefit. In this study, we demonstrated that under arginine scarcity, cancer cells with low ASS1 expression resort to aberrant mRNA translation, characterized by ribosomal frameshifts and amino acid misincorporations. Although aberrant proteins originated from most arginine codons, the predominant effect was observed at AGA. This codon preference was caused by a selective decrease in tRNAArg (UCU) levels following arginine deprivation, linked to methyltransferase-like 1 (METTL1)-mediated tRNA modification. Proteomics and immunopeptidomics analyses validated that arginine shortage induced aberrant protein production at the endogenous level. T-cell receptor (TCR) T cells that specifically recognize these HLA-presented mistranslated peptides efficiently killed cancer cells after arginine deprivation. These results lay the foundation for improved cancer therapies by combining systemic arginine-depleting strategies with TCR-based targeting of nonclassical neoantigens. Aberrant protein production induced by arginine deprivation in ASS1-low cancers leads to production of neoantigens that represent promising targets for TCR-T cell therapies.

Analysis of tumors using single-cell and spatial modalities is critical to advance our understanding of cancer. The growth of technologies that enable these studies provides an increasing number of single cell datasets. Integrating such data across studies will increase the impact of individual studies and speed cancer research. Most existing integration approaches are tailored to transcriptomic data and assume large sets of shared features, an assumption that fails for lower dimensional proteomic measurements. Here, we developed CellFuse, a deep learning-based integration framework that unifies antibody-based proteomic datasets, including high-dimensional cytometry, cellular indexing of transcriptomes and epitopes by sequencing, and spatial proteomics data. Leveraging supervised contrastive learning, CellFuse learned a shared embedding space that enabled accurate cross-modality cell-type prediction and robust label transfer across tumor samples and experimental conditions. Applied to datasets spanning peripheral blood, bone marrow, and lymphoma, CellFuse consistently outperformed existing approaches in recovering clinically relevant populations, including rare malignant and immune subsets. In solid tumors, it reconstructed spatially resolved microenvironments, capturing interactions between malignant, stromal, and immune cells that correlated with treatment response. By enabling scalable, modality-agnostic integration, CellFuse provides a powerful tool to uncover prognostic cell states and delineate the architecture of the tumor-immune ecosystem with translational relevance, driving cancer discoveries. CellFuse is a supervised contrastive learning framework enabling accurate, scalable integration of single-cell proteomic and transcriptomic data, overcoming sparse marker overlap and differing distributions to improve analysis of single-cell data in cancer.

Extracellular tumor-derived DNA (tDNA) has emerged as an important biomarker for cancer diagnosis and monitoring. A better understanding of the mechanisms controlling the abundance of tDNA could help improve biomarker and treatment strategies. In this study, we identified oncogenic KRAS as a critical regulator of tDNA levels. Mutant KRAS promoted tDNA clearance by inducing the tetraspanin CD9, which recruited FXR1 to remodel the actin cortex, lower plasma membrane tension, and promote endocytic uptake of extracellular tDNA. The reduction in tDNA dampened ZBP1-dependent DNA sensing in tumor-associated macrophages (TAM), shifting them toward an immunosuppressive state. Blockade of CD9 restored extracellular tDNA and DNA sensing, reprogrammed TAMs, and synergized with PD-1 blockade in KRAS-mutant cancer models. These findings delineate a KRAS-CD9-FXR1 pathway that couples membrane mechanics to extracellular DNA clearance and immune evasion, providing a strong rationale for targeting CD9 to augment the efficacy of immune checkpoint blockade therapy. KRAS activates CD9-FXR1 signaling that reduces membrane tension to promote extracellular tumor DNA uptake and reduce innate DNA sensing, reshaping the immune landscape and opening opportunities for KRAS-mutant cancer diagnosis and treatment. See related commentary by McAndrews, p. 3371.

A major limitation in applying chimeric antigen receptor (CAR) T cells to solid tumors is toxicity in healthy tissues caused by a lack of tumor-specific targets. A promising strategy to overcome this deleterious cytotoxicity is to engineer control into CAR T cells beyond that conferred by antigen recognition alone. In a recent issue of Nature Chemical Biology, Scheller and colleagues report the development of a CAR that is inactivated through introducing the small molecule, venetoclax, which is a clinically approved targeted Bcl-2 inhibitor. The authors design venetoclax-dependent release of the CAR extracellular binding domain, thereby disrupting T-cell contact with tumor cells and suppressing cytotoxicity. Furthermore, they demonstrate the reversibility of this approach as withdrawal of the drug restores CAR T-cell function. This work establishes a foundation for clinically translatable remote-controlled CAR T-cell therapy for solid tumors.

MicroRNA (miRNA) is an endogenously expressed non-coding RNA, and the sensitive monitoring of low-abundance miRNAs holds significant importance for early therapeutic intervention in cancer. Consequently, developing a simple yet highly sensitive miRNA-21 detection method is of substantial importance. This study employs a new Y-shaped DNA-facilitated photoelectrochemical (PEC) sensing interface constructed on a yolk-shell CuO-Co3O4@Co3O4 photocathode, supplemented by a gold nanoparticle-assisted dissolved oxygen cycle for detecting miRNA-21. A yolk-shell CuO-Co3O4@Co3O4 nanocomposite was prepared by encapsulating CuO-Co3O4 within hollow Co3O4 nanocubes, whose unique structure causing an elevate increased specific surface area. Firstly, due to the bandgap matching between Co3O4 and CuO, a type II heterojunction formed in the yolk region with an intrinsic electric field, enabling higher carrier separation efficiency. Under xenon lamp illumination, the photocurrent intensity of the CuO-Co3O4@Co3O4 electrode surpasses that of standalone Co3O4 and CuO electrodes. Furthermore, catalytic hairpin assembly driven by miRNA-21 promotes the generation of Y-shaped DNA structures embedded with gold nanoparticles. Gold nanoparticle-assisted dissolved oxygen cycling amplified the detection signal, allowing for ultra-sensitive determination of miRNA-21. The photocurrent response is positively correlated with the number of gold nanoparticles anchored on the electrode surface. This biosensor exhibits favourable linearity and selectivity for the detection of miRNA-21, and the detection limit is 47.464 fM, demonstrating excellent stability and sensitivity. The constructed PEC biosensor holds practical application worth for the early diagnosis and management of miRNA-21 associated disorders.

Single nucleotide variants (SNVs), including single-nucleotide polymorphisms, are key biomarkers for numerous human diseases, including cancer, and their accurate detection at single loci remains a major diagnostic challenge. Imaging-based approaches for in situ nucleic acid analysis further require highly specific, non-fluorescent readout strategies. Here, we report a Raman spectroscopy-based method for single-nucleobase discrimination using alkyne-tagged oligonucleotides. Ab initio calculations predict, and Raman measurements confirm, that the alkyne stretching vibrations of 5-ethynyluracil is highly sensitive to hydrogen-bonding interactions with its complementary nucleobase. Incorporation of this moiety as 5-ethynyl-2'-deoxyuridine into oligonucleotide probes enables detection of specific nucleobases within target DNA strands by high-resolution Raman spectroscopy. Distinct Raman shifts allow discrimination of single nucleotide variants, including the clinically relevant BRAF V600E mutation. These results demonstrate that alkyne-tagged base discriminating probes provide a robust Raman readout for SNV detection at single nucleotide resolution, establishing a foundation for future nucleic acid diagnostics and imaging applications.

KRAS mutations are among the most prevalent oncogenic alterations in colorectal, lung and pancreatic cancer, yet their detection remains analytically challenging in the presence of an overwhelming wild-type (WT) background. Here, we report a photoelectrochemical (PEC) genotyping platform that integrates clamp-inhibited loop-mediated isothermal amplification (C-LAMP) with enzyme-free singlet oxygen (1O2)-driven PEC transduction for mutation-selective KRAS detection. Locked nucleic acid (LNA) clamp probes selectively suppress WT amplification during isothermal amplification, enriching mutant alleles and enabling single-nucleotide variant (SNV) discrimination with high selectivity. Amplified products are magnetically captured and transduced into photocurrent via visible-light-induced 1O2 redox cycling, eliminating enzymatic reporters and reducing background interference. The C-LAMP/PEC platform achieves a limit of detection of 35 copies μL-1 (58 aM) and a minimum detectable variant allele frequency (VAF) of 4.8% in heterogeneous mutant/WT genomic DNA mixtures. Analytical performance was validated in cancer cell lines and in patient-derived fresh frozen tissues, showing complete concordance with Nanopore sequencing and droplet digital PCR (ddPCR) within the evaluated cohort (n = 16). This work introduces a robust and modular PEC biosensing strategy that combines molecular WT suppression with enzyme-free photoelectrochemistry, offering an economically competitive and instrumentation-simplified approach for clinically relevant KRAS mutation analysis toward decentralized testing.

The concept of nuclear receptor (NR) coregulation was proposed nearly 2 decades before it was experimentally validated. According to this model, NRs-executors of a vast array of transcriptional programs-do not act independently but are governed by a network of regulatory proteins that either activate or repress their biological function. Nuclear receptors identify the genes to be regulated. However, coregulators ultimately serve as the true controllers of transcriptional outcomes. They recruit cofactors and coordinate the activity of transcriptional complexes, thereby shaping NR-mediated gene expression beyond NRs' intrinsic functionality. The steroid receptor coactivator (SRC) family is the most extensively studied and functionally dominant group of NR coregulators, modulating nearly all gene activities. The pleiotropic biological roles of SRCs-spanning key physiological and pathological processes-make them compelling therapeutic targets. Here, we outline the evolution of the coregulation concept, which reached a critical milestone with the discovery of the SRC family. We highlight the central roles of SRCs in both physiology and oncology and trace the development of therapeutic strategies aimed at targeting these proteins.

Death receptors (DRs) are a subset of the tumor necrosis factor receptor (TNFR) superfamily with a protein interaction motif called the death domain. DRs, particularly TNFR1, FAS and TRAIL-Rs, mediate critical cellular outcomes, including apoptosis, necroptosis, inflammation, survival, and proliferation. Phosphorylation is a rapid, reversible post-translational modification, and therefore might alter the downstream fate decisions in signal transduction pathways. Like most cytokines, death ligands activate a wide range of tyrosine kinases. However, our understanding of whether these tyrosine kinases phosphorylate DRs and modulate downstream signaling is limited. Depending on the cell type, several different tyrosine kinases, including Src family members, JAKs and EGFR have been demonstrated to be activated upon death ligand exposure. All three of these kinase families have the potential to phosphorylate DRs, as reviewed here, and these phosphorylation events create a bias towards proinflammatory and proliferative pathways over cell death. Since these kinases are frequently hyperactivated in cancer, this bias may sustain tumor cell survival and resistance to death receptor-targeted therapies, marking the responsible kinases and their opposing phosphatases as candidate therapeutic targets.

Surgical resection inadvertently promotes metastasis via platelet activation and tumor cell-platelet aggregation. Conventional postoperative therapies fail to address this early pro-metastatic window and lack precise local modulation. Herein, we develop an injectable self-healing hydrogel (CS@Art-2CHO/Cil gel) co-delivering artesunate (Art) and integrin inhibitor Cilengitide (Cil) for local application. Interestingly, we employ a "drug-as-structural-unit" design, where chemically engineered Art derivatives act as both active agents and intrinsic crosslinkers. They form a dynamic chitosan (CS) based network via Schiff base bonds, eliminating exogenous crosslinkers. By tuning aldehyde groups on Art, we precisely tailor crosslinking density and mechanical strength to a "mild yet stable" modulus suitable for the postoperative site. In this system, Art directly kills residual tumor cells, while Cil blocks integrin αvβ3 to inhibit platelet-tumor cell interactions. This dual "local clearance-systemic blockade" strategy synergistically prevents metastasis. This work presents a novel hydrogel paradigm that integrates precise mechanical adaptation with therapeutic synergy, offering a promising strategy against surgery-driven metastatic relapse.

Lysosomes serve as central degradative hubs in cells, playing critical roles in maintaining protein homeostasis, clearing damaged organelles, and regulating metabolic signaling. Tumor cells heavily rely on lysosomal functions during proliferation, invasion, and drug resistance, a dependency that concurrently endows them with inherent susceptibility to lysosomal membrane permeabilization (LMP). Current cancer therapies rely heavily on surgical resection for early-stage disease, and chemotherapy or radiotherapy for advanced-stage cancers, but these modalities are limited by poor efficacy, severe side effects, and drug resistance. Therefore, targeting LMP to induce lysosome-dependent cell death (LDCD) represents a promising breakthrough. This review systematically summarizes the molecular mechanisms underlying LMP initiation and execution, as well as the regulatory pathways of LDCD modalities, including apoptosis, necroptosis, ferroptosis, pyroptosis, immunogenic cell death, and autophagy-dependent death. It further highlights the dual roles of lysosomes and LDCD in the tumor microenvironment and their core functions in tumor progression. Additionally, we outline classic therapeutic strategies targeting LMP and novel lysosome-targeting technologies, and discuss combination therapy regimens based on lysosomal modulation. These advances provide comprehensive theoretical foundations and new insights for the development of broad-spectrum lysosome centered anticancer drugs.

Immune checkpoint inhibitors have revolutionized cancer therapy, yet a substantial proportion of patients exhibit primary or acquired resistance. Immunocytokines offer a strategy to enhance antitumor immunity by delivering cytokine signals selectively to the tumor microenvironment. Here, we describe the immunoconjugate anti-programmed cell death protein 1 (PD-1)-interleukin (IL)-18 (aPD1-IL18mut), designed to couple PD-1-blockade with localized IL-18-mediated immune activation. aPD1-IL18mut was generated by chemically conjugating the anti-human PD-1 antibody Lipustobart to an IL-18 variant engineered to evade IL-18 binding protein. Its mechanism of action was characterized using human PD-1 transgenic mouse models. To assess the translational relevance of these in vivo findings, complementary in vitro assays were conducted on human tumor samples, alongside analyses of publicly available single-cell RNA sequencing datasets. In vitro, PD-1 engagement enhanced functional IL-18 activity, preserving interferon (IFN)-γ secretion even under IL-18BP pressure. In MC38 tumors, aPD1-IL18mut induced robust CD8+ T cell-driven tumor control, accompanied by intratumoral accumulation of activated CD8+ T cells and a pronounced type 1-associated cytokine response. In anti-PD-1-resistant YUMM1.7 tumors, therapeutic efficacy instead relied predominantly on Th1-like CD4+ effector T cells, and aPD1-IL18mut stimulation was associated with enhanced activation, proliferation, IFN-γ, and granzyme B expression in PD-1+IL-18Rɑ+ CD4+ T cells. In human cancer digest cultures, aPD1-IL18mut elicited a dominant IL-18/IFN-γ-driven cytokine signature and enhanced tumor cell killing. Importantly, analyses of the immune infiltrate across multiple human solid tumor types identified analogous PD-1+IL-18Rɑ+ CD4+ Th1-like and CD8+ effector-like T-cell subsets with effector-associated and tumor-reactive transcriptional and protein signatures. aPD1-IL18mut activates T-cell populations with tumor-reactive features and promotes IFN-γ-driven inflammation across distinct tumor immune contexts. Its ability to activate CD8+ and CD4+ Th1-like effector cells, together with the presence of analogous subsets in multiple human cancers, provides mechanistic and translational support for PD-1-targeted IL-18 therapy.

Extracellular ATP (eATP) is an immunogenic danger signal. In this issue of Immunity, Hu et al. report that in the tumor microenvironment, eATP signals through the receptor P2RY2 on cancer cells, triggering the COX-PGE2 pathway and thereby immune suppression and immunotherapy resistance. These findings identify eATP as the upstream signal driving PGE2 expression, with therapeutic implications.

Non-canonical metabolic functions of signaling molecules contribute to cancer plasticity and metastatic progression. Here, we demonstrated that PDE5a inhibitors, including sildenafil (Viagra), induced lysosomal cholesterol accumulation across multiple mouse and human cancer models, reducing cholesterol bioavailability and impairing cancer cell migration and metastasis. Cancer cells exhibited heightened sensitivity due to reduced lysosomal gene expression, rendering them particularly vulnerable to disrupted cholesterol trafficking. Mechanistically, elevated cGMP bound the lysosomal cholesterol transporter NPC1, impairing cholesterol export and phenocopying Niemann-Pick type C pathology. The resulting cholesterol depletion disrupted membrane lipid rafts and mitochondrial bioenergetics, thereby limiting metastatic capacity and triggering compensatory SREBP2 activation with increased cholesterol synthesis. Combining sildenafil with statins yields additive antimetastatic effects by concurrently blocking lysosomal cholesterol export and cholesterol biosynthesis. Consistently, analysis of digital health records demonstrated significantly improved survival among sildenafil users, with a dose-dependent additive benefit observed when combined with statins. Together, these findings identify increasing cGMP levels through PDE5a inhibition as a potential strategy to restrict metastasis and offer a potential mechanistic basis for the beneficial effects of sildenafil.

The proto-oncogene MYC acts as a master regulator of diverse metabolic programs that fuel tumor initiation and progression, but the exact programs it activates depend heavily on context, tissue type, accompanying mutations, and the tumor microenvironment (TME). Furthermore, extrinsic influences, including diet and systemic metabolism, interact with MYC to reshape tumor and stromal cell metabolism, revealing a multifaceted metabolic landscape governed by MYC. This review focuses on MYC-driven in vivo metabolic addictions and discusses tissue-specific programs, TME interactions, and systemic influences to illustrate how MYC-driven metabolic reprogramming creates context-dependent vulnerabilities. These insights may uncover actionable targets and guide precision therapeutic strategies for MYC-driven cancers.

Tobacco use remains the leading preventable cause of cancer mortality worldwide, with marked disparities in cancer incidence and outcomes across racial and ethnic groups. This study investigates the genetic determinants of tobacco carcinogen metabolism among smokers, focusing on ancestry-informative markers (AIMs) near genes involved in metabolic activation (Phase I), detoxification (Phase II), and transport (Phase III) of tobacco-specific carcinogens. Using data from 274 smokers recruited in the United States and the Caribbean, we measured urinary NNAL (a metabolite of NNK) to classify metabolic phenotypes and performed genotyping with a customized array enriched for AIMs. Most participants (94%) were classified as poor metabolizers of NNAL, with a higher prevalence among Black/African American smokers. Multivariable analyses, after adjusting for age, sex, assay batch, and number of cigarettes smoked per day, identified 14 AIMs near Phase I, II, and III pathway genes significantly associated with NNAL metabolizer phenotype, with African ancestry alleles conferring increased odds of poor metabolism. Functional annotation revealed that most significant AIMs overlapped regulatory regions, and the CYP3A5 rs776746 variant demonstrated structural disruption likely to impair enzymatic activity. Majority of participants had high African ancestry and were poor NNAL metabolizers, consistent with the observed association between African ancestry and poor NNAL metabolism. These findings highlight the biological basis for tobacco-related cancer disparities and underscore the importance of integrating genetic ancestry and metabolic phenotyping in risk assessment and personalized prevention strategies for tobacco-associated cancers.

The textbook view of the regulation of calcium homeostasis centers on parathyroid hormone, calcitriol, and calcitonin. While this paradigm has long dominated the field, it represents only part of a broader evolutionary and physiological landscape. This article focuses on Stanniocalcin 1 (STC1), a calciotropic hormone originally discovered in bony fish. Since its discovery in the 1970s, our understanding of STC1 has evolved from a "fish-specific" hormone to a member of a multifunctional glycoprotein family with deep roots in metazoan evolution. Recent studies in zebrafish show that STC1 inhibits calcium uptake through two mechanisms: it reduces TRPV6-dependent calcium influx and limits the proliferation of calcium-transporting ionocytes. Mechanistically, STC1 suppresses local insulin-like growth factor (IGF) signaling by binding pappalysin-family IGF-binding protein (IGFBP) proteases, thereby modulating IGFBP processing and downstream IGF activity. This STC1-Pappalysin-IGFBP-IGF axis has recently been positioned as a central node in an organ-brain communication pathway that links calcium uptake to brain function and animal behavior. In humans and other mammals, STC1 is expressed in many tissues locally and exerts local actions in a paracrine fashion. Dysregulation of STC1 expression has been associated with chronic kidney diseases and multiple cancer types. Recent studies have uncovered a novel role for STC1 in tumor immune evasion as a phagocytosis checkpoint and a "don't eat me" signal. This review integrates recent findings on the multifunctional roles of STC1 in regulating calcium homeostasis, IGF signaling, and cancer immunity, and to highlight the therapeutic potential of targeting STC1 in kidney diseases and cancer immunotherapy.

For many decades, interest in the medical application of boron chemistry focuses mainly on the boron neutron capture therapy (BNCT) approach to cancer treatment, which is a transformative binary-targeted radiotherapeutic modality, yet its global clinical translation is limited by the lack of high-performance boron delivery agents. Icosahedral carboranes hold unparalleled potential for the development of BNCT agents due to their exceptional boron density, excellent metabolic stability, and unique 3D architecture, but their biomedical application is hampered by intrinsic superhydrophobicity, poor tumor targeting, and limited theragnostic modifiability. Herein, we report a class of boryl-carboranes as multifunctional boron-bearing building blocks that directly address these long-standing limitations. The pendant borane moiety is readily converted to B(OH)2 for tumor targeting and enhanced aqueous solubility, and to BF3K for 18F-PET theragnostic integration. To enable scalable synthesis, we develop the electron donor-acceptor (EDA) complex-based strategy for the controllable generation of boron-centered carboranyl radicals from N-hydroxyphthalimide (NHPI) carboranyl carboxyl esters. This additive-free, room-temperature photochemical strategy enables boron-centered carboranyl radical-mediated unsymmetrical B-B bond construction, with decagram-scale production achieved via a continuous-flow system. Mechanistic studies confirm the EDA complex-mediated radical pathway. In vitro and in vivo studies demonstrate that these boryl-carboranes exhibit excellent biosafety, favorable aqueous solubility, and better tumor boron uptake and accumulation than clinically established agents, rendering this platform potential for next-generation BNCT therapeutics.

Transcription factors (TFs) occupy a central position in cancer biology, functioning as master regulators that translate genetic, epigenetic and environmental cues into cell fate decisions, proliferation, survival, and therapy response. Historically deemed "undruggable" owing to their lack of catalytic sites, conformational flexibility, and engagement in broad protein-DNA and protein-protein interfaces, TFs were long considered beyond the reach of conventional pharmacology. Over the past decades, advances in structural biology, chemical biology, epigenetics, and nucleic acid therapeutics have begun to overcome these challenges, revealing actionable vulnerabilities within TF networks. This review synthesizes current understanding of TF function in tumorigenesis, moving from mechanistic insights at the level of individual TFs to the higher-order organization of transcriptional regulatory networks. It further assesses emerging therapeutic strategies aimed at perturbing aberrant TF activity, encompassing direct inhibition, targeted protein degradation, modulation of TF-cofactor interactions, and nucleic acid-based interventions. We further highlight exemplary TFs and their typical targeting strategies, including Myelocytomatosis oncogene (MYC), Signal transducer and activator of transcription 3 (STAT3), Catenin beta-1 (β-catenin), Yes-associated protein/Transcriptional coactivator with PDZ-binding motif/Transcriptional enhanced associate domain (YAP/TAZ/TEAD), Estrogen receptor/Androgen receptor (ER/AR), and Phosphatase and tensin homolog/Protein kinase B/Forkhead box O (PTEN/AKT/FOXO), which illustrating mechanistic understanding of transcriptional regulation drives therapeutic development and enables genomics-guided precision oncology. By unifying mechanistic insight with pharmacological innovation, we aim to provide a conceptual framework for targeting the transcriptional architecture of cancer and charting paths toward next-generation transcription-directed therapies.

CD4+ T helper (Th) cells contribute to tumor immunity, yet the subsets and differentiation programs involved remain unclear. Here, we show that the transcription factor Eomesodermin (Eomes) is essential for Th-mediated anti-tumor immunity. Eomes orchestrated the differentiation and maintenance of an exhausted-like Th cell lineage, transcriptionally and functionally distinct from conventional effector or memory Th subsets. This Eomes-dependent program was enhanced by 4-1BB stimulation and promoted effective Th-cell-mediated tumor control. The progenitor subset of this lineage (pTh) expressed stemness-associated transcription factors, displayed self-renewal capacity, and seeded effector subsets capable of controlling tumor growth. At the transcriptional level, Eomes supported the survival, metabolic fitness, and apoptotic resistance of this lineage. Eomes⁺ pTh cells exhibited conserved transcriptional features in humans across multiple tumor types. As the most expanded Th cell population upon immune checkpoint inhibitor therapy, targeting these cells has potential to improve current immunotherapies.

Tumor resistance to immunotherapy is driven by several mechanisms, including those imposed by myeloid populations. Neutrophils are prominent within this landscape and display functional heterogeneity. Here, we investigated the contextual role of neutrophils, and using neutropenic mice, we found that the dominating function was to block the response when targeting T cells or myeloid cells. We found that neutrophils upregulated programmed death ligand-1 (PD-L1) in response to the treatment and, using this as a target, depleted this population. The upregulation of PD-L1 was dependent on interferon-γ (IFN-γ) produced by cytotoxic lymphocytes. Specific genetic deletion of cd274 or Ifngr1 on neutrophils showed that this was cell intrinsic. Moreover, in the absence of the capacity for specific IFN-γ-driven suppression, neutrophils changed their phenotype to support immunotherapy. Thus, we find that the type II interferon, IFN-γ, is key in determining whether neutrophils will support or block immunotherapy for cancer.

Exercise prevents disease and promotes health, yet the cellular and molecular drivers of these benefits remain incompletely resolved. A principal mechanism is immune modulation. This review synthesizes recent evidence from healthy individuals and disease contexts, focusing on cancer and autoimmunity, detailing how acute bouts and consistent training reshape the mobilization, fate, and function of innate and adaptive immune cells. We also delineate how exercise reprograms non-immune tissues, including skeletal muscle, adipose tissue, vasculature, and the gut microbiota, to shape immunity via exercise-induced signaling factors ("exerkines")-including cytokines, metabolites, catecholamines, and microbiota-derived products-that, together with hemodynamic cues, rewire leukocyte trafficking, phenotype, and effector activity. Recognizing exercise as a potent, context-dependent immunomodulator, we argue that deeper mechanistic insight will enable dosable, immune-informed exercise prescriptions and the development of exerkine-based therapeutics for patients unable to train.

The CRISPR/Cas12a system holds great promise for nucleic acid detection, but its strict dependence on the protospacer adjacent motif (PAM) severely limits its application in gene point mutation analysis, with fewer than 2% of known mutation sites naturally harboring adjacent PAM sequences. Herein, we developed a PAM-free Cas12a system with double-stranded substrate positioning-unwinding (dsPU-Cas12a), wherein "bubble" structures formed by unpaired base pairs release partial single-stranded target strand as a toehold, and excess auxiliary strands induce local unwinding of double-stranded DNA to facilitate R-loop formation. After optimization, the dsPU-Cas12a system achieved an ultra-low limit of detection of 0.013% for gene point mutations, with excellent linearity over the mutation abundance range of 0-10%. Furthermore, it exhibited robust feasibility and accuracy in detecting the JAK2 V617F mutation in blood samples from patients with myeloproliferative neoplasms. This simple and universal strategy overcomes the sequence limitation of Cas12a, providing a high-performance tool for clinical gene point mutation detection.

While cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) are well-recognized toxicities of CAR T-cell therapy, immune effector cell-associated hemophagocytic lymphohistiocytosis-like syndrome (IEC-HS) remains poorly characterized. We aimed to describe the incidence, clinical features, management, and outcomes of IEC-HS using real-world data from the DESCAR-T registry (NCT04328298). We conducted a multicenter retrospective study of patients registered in DESCAR-T who received standard-of-care CD19 CAR T-cell therapy for relapsed or refractory B-cell non-Hodgkin lymphoma (B-NHL) or B-cell acute lymphoblastic leukemia (B-ALL) and fulfilled ASTCT criteria for IEC-HS. We analyzed 42 patients who developed IEC-HS. Most had a high CAR-HEMATOTOX score (≥2, n = 36) and prior high-grade CRS (grade ≥3, n = 20). IEC-HS was characterized by marked hyperinflammation, with high median ferritin (18,561 µg/L), lactate dehydrogenase (825 IU/L), and triglycerides (4.5 g/L), and low fibrinogen (0.9 g/L). Median time to IEC-HS onset was 9 days (IQR 6-21). Patients received a median of two treatment lines. Observed response rates were 77% with etoposide, 70% with corticosteroids, 70% with corticosteroids plus anakinra, and 50% with tocilizumab, with combination therapy more frequently used in severe cases. After a median follow-up of 24.1 months, overall mortality was 81%. One-year overall survival was 15.6% for B-NHL and 47.1% for B-ALL. In this real-world cohort, IEC-HS was a rare but life-threatening complication of CD19 CAR T-cell therapy, often occurring after severe CRS and associated with poor outcomes. Despite initial inflammatory control, mortality remained high, underscoring the need for improved risk stratification and management strategies.

Quadruplets incorporating anti-CD38 monoclonal antibody have led to improved outcomes, potentially allowing limited-duration therapy and reduction in steroid doses. We designed this trial to examine the efficacy of the quadruplet regimen containing daratumumab, ixazomib, lenalidomide, and dexamethasone (Dara-IRd) given for fixed duration and to evaluate early discontinuation of steroids. Patients with untreated MM were enrolled, irrespective of their transplant eligibility. The primary objective was to determine the ≥complete response (CR) rate. Treatment involved 28-day cycles of ixazomib 4 mg days 1, 8, 15; lenalidomide 25 mg days 1-21, dexamethasone 40 mg, weekly and daratumumab 16 mg/kg, weekly for two cycles, every other week during cycles 3-6 and every 4-weeks thereafter during induction (12-cycles) followed by daratumumab and ixazomib maintenance (24-cycles). Seventy-eight patients were enrolled into two sequential cohorts: cohort A (n = 38) and B (N = 40), with dexamethasone given only for two cycles in Cohort B. At the time of the data cut-off, all patients had completed the treatment. The overall response rate was 96%, including a ≥CR rate of 32%, similar in both cohorts. Responses deepened over time; 32% achieved a marrow MRD negative status and 29% MRDneg-CR. After a median follow-up of 37.9 months, the median PFS or OS has not been reached. A grade ≥3 adverse event, at least possibly attributed to the study drugs, was seen in 55% of patients. The most common toxicities included fatigue, neutropenia, lymphopenia, peripheral neuropathy, diarrhea, and nausea. Stem cells were collected in 55 patients; the median sCD34+ cell yield (range) was 7.8 (2.8-15.9) × 106/kg. Dara-IRd is an active regimen in newly diagnosed MM, with high overall response rate as well as deep responses. Nearly a third of the patients attained MRDneg status, which improved over time. The PFS with finite-duration therapy is comparable to other studies in NDMM. Early discontinuation of dexamethasone did not impact efficacy.

The introduction of venetoclax (a BCL2 inhibitor) and targeted therapies, including inhibitors of CD33, FLT3, IDH1, IDH2, and menin, has expanded treatment options for newly diagnosed acute myeloid leukemia (AML). In younger, fit patients, the primary goal remains long-term survival, which in most cases is secured through allogeneic stem cell transplant. Transplant in first complete remission is recommended for FLT-ITD, TP53 mutated, KMT2A rearranged, AML with other adverse genetic abnormalities, and is considered in most intermediate-risk patients. It is also recommended in relapsed/refractory disease or persistent measurable residual disease (MRD). The role of intensive chemotherapy, such as cytarabine (7) plus anthracycline (3), is limited to patients with core-binding factor AML, NPM1 mutation, CEBPA bZIP mutation and those with intermediate-risk disease. Intensive regimens such as FLAG-IDA and CLIA plus venetoclax have shown impressive long-term outcomes, but their use is not widespread. In FLT3 mutated AML, 7 + 3 plus an FLT3 inhibitor (midostaurin or quizartinib) remains a standard, with venetoclax-hypomethylating agent-FLT3 inhibitor triplets emerging as an alternative. Similarly, in IDH1/2 mutated AML, venetoclax-hypomethylating agent with or without IDH1/2 inhibitor combinations challenge intensive chemotherapy approaches. Patients with TP53 mutations or other adverse-risk features, where intensive chemotherapy is known to be less effective, should be referred for clinical trials. There remains ongoing debate regarding optimal management of fit patients with newly diagnosed AML without targetable mutations, as emerging data suggest that venetoclax- hypomethylating agents may be comparable to intensive chemotherapy in selected patients proceeding to transplant. Accordingly, treatment decisions should be individualized to maximize remission while minimizing toxicity.

TP53 gene alterations remain one of the major determinants of poor prognosis in multiple myeloma (MM). However, p53 dysfunction can also arise through non-genetic mechanisms, like post-translational modifications (PTMs). Indeed, phosphorylation and acetylation play essential roles in modulating p53 stability and activity, but their functional relevance in primary MM samples has not been systematically explored. Here, we quantified the expression of four key p53 PTMs, phosphorylation at S15, S20, and T55, and acetylation at K382, using capillary nanoimmunoassay, in 127 newly diagnosed MM patients. We also evaluated the DNA damage marker γ-H2AX, the kinase Chk2, and the phosphatases PP1A and PP2A-C. We found that p53 PTM expression was highly heterogeneous among patients. p-p53 T55 and Ac-p53 K382 were the most frequently expressed, detected in more than 70% of samples. Patients with double-hit TP53 showed significantly lower p-p53 S20 and p-p53 T55 levels compared with those with normal TP53. Low p53 phosphorylation at S20, S15 and T55 was correlated with higher progression risk (HR = 2.39, p = 0.02; HR = 2.14, p = 0.04; and HR = 2.13, p = 0.005, respectively). We also identified deregulated expression of Chk2 and phosphatases, suggesting upstream regulatory imbalances. This is the first study to quantify p53 PTMs in a large set of MM patients, revealing that post-translational dysregulation contributes to p53 impairment independently of TP53 gene alterations. Characterizing p53 PTMs sheds light on the mechanisms underlying its dysfunction, particularly in patients with an intact TP53 gene, but dysfunctional p53 protein regulation.

Multiple myeloma (MM) is a plasma cell malignancy associated with significant morbidity and mortality. Recent advances in the treatment modalities have improved survival, but less is known about MM incidence and associated demographic disparities across sex, age, and racial/ethnic groups. We conducted a retrospective population-based analysis using the Centers for Disease Control and Prevention Wide-ranging Online Data for Epidemiologic Research (CDC WONDER) database from 1999 to 2022. Annual crude and age-adjusted incidence rates (per 100,000 population) were analyzed overall and stratified by age, sex, race, and ethnicity. Temporal trends were assessed using Joinpoint regression to estimate annual percent change (APC) with 95% confidence intervals. Age-specific incidence patterns were evaluated using crude rates, and differences between consecutive age groups were assessed descriptively. Between 1999 and 2022, the age-adjusted incidence of MM increased by approximately 23%. Incidence was higher in males than females (APC: males 1.19%, females 1.19%). Black or African American individuals had the highest incidence, with greatest annual increase (APC 1.40%), whereas White individuals had a more gradual rise (APC 1.07%). Non-Hispanic individuals had higher incidence compared with Hispanic individuals. Incidence increased markedly with advancing age, peaking at 80-84 years before declining significantly among those aged ≥85 years. MM incidence has increased steadily in the United States over the past two decades, with males, older adults, Black or African American individuals, and non-Hispanic populations bearing a disproportionate disease burden. This reflects the need for further research into demographic disparities in multiple myeloma incidence. Clinical Trial Registration: This study was not a clinical trial and therefore was not registered in a clinical trial registry.

Clonal haematopoiesis of indeterminate potential (CHIP), a recently recognized, age-related cardiovascular risk factor, results from acquired, somatic mutations in a subset of known leukaemia driver genes in haematopoietic stem or progenitor cells. While prior work suggested that screening for these somatic mutations, which requires DNA sequencing, was premature due to lack of actionable evidence-based interventions, this review proposes that in light of the rapidly accumulating knowledge regarding CHIP, selective clinical testing by cardiovascular clinicians is now justified for specific high-risk individuals to improve cardiovascular risk stratification and inform preventive care. By identifying distinct mutations and estimating their burden (variant allele fraction), clinicians can understand better an individual's CHIP-related cardiovascular risk and tailor surveillance and management strategies, even before validation of targeted therapies. This review advocates a biologically driven approach to selective assessment of CHIP that leverages the advances in understanding this condition and its clinical consequences.

Peginterferon‑α is useful to treat myeloproliferative neoplasms (MPNs) but can precipitate broad autoimmunity. By promoting beta-cell, thyroid, and systemic immune injury, it can lead to Type 1 diabetes, autoimmune thyroiditis, systemic lupus erythematosus, or Sjögren's syndrome. Onset typically occurs within the first months but may be delayed until after treatment cessation. Type 1 diabetes is usually irreversible and requires lifelong therapy, whereas thyroiditis and lupus more often improve after interferon withdrawal, though autoantibodies frequently persist. In this narrative review, we provide an overview of how peginterferon can induce autoimmunity in genetically susceptible individuals with pre-existing subclinical autoimmunity. Pre-treatment risk assessment, including personal/family history of autoimmune disease and consideration of baseline thyroid function and anti-GAD antibodies in high-risk patients, may identify those at elevated risk for irreversible complications, particularly Type 1 diabetes. Targeted clinical and laboratory monitoring throughout therapy and for 12-24 months post-cessation can enable early detection and appropriate intervention.

There is an unmet need for effective, off-the-shelf therapies for relapsed or refractory aggressive B cell non-Hodgkin lymphoma (B-NHL). Part 2 of the current study was an open-label, nonrandomized, phase 1 study of escalating doses of the CD19-4-1BBL co-stimulatory molecule, englumafusp alfa, in combination with glofitamab in patients with relapsed or refractory B-NHL. Obinutuzumab pretreatment was administered 7 days before the first glofitamab dose. Glofitamab step-up dosing in cycle 1 was followed by 11 cycles of glofitamab plus englumafusp alfa. Englumafusp alfa was administered at escalating doses, with the initial dose on cycle 2 day 8 (C2D8) or cycle 1 day 10 (C1D10). Primary objectives were to establish the maximum tolerated dose, and safety and tolerability. A total of 134 patients were enrolled, including 109 with aggressive B-NHL and 25 with indolent B-NHL. The maximum tolerated dose of englumafusp alfa was not reached; one dose-limiting toxicity occurred (grade 5 Pneumocystis jirovecii pneumonia). Adverse events were reported in 98.5% of all patients, with grade 3/4 adverse events in 59.0%. Grade 5 adverse events occurred in ten patients. In the subgroup of C2D8 patients with aggressive B-NHL (n = 83), overall response and complete metabolic response rates were 68.7% and 56.6%, respectively; among those without previous exposure to chimeric antigen receptor T cell therapy (n = 41), the corresponding rates were 73.2% and 65.9%. Pharmacodynamic changes following englumafusp alfa administration supported its co-stimulatory mode of action. These data demonstrate that the addition of englumafusp alfa to glofitamab is associated with encouraging efficacy and robust pharmacodynamic modulation, as well as a safety profile consistent with glofitamab monotherapy, in patients with relapsed or refractory B-NHL. CTIS identifier: 2022-502616-37-00 ; ClinicalTrials.gov identifier: NCT04077723 .

Talicabtagene autoleucel (tali-cel) is the first humanized CD19-directed CAR-T cell therapy approved in India for relapsed/refractory (r/r) B-cell acute lymphoblastic leukemia (B-ALL) and B-cell non-Hodgkin lymphoma (B-NHL). Here, we integrated clinical and manufacturing operations through a centralized coordination unit (CCU) and evaluated tali-cel implementation. Patients with r/r B-ALL (n = 105) and r/r B-NHL (n = 145) who underwent leukapheresis between 15 November 2023 and 31 January 2025 across 56 treatment centers were included. The CCU enabled timely delivery, with manufacturing slot allocation within one week of slot request and a median vein-to-vein time of 29 days (range, 16-102), independent of geographic location. In r/r B-ALL patients, the median follow-up was 14 months, and the median overall survival (OS) not reached, and progression-free survival (PFS) was 18 months (range: 9-NR). The 12-month OS and PFS were 64% (95% CI: 53-72) and 55% (95% CI: 45-65). In r/r B-NHL, the median follow-up was 13 months and the median PFS was 11 months (range:7-16) and OS was not reached. The 12-month OS and PFS were 63% (95% CI: 54-71) and 47% (95% CI: 38-56). No patients underwent consolidative stem cell transplantation. Grade 3/4 cytokine release syndrome (CRS), immune effector cell-associated neurotoxicity syndrome (ICANS), and Immune Effector Cell-associated Hemophagocytic Lymphohistiocytosis-like Syndrome (IEC-HS) occurred in 6%, 4%, and 23% of r/r B-ALL and 6%, 3%, and 18% of r/r B-NHL patients, respectively. These findings demonstrate the successful implementation of tali-cel across multiple centers in a large cohort, which might contribute towards improving CAR-T access globally.

Non-Hodgkin lymphomas (NHLs) are among the most common cancers in the US, yet their etiology is mostly unknown. Epstein-Barr virus (EBV) is a known driver of rare NHLs, but its role in common subtypes is unclear. We evaluated associations between antibodies to 16 EBV antigens, including 12 emerging markers (anti-: BARF1, BBRF1, BFRF1, BGLF2, BHRF1, BORF1, BPLF1, BRLF1, BXLF1, LF2, LMP1, VCAp40) and NHL risk in prediagnostic blood samples from 2134 case-control sets from 4 prospective cohort studies. Conditional logistic regression estimated odds ratios (ORs) and 95% confidence intervals (CIs). Associations across multiple antibodies were summarized using a β‑weighted score (EBVAb score). Eight markers were positively associated with NHL risk. Anti-LMP1 was associated with diffuse large B-cell lymphoma (DLBCL; OR = 2.83, 95% CI: 1.12-7.19) only. Six lytic markers (anti-: BGLF2, VCAp40, BFRF1, BARF1, BHRF1, BXLF1) were associated with a 2-3 times higher risk of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). Lytic marker associations for DLBCL and follicular lymphoma (FL) were weaker, with exceptions: anti-BXLF1 and DLBCL: OR = 2.25, 95% CI: 1.33-3.81; anti-BHRF1 and FL: OR = 2.75, 95% CI: 1.22-6.18. Anti-EBV scores > 0 were associated with ~50% higher NHL risk, strongest for CLL/SLL (Tertile 3: OR = 2.43, 95% CI: 1.30-4.55). Select EBV antibodies may serve as markers of common NHL risk.

Intensified chemotherapy regimens have improved outcomes in adults with Philadelphia chromosome-negative (Ph-) B-cell acute lymphoblastic leukemia (B-ALL), yet relapse remains a major cause of treatment failure and death. Blinatumomab is a T-cell engager that demonstrated marked activity in relapsed and measurable residual disease-positive (MRD+) B-ALL, as well as in frontline consolidation for MRD- patients. The phase 2 GRAALL-2014/B-QUEST substudy evaluated the integration of blinatumomab into consolidation and maintenance therapy for adults with high-risk (HR) B-ALL. Between 2015 and 2020, 489 adults aged 18 to 59 years with newly diagnosed Ph- B-ALL were enrolled, and among these 259 were classified as HR (presence of KMT2A-r, IKZF1 deletion, or end-of-induction MRD ≥ 10-4). A total of 94 patients with HR were enrolled in the QUEST study (2018-2020) and received up to 5, 28-day cycles of blinatumomab during consolidation and maintenance. In addition, 90 patients with HR who were treated before QUEST activation without blinatumomab served as controls in a post hoc analysis. Baseline characteristics were comparable between groups. Blinatumomab consolidation significantly improved MRD clearance, reduced relapse, and prolonged survival. At 5 years, cumulative incidence of relapse, disease-free survival (DFS), and overall survival were 23%, 68%, and 79% in the blinatumomab group compared with 49% (P = .001), 42% (P = .001), and 60% (P = .03) in controls. Patients eligible for allogeneic hematopoietic stem cell transplantation (alloHSCT) derived overall benefit. However, no clear additional DFS advantage was observed among those who ultimately underwent transplantation. These findings support the frontline integration of blinatumomab and warrant prospective evaluation of transplantation strategies in this setting. This trial was registered at www.clinicaltrials.gov as #NCT03709719.

Malignant histiocytic neoplasms (MHNs) are rare tumors derived from the mononuclear phagocyte system (MPS), encompassing histiocytic sarcoma, Langerhans cell sarcoma, interdigitating dendritic cell sarcoma, and other high-grade MPS lineage tumors. Despite advances in understanding histiocytic neoplasms, MHNs remain diagnostically challenging and lack standardized treatment algorithms. Current classification systems differ in lineage framing and fail to address mixed or ambiguous phenotypes, contributing to diagnostic uncertainty and inconsistent care. To address these gaps, the Histiocyte Society convened an international working group of pathologists and oncologists, including World Health Organization and International Consensus Classification contributors, to harmonize nomenclature, define minimum diagnostic criteria, and develop pragmatic treatment recommendations. Using a modified Delphi process and case-based review, the group formulated >40 consensus statements spanning classification, pathology, molecular testing, clinical evaluation, and therapeutic strategies. Key recommendations include the adoption of a unified MHN designation, use of a minimum immunophenotypic panel, integration of broad molecular profiling, and the documentation of previous hematopoietic malignancy. Treatment algorithms emphasize surgical resection for unifocal disease and targeted therapy or immune checkpoint inhibition for multifocal disease when actionable mutations or programmed death ligand 1 expression are present. These consensus recommendations aim to reduce diagnostic ambiguity, standardize reporting, and improve outcomes for both pediatric and adult patients. Future priorities include international registries to refine risk stratification and biomarker-driven trials exploring targeted therapy, immunotherapy, and combination approaches.

Despite great progress in understanding the genomic basis of immature T-cell acute lymphoblastic leukemia/lymphoblastic lymphoma (T-ALL) and acute leukemias of ambiguous lineage, there are still cases that lack defining genetic markers, complicating risk stratification, and limiting targeted therapeutic options. Recent studies have shown that enhancer hijacking drives oncogene activation in approximately half of T-ALL cases, with the BCL11B (BCL11 transcription factor B) enhancer frequently involved. Here, we describe a subtype of leukemia with a distinct gene-expression signature, and immunophenotype characterized by positivity for immature (CD38), myeloid (CD13), T-lymphoid (cytoplasmic [c]CD3, CD7), and B-lymphoid markers (CD19, CD79a, CD10). This subtype is defined by the t(14;16)(q32;q24) translocation, which places the FOXF1 gene and its antisense long noncoding RNA gene FENDRR under the regulatory control of the BCL11B enhancer, leading to their ectopic transcriptional activation. Common concomitant genetic lesions are loss-of-function alterations of GATA3, CDKN2A/CDKN2B deletion and activating JAK/STAT and NOTCH1 pathway mutations. Patients were predominantly children and adolescents/young adults and experienced poor treatment outcome. High-throughput drug screening of 176 compounds demonstrated efficacy of combined BCL2 apoptosis regulator family proteins and JAK/STAT signaling inhibitors. Additionally, the clinical use of tyrosine kinase inhibitors in some of these patients showed therapeutic efficacy. Collectively, these findings identify BCL11B-enhancer-mediated deregulation of FOXF1/FENDRR as a hallmark of a subtype of high-risk lineage ambiguous leukemia that is potentially amenable to targeted therapeutic intervention.

The molecular classification of T-cell acute lymphoblastic leukemia (T-ALL) remains incomplete, limiting risk stratification and the development of targeted therapies. Enhancer hijacking is a critical oncogenic mechanism that deregulates proto-oncogenes by repositioning cisregulatory regions via structural variants. Here, we performed an integrated analysis of pediatric and adult T-ALL and mixed-phenotype acute leukemias (MPALs), using whole-genome and whole-transcriptome sequencing. This analysis identified a group of 14 patients with predominantly T-lineage neoplasms driven by a t(14;16)(q32;q24) translocation, harboring universal GATA3 mutations and CDKN2A/B deletions. Mechanistically, this translocation repositions the ThymoD locus downstream of BCL11B, causing monoallelic, ectopic overexpression of FENDRR and mesenchymal transcription factor genes FOXF1 and FOXC2 and activating epithelial-mesenchymal transition transcription signatures. Immunophenotypic and single-cell RNA sequencing analyses revealed marked lineage ambiguity with myeloid and B-cell differentiation potentials specific to this subtype. Furthermore, functional analyses in CD34+ cord blood cells demonstrated that FOXF1 overexpression promotes myeloid differentiation while suppressing T-cell differentiation, serving as a key factor for lineage specification. Clinically, this subtype was detected in 0.15% to 4.0% of T-ALL/MPAL cases depending on the cohort, showing a median age of 15 years and enrichment in adolescents and young adults. Importantly, patients with t(14;16)(q32;q24) have an extremely poor prognosis, showing a trend toward worse outcomes than high-risk groups such as KMT2A-rearranged early T-cell progenitor-like, SPI1-rearranged, and LMO2 γδ-like T-ALLs. The unique molecular landscape and poor prognosis of patients with the t(14;16)(q32;q24) translocation underscore the need for the development of novel subtype-specific therapeutic approaches.

This study assessed real-world effectiveness and safety of lisocabtagene maraleucel (liso-cel) in patients with relapsed/refractory (R/R) large B-cell lymphoma (LBCL), including those with high-risk disease, secondary central nervous system (sCNS) involvement, comorbidities, and poor fitness, using data in the Center for International Blood and Marrow Transplant Research Registry from 5 February 2021 to 4 February 2025. Eligible patients (N = 1116) received liso-cel and had ≥1 effectiveness and safety assessment after infusion, including 195 in the second-line setting, 71 with sCNS, and 257 with transformed LBCL. Median age was 71.1 years (range, 21.5-91.2), with 72.3% aged ≥65 years. Within the overall population, 6.6% had an Eastern Cooperative Oncology Group performance status of ≥2, 53.4% had ≥1 comorbidity, and the median number of previous lines of therapy was 3 (range, 1-16). Median study follow-up was 12.6 months (95% confidence interval [CI], 12.5-12.8). Among effectiveness-evaluable patients (n = 1109), objective response rate was 81.2% and complete response rate was 71.3%. Duration of response, progression-free survival, and overall survival rates at 12 months were 60.2% (95% CI, 56.4-63.9), 51.2% (95% CI, 48.0-54.4), and 67.6% (95% CI, 64.5-70.6), respectively. Cytokine release syndrome was reported in 51.0% of patients, with grade ≥3 events in 2.5%. Immune effector cell-associated neurotoxicity syndrome was reported in 26.6% of patients, with grade ≥3 events in 9.2%. The 12-month nonrelapse mortality rate was 6.1% (95% CI, 4.6-7.8). These real-world data reinforce the effectiveness and safety of liso-cel in this broad population of patients with R/R LBCL, including younger patients and those with high-risk disease features.

In clonal hematopoiesis (CH), hematopoietic stem cells (HSCs) with mutations conferring a fitness advantage preferentially expand under evolutionary constraints. Over the last decade, cancer therapy has emerged as a selective pressure contributing to high rates of CH in patients with hematological and nonhematological cancers. Distinct chemotherapies, radiotherapies, immunotherapies, and targeted cancer therapies shape somatic evolution by inducing de novo mutations and by conferring a selective advantage to pre-existing CH clones. Such expansion may influence treatment response and toxicity, and is linked to the development of therapy-related myeloid neoplasms. CH prevalence will continue to increase as treatment options expand, cancers are detected earlier, and patients live longer. Managing CH and its adverse effects has the potential to improve cancer outcomes and survivorship.

Chimeric antigen receptor (CAR) T-cell therapy has significantly improved outcomes in patients with relapsed/refractory aggressive large B-cell lymphoma (R/R LBCL). Although thrombosis and bleeding complications have been increasingly reported, their clinical relevance and risk factors remain unclear, also due to heterogeneity between reporting studies. We investigated the incidence of thrombosis and bleeding in a more homogeneous real-world multicenter cohort by analyzing 250 adults with R/R LBCL who were included in the Dutch "Follow that CAR!" multicenter registry, and all received axicabtagene ciloleucel (axi-cel) CAR T-cell therapy. We assessed the impact of thrombosis and bleeding, incorporated as time-dependent covariates on overall survival and explored associations with cytokine release syndrome (CRS), immune effector cell-associated neurotoxicity syndrome (ICANS), anticoagulant therapy, and laboratory parameters. One-year cumulative incidences of thrombosis and bleeding were 6.3% and 11.0%, respectively. Median time to thrombosis or bleeding was 28 days after infusion for both events. Bleeding was associated with poorer overall survival (hazard ratio [HR] 3.77, 95% CI 1.96-7.27). Patients with a CAR-HEMATOTOX score ≥2 before CAR T-cell therapy (HR 3.62, 95% CI 1.23-10.69), therapeutic anticoagulation at time of infusion (HR 3.74, 95% CI 1.33-10.49), or thrombocytopenia grade ≥3 (platelet count < 50 × 109/L) at time of infusion (HR 3.55, 95% CI 1.12-11.26, P = 0.03) had a significantly higher risk of bleeding. Our study confirms the high risk and clinical relevance of bleeding in patients receiving CAR T-cell therapy. Prospective studies are necessary to confirm these results and to guide recommendations regarding modifiable risk factors such as the use of anticoagulation or transfusion thresholds during CAR T-cell therapy.

Paroxysmal nocturnal hemoglobinuria (PNH) is an ultra-orphan disease. In 2026, approved complement inhibitors (CIs) for PNH include three C5 inhibitors (C5i) and three proximal inhibitors (PIs). Clinical trials for the approved PI pegcetacoplan, iptacopan, and C5i + danicopan had clear protocols for changing from terminal to PI, extrapolated into real-world practice. There is no guidance for patients changing from a PI to a different CI. We present the largest international cohort to date. The inclusion criteria were as follows: patients established on a PI, who have changed treatment to a different CI. Sixty-five patients with a median age at PNH diagnosis of 40 years were included. Indications for CI switch were as follows: hemolysis (extravascular/recurrent breakthrough/not specified), side effects, trial termination, patient choice, and other. CI changes (149) were classified as (1) terminal-to-proximal (75/149), 6 hemolytic events reported within 14 days of CI change; (2) proximal-to-proximal (45/149), 2 hemolytic events reported within 14 days of CI change; and (3) proximal-to-terminal (29/149), 13 hemolytic events within 14 days of CI change. This is the largest cohort of PNH patients facing multiple CI changes. With the increasing availability of different CI classes, patients may experience several treatment changes over their disease course. CI should not be interrupted. The cohort would suggest that overlapping treatment for patients changing from PI to another CI is not required. Tapering of treatment does not seem to be effective at preventing hemolysis complications. Patients should be monitored closely for hemolysis, especially when changing from proximal-to-terminal inhibition. Prospective clinical/laboratory analysis is recommended for further clarification management for this patient group.

Relapse following B-cell maturation antigen (BCMA)-directed therapies represents a critical challenge in relapsed/refractory multiple myeloma (R/R MM). We performed a systematic review and meta-analysis of 34 studies (n = 1280) published between 2020 and 2025 to evaluate salvage CAR-T versus engineered antibody therapies post-BCMA exposure. The pooled overall response rate (ORR) was 60%. CAR-T therapy achieved significantly superior ORR compared to antibody-based approaches (77% vs 52%; p < 0.0001), with comparable complete response rates. While incidences of cytokine release syndrome and immune effector cell-associated neurotoxicity syndrome were similar, grade ≥3 cytopenias were more frequent with CAR-T. Notably, GPRC5D-targeted CAR-T yielded higher ORRs than BCMA-targeted constructs (86% vs 66%; p = 0.0006). Furthermore, patients with prior exposure to BCMA bispecific antibodies exhibited lower response rates compared to those with prior CAR-T exposure (71% vs 86%; p = 0.0404). In addition, studies with a median interval of ≥10 months demonstrated significantly higher ORRs than those with <10 months (70% vs 50%; p = 0.0146). In conclusion, GPRC5D CAR-T demonstrates superior efficacy to engineered antibodies in the post-BCMA setting. These findings support prioritising earlier CAR-T use and underscore the importance of antigen selection in optimising treatment sequencing.

Disease recurrence is the main cause of treatment failure after CD19-directed CAR T cells, often due to CD19 antigen loss, stability and/or coverage. To overcome single-antigen escape, we evaluated a trispecific CAR targeting CD19, CD20, and CD22 with OX40 co-stimulatory domain. Preclinical studies demonstrated potent, antigen-specific cytotoxicity in in vitro and in vivo lymphoma models. We then conducted a first-in-human phase I trial in patients with relapsed/refractory B-cell malignancies. Sixteen patients received infusions at a median vein-to-vein time of 7 days, at doses of 0.5-2×10⁶ cells/kg. No severe cytokine release syndrome nor neurotoxicity occurred. Overall response rate was 50%, including complete responses in 83% of lymphoma patients. One-year overall survival rate was 61%, with durable remissions observed in lymphoma. CAR T expansion did not correlate with dose or response. T-cell exhaustion in apheresis cells correlated with progressive disease. Trispecific CAR T cells are safe and potentially active in lymphoma.

Developing cancer therapies that induce specific death of malignant cells is critical for preventing relapse. Highly effective strategies, such as immunotherapy, exemplify this principle. Here, we provide the mechanistic basis for a small-molecule approach that leverages chemically induced proximity (CIP) to kill diffuse large B cell lymphoma, the most common non-Hodgkin lymphoma. We developed lysine acetyltransferase (KAT)-based TCIPs (transcriptional/epigenetic chemical inducers of proximity), or KAT-TCIPs, which redirect p300/CREB-binding protein (CBP) to activate cell-death networks repressed by the oncogenic driver BCL6. Our lead KAT-TCIP reprograms the epigenome to initiate apoptosis. The crystal structure of the chemically induced p300-BCL6 complex reveals how chance protein-protein interactions may be exploited to confer the potency and selectivity of KAT-TCIPs. Thus, oncogenic drivers can be co-opted to activate robust cell death. Consistent with their gain-of-function mechanism, TCIPs recruiting different transcriptional activators-p300, BRD4, or CDK9-produce distinct genomic responses, suggesting specialized therapeutic uses.

Monoclonal gammopathy (MG) of undetermined significance (MGUS) is the earliest and most prevalent stage of MG that may progress to symptomatic multiple myeloma (MM) or Waldenström's macroglobulinemia (WM) through intermediate smoldering forms of both diseases. We used next-generation flow cytometry (NGF) to identify and purify bone marrow (BM) clonal plasma cells (cPC) and B-cells/lymphoplasmacytic cells (cB-cells) from 25 asymptomatic individuals with MG diagnosed within the population-based iStopMM screening program for further molecular analysis by next-generation sequencing (NGS). IgM-MGUS cases consistently exhibited WM-like phenotypes by NGF, and MYD88 p.L265P and/or CXCR4 mutations in the coexisting cPC and cB-cells. Conversely, cPC from non-IgM MGUS and smoldering MM (SMM) displayed a MM-like phenotype associated with genetic drivers -i.e., t(11;14)-, frequently coexisting with genetically unrelated cB-cells. Interestingly, recurrent RUNX1 single nucleotide variants were detected in cPC, but not coexisting cB-cells, from WM-like cases, suggesting a role for RUNX1 in PC differentiation. In summary, NGF enabled detailed immunophenotypic characterization of BM cPC and cB-cell populations in all cases analyzed, subdivided in IgM vs non-IgM cases with WM-like or MM-like phenotypic and genetic profiles, respectively. These findings underscore the potential contribution of NGF and NGS for accurate classification or early-stage disease, beyond conventional criteria.

In this issue of Molecular Cell, Budinich et al. reveal a role for the local production of RNA in mutant ENL condensate formation, chromatin occupancy, transcriptional regulation, and leukemogenesis.

Aberrant chromatin-associated condensates have emerged as drivers of transcriptional dysregulation in cancer, yet how extrinsic factors modulate their assembly and function remains poorly understood. Gain-of-function mutations in the chromatin reader ENL ("Eleven-nineteen-leukemia") drive oncogenesis by inducing condensate formation at select target loci. Here, we demonstrate that locally produced transcripts reinforce the nucleation, chromatin engagement, and oncogenic activity of mutant ENL condensates. Mutant ENL binds to RNA in part through a basic patch within its YEATS domain, and this interaction enhances condensate formation in vitro and in cells. Using a chemically inducible condensate displacement and renucleation system, we show that blocking ENL-RNA interactions or transcription impairs condensate reformation at endogenous targets. RNA binding preferentially enhances mutant ENL occupancy and transcriptional bursting at condensate-permissive loci. In mouse models, disrupting RNA binding suppresses mutant ENL-driven oncogenic transcription and leukemogenesis. These findings reveal how chromatin-associated oncogenic proteins hijack local transcripts to reinforce condensate nucleation and drive tumorigenesis.

Polyadenylation is essential for mRNA stability and translational efficiency. Although poly(A) tail length is dynamically regulated under physiological conditions, its dysregulation and functional importance in cancer remain poorly understood. Here, we identify widespread poly(A) tail elongation and aberrant upregulation of poly(A) polymerase alpha (PAPOLA) in acute myeloid leukemia (AML), with high PAPOLA expression associated with poor clinical outcomes. Using primary AML samples, leukemia cell lines and multiple mouse models, we demonstrate that PAPOLA-driven hyperactive polyadenylation promotes leukemogenesis and sustains leukemia stem cell maintenance. Mechanistically, PAPOLA enhances metabolic reprogramming by upregulating glutathione S-transferase mu 2 (GSTM2), which activates the 4-hydroxynonenal (HNE)-dihydrolipoamide dehydrogenase (DLD) axis to drive AML progression. Notably, pharmacological inhibition of PAPOLA with cordycepin suppresses metabolic reprogramming and impairs leukemogenesis. Overall, our findings establish hyperactive polyadenylation as a core oncogenic mechanism linking RNA processing to cancer metabolism in AML, highlighting the PAPOLA-GSTM2-HNE-DLD axis as a promising therapeutic target.

On the basis that T-cell acute lymphoblastic leukemia (T-ALL) cells overexpress IL7 receptor (IL7R), which promotes resistance to chemotherapy and disease relapse, here we develop IL7Rα (CD127)-targeted chimeric antigen receptor (CAR) T cells with low- and high-affinity single-chain variable fragments. We establish the antitumor efficacy of CD127 CAR against T-ALL cells in vitro, in female mouse models of T-ALL, and against blasts from patients with T-ALL using the patients' own T cells transduced with CD127 CAR. Antitumor efficacy is higher with low-affinity CAR T cells than high-affinity CAR T cells, albeit with fratricide of CAR T cells following eradication of CD127-overexpressing blasts. CRISPR-Cas9 knockout of CD127 eliminates fratricide at the risk of prolonged lymphopenia in vivo. To overcome fratricide, we investigate short-term ( < 7 days) co-culture with or without dasatinib, a tyrosine kinase inhibitor, versus a natural selection method (10 days) and demonstrate that co-culturing with dasatinib facilitates higher CAR T-cell yield, improved fitness, and preserved functionality. In vivo, dasatinib can be used to temporarily and reversibly suppress CAR T-cell activity. With this supporting translational data, we are initiating a trial with low-affinity CD127 CAR T cells for adult and pediatric patients with relapsed or refractory T-ALL.

Multiple myeloma (MM) is a plasma cell malignancy characterized by genomic instability and a high dependency on proteasome activity. Accordingly, DNA-damaging agents and proteasome inhibitors (PIs) are frontline therapies; however, therapeutic resistance inevitably develops. RAD23A, a dual-function protein involved in nucleotide excision repair and proteasome-mediated protein degradation, is highly expressed in MM and is associated with poor patient survival. Here, we demonstrate that depletion of RAD23A exerts potent anti-MM activity both in vitro and in vivo. Inducible CRISPR/Cas9 knockout (iKO) or shRNA-mediated knockdown of RAD23A in MM cell lines, including PI-resistant models, markedly suppressed cell proliferation. RAD23A loss impaired DNA repair capacity, triggered activation of the stress-associated eIF2α-ATF4 axis and promoted caspase-dependent apoptosis. Quantitative proteomic profiling of RAD23A-iKO cells confirmed upregulation of apoptotic and stress-response pathways, accompanied by downregulation of DNA repair and deubiquitination pathways. In an MM xenograft model, RAD23A depletion significantly reduced tumor growth and prolonged survival. Collectively, these findings identify RAD23A as a critical survival factor in MM and highlight its potential as a novel therapeutic target.

Chimeric Antigen Receptor T-cell (CAR-T) therapies represent a powerful modality for treating a variety of hematological cancers. However, limited efficacy in broader disease contexts, particularly solid tumors, underscores the need for improved CAR design to enhance potency, persistence, and safety. Key design features of CARs have been profoundly informed by extensive knowledge of the T-cell receptor (TCR) and its interactome. To advance CAR-T therapies, new functionally relevant proteins are needed to support engineering efforts. Until now, the actual molecular microenvironment surrounding CARs has remained poorly defined, chiefly due to the lack of a characterization method with the required precision and sensitivity. Herein, we introduce μMap-CAR, a high-resolution photocatalytic proximity labeling platform featuring key methodological optimizations that enable direct elucidation of the CAR interactome on live T-cell surfaces. The platform performs robustly in a model CAR-T cell system under both resting and simulated activation conditions. The high sensitivity of μMap-CAR allows interrogation of interactome differences upon CAR endodomain alterations, linking perturbed signaling networks directly to CAR components. We further deliver the first high-resolution intrinsic CAR interactome in primary T-cells, defined by shared interactors across donors, and validate candidates via super-resolution microscopy and CAR-T activation perturbation. This platform constitutes a powerful, broadly applicable tool for CAR interactome profiling while also providing actionable targets for proximity-guided CAR engineering applications.

Mutant spliceosome proteins (e.g., U2AF1S34F, SF3B1K700E, or SRSF2P95H) alter RNA splicing in myeloid neoplasms, leading to increased production of nonsense transcripts. Inhibiting the nonsense-mediated RNA decay (NMD) pathway, which is responsible for the degradation of nonsense transcripts, preferentially kills cells expressing mutant spliceosome proteins in vitro. In this study, we used an inhibitor of the kinase SMG1, a key regulator of NMD, to provide in vivo evidence that NMD is a therapeutic vulnerability for splicing factor mutant myeloid neoplasms. Primary mouse acute myeloid leukemia cells and human K562 leukemia cell lines expressing splicing factor mutants were more sensitive than wild-type (WT) cells to in vivo inhibition of SMG1 (SMG1i). Disruption of NMD activity by SMG1i led to increased R-loop levels in spliceosome WT cells, which were further increased in treated U2AF1S34F cells. This R-loop accumulation was accompanied by an increase in DNA damage. Degradation of R-loops with RNase H1 rescued spliceosome mutant cells from NMD inhibition-induced cell death. In U2AF1S34F cells, SMG1i increased NMD transcript isoforms (with reduced but detectable protein levels), which were enriched for DNA repair genes, including ATR and RAD51. Consequently, SMG1i-induced cell death in splicing factor mutant leukemias could be further enhanced by the inhibition of ATR or RAD51. This study shows that in vivo targeting of NMD is a therapeutic strategy to treat myeloid neoplasms with aberrant splicing. Cells with spliceosome mutations are sensitive to SMG1 inhibition in vivo due to R-loop accumulation, suggesting that nonsense-mediated decay targeting, alone or with DNA repair inhibitors, could be effective in splicing-mutant myeloid neoplasms.

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.

Chronic inflammation and aging skew hematopoiesis toward myelopoiesis at the expense of lymphoid output. We screened type 2 and anti-inflammatory cytokines to identify extrinsic signals capable of restoring lymphoid lineage commitment in hematopoietic stem and progenitor cells (HSPCs). Interleukin 4 (IL-4) specifically inhibited inflammation-induced myelopoiesis and shifted multipotent progenitor (MPP) differentiation toward the lymphoid lineage. IL-4 activated a signal transducer and activator of transcription 6 (STAT6)-dependent transcriptional program in MPPs, increasing the expression of lymphoid-specific genes. Mechanistically, the receptor tyrosine kinase FMS-like tyrosine kinase 3 (FLT3), which is highly expressed in MPPs, interacted with IL-4Rα to facilitate STAT6 activation. In vivo, IL-4 reversed inflammation-induced hematopoietic imbalance and accelerated lymphoid recovery. In aged mice, IL-4 administration shifted the MPP composition toward a lymphoid bias and restored B and T lymphocyte output. Long-term IL-4 treatment in aged mice improved immune, metabolic, physical, and cognitive functions; these rejuvenating effects were recapitulated by transplantation of IL-4-treated HSPCs. Promoting IL-4 signaling in MPPs may enable correction of hematopoietic dysregulation in inflammatory and aging-related conditions.

Hepatocellular carcinoma (HCC) exhibits substantial interpatient heterogeneity, leading to markedly variable outcomes and survival even among patients with similar stages and imaging phenotypes. Mainstream staging systems remain suboptimal, whereas pathology-dependent factors and high-cost genomic assays are neither scalable nor timely for clinical decision-making. Existing algorithms provide coarse risk stratification or static binary predictions, failing to capture the time-varying risk of death. We developed and externally validated TEMPO-HCC, a multimodal deep survival model with hierarchical interpretability, to estimate individualized overall survival risk trajectories after curative-intent resection. We curated a six-center cohort of 1475 patients and integrated multiphasic MRI, postoperative H&E whole-slide images, and perioperative predictors. A discrete-time survival head generated probabilities at 1, 2, 3, and 5 years after surgery. TEMPO-HCC outperformed unimodal models and guideline-based staging systems, achieving C-index of 0.751 and time-dependent AUCs of 0.836, 0.781, 0.812, and 0.680 at 12, 24, 36, and 60 months, respectively, in the external validation cohort. TEMPO-HCC represents a paradigm shift from static binary classification toward clinically actionable temporal risk prediction. Its hierarchical interpretability provides an auditable evidence chain linking macro-scale radiologic phenotypes to micro-scale histopathologic patterns. Importantly, augmenting guideline staging with TEMPO-HCC improved discrimination, enabling personalized postoperative surveillance and risk-adapted clinical management.

Alcohol-associated liver disease (ALD) is a major driver of morbidity and mortality globally, especially in its most severe phenotypes, including alcohol-associated hepatitis (AH), decompensated cirrhosis, and hepatocellular carcinoma. With no US Food and Drug Administration-approved therapies to improve ALD, there is an unmet clinical need to better define the pathophysiology of early and severe liver disease. Recent improvements in preclinical models have furthered our understanding of the mechanisms of disease in advanced ALD. In this review, our focus is on current progress in identifying therapeutic targets utilizing preclinical models and reporting the rationale for and current status of ongoing clinical trials that aim to develop effective therapies to slow the progression of earlier stages of ALD and especially improve survival in severe AH.

Tertiary lymphoid structures (TLS) are associated with heterogeneous outcomes in hepatocellular carcinoma (HCC), but whether their anatomical context determines clinical impact remains unknown. We hypothesized that spatial compartmentalization of TLS influences their prognostic significance. We developed SpatialDecoder, a deep learning pipeline that simultaneously segments TLS, classifies three maturation subtypes (Agg, Fol I, Fol II), and assigns spatial compartments (intratumoral, peritumoral, capsular), achieving accurate tissue segmentation (mean Dice similarity coefficient (DSC)=0.86), TLS segmentation (DSC=0.8609), and subtyping (macro Area Under the Receiver Operating Characteristic Curve (AUROC)=0.8810). Applied to 1188 resected HCC patients, it enabled large-scale spatial TLS mapping. Spatial mapping revealed a prognostic dichotomy: higher intratumoral TLS density independently predicted prolonged overall survival, whereas higher extratumoral (peritumoral and capsular) TLS density correlated with increased recurrence risk. Based on TLS distribution, we defined four spatial immune phenotypes: TLS-Enriched (high intra/low extra), TLS-Balanced (high/high), TLS-Excluded (low intra/high extra), and TLS-Deficient (low/low). These phenotypes stratified patients into a survival gradient (log-rank p<0.0001): TLS-Enriched conferred the best outcome (HR = 0.48), TLS-Excluded the worst (HR=1.80), with intermediate prognosis for the other two phenotypes. Multivariable analysis indicated independent prognostic value. Transcriptomic profiling uncovered distinct immune mechanisms across the four phenotypes. This study establishes TLS spatial context as a determinant of their dualistic role in HCC immunity, providing a large-scale demonstration of opposite prognostic roles based on TLS location. The SpatialDecoder pipeline and four-phenotype framework transform TLS assessment from a binary metric into a spatially informed approach for postoperative risk stratification.

Glypican-3 (GPC3) is highly expressed in hepatocellular carcinoma (HCC), making it an attractive target for chimeric antigen receptor (CAR) T cell therapy; however, this approach has previously shown limited clinical efficacy, potentially owing to high levels of transforming growth factor-β (TGFβ) in the tumour microenvironment1-4. We therefore engineered CAR T cells with a dominant-negative TGFβ receptor II, which showed enhanced antitumour activity in preclinical studies5. Here we report findings from a first-in-human trial evaluating the safety and efficacy of C-CAR031 in patients with advanced, treatment-refractory HCC ( NCT05155189 ). Thirty-six patients received CAR T infusions at four dose levels (from 0.75 × 106 to 4.0 × 106 cells per kg). Cytokine release syndrome was reported in 34 patients, of which two cases were grade 3. Nine patients had non-haematological adverse events of grade 3 or higher. Tumour regression was observed in 32 patients, with a median best tumour reduction from baseline of 41.6% (range: 3.4-94.4%) in target lesions. The objective response rate was 44.4%, and the median duration of response was 4.4 months (95% confidence interval: 2.9-7.4). Median progression-free survival and overall survival were 4.2 months (95% confidence interval: 2.9-4.8) and 14.2 months (95% confidence interval: 10.1 to not evaluable), respectively. High-throughput analyses of tumour samples and functional validation suggested that GPC3 antigen loss and increased TGFβ levels may contribute to C-CAR031 resistance. Collectively, these results indicate that C-CAR031 has a manageable safety profile and encouraging antitumour activity in heavily pretreated patients with advanced HCC.

Hepatocellular carcinoma (HCC) typically develops in a clinically silent manner, and the suboptimal sensitivity and specificity of currently available diagnostic biomarkers remain significant obstacles to its accurate and early detection. To improve molecular diagnostic performance, we developed a dual-function CRISPR/Cas12a assisted strand displacement reaction (dCas12a-SDR) with hexaammine ruthenium(III) chloride (RuHex)-loaded DNA condensate for ultrasensitive and highly specific detection of HCC-associated mRNAs. Upon target recognition, the previously sequestered Cas12a activation site within the electrode-immobilized capture probe is exposed, thereby inducing hybridization between the accessible single-stranded domains retained in RuHex-loaded DNA condensates (RuDC) and the displacement strand (Ds), which ultimately leads to the release of electroactive Ds-RuDC condensates and the effective activation of Cas12a. Activated Cas12a then removes the activation site via cis-cleavage, releasing the target to enter subsequent reaction cycles; concurrently, the activated system initiates trans-cleavage of adjacent capture probes and Ds-RuDC assemblies on the electrode that harbor trans-cleavage motifs, thereby promoting the release of RuDC from the electrode interface. This cascade ultimately leads to a pronounced reduction in the electrochemical signal. Owing to this target-triggered dual cis- and trans-cleavage mechanism mediated by Cas12a, the biosensor achieves highly efficient signal amplification. The platform affords a limit of detection as low as 39.2 aM for PD-L1 mRNA and exhibits excellent specificity, stability, and reproducibility. Moreover, by jointly detecting a panel of HCC-associated mRNAs (PD-L1, GPC3, EpCAM, and FGA), the platform successfully discriminated healthy individuals from patients with early-stage hepatocellular carcinoma in clinical serum samples. Collectively, this platform provides a powerful tool for molecular diagnosis of hepatocellular carcinoma.

Metabolic dysfunction-associated steatotic liver disease (MASLD) progresses along a continuum from simple steatosis to steatohepatitis, fibrosis, cirrhosis and hepatocellular carcinoma. However, current clinical and research frameworks rely primarily on static, histology-defined stages that fail to capture the continuous nature of disease progression. Here, we present a data-driven framework that reconstructs MASLD progression as a continuous molecular trajectory from cross-sectional liver transcriptomic profiles. By positioning patients along this trajectory, we move beyond conventional stage-based classifications and resolve the ordered activation of regulatory programmes, signalling pathways and cellular remodelling processes underlying disease progression. To enable non-invasive patient stratification, we integrate the inferred molecular trajectory with paired liver-plasma proteomics data and identify a 57-gene plasma-accessible biomarker panel that accurately predicts advanced fibrosis and continuously positions patients along the disease trajectory across independent cohorts, outperforming established non-invasive clinical scores. Together, this work establishes a generalizable trajectory-based framework for understanding MASLD pathophysiology and provides a foundation for mechanistically informed biomarker discovery, precision staging and stage-aware therapeutic prioritization.

Recurrence after tumor resection and liver transplantation remains a major clinical challenge in hepatocellular carcinoma (HCC). We examined tumor samples from HCC liver transplant patients to identify drivers of disease recurrence. Integration of proteomic profiling of human HCC samples with or without recurrence and T cell killing assays linked the cholesterol esterification enzyme SOAT1 to immune evasion and cancer recurrence. Genetic or pharmacological inhibition of SOAT1 sensitized liver cancer cells to CD8+ T cell-mediated immunosurveillance, anti-programmed cell death 1 (anti-PD-1) therapy, and chimeric antigen receptor (CAR)-T cell therapy and increased intratumoral CD8+ T cell infiltration. Mechanistically, the inhibition of SOAT1-mediated cholesterol esterification disrupted intratumoral cholesterol and lipid metabolism and reduced unsaturated fatty acids and prostaglandin E2 production; this impaired the antioxidant capacity and metabolic resilience of cancer cells under immune attack. Accordingly, SOAT1 inhibition enhanced immunotherapy efficacy in obesity-associated tumors and under immunosuppressant-induced conditions. Thus, cholesterol esterification supports redox fitness and cancer cell resilience upon immune attack, suggesting a strategy to prevent recurrence and improve immunotherapy outcomes.

Liver diseases constitute a major global health challenge, affecting over 25% of the world's population, with metabolic dysfunction-associated steatotic liver disease (MASLD) emerging as the most prevalent chronic liver disorder and a leading cause of cirrhosis, hepatocellular carcinoma, and liver-related mortality. Despite its increasing burden, effective and targeted pharmacological therapies remain limited, highlighting a critical need to identify novel molecular targets and mechanistic pathways for therapeutic intervention. The endocannabinoid system, comprising cannabinoid receptors, endogenous ligands, and the enzymes involved in their synthesis and degradation, has gained significant attention as a regulator of hepatic physiology and disease. Although cannabinoid receptor 1 (CB1) has been extensively characterized, cannabinoid receptor 2 (CB2), previously considered predominantly immunomodulatory, is now recognized as a key regulator of hepatic lipid metabolism, inflammatory signaling, and fibrogenesis. Metabolic dysfunction induces hepatic lipid accumulation, oxidative stress, and endoplasmic reticulum stress, triggering inflammatory cascades that drive disease progression from simple steatosis to MASLD, metabolic dysfunction-associated steatohepatitis (MASH), and ultimately fibrosis. Emerging evidence indicates that CB2 activation exerts anti-steatotic, anti-inflammatory, and anti-fibrotic effects through cell-specific mechanisms in hepatocytes, Kupffer cells, and hepatic stellate cells. This review revisits CB2 as a central regulator of hepatic pathophysiology, integrating mechanistic insights with preclinical and clinical evidence. Collectively, targeting CB2 signaling presents a promising therapeutic strategy to mitigate steatosis, inflammatory cascades, and fibrotic progression in chronic liver diseases.

Combination therapies involving vascular targeting drugs have shown promise in overcoming resistance to immunotherapy. However, the prerequisite for vascular modulation to evoke an effective antitumor T-cell response remains elusive. Tracing the transcriptional response of liver metastasis-associated peritumoral and tumor endothelial cells (TEC) to T-cell intervention, we discovered an immunomodulatory TEC subpopulation that highly expressed lipoprotein lipase (LPL). LPL+ TECs facilitated intratumoral homing of activated antitumor CD8+ T cells driving liver metastatic regression. Mechanistically, LPL enhanced MHC-I-dependent cross-presentation of tumor antigens on TECs for T-cell trafficking. Consequently, LPL+ TECs were recognized and targeted by T cells, further aiding antitumor response. Corresponding analyses of human liver metastasis samples identified a significant correlation between the presence of intratumoral LPL+ blood vessels and the accumulation of T cells. Altogether, the study identifies a decisive role of TECs in orchestrating an effective T-cell response by overcoming tumor's intrinsic insufficient antigen presentation. The study identifies LPL+ TECs as orchestrators of activated CD8+ T-cell homing into immunologically cold tumors with low baseline MHC-I expression. Enhancing tumor antigen exposure by MHC-I cross-presentation in TECs presents a promising approach to compensate the intrinsic inability of tumor cells and boost antitumor immunotherapy.

Human cytoplasmic leucyl-tRNA synthetase (LARS) is known to catalyze the ligation of leucine to tRNALeu during protein biosynthesis. However, LARS also acts as a nutrient sensor in a non-canonical activity to regulate cell growth. In this investigation, LARS was determined to be expressed at high levels in human liver cancer, which correlated with poor clinical outcomes in patients. Knockdown of LARS in HepG2 liver cancer cells suppressed cell proliferation and growth, yet promoted cell migration, without affecting global protein translation. Using data from RNA sequencing, differentially expressed genes in response to LARS knockdown were significantly clustered in the cellular senescence pathway. Elevated p21 and p16 expression with increased senescence-associated β-galactosidase activity was observed in LARS knockdown HepG2 cells. In addition, autophagy with increasing autophagic flux was triggered in LARS knockdown HepG2 cells. Furthermore, the depletion of LARS induced oxidative stress with increased production of reactive oxygen species and decreased mitochondrial membrane potential. Importantly, re-expression of shRNA-resistant LARS largely rescued the senescence, autophagy, and oxidative stress caused by LARS knockdown, confirming the specificity of these effects. These findings suggest that LARS plays a role in regulating liver cancer cell proliferation via multiple cellular responses and may serve as a promising therapeutic target in liver cancer.

Intrahepatic cholangiocarcinoma (ICC) is a lethal hepatic malignancy characterized by a prominent desmoplastic stroma. Here, we demonstrated that ARID1A, a core component of the SWI/SNF protein complex, is upregulated in ICC and plays an oncogenic role. Mechanistically, ARID1A stabilized NICD1 protein by inhibiting AMPK-dependent autophagy and lysosomal degradation, thereby sustaining Notch signaling. TLL1, a metalloproteinase that promotes collagen maturation, was found to be a downstream effector of ARID1A/Notch signaling. Specifically, TLL1 derived from ICC cells activated hepatic stellate cells (HSCs) and thus promoted tumor progression. Accordingly, ablation of TLL1 attenuated cancer-associated fibroblast (CAF) infiltration, collagen accumulation, and tumor progression. Moreover, virtual screening of a bioactive compound library identified acarbose, an effective medicine for glycemic control, as a potent inhibitor for TLL1. Pharmacological inhibition of TLL1 with acarbose suppressed HSC activation, collagen deposition, and tumor development. Collectively, these results establish a tumor-promoting ARID1A/Notch/TLL1 axis in ICC and reveal acarbose as a potential precision therapy for ARID1A-high ICC patients.

Type 1 innate lymphoid cells (ILC1) are abundant in the adult liver and are pivotal for immune surveillance and modulation, but the regulation of their maintenance and functionality remains underexplored. Here, we re-analyze published single-cell RNA- sequencing data and find increased expression of Asb2 in ILC1s from adult mouse livers. Conditional ablation of Asb2 in NKp46+ cells, depleting ASB2 in ILC1s and NK cells, in mice impairs ILC1 survival and reduces adult liver ILC1 numbers. Proteomics and bulk RNA-sequencing reveal enriched lipid metabolism pathways in ASB2-deficient ILC1s, concomitant with increased lipid storage. Importantly, pharmacological inhibition of lipid synthesis prevents the apoptosis of ASB2-deficient ILC1s in vitro. In a mouse model of colorectal cancer liver metastasis, we find increased ILC1 lipid storage, and conditional Asb2 deficiency in ILC1 cells exacerbates liver metastasis progression. Conversely, inhibiting lipid accumulation in wild-type colorectal cancer-bearing mice prolongs animal survival, potentially via promoting ILC1-mediated anti-tumor immunity. Thus, our study uncovers ASB2-regulated lipid metabolism as a gatekeeper for ILC1 homeostatic fitness and tumor surveillance, highlighting potential ILC1-based therapeutic strategies against liver tumors.

The prevailing notion is that effector T cell activation mediates anti-PD-1 efficacy in cancer. Here, we conducted a mechanistic study parallel to our phase 2 trial of perioperative anti-PD-1 therapy in patients with resectable recurrent hepatocellular carcinoma (HCC) (NCT04615143) to study its mechanism of action. Late-recurrence patients present two distinct subtypes characterized by T cell or B cell dominant responses in the tumor microenvironment by dynamic single-cell multi-omics analysis. Clonal antibody repertoire analysis and spatially paired scRNA-seq/BCR-seq reveal somatic hypermutation promoting antibody binding against hepatitis B virus core antigen (HBcAg) within tumor tertiary lymphoid structures (TLSs) in these type B-late recurrence patients. Mechanistically, HBcAg is exported into the extracellular space, triggering local B cell and antibody responses and complement activation. In mice, these high-affinity HBcAg-reactive antibodies lead to complement-mediated antitumor activity with enhanced anti-PD-1 efficacy. Thus, we uncover enhanced anti-virus B cell immunity within the TLS as a mechanism to anti-PD-1 in HCC.

Hepatocellular carcinoma (HCC) remains one of the most lethal malignancies, characterized by a highly complex and dynamic tumor microenvironment in which tumor-associated macrophages (TAMs) are particularly abundant. TAMs regulate tumor initiation, progression, metastasis, and therapy response, making them attractive targets. However, targeting TAMs remains challenging due to their remarkable plasticity and functional heterogeneity. Recent high-throughput multi-omics studies have revealed that the HCC microenvironment consists of distinct regions defined by cellular architecture, such as the fibrotic invasive margin, perivascular niche, poorly vascularized hypoxic/necrotic areas, and cancer stem cell niche, each associated with unique protumorigenic properties. This review summarizes TAM heterogeneity across liver niches, disease stages, and treatment contexts, and evaluates TAM-targeted strategies, emphasizing key unresolved challenges to improving HCC outcomes.

Tertiary lymphoid structures (TLSs) are ectopic lymphoid structures in chronic inflammation or tumors in peripheral tissues. TLS maturation is associated with clinical advantages of tumor immunotherapy. However, the underlying mechanisms of TLS formation remain elusive. Here we investigated the roles of SETDB1 in TLS formation in hepatocellular carcinoma (HCC). Setdb1 was knocked out in multiple HCC models, including Akt/NRAS and Myc/CTNNB1 hydrodynamic models, Hepa1-6 orthotopic and subcutaneous models. Spatial transcriptomics and multiplex staining were used for TLS verification. real-time RT-PCR and western blotting were used for mechanistic investigation. Low SETDB1 expression is associated with high TLS level in clinical samples, and Setdb1 depletion induces TLSs in multiple HCC models. Setdb1 depletion generates cytosolic DNA and activates cGas pathway, which increases Cxcl12 and Zbp1 expression. Cxcl12 is up-regulated in hepatocytes and cancer cells, and contributes to the recruitment and maturation of B cells. Zbp1 is activated in malignant cells and induces necroptosis in TLSs. Setdb1 depletion also increases the expression of ERVs, which may act as neoantigens for B cell maturation. Our study has revealed a critical role for Setdb1 in TLS formation and suggests a potential role for Setdb1 as a therapeutic target for enhancing the effects of immunotherapy in HCC. The current study found that Setdb1 depletion induces the formation of Tertiary lymphoid structures (TLSs) in hepatocellular carcinoma (HCC). TLSs are ectopic lymphoid structures in chronic inflammation or tumors in peripheral tissues, and mature TLSs are associated with clinical advantages of tumor immunotherapy. The study discovers a new pathway involving cGas/Cxcl12 for TLS formation, which will benefit the investigations about the mechanistic regulation of anti-tumor immunity. It also provides a promising strategy to combine SETDB1 inhibition with immunotherapy in drug design and HCC treatment.

Imaging reporter genes are essential tools in cancer research for monitoring tumor evolution, cell dissemination, gene activation and treatment response. However, germline reporter models typically rely on single imaging modalities operating over limited spatial scales. To address these limitations, we developed an inducible triple-reporter mouse model (Rosa26LSL-NRL) that integrates three reporters for complementary imaging modalities, fluorescence, bioluminescence and positron emission tomography (PET), along with inducible Cre-lox functionality for precise spatiotemporal control of reporter expression in genetically engineered mouse models. Using a multiscale, multimodal approach, we visualize deep tissue oncogenesis in models of hepatocellular carcinoma and lung adenocarcinoma and, guided by whole-body imaging with bioluminescence and [18F]tetrafluoroborate PET/magnetic resonance imaging (MRI), resolve cell-cell interactions within tumor microenvironments using in situ microscopy. This triple-reporter system enables multiscale investigation of biological processes within whole animals, facilitating sensitive, tissue-specific, in vivo cell tracking from whole-body to cellular resolution.

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

mRNA nuclear export is an essential biological process that is mediated primarily by nuclear export factor 1 (NXF1). NXF1 exerts its mRNA transport activity by binding to adaptor proteins. In an effort to identify novel NXF1 adaptors involved in hepatocellular carcinoma (HCC) development, we discovered that C7orf25, a previously uncharacterized protein named proliferation-associated NXF1 adaptor (PANAD), interacts with NXF1 in the nucleus. Specifically, the R122/E125/G133/D139 residues of PANAD bound to the Q20/K22/K23 residues in the RBD of NXF1. This binding disrupted the intramolecular interaction between the RBD and NTF2L domains of NXF1, thereby enhancing the association of NXF1 with target mRNAs. The mRNAs whose nuclear export was impaired upon PANAD knockdown were highly enriched in cell proliferation pathways. Moreover, PANAD directly bound to CDK6 mRNA. Gain- and loss-of-function analyses in cellular and mouse models revealed that PANAD accelerated G1/S transition and tumor cell proliferation, promoted colony formation and hepatoma xenograft growth, and acted through interactions with NXF1 to increase the nuclear export and translation of mRNAs encoding proliferation stimulators, including CDK6. PANAD and CDK6 were both upregulated in HCC tissues and were positively correlated. Furthermore, PANAD-overexpressing hepatoma cells exhibited increased sensitivity to the CDK4/6 inhibitor palbociclib, whereas PANAD-silenced cells exhibited reduced sensitivity. These findings identify PANAD as a novel NXF1 adaptor; delineate its critical regulatory roles in mRNA nuclear export, G1/S transition and tumor development; and highlight its potential as both a therapeutic target and a predictive biomarker for CDK4/6-targeted therapy.

N-acyl amino acids (N-acyl AAs) are potential bioactive compounds in fermented soybean (FS), but their accurate annotation is challenging due to their structural diversity, the existence of various isomers and wide polarity differences. Herein, we established a novel One-pot Micro Synthesis-Chemical Derivatization (OMSCD)-LC-MS analytical approach for comprehensive profiling of N-acyl AAs. First, the rapid synthesis of 142 N-acyl AA standards combined with in situ DIAAA derivatization in a one-pot manner significantly enhanced ionization and separation efficiency, particularly enabling excellent discrimination of isomers. Next, using this method, 222 N-acyl AAs were rapidly identified in FS, of which 200 were reported for the first time. Among them, N-oleoyl AAs, present at high contents in FS, demonstrated significant anti-liver cancer activity in a cell model, with MAPK1 acting as a promising candidate target. Overall, the OMSCD-LC-MS strategy facilitates the N-acyl AAs profiling, offering new preliminary insights into the functional components of FS.

Navitoclax (ABT-263) exhibits strong cytotoxic effects against hepatocellular carcinoma cells in vitro; however, its therapeutic efficacy in vivo is limited by insufficient local drug accumulation and compensatory upregulation of the anti-apoptotic protein myeloid cell leukemia-1 during long-term treatment. In addition, effective immune activation in hepatocellular carcinoma remains challenging due to the immune-privileged hepatic microenvironment. This study aimed to overcome these limitations through a tumor-targeted nanotherapeutic strategy. Dual pH-responsive copolymeric nanoparticles modified with folic acid and co-loaded with navitoclax and indocyanine green (FPNAI) were developed. FPNAI preferentially accumulated at hepatocellular carcinoma sites through folate receptor-mediated targeting and released navitoclax in response to the acidic tumor microenvironment, thereby inducing mitochondria-mediated apoptosis. Upon near-infrared irradiation, indocyanine green generated excessive reactive oxygen species, leading to downregulation of myeloid cell leukemia-1 and further enhancement of apoptotic signaling. The combined effects of navitoclax and reactive oxygen species synergistically promoted tumor cell apoptosis. Moreover, apoptosis induced by FPNAI triggered immunogenic cell death, characterized by dendritic cell maturation and activation of cytotoxic CD8⁺ T cells. In vivo studies demonstrated that FPNAI with near-infrared irradiation achieved significantly improved antitumor efficacy compared with navitoclax monotherapy, while maintaining favorable tumor-targeting capability and biosafety. This study demonstrates that FPNAI enhances the antitumor activity of navitoclax by integrating targeted drug delivery, reactive oxygen species-mediated mitochondrial apoptosis, and immunogenic cell death induction. The proposed nanotherapeutic platform represents a promising approach for improving hepatocellular carcinoma treatment outcomes.

Hepatocellular carcinoma (HCC) cells sustain viability and radioresistance by actively countering oxidative stress. Understanding the mechanisms regulating reactive oxygen species (ROS) homeostasis is therefore crucial for developing novel therapies. Using integrated genome-wide CRISPR-Cas9 screening coupled with transcriptomic and metabolomic profiling, we identified DDB1 and CUL4-associated factor 4 (DCAF4) as an essential regulator of oxidative stress resistance in HCC. Mechanistically, DCAF4 functions as a CRL4 E3 ligase adapter that promotes KEAP1 ubiquitination and degradation. Notably, under oxidative stress, cytoplasmic stress granules (SGs) form a localized platform that facilitates the DCAF4-KEAP1 interaction, accelerating KEAP1 degradation and leading to NRF2 activation and upregulation of antioxidant genes. We further identified that the transcription factor XBP1 enhances DCAF4 expression. Targeting this axis, we performed computational screening to identify a small-molecule inhibitor that disrupts the DCAF4-KEAP1 interaction. This compound effectively enhanced brachytherapy (BT) sensitivity and inhibited tumor growth in preclinical HCC models.

Hepatocellular carcinoma (HCC) recurrence following microwave ablation poses a pressing clinical challenge, driven by metabolically adapted residual cells that establish an immunosuppressive tumour microenvironment. Our study identifies the post-ablation upregulation of fumarylacetoacetate hydrolase (FAH) as a crucial "metabolic checkpoint" promoting relapse. Elevated FAH expression results in increased fumarate levels within residual HCC cells, supporting tumour cell survival by enhancing energy metabolism while concurrently impairing CD8+ T cell function. Mechanistically, fumarate binds to and stabilises heat shock protein 70 (HSP70), establishing a thermal ablation induced FAH-fumarate-HSP70 axis that drives immunosuppression. To counteract this pivotal axis, we engineered a gallium-based functionalized nanoplatform. This system incorporates a lactate oxidase shell that responds to the lactate-rich tumour microenvironment, enabling the site-specific co-release of Ga3+, FAH-silencing plasmids and the glycolysis inhibitor 2-deoxy-D-glucose. Our nanoplatforms disrupt the FAH-fumarate-HSP70 axis to eradicate residual HCC, activate CD8+ T cells, and restore immunity, targeting recurrence dysregulation while integrating metabolic blockade with immunomodulation to prevent post-ablation relapse with clinical translation potential.

Progression-free survival (PFS) is a widely accepted primary end point in metastatic breast cancer (mBC) trials; however, it remains susceptible to assessment bias. To reduce this risk, blinded independent central review (BICR) is often incorporated into trial designs, but emerging evidence has suggested discordance between investigator- and BICR-assessed PFS. To compare BICR- and investigator-assessed PFS of patients with hormone receptor-positive/ERBB2-negative mBC. PubMed, Embase, CENTRAL, and major conference proceedings were searched from database inception to November 27, 2025, to identify all potential eligible studies. Phase 2 or 3 randomized clinical trials (RCTs) including patients with hormone receptor-positive/ERBB2-negative mBC with progression following CDK4/6 inhibitor therapy. Eligible studies required or permitted prior CDK4/6 inhibitor use and reported PFS assessed by both local investigators and BICR. Data from the eligible RCTs were extracted by 2 reviewers (L.V.R. and Z.B.). A trial-level random-effect meta-analysis was performed. Outcomes were the discrepancy index (DI)-the ratio of hazard ratios for BICR- vs investigator-assessed PFS-and the absolute difference in median PFS between the 2 assessments. Of 4989 identified records, 21 RCTs with 9165 patients met inclusion criteria. Eleven RCTs used BICR as the primary assessment, 13 were open label, and 7 required prior treatment with CDK4/6 inhibitors as an inclusion criterion. No statistically significant difference was observed between investigator- and BICR-assessed PFS overall, with a DI of 1.02 (95% CI, 0.95-1.10; P = .57), which was consistent across meta-regression analyses. The pooled PFS difference was 0.26 months (95% CI, -0.16 to 0.68 months; P = .22) in interventional arms and 0.44 months (95% CI, 0.09-0.78 months; P = .01) in control arms. Results of this systematic review and meta-analysis show that despite a trend toward longer median PFS with BICR in recent RCTs of patients with hormone receptor-positive/ERBB2-negative mBC, there was no statistically significant DI between BICR- and investigator-assessed PFS. Results were consistent across meta-regression analyses and in outlier trials, suggesting that both approaches yield comparable PFS estimates.

Centrosome abnormalities (CA) are a hallmark of epithelial cancers, yet their spatial complexity and phenotypic heterogeneity remain poorly understood due to limitations in conventional image analysis. Here we present CenSegNet (Centrosome Segmentation Network), a modular deep learning framework for high-throughput segmentation of centrosomes and epithelial architecture, enabling accurate and generalisable centrosome phenotyping at spatial and single-cell resolution across imaging modalities and tissue contexts. Applied to tissue microarrays comprising 911 breast cancer cores from 127 patients, CenSegNet enables large-scale, spatially resolved quantification of numerical and structural CA. We show that these CA subtypes are mechanistically uncoupled, exhibiting distinct spatial distributions, age-dependent dynamics, and associations with tumour grade, hormone receptor status, genomic alterations and nodal involvement. Structural CA are associated with overall survival, whereas discordant CA profiles at tumour margins correlate with local tumour aggressiveness and stromal remodelling. These findings establish CenSegNet as a scalable platform for spatially resolved centrosome phenotyping, enabling systematic investigation of centrosome biology and its dysregulation in cancer and other epithelial diseases.

While breast cancer (BC) has always been considered an immunological 'cold' tumor type, anti-programmed cell death protein 1 (PD-1) has recently changed the treatment landscape of triple-negative breast cancer, and neoadjuvant Phase III trials for high-grade estrogen receptor-positive breast cancer have reported promising results. However, a substantial number of patients do not benefit and are in need of novel combination strategies. In addition, since anti-PD-1 is added to combination chemotherapy, there is a growing issue of overtreatment for patients with an already excellent outcome. In this review, we discuss the status and challenges of neoadjuvant immunotherapy in BC, including the need for prognostic and predictive biomarkers to guide treatment individualization and optimize the balance between the efficacy and toxicity of immunotherapy.

Triple-negative breast cancer (TNBC) is characterized by aggressive metastatic behavior and limited therapeutic options. Although metabolic reprogramming is increasingly recognized as a hallmark of TNBC, the mechanisms by which specific metabolic enzymes and intermediates drive metastasis remain poorly defined. We performed untargeted metabolomic profiling on TNBC tumors and matched normal tissues to identify dysregulated metabolic pathways. Functional assays, chemoproteomic succination profiling, molecular interaction analyses, and in vivo cancer metastasis models were used to define the mechanistic and biological consequences of altered metabolism. In vitro experiments validated the effects of N-acetylcysteine (NAC) in TNBC cell lines. Metabolomic analyses revealed aberrant activation of the alanine-aspartate-glutamate axis and upregulation of adenylosuccinate lyase (ADSL) in TNBC. ADSL promoted tumor cell proliferation and metastasis by generating fumarate, which accumulated primarily through covalent protein succination. Chemoproteomic profiling identified the cell polarity regulator SCRIB as a critical fumarate target. Fumarate-mediated succination of SCRIB impaired its membrane localization, promoted epithelial-mesenchymal transition, and facilitated aberrant interaction with the mTORC2 component RICTOR, leading to activation of AKT/mTOR signaling. Genetic disruption of SCRIB succination abrogated these effects. Importantly, pharmacological perturbation of fumarate using NAC reduced SCRIB succination and attenuated the malignant phenotypes of TNBC cells in vitro. These findings identify an ADSL-fumarate-SCRIB signaling axis that links metabolic reprogramming to the loss of cell polarity and activation of pro-metastatic signaling in TNBC. Targeting fumarate-mediated protein succination represents a previously unrecognized and experimental vulnerability in TNBC.

Photothermal therapy (PTT) has emerged as a promising strategy for tumor treatment, owing to its exceptional treatment outcomes and spatiotemporal control. However, persistent challenges, such as high-energy laser-induced damage to surrounding tissue and induces a cascade of heat shock responses (HSRs) in tumor cells, collectively contribute to suboptimal therapeutic outcomes. In addition, single-modality treatment approaches often fail to attain optimal therapeutic efficacy. In this study, we designed a gas/PTT nanocomposite material (PTZSCN-NO NPs) that integrates the photothermal molecule phenothiazin-thiophen-dimethylfuran (PTZSCN) with a thermosensitive nitric oxide (NO) donor (S-nitroso-N-acetylpenicillamine, SNAP). Upon laser irradiation, PTZSCN generates heat for PTT and promotes the release of NO from SNAP, can inhibit the expression of multiple HSPs, thus enhancing the efficacy of PTT. Furthermore, upon light exposure, PTZSCN-NO NPs generate a photothermal effect, leading to mitochondrial damage, caspase-1 activation, and gasdermin D cleavage. These events ultimately induce pyroptosis, achieving a synergistic gas/photothermal therapeutic effect and suppressed orthotopic breast tumor growth and metastasis in a tumor-bearing mice model. This approach optimizes conventional PTT and providing a repeatable, noninvasive treatment process.

Therapeutic antibodies and antibody-drug conjugates are limited by large molecular size, poor tumor penetration, and signaling-adaptive resistance. We introduce an engineering strategy for radiotherapeutic antibody fragments in which an albumin-binding domain (ABD) rebalances molecular size and pharmacokinetics. Using human epidermal growth factor receptor 2 (HER2) as a proof of concept, we developed 177Lu-DOTA-Fab-ABD, which integrates rapid tumor penetration with albumin-mediated extended circulation, improving tumor dosimetry while reducing off-target radiation. In HER2 tumor models, 177Lu-DOTA-Fab-ABD showed strong HER2 and albumin binding, reduced off-target retention, and substantially lower hematologic toxicity than full-length 177Lu-DOTA-pertuzumab while maintaining potent β-particle tumor control. It also overcame intrinsic trastuzumab resistance by bypassing receptor blockade and inducing DNA double-strand breaks and displayed mechanistic complementarity with trastuzumab. Overall, 177Lu-DOTA-Fab-ABD represents a generalizable framework for engineering fragment-based radiotherapeutics.

Neutrophils are protumorigenic and prometastatic in tumor-bearing mice. Comprehensive single-cell RNA sequencing analyses on various tissues of naïve and tumor-bearing MMTV-PyMT (mouse mammary tumor virus-polyoma middle T antigen) mice unraveled a neutrophil reprogramming process and uncovered a strategy to selectively target reprogrammed neutrophils. Transcriptional reprogramming of neutrophils in tumor-bearing mice is initiated in the bone marrow (BM) by granulocyte colony-stimulating factor (G-CSF). Among the seven neutrophil clusters in the BM, the abundance of one cluster is markedly increased whereas another cluster is diminished in tumor-bearing mice. This reprogramming endures across tissue types including blood, tumor microenvironment, and the metastatic niche in the lungs. Thus, in all tissues tested, neutrophils derived from tumor-bearing mice are transcriptionally distinct from those derived from naïve mice. Among the genes that are markedly induced are members of the Ifitm family. We leverage this facet to selectively target reprogrammed neutrophils through treatment with Rapalink-1-an mTOR (mechanistic target of rapamycin) inhibitor-whose intake is greatly enhanced by IFITM proteins. Reprogrammed neutrophils selectively uptake Rapalink-1, which selectively eradicate them in vivo and normalized the neutrophil-to-lymphocyte ratio in tumor-bearing mice without affecting naïve neutrophils. Last, Rapalink-1 inhibited metastasis of a Rapalink-1-resistant tumor. Thus, Rapalink-1 not only could affect the primary tumor but also selectively eliminates tumor-reprogrammed neutrophils to diminish metastasis.

Microbiome and transcriptome analyses revealed that Fusobacterium nucleatum (F.n) in clinical samples is associated with immune suppression and poor prognosis in triple-negative breast cancer spinal metastasis. However, its preferential localization in hypoxic tumor regions limits the efficacy of conventional antimicrobial therapies, which poorly penetrate solid tumors and function suboptimally under anaerobic conditions. Developing strategies that enable deep tumor penetration, eliminate anaerobic bacteria, and induce immunogenic cell death remains a major challenge. In this study, a novel charge-enrichment and light-activated biomimetic nanosystem, designated as polyion liquid-bridged eosin Y (PIL-BEY), was developed. On one hand, interionic hydrogen bonding and dynamic electrostatic interactions within polyionic liquids reduce the surface energy of the nanoprobe and synergistically remodel the dense tumor stromal microenvironment via photodynamic therapy, thereby facilitating the deep intratumoral penetration and accumulation of PIL-BEY. On the other hand, the novel photosensitizer BEY generates reactive oxygen species via electron transfer under hypoxic conditions, thereby effectively eradicating bacteria within hypoxic tumor regions. The resulting pathogen-associated molecular patterns, together with damage-associated molecular patterns, activate dendritic cells, promote cytotoxic T lymphocyte infiltration, trigger immunogenic cell death, and induce systemic antitumor immune responses with durable immune memory. This oxygen-independent, dual-functional nanoplatform offers a promising strategy for treating invasive metastatic tumors.

Triple-negative breast cancer (TNBC) exhibits iron homeostasis that supports tumor growth and proliferation, yet the regulatory mechanisms controlling iron flux remain poorly defined. Here, we identify a ferritinophagic cargo receptor NCOA4 as a novel substrate of the lysine methyltransferase SUV39H2, uncovering a previously unrecognized mechanism that regulates ferritinophagy and ferroptosis. SUV39H2 directly binds and mono-methylates NCOA4 at lysine 356, a modification that reduces NCOA4 stability. Mechanistically, K356 methylation enhances NCOA4 interaction with the E3 ligase HERC2, promoting its ubiquitination and proteasomal degradation. This degradation increases FTH1 stability, suppresses ferritinophagic flux, limits iron release, maintains the high-risk iron homeostasis and ferroptosis resistance, ultimately promoting tumor proliferation and chemoresistance. Conversely, genetic or pharmacologic inhibition of SUV39H2 (OTS186935) inhibit NCOA4 methylation, stabilizes NCOA4 protein, enhances ferritinophagy, and triggers ferroptosis. Furthermore, SUV39H2 inhibition sensitizes TNBC cells to chemotherapy in vitro and in vivo, indicating OTS186935 treatment is a feasible therapeutic strategy. Collectively, the SUV39H2-NCOA4-HERC2 axis as a critical regulatory pathway in iron metabolism and ferroptosis, and highlight inhibition of NCOA4 K356 methylation as a promising therapeutic target in TNBC.The mechanistic scheme of SUV39H2 depletion to facilitate ferroptosis in TNBC. A SUV39H2 binds and methylates NCOA4, which enhanced the interaction between NCOA4 and the E3 ubiquitin ligase HERC2, leading to NCOA4 ubiquitination and proteasomal degradation. As a result, iron metabolism was reprogramed via enhancing ferritinophagy, leading to an increase in the level of iron, ultimately triggering ferroptosis. B Cell-state transitions induced by SUV39H2.

Guanine-rich nucleic acid sequences can fold into G-quadruplex (G4) structures that regulate DNA replication, transcription, and translation. Fanconi anemia group J helicase (FANCJ) resolves G4 structures at stalled replication forks. Despite its central role in genome maintenance, the molecular basis of G4 recognition and unwinding by FANCJ has remained unclear. Here, we report cryo-EM structures of human FANCJ bound to a G4-containing DNA substrate and ATPγS. The structures reveal direct engagement of the G4 by the Fe-S domain. Structure-guided mutagenesis demonstrates that this interface is essential for G4 binding and unwinding. The structures further capture open and closed conformational states linked to ATP hydrolysis, providing a mechanism for directional translocation along 5' ssDNA and progressive G4 unwinding. Together, these findings establish the structural basis of G4 recognition by FANCJ and provide mechanistic insights into how disease-associated mutations linked to Fanconi anemia and breast cancer impair helicase function.

Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors, such as ribociclib (RB), limits breast cancer therapy. Although PEG10 siRNA (siPEG10) can counteract this resistance, achieving targeted, synchronized co-delivery to reshape the immunosuppressive tumor microenvironment (TME) remains a formidable challenge. By loading RB and siPEG10 into PEG-β-CD-modified mesoporous silica nanoparticles (PMSNs) and coating them with breast cancer cell membranes (CM), the nanocomposite RB/siPEG10@PMSNs-CM was synthesized. The nanomaterials were characterized for their size, charge, drug loading, and pH-responsive release. Therapeutic efficacy, immunotherapy-related potentials, and underlying mechanisms were evaluated using in vitro functional assays, an anoikis resistance model, and in vivo breast cancer models. Toxicity was assessed in liver and kidney tissue. The nanodrug RB/siPEG10@PMSNs-CM (<200 nm) exhibited enhanced cellular uptake and pH-dependent drug release. The zeta potential of the RB/siPEG10@PMSNs-CM was negative. The nanodrug significantly inhibited tumor cell growth, migration, invasion, and metastasis, while inducing apoptosis. Crucially, the nanodrug overcame anoikis resistance, upregulated cleaved caspase-3 and cleaved PARP, reversed epithelial-mesenchymal transition (EMT) markers (increased E-cadherin and decreased Vimentin), and inhibited p-STAT3 and p-ERK signaling. Furthermore, the nanodrug exhibited lower toxicity to liver and kidney than RB. In Conclusion, the developed biomimetic nano-delivery system RB/siPEG10@PMSNs-CM successfully co-delivers RB and siPEG10 and exhibits a favorable safety profile. This platform exhibits anti-primary tumor and anti-metastatic efficacy and effectively overcomes anoikis resistance in breast cancer, which correlated with the suppression of the MEK/ERK and STAT3 signaling pathways, offering a promising combination strategy for advanced breast cancer immunotherapy and targeted treatment.

In this issue of Neuron, Yu et al. show that 6-hydroxydopamine (6-OHDA) suppresses tumor growth beyond its established sympathetic neurotoxic activity by inducing a macrophage-TH1 immune axis, uncovering an unexpected immunomodulatory role with potential therapeutic implications for breast cancer.

Spatial transcriptomics (ST) is a powerful assay for capturing gene expression in tissues. However, due to inherent limitations of spatial resolution, most existing ST datasets remain at a multicellular resolution, which hinders a comprehensive understanding of spatial organization. We propose SpatioCell, a computational algorithm to automatically extract both cell type and expression information at single-cell resolution from ST data, through a morpho-transcriptomic spatial reconstruction framework solved via dynamic programming, integrating morphological and transcriptomic information. This framework enables deterministic single-cell spatial reconstruction, assigning cell identities to precise locations rather than inferring the spot-level cell type composition and expression. It further uncovers overlooked microenvironmental features while correcting deconvolution errors. Using ST data from triple-negative breast cancer as an example, SpatioCell reveals the significance of the distance between cancer-associated fibroblasts (CAFs) and tumor or immune cells in terms of disease progression. Thus, the establishment of SpatioCell will broaden the biomedical applications of ST and facilitate investigations of single-cell spatial organization.

Doxorubicin (DOX) chemotherapy for breast cancer is constrained by systemic toxicity and limited tumor selectivity, underscoring the need for polysaccharide-based carriers in which pH responsiveness arises from intrinsic polymer properties rather than chemical modification. In this study, a carbohydrate-centered biomimetic nanocarrier was developed using chitosan as the primary structural and pH-responsive matrix, integrating zinc oxide nanoparticles (ZnO) and carbon quantum dots (CQDs) through a W1/O/W2 double-emulsion process, followed by surface association with bone marrow-derived mesenchymal stem cell (BM-MSC) exosomal membranes. The resulting CS/ZnO/CQDs@DOX nanocarriers exhibited an average hydrodynamic diameter of ∼180 nm, which increased to ∼205 nm after exosome association, accompanied by partial surface charge shielding. Physicochemical analyses supported a predominantly non-covalent assembly without evidence of alteration of the chitosan backbone, yielding a high encapsulation efficiency (88.75 ± 2.1%) and a drug loading of 6.8 ± 0.5 wt%. In vitro studies revealed pronounced pH-dependent doxorubicin release and enhanced cytotoxicity toward MCF-7 cells (IC₅₀ = 0.8 ± 0.1 μM), while maintaining minimal toxicity toward normal cells. Overall, these findings demonstrate that chitosan-driven pH responsiveness, complemented by inorganic components and biomimetic surface camouflaging, provides an effective and chemically conservative strategy for carbohydrate-based drug delivery design.

Brain metastasis in patients with breast cancer represents a terminal disease stage, with a median survival typically measured in months. Tumors that colonize the brain must adapt to its unique microenvironment, such as high acetate levels. Primary brain tumor cells enhance acetate conversion to acetyl-CoA through phosphorylation of acetyl-CoA synthetase 2 (ACSS2) by cyclin-dependent kinase 5 (CDK5), a process regulated by the nutrient sensor O-GlcNAc transferase (OGT). In this study, we showed that brain-metastatic breast cancer cells exhibited elevated O-GlcNAc, OGT, and phosphorylated ACSS2 (Ser267) compared with their parental counterparts. Both OGT and CDK5 were essential for in vivo tumor growth in the brain, and ACSS2 and a phosphomimetic S267D mutant drove progression of brain-metastatic breast cancer. Mechanistically, ACSS2 supported tumor cell survival by suppressing ferroptosis through early region 2-binding transcription factor (E2F1)-dependent transcription of the antiferroptotic protein solute carrier family 7 member 11 (SLC7A11). Treatment with brain-penetrant ACSS2 inhibitor AD-5584 induced ferroptosis and significantly suppressed breast cancer brain metastatic growth ex vivo and in vivo. Together, these findings identify ACSS2 as a key metabolic regulator of brain-metastatic breast cancer survival and a promising target for ferroptosis-inducing therapies. Targeting ACSS2 with brain penetrant inhibitors induces ferroptosis in brain metastatic breast cancer by perturbing an E2F1/SLC7A11 axis, providing an efficacious strategy for treating brain metastatic tumors.

Breast cancer remains the second leading cause of cancer-related mortality among women, with triple-negative breast cancer (TNBC) exhibiting a particularly poor 5-year prognosis. In this study, we have demonstrated that among genetic and pharmacologic perturbations targeting DNA replication, suppression of the DNA polymerase epsilon catalytic subunit (POLE) induced a potent, TNBC-specific gene expression signature enriched in inflammatory cytokines that are transcriptional targets of NF-κB. TNBC cells exhibited markedly higher levels of DNA damage and canonical NF-κB activation compared with luminal breast cancer cells. Notably, NF-κB activation in this context depended on the canonical component RELA but not on the noncanonical component RELB. Mechanistically, ataxia-telangiectasia mutated, stimulator of IFN genes, and retinoic acid-inducible gene I each contributed to NF-κB activation following POLE suppression. POLE suppression in an in vivo murine TNBC model led to cancer cell-intrinsic elimination of tumor burden and increased immune cell infiltration. Together, these findings support a model in which replication stress from POLE inhibition triggers robust NF-κB-mediated inflammation and immune microenvironment remodeling in TNBC and can independently trigger tumor eradication. These results suggest a potential therapeutic avenue for targeting POLE in TNBC. DNA replication protein POLE can be selectively targeted in triple-negative breast cancer to activate signaling pathways that increase expression of inflammatory genes dependent on canonical NF-κB signaling and to eradicate tumors.

Rapid assessment of chemotherapeutic response is essential for precision oncology but remains hindered by tumor heterogeneity and complex biological matrices. Here, we develop MetaRing, a programmable coffee-ring-derived plasmonic biosensor fabricated through dual regulation of nanoparticle concentration and evaporation temperature. This strategy enables deterministic nanoassembly, generating hierarchical structures with dense and stable nanogaps and conferring exceptional matrix robustness in water, PBS, protein-rich buffers, and complex cell lysates. MetaRing enables rapid, label-free surface-enhanced Raman spectroscopy (SERS) profiling of paclitaxel (PTX) response using minimal biological material. Distinct PTX-sensitivity fingerprints are consistently identified across drug-resistant breast cancer cell lines, xenograft tumors, and patient-derived biopsy tissues. Metabolomic analysis reveals that these spectral signatures originate from metabolic reprogramming involving arginine and methionine-cysteine pathways, providing mechanistic insight into chemoresistance. Integration with a lightweight one-dimensional convolutional neural network enables accurate classification of PTX sensitivity within 10 min without labeling or culture expansion, achieving >92% accuracy in clinical cohorts. Collectively, MetaRing establishes a robust and scalable plasmonic platform for rapid phenotypic drug response profiling with strong translational potential.

The sympathetic nervous system has emerged as a critical regulator of cancer progression, yet the underlying mechanisms remain unclear. Here, we compare genetic, pharmacological (6-hydroxydopamine [6-OHDA]), and surgical denervation in mouse breast cancer models. While all methods deplete sympathetic nerves, only 6-OHDA suppresses tumor growth, revealing a disconnect between sympathetic ablation and antitumor effect. Mechanistic investigations reveal that 6-OHDA suppresses tumor growth through immune activation rather than sympathetic ablation. 6-OHDA induces cancer cell interferon (IFN)-β production, which promotes monocyte differentiation into pro-inflammatory macrophages characterized by interferon-stimulated gene (ISG) expression. These ISG+ macrophages are essential for the expansion of type 1 T helper (TH1) cells, which mediate prolonged tumor suppression. By contrast, sympathetic ablation alone does not affect macrophage differentiation or tumor growth. Our findings uncover an immunomodulatory function of 6-OHDA beyond its established neurotoxic activity and suggest the therapeutic potential of harnessing the macrophage-TH1 axis for breast cancer.

The spontaneous leakage and constitutive activity of the conventional fluorescent dye-labeled entropy-driven catalytic (EDC) circuit severely compromise the accuracy of low-abundance intracellular microRNAs (miRNAs) analysis. Herein, a Hairpin fuel-Driven Fluorogenic DNA Aptamer (HDFA) amplifier has been engineered for high-contrast imaging of miRNA-21 (miR-21) in living cells by integrating a hairpin fuel strand and a fluorogenic DNA aptamer into the EDC circuit. The pre-locked fuel strand effectively suppresses spurious strand displacement reactions, whereas target-induced unlocking restores the fuel strand activity and initiates the EDC circuit amplification cascade. Concurrently, the fluorogenic DNA aptamer-mediated activation of fluorescent dyes ensures an ultralow background signal, thereby significantly enhancing the signal-to-noise ratio and imaging contrast. The exceptional sensitivity, specificity and stability of the HDFA amplifier enable the detection of miR-21 expression differences in living cells, yielding images with low background and high visual contrast. Furthermore, the HDFA amplifier has been applied to monitor dynamic changes in miR-21 expression levels in MCF-7 cells upon treatment with various anticancer drugs, demonstrating its utility in drug screening applications. Therefore, the proposed HDFA amplifier provides a robust and versatile platform for high-contrast imaging of low-abundance miRNAs and cancer-related drug screening.

Pre-metastatic niche formation in the lung creates a permissive microenvironment for breast cancer metastasis, characterized by metabolic reprogramming of resident cells, recruitment of suppressive neutrophils, and vascular remodeling. However, the role of lipids in regulating neutrophil-endothelial interactions, particularly in facilitating tumor cell extravasation, remains largely undefined. Here, we showed that triple-negative breast cancer established a palmitic acid-enriched lung microenvironment that drove tumor cell extravasation and colonization. Pulmonary endothelial cells were a major source of palmitic acid, which activated neutrophils to produce lipocalin-2 (LCN2) via the Toll-like receptor 4 (TLR4)-NF-κB pathway. Neutrophil-derived LCN2 disrupted endothelial tight junctions, compromised vascular integrity, and facilitated tumor cell extravasation. Targeting endothelial fatty acid synthesis using glucagon-like peptide-1 receptor agonists preserved vascular integrity and suppressed lung metastasis. These findings uncover a lipid-driven mechanism underlying metastatic organotropism and highlight metabolic intervention as a potential therapeutic strategy against breast cancer lung metastasis.

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

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal malignancies worldwide, and accurate prognostic prediction remains highly challenging due to its marked biological heterogeneity and complex tumor microenvironment. To address this challenge, a histopathomics-based survival prediction system (HPSurv) was developed using histopathological whole-slide images (WSIs) for individualized overall survival (OS) prediction. Within this framework, pathological tissue classification, quantitative characterization of tumor spatial heterogeneity, and a survival Transformer were integrated to enable multi-level representation learning from histopathological data. The system was developed and evaluated in 1020 patients across five independent cohorts. Compared with conventional clinicopathological indicators, significantly improved prognostic performance was achieved across multicenter cohorts (p < 0.05), with a mean C-index of 0.761 and time-dependent AUCs of 0.936, 0.877, and 0.772 for predicting 6-month, 2-year, and 3-year survival, respectively. Subgroup analyses further supported its role as an independent prognostic factor and suggested its potential utility in stratifying patients with respect to ACT-related outcomes. In addition, significant associations with key PDAC molecular pathways were observed, providing biological insights into the model predictions and supporting interpretability. In the study, an interpretable and high-performing artificial intelligence (AI) framework for quantitative modeling of PDAC was established. Objective characterization of tumor heterogeneity and accurate postoperative survival prediction are enabled, with potential value for personalized management in PDAC.

Nanoliposomal irinotecan (nal-IRI) has become an established therapy for advanced pancreatic ductal adenocarcinoma (PDAC), both in the post-gemcitabine setting and, more recently, in the first-line NALIRIFOX regimen. Concerns remain regarding cross-resistance in patients previously exposed to conventional irinotecan. We performed a systematic review and meta-analysis to evaluate the impact of prior irinotecan exposure on outcomes with nal-IRI. We conducted a systematic literature search of PubMed, Embase, and the Cochrane Library through July 2025 following Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. Eligible studies included patients with advanced PDAC treated with nal-IRI-containing regimens and reported outcomes stratified by prior irinotecan exposure. Data extraction and risk of bias assessments were performed independently by two reviewers. Hazard ratios (HRs) for overall survival (OS) and progression-free survival (PFS) were pooled using random-effects models. Sensitivity and publication bias analyses were conducted. Thirteen retrospective studies comprising 2271 patients were included. One-third (33.1%) had received conventional irinotecan before nal-IRI. Prior irinotecan exposure was associated with inferior outcomes: pooled unadjusted and adjusted HRs for OS were 1.48 [95% confidence interval (CI) 1.24-1.76, P = 0.001] and 1.42 (95% CI 1.03-1.96, P = 0.030), respectively. For PFS, pooled unadjusted and adjusted HRs were 1.45 (95% CI 1.28-1.64, P < 0.001) and 1.72 (95% CI 1.16-2.54, P = 0.007), respectively. Patient-level subgroup analyses indicated that patients discontinuing prior irinotecan due to progression had significantly worse OS compared with those discontinuing for other reasons (HR 1.97, 95% CI 1.10-3.52). Most studies were high quality, and sensitivity analyses confirmed the robustness of the findings. Prior exposure to conventional irinotecan, particularly discontinuation for progression, reduced clinical benefit from subsequent nal-IRI therapy in advanced PDAC in observational cohorts. These findings support caution when considering nal-IRI soon after irinotecan progression and highlight the need for prospective sequencing studies.

Pancreatic ductal adenocarcinoma (PDAC) arises from precursor lesions over a decade-plus, offering a window for interception in high-risk individuals, but current surveillance detects a minority of precursors. Mutant KRAS (mKRAS) is present in most PDACs and their precursors, making it an appealing target for immune-based interception. We conducted a phase I, first-in-human study of a peptide vaccine targeting six common KRAS mutations (mKRAS-VAX) in 20 individuals with hereditary PDAC predisposition and a radiographic pancreatic abnormality (NCT05013216) to assess safety, immunogenicity, and T cell persistence. Adverse events were grade 1-2. Vaccination elicited a significant mKRAS-specific T cell response in 18/20 participants (90%). Longitudinal TCR sequencing demonstrated persistence of vaccine-induced mKRAS-specific clonotypes for up to 2 years. Over a median follow-up of 16.5 months, no participants developed PDAC. These findings demonstrate that mKRAS-VAX is safe and generates durable T cell responses, which support the advancement of mKRAS-targeted vaccination for PDAC interception.

Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal malignancy due to its aggressive biology and therapeutic resistance. Lysine-specific demethylase 1 (LSD1), an epigenetic regulator, is overexpressed in PDAC and linked to poor prognosis, yet its context-dependent roles in metabolic subtypes and chemoresistance remain undefined. Here, we show that LSD1 knockdown has opposing, subtype-specific effects on chemotherapeutic responses: it sensitized RSK-subtype cells (L3.6pl, PANC-1) to chemotherapy but induced resistance in KRAS-subtype cells (BxPC-3, TBO368). Integrated analyses revealed mitochondrial dysfunction and defective mitophagy as hallmarks distinguishing KRAS- from RSK-subtype PDAC. Critically, mitochondrial targeting through respiratory modulation or mitophagy manipulation overrides LSD1-mediated subtype-specific chemoresistance, establishing mitochondrial fitness as the mechanistic determinant. Mechanistically, LSD1 transcriptionally regulates GLS2 to drive glutamine metabolic reprogramming, promoting reductive carboxylation in KRAS-subtype cells and oxidative metabolism in RSK-subtype cells. Our work establishes the LSD1-GLS2 axis as a metabolic switch controlling PDAC chemosensitivity and provides a framework for subtype-specific therapeutic strategies.

Chemoresistance in pancreatic ductal adenocarcinoma (PDAC) is partly driven by pathological stromal remodeling, yet the underlying mechanisms remain poorly understood. Here, we show that gemcitabine treatment induces tumor cell senescence and activates cancer-associated fibroblasts via the senescence-associated secretory phenotype, leading to progressive fibrotic matrix stiffening. This biomechanical reprogramming engages the mechanosensitive ion channel Piezo1, triggering metabolic rewiring that renders BRG1-positive tumor cells increasingly dependent on NRF2-mediated antioxidant defenses. Piezo1 signaling promotes NRF2 nuclear translocation and its chromatin-remodeling cooperation with BRG1, thereby upregulating SLC7A11-dependent antioxidant programs and suppressing ferroptosis. Notably, the combination of the senolytic agent ABT-263 with the ferroptosis inducer Erastin effectively dismantles BRG1-NRF2-driven gemcitabine resistance, alleviates stromal fibrosis, enhances T-cell infiltration, and suppresses tumor growth in vivo. This senolytic-ferroptosis approach exploits metabolic vulnerabilities in chemotherapy-aged PDAC and provides a mechanistic rationale for stroma-targeted combination therapies.

Pancreatic cancer (PC) remains a highly lethal malignancy presenting formidable therapeutic challenges, primarily attributable to the substantial barriers posed by its dense desmoplastic stroma, profoundly immunosuppressive microenvironment, and poor drug bioavailability. Nanotechnology offers an effective strategy to enhance therapeutic efficacy through improved targeting, optimized pharmacokinetics, enhanced tissue penetration, increased biosafety, and high clinical translational potential. Consequently, this nanotechnology-based platform has been extensively investigated over the past decade for treating PC. This review examines a comprehensive overview of nanoparticle-mediated delivery systems for targeted PC therapy, critically evaluating recent developments and ongoing hurdles along the bench-to-bedside pathway. We discuss applications across chemotherapy, immunotherapy, gene therapy, and other modalities employed as monotherapies or combination regimens.

Pancreatic ductal adenocarcinoma (PDA) is an aggressive cancer that frequently presents with disseminated disease. The PDA metastatic microenvironment imposes distinct metabolic stressors, potentially generating context-dependent vulnerabilities. Therefore, we employed CRISPR-based genetic screening in a model of PDA liver metastasis to identify novel and possibly targetable liabilities. Remarkably, ferritin heavy chain (FTH1) emerged as the most prominent liver-specific dependency - loss of FTH1 suppressed tumor growth specifically in the liver microenvironment. FTH1 deletion and subsequent disruption of iron handling triggers mitochondrial dysfunction and ionic imbalance, including cytosolic calcium overload. These perturbations result in the activation of a transcriptional program that triggers anti-tumor immunity mediated by immunostimulatory cytokine IL36G. Mechanistically, FTH1 deletion and subsequent ionic imbalance causes decreased protein levels of the tumor suppressor Stk11 (LKB1) which we propose to be mediated by an RNA G-quadruplex located in the 5'-UTR of LKB1. The loss of LKB1 protein levels alters signaling cascades resulting in reduced SIK signaling and inhibition of nonsense mediated decay, ultimately leading to Il36g mRNA stabilization. Taken together, this work elucidates novel ionic disruptions that regulate the translation of LKB1 through a previously undescribed quadruplex in the 5'UTR, altering signaling axes that can be targeted to generate an anti-tumor immune response in PDA.

Pancreatic ductal adenocarcinoma (PDAC) is largely refractory to immune checkpoint blockade, owing to its immunosuppressive tumour microenvironment. Neutrophil extracellular traps (NETs) accumulate in PDAC and correlate with disease progression, yet whether NETs reprogram cancer-associated fibroblast (CAF) heterogeneity and the upstream tumour-intrinsic signals that sustain pathological NETosis remain undefined. To delineate how NETs instruct CAF subtype specification and immunosuppression in PDAC, identify upstream NETosis drivers and evaluate combinatorial therapeutic strategies targeting this axis. Quantitative spatial analysis of human PDAC specimens, NETs-pancreatic stellate cell and patient-derived CAF cocultures, biotinylated DNA pull-down with liquid chromatography-tandem mass spectrometry, live-cell integrin-blocking assays, Cleavage Under Targets and Release Using Nuclease (CUT&RUN) sequencing, orthotopic and hepatic colonisation models, CD8+ T-cell functional assays and single-cell RNA sequencing of an eight-arm therapeutic study were employed. NETs-DNA directly engaged integrin α5β1 (ITGA5) on fibroblasts via its N-terminal domain, initiating a FAK (focal adhesion kinase)/SRC (Src family tyrosine kinase)-YAP (Yes-associated protein)-IL-6 (interleukin-6)-JAK (Janus kinase)/STAT3 (signal transducer and activator of transcription 3) autocrine cascade specifying inflammatory CAF (iCAF) differentiation. Tumour-derived neutrophil gelatinase-associated lipocalin (NGAL), induced by IL-17, drove extracellular signal-regulated kinase (ERK)-reactive oxygen species-mediated NETosis. NETs-reprogrammed iCAFs accelerated tumour growth, hepatic colonisation and CD8+ T-cell exhaustion, with exhausted T cells spatially enriched in iCAF-rich regions. A composite NETs/ITGA5 signature stratified overall survival in patients with PDAC. Single-cell transcriptomics demonstrated that triple therapy (anti-IL-17+AV3+anti-PD-1) shifted fibroblasts from iCAF towards myofibroblastic CAF dominance, restored CD8+ T-cell effector programmes and achieved the greatest tumour suppression and survival benefit. We define an IL-17/NGAL/NETs/ITGA5 axis linking neutrophil-derived extracellular DNA to iCAF specification and immunosuppression in PDAC. Cotargeting IL-17 and ITGA5 synergises with PD-1 blockade, providing a rationale for combinatorial immunotherapy.

For decades, we have viewed pancreatic fibrosis as a passive scar-the end-stage wreckage of chronic inflammation or a late accomplice of growing tumors. A recent study by He Ren and colleagues sheds critical light on the origins of the phenomenon tracing it to an unexpected cellular culprit: a rare subset of pancreatic epithelial cells that express FOXP3-a transcription factor predominantly associated with regulatory T cells. These epithelial FOXP3 (E-FOXP3) positive cells do not need oncogenic Kras or overt inflammation to ignite fibrosis. On their own, they orchestrate a glycosylation-dependent IL-6 switch that turns quiescent pancreatic stellate cells into a self-amplifying fibrotic machine. In discovering a proto-fibrogenic "trigger cell" that acts silently, long before any symptom or radiological sign, the work redefines the cellular hierarchy of pancreatic fibrocarcinogenesis. Further, the paper identifies a post-translational "sugar code" as a therapeutic foothold, and opens a long-sought preemptive window for intercepting pancreatic cancer at its most curable stage-before the desmoplastic fortress is ever built.

The biological significance of forkhead box A1 (FOXA1) in non-steroid-driven malignancies, such as pancreatic ductal adenocarcinoma (PDAC), has garnered increasing recognition. It assumes a pivotal role in regulating critical processes such as PDAC cell lineage, metabolism, and metastasis. However, its regulatory mechanisms remain elusive. Here, we demonstrate that histone deacetylase 5 (HDAC5) mediates the deacetylation of FOXA1 at lysine residue 270 (K270), leading to repression of FOXA1's global chromatin occupancy. In HDAC5-loss PDAC, K270 hyper-acetylated FOXA1 is reprogrammed to the transcription start sites (TSSs) of HIF1α-targeted genes, functioning as a pioneer factor of HIF1α signaling. Additionally, we show that HDAC5 antagonizes LSD1-mediated FOXA1 activation by converging on the dynamic equilibrium of acetylation-methylation transition at K270. Pharmacological inhibition of HIF1α/LSD1 suppresses the growth and progression of HDAC5-deficient PDAC in in vivo and in vitro models. Our study reveals the role of FOXA1 as a pioneer factor of HIF1α in PDAC, providing potential therapeutic strategies for HDAC5-deficient PDAC.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal cancers due to its aggressive biology and resistance to existing therapies. Oncofetal chondroitin sulfate (ofCS) is a tumour-restricted glycosaminoglycan broadly expressed across solid cancers but largely absent from normal adult tissues. We developed C9-based chimeric antigen receptor (CAR)-T cells targeting ofCS to overcome poor antigen specificity and the immunosuppressive tumour microenvironment (TME). To optimise ofCS-targeted C9 CAR-T cell therapy for PDAC through integrated CAR design optimisation, metabolic enhancement and TME reprogramming. C9 and charge-optimised C9-66 CAR-T cells were engineered using a humanised ofCS-binding single-chain antibody fragment. Antitumour efficacy, functional durability and metabolic fitness were assessed in murine and patient-derived PDAC models. Enhancement strategies included inosine-mediated metabolic reprogramming, Nr5a2 overexpression and sequential TME remodelling using GLP-1R modulation, CSF-1R blockade and programmed cell death protein-1 inhibition. C9 CAR-T cells exhibited potent cytotoxicity, delayed tumour progression and extended survival in PDAC models. Compared with the parental construct, charge-optimised C9-66 CAR-T cells showed reduced exhaustion and more sustained activity in vivo. Inosine enhanced cytokine production, promoted central-memory differentiation and mitigated exhaustion, whereas Nr5a2 overexpression increased mitochondrial respiration and cytotoxicity. Sequential GLP-1R on-off modulation with macrophage and checkpoint blockade enhanced intratumoural CAR-T cell activity and prolonged survival. Human C9-66 CAR-T cells retained specific ofCS recognition and lysed patient-derived PDAC cells in vitro and in vivo. C9-66 CAR-T cells with metabolic optimisation and TME reprogramming represent a tumour-specific and clinically translatable immunotherapeutic strategy for PDAC and other ofCS-expressing solid tumours.

Gemcitabine remains a cornerstone treatment for pancreatic ductal adenocarcinoma (PDAC), yet the emergence of resistance constitutes a major clinical challenge with poorly understood epigenomic mechanisms. Here, we identified the pioneer transcription factor Foxa1 as a master regulator of gemcitabine resistance through multi-omics analysis. Mechanistically, Foxa1 drives widespread super-enhancer (SE) reprogramming and 3D genome remodelling in resistant cells, which coordinately activates the expression of key resistance genes, notably Rrm1 and Cdadc1. This is accompanied by increased chromatin accessibility, elevated H3K27ac enrichment at SEs, and enhanced Foxa1 binding at regulatory elements. Moreover, post-translational stabilization of Foxa1 via USP7-mediated deubiquitination sustains this epigenomic program. Genetic ablation of Foxa1 or specific SE regions near Rrm1 resensitizes resistant cells to gemcitabine. Building upon this mechanism, we demonstrate that bromodomain and extraterminal (BET) inhibitors, which disrupt SE function, potently reverse resistance. Notably, the clinical-stage BET inhibitor AZD5153, in combination with gemcitabine, achieves robust tumor suppression and overcomes resistance in cell-derived xenograft (CDX) models by dismantling the Foxa1-mediated resistant transcriptome and reinvigorating drug sensitivity. Our findings establish Foxa1-orchestrated enhancer reprogramming as a fundamental mechanism of gemcitabine resistance and unveil a promising epigenetic therapy to restore treatment efficacy in PDAC.

Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer death in the United States, driven by its aggressive biology and high metastatic incidence at diagnosis. With a 5-year survival rate of just 8%, PDAC remains one of the most lethal cancers. Mutant KRAS, present in more than 90% of cases, serves as a key driver of tumorigenesis and metabolic reprogramming. In this issue of Cancer Research, Thakur and colleagues uncover a novel metabolic adaptation that PDAC cells use to survive therapeutic stress. Their integrated metabolomic and lipidomic analyses show that ERK inhibition-targeting a key KRAS pathway effector-not only disrupts glycolysis and glutamine metabolism but also triggers a compensatory increase in fatty acid oxidation (FAO). This shift occurs through lipophagy, a lysosome-mediated lipid degradation process, rather than cytosolic lipolysis. Mechanistically, ERK inhibition promotes the nuclear translocation of TFEB, which drives the upregulation of FAO and lipophagy genes. This metabolic reprogramming enables PDAC cells to survive KRAS pathway blockade. Importantly, cotargeting FAO alongside ERK or KRAS inhibitors elicits a potent synergistic antitumor effect in vivo. This dual-target strategy holds promise for overcoming PDAC resistance to KRAS-targeted therapies, laying the groundwork for novel combination treatments. See related article by Thakur et al., p. 3519.

The tumor microenvironment (TME) actively contributes to pancreatic ductal adenocarcinoma (PDAC) pathogenesis through dynamic bidirectional tumor-stroma interactions. In this study, we demonstrated that ATM-deficient tumor epithelium reprograms the TME in a genotype-specific manner to enhance cancer aggressiveness. In genetically engineered mouse models, pancreatic stellate cell and cancer-associated fibroblast (CAF) coculture systems, single-nucleus multiomics, and human PDAC models, tumoral loss of ATM serine/threonine kinase drove CAFs toward αSMA+ myofibroblastic (myCAF) differentiation, independently of p53 status. The myCAFs, in turn, promoted cancer aggressiveness and chemoresistance. Mechanistically, ATM deficiency increased reactive oxygen species and contractility signaling, enhancing TGFβ1 secretion. Pharmacologic TGFβ inhibition reversed myCAF differentiation, sensitized tumors to chemotherapy, and impaired tumor progression in both murine and human ATM-null models. These findings reveal that ATM-deficient tumors shape a cancer-promoting niche via TGFβ signaling and identify dual targeting of intrinsic and extrinsic vulnerabilities as a promising precision oncology strategy. TGF-β-driven myofibroblastic stromal differentiation in ATM-deficient pancreatic cancer generates a genotype-specific tumor microenvironment, providing a targetable axis and highlighting the need to integrate epithelial genotype and stromal context in pancreatic cancer therapy.

Pancreatic ductal adenocarcinoma (PDAC) carries an extremely poor prognosis, in part resulting from cellular heterogeneity that supports overall tumorigenicity. Cancer-associated fibroblasts (CAF) are key determinants of PDAC biology and response to systemic therapy, and multiple CAF subtypes have been defined. However, defining the effects of patient-specific CAF heterogeneity and plasticity on tumor cell behavior is required to better characterize the role of CAFs in PDAC. In this study, we used multiomic analyses to characterize the tumor microenvironment (TME) in tumors from patients undergoing curative-intent surgery for PDAC. In these same patients, matched tumor organoid and CAF lines were established to functionally validate the impact of CAFs on the tumor cells. CAFs promoted epithelial-mesenchymal transition and a switch in tumor cell classification from classical to basal subtype. Furthermore, CAF-specific interleukin 8 functioned as a modulator of tumor cell subtype. Finally, neighborhood relationships between tumor cells and T cell subsets were defined, demonstrating a distinct spatial coordination among CAF and tumor cell subtypes. Overall, this study provides data supporting CAF signaling as a regulator of the cellular and behavioral heterogeneity in the PDAC TME. These findings can be used to explore rational approaches to improve therapies for this difficult-to-treat disease. Multidimensional analyses highlight the diverse role of cancer-associated fibroblasts in influencing cells in the tumor microenvironment and provide a platform for evaluating emerging therapeutic approaches and studying mechanisms dictating tumor behavior.

Pancreatic ductal adenocarcinoma (PDAC) is characterized by frequent KRAS mutations, which activate the MAPK pathway to promote PDAC progression. In this study, we explored metabolic vulnerabilities of PDAC by assessing initial metabolic reprogramming upon ERK inhibition using metabolomics, lipidomics, and isotope-tracing experiments. ERK inhibition enhanced lipid turnover and fatty acid (FA) oxidation while inhibiting glycolysis, glucose oxidation, and glutamine metabolism in PDAC cells. Moreover, lipophagy, but not cytosolic lipolysis, was responsible for the increased lipid turnover and FA oxidation upon ERK inhibition. Lipophagy and lipophagy-fueled FA oxidation were induced by increased nuclear translocation and activity of the transcription factor TFEB. Pharmacologic inhibition of FA oxidation in combination with KRASG12D/MEK/ERK inhibitors synergistically decreased the growth of PDAC cell lines and organoids. The combination decreased tumor burden and improved survival in orthotopic cell line and patient-derived xenograft PDAC models. Overall, this study provides mechanistic insights into the development of metabolic resistance to KRAS signaling inhibition and demonstrates that FA oxidation is a metabolic vulnerability following KRAS signaling inhibition that can be utilized as an effective therapeutic target to treat PDAC. Treating pancreatic cancer with inhibitors that target the KRAS pathway rewires metabolism by increasing lipophagy and fatty acid oxidation, which can be targeted to sensitize tumors to KRAS signaling inhibition. See related commentary by Delgado Herrera and Ferrer, p. 3374.

The output signal of the sensor is closely correlated with its detection sensitivity, while the selection of luminescent materials and the presence of interfering substances in the detection medium can both affect the output signal. In this study, a bifunctional peptide (HWRGWVDEDKDKDKC) with antibody targeted fixation and interface anti-adsorption properties was designed by integrating recognition (HWRGWV), charge neutralization (DE), antifouling (DKDKDKDK) and anchoring (C) fragments, which endowed the constructed biosensing interface with high target capture efficiency and good antifouling ability. At the same time, AuAgTb-MOF with multiple synergistic amplification effects was developed as luminophore. Concretely, the antenna effect of 3,5-dicarboxyphenylboronic acid to Tb3+, the catalysis of Ag NPs for S2O82- and the conductivity of Au NPs significantly prompted the luminescence of AuAgTb-MOF. The design of intramolecular self-enhanced AuAgTb-MOF not only reduced energy transfer losses, but also simplified the construction of the biosensing platform. On this basic, the constructed biosensor had a wide detection range of 10-4 to 103 U/mL and a low limit of detection of 3.38 × 10-5 U/mL (IUPAC standard), realizing the sensitive and precise detection for pancreatic cancer markers, thus providing a positive reference for the clinical diagnosis of pancreatic cancer.

Cancer-associated fibroblasts (CAFs) are principal determinants of pancreatic ductal adenocarcinoma (PDAC) progression. CAFs can shape tumor behavior via multiple pathways, underscoring the need for a complete understanding of the regulatory mechanisms that govern CAF function. Here, we identified N6-methyladenosine (m6A) remodeling as a hallmark of CAF activation and defined a critical role for the m6A demethylase ALKBH5 in PDAC metastasis. Activated CAFs exhibited a global reduction in m6A abundance, with ALKBH5 emerging as a key regulator of the CAF epitranscriptome. Functionally, CAF-derived ALKBH5 enhanced pancreatic cancer cell migration and invasion in vitro and promoted epithelial-mesenchymal transition-associated gene expression in tumor cells in an m6A-dependent manner. Orthotopic co-implantation models and host genetic ablation models demonstrated that ALKBH5 plays a critical role in metastatic dissemination, with minimal impact on primary tumor growth. Mechanistically, ALKBH5 enhanced the m6A-dependent translation of HSF1 in CAFs, at least in part by relieving IGF2BP3-associated translational constraints. Elevated HSF1 subsequently activated LIF transcription through distal enhancer elements, establishing an ALKBH5-HSF1-LIF signaling axis that mediated the pro-metastatic CAF-tumor cell communication. Clinically, enrichment of ALKBH5⁺HSF1⁺ CAFs independently predicted poor prognosis and was preferentially observed in metastatic PDAC. Collectively, these findings uncover a CAF-intrinsic epitranscriptomic program that drives PDAC metastasis and highlight stromal m6A regulation as a potential therapeutic vulnerability.

Multi-task DL for predicting EGFR mutation status Epidermal growth factor receptor (EGFR) mutation status is a critical biomarker in the management of non-small cell lung cancer (NSCLC), playing an essential role in selecting patients for EGFR-targeted treatment. With advancements in deep learning (DL), there is a growing interest in developing non-invasive methods for predicting EGFR mutation status. In this study, we present a multi-task deep learning (MTDL) model that utilizes CT images to predict EGFR mutation status (ChiCTR2400083082 in the WHO International Clinical Trials Registry). Our MTDL model achieved promising performance in accurately predicting EGFR mutation status. Additionally, the MTDL score was significantly associated with survival in patients receiving EGFR-targeted treatment, as well as relevant gene expression patterns and tumor microenvironment. These findings suggest that our method has the potential to serve as an accurate and non-invasive biomarker for predicting EGFR mutation status, thereby facilitating personalized treatment decisions for NSCLC patients.

Genetic testing is standard of care in Non-small Cell Lung Cancer(NSCLC). Few studies evaluate concurrent Endobronchial Ultrasound-Transbronchial Needle Aspiration(EBUS-TBNA) and liquid biopsy for Next Generation Sequencing(NGS). Compare diagnostic yield differences for detecting clinically relevant mutations via NGS in concurrent EBUS-TBNA and liquid biopsies. Prospective observational cohort study of NSCLC patients undergoing concurrent biopsies. Yield differences, receiver operating characteristic (ROC) and kappa statistics were calculated for clinically relevant mutations. Of 199 subjects, diagnostic yield for EGFR was 9.5% in EBUS-TBNA and 7.0% in blood with yield difference 2.5%(95% CI: 3.4%, 8.5%, p = 0.36). For clinically relevant targets, there were 79(39.7%) from EBUS-TBNA and 49(29.6%) from blood with yield difference 15.1%(95% CI: 5.5%, 24.2%, p = 0.001). ROC statistics for detection of clinically relevant mutations (where tissue results were the gold standard) included: sensitivity 0.53 (95% CI: 0.42, 0.64), specificity: 0.95(95% CI: 0.90,0.98), positive predictive value: 0.89 (95% CI: 0.77, 0.96), negative predictive value: 0.74 (95% CI: 0.65, 0.81), and kappa statistic: 0.51 (95% CI: 0.39, 0.63). There were 37 (18.6%) participants with mutations in tissue only and 7(3.5%) with mutations in blood only, while 42(21.1%) had mutations reported from both sources. EBUS-TBNA was more likely to detect mutations than blood. However, some patients had mutations in blood only, suggesting blood testing is complementary. Strategies are needed to improve NGS turnaround time(TAT) and QNS definition.

How cancer-associated fibroblasts (CAFs) dictate CD8+ T cell dysfunction during lung cancer remains unclear. Through single-cell analysis of 84 human NSCLC samples, we identified insulin-like growth factor 2 (IGF2) as a key immunosuppressive factor in a specific CAF subset. IGF2+ CAF infiltration correlated with CD8+ T cell dysfunction. Fibroblast-specific IGF2 knockout enhanced antigen presentation cell-autonomously via major histocompatibility complex class I (MHC-I), boosting CD8+ T cell effector function, tumor suppression, survival, and PD-1 blockade synergy. Mechanistically, IGF2 sustained MYC signaling to up-regulate DNA methyltransferase 1 (DNMT1), which methylated the STAT1 promoter, epigenetically silencing STAT1. This impaired MHC-I presentation and CD8+ T cell activation. Clinically, high IGF2 levels associated with reduced CD8+ T cell cytotoxicity and poor outcomes. An IGF2+ fibroblast signature predicted worse response to immunotherapy in multiple cohorts. Our findings establish IGF2 as a central regulator of CAF-mediated immunosuppression and a stromal target for enhancing immunotherapy in lung cancer.

Optimal cancer prognostication combines multimodal data including biopsy results, CT scan, patient characteristics, and clinical trajectory thus far. However, many patients do not have all modalities of data available, so requiring physicians to have all patient data modalities to use an AI prediction algorithm limits the tool’s clinical utility. To increase the clinical potential of these AI algorithms, Ruffini et al. developed a “missing data aware” survival prediction approach that is able to handle inputs with missing data for risk stratification in non-small cell lung cancer patients.

Perioperative immunotherapy has improved clinical outcomes for patients with early-stage non-small cell lung cancer (NSCLC). The influence of immune checkpoint inhibitor in combination with chemotherapy on surgical outcomes remains to be explored. To evaluate perioperative toripalimab in combination with chemotherapy on surgical outcomes. This multicenter, double-blind, placebo-controlled phase 3 randomized clinical trial (Neotorch study) enrolled patients with resectable stage III NSCLC and took place at 50 centers in China. Patients who had histologically confirmed resectable stage IIIA or IIIB NSCLC were eligible. These data were analyzed from July 2024 to March 2026. Patients were randomized (1:1) to receive toripalimab (240 mg) plus platinum-based chemotherapy or placebo plus platinum-based chemotherapy for 3 cycles before surgery and 1 cycle after surgery, followed by maintenance with toripalimab or placebo alone for 13 cycles. Surgical outcomes, including perioperative complications, tumor downstaging, and lymph node downstaging, and their association with event-free survival (EFS), were studied in this post hoc analysis. Among 404 patients enrolled, 314 patients (median [SD] age 60 [6.82] years; 90% of patients were male and 10% were female) underwent surgery (166 in toripalimab group and 148 in placebo group). Percentage of patients canceling surgery (17.8% vs 26.7%; P = .03) was significantly lower in the toripalimab group. Proportions of minimally invasive surgery, R0 resection, and lobectomy were slightly higher in the toripalimab group. Surgical complications were similar between the 2 groups. Rates of postsurgery tumor (80.7% vs 50.7%; P < .001) and lymph node downstaging (67.5% vs 48.6%; P = .001) were both significantly higher with toripalimab than placebo. With a median follow-up of 18.3 months, a better EFS was noticed in the toripalimab group. Tumor and lymph node downstaging in the toripalimab group were both associated with better EFS than nondownstaging (median EFS, not estimable [NE] vs 17.5 months; P = .004 and NE vs 19.2 months; P = .001, respectively), and were also associated with even better EFS than tumor and lymph node downstaging in the placebo group (median EFS, NE vs 22.0 months; P = .002 and NE vs NE; P = .009, respectively). In this study, perioperative toripalimab plus chemotherapy showed comparable perioperative outcomes, as with chemotherapy alone without new safety signals, and could help improve survival through effective tumor downstaging in patients with resectable stage III NSCLC. ClinicalTrials. gov Identifier: NCT04158440.

Accurately assessing the risk of lung cancer metastasis is crucial for improving patient prognosis and quality of life. In this study, we developed a rapid, homogeneous fluorescence detection system (DNA-N@Ag+@PIX) for lung cancer mesenchymal circulating tumor cells (M-CTCs) based on a molecular filling strategy and metal ion-mediated DNA self-assembly. By introducing small molecules such as pixantrone dimaleate (PIX) as molecular fillers, the stability of the DNA nanomaterials and the detection sensitivity of the detection system were significantly improved. The constructed DNA-N@Ag+@PIX system employed vimentin as a marker. Specific target-aptamer binding triggered the depolymerization of the DNA nanostructure and the release of Ag+, which was subsequently transduced into a fluorescence signal using CdTe quantum dots (QDs), thereby enabling ultrasensitive detection of M-CTCs in the peripheral blood of patients with non-small cell lung cancer. The system completed detection within 30 min, and the detection limit reached the single cell level. Analysis of 60 clinical samples demonstrated that the system could effectively distinguish non-metastatic and metastatic patients, achieving high specificity (91.9%) and sensitivity (95.7%), and the results were consistent with clinical diagnosis. This study provided an effective liquid biopsy strategy for monitoring lung cancer metastasis risk.

The gut microbiome is increasingly recognized as a key modulator of cancer immunotherapy efficacy. Given that diet is one of the most important determinants of the gut microbiome composition and function, nutritional strategies have emerged as promising tools to modulate anti-tumor immune responses. Here, we demonstrate that dietary supplementation with inulin reduces tumor growth and enhances αPD-1 efficacy in mice. These effects were associated with increased frequencies of intra-tumoral CD8⁺ and CD4⁺ T cells, particularly CCR9⁺CXCR3⁺ subsets, and enrichment of beneficial taxa such as Akkermansia and Lachnospiraceae, alongside elevated short-chain fatty acids (SCFA) levels. Among the SCFA, butyrate alone recapitulated the anti-tumor effect of inulin and had an additive effect when combined with αPD-1 therapy in a CD8⁺ T cell-dependent manner. Butyrate exerted its anti-tumor effects by transcriptional changes in CD8⁺ T cells involving activation of proliferation, trafficking, and metabolic pathways. In a cohort of 117 non-small cell lung cancer (NSCLC) patients amenable to immunotherapy, the median dietary fiber intake was lower than previously published studies but correlated with enrichment of Faecalibacterium praunitzii and metabolic pathways related to sucrose degradation and tryptophan biosynthesis. Collectively, our findings highlight the therapeutic potential of targeting diet-microbiome-immune system interactions to improve cancer immunotherapy outcomes.

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

Despite the success in infectious diseases, mRNA vaccines have inherent limitations in stability and immunogenicity. Non-small cell lung cancer remains the leading cause of cancer-related death worldwide, with poor prognosis in advanced stages, underscoring the urgent need for novel therapeutic strategies. Previously, we have developed a scarless circular RNA synthesis platform NeoAna that exhibited enhanced stability and long-lasting antigen expression with minimal cytotoxicity compared to mRNA. In this study, we construct circRNA vaccine using NY-ESO-1 as the antigen and deliver circRNA vaccine with lipid nanoparticles. The circRNA vaccine elicited robust innate and adaptive immune responses both in vitro and in vivo. It demonstrated superior antitumor efficacy relative to mRNA vaccine, achieving 93.1% tumor inhibition when combined with immune checkpoint blockade. Prophylactic vaccination prevented tumor establishment, while adjuvant therapy conferred protection against tumor re-challenge, highlighting durable immune memory. Mechanistically, circRNA vaccine promoted dendritic cell maturation and enhanced tumor infiltration of T cells, and mediated antitumor immunity in a CD8+ T cell-dependent manner. Additionally, single-nucleus RNA sequencing revealed that it remodeled the tumor microenvironment toward a pro-inflammatory and antitumor phenotype. Following circRNA vaccination, the proportions of cytotoxic T cells, pro-inflammatory macrophages, and antigen-presenting cancer-associated fibroblasts were all increased within the tumor microenvironment, accompanied by activation of immunity-related signaling pathways. In conclusion, our findings demonstrate the immunogenicity and therapeutic potential of the NeoAna circRNA-based therapeutic vaccine, supporting its further evaluation in clinical trials.

Fibroblast growth factor 21 (FGF21) is upregulated in lung squamous cell carcinoma (LUSC) tissues, cell lines, and tumor-conditioned media. Functional studies using LUSC cell lines (H1703, H520) demonstrate that FGF21 promotes cancer cell proliferation, migration, invasion, and tumor sphere formation in vitro. This pro-tumorigenic effect was validated in vivo through xenograft models, where intra-tumoral FGF21 administration accelerated tumor growth. Mechanistically, FGF21 activates calcium/calmodulin-dependent protein kinase II (CaMKII), which phosphorylates dynamin-related protein 1 (DRP1, encoded by DNM1L) at Ser616. Phosphorylated DRP1 translocates to mitochondria, inducing mitochondrial fission. Metabolic analyses revealed that FGF21 remodels cellular energetics in a cell type-specific manner, enhancing glycolysis in H520 cells and oxidative phosphorylation (OXPHOS) in H1703 cells, consistent with their inherent predominant metabolic states. Knockdown of DRP1 or pharmacological inhibition of CaMKII (KN93) abolished FGF21-driven mitochondrial fission, metabolic reprogramming, and tumor-promoting effects. Collectively, FGF21 acts as a tumor-promoting factor in LUSC by activating the CaMKII/DRP1-Ser616 axis to induce pathological mitochondrial fission and metabolic reprogramming, identifying this pathway as a potential therapeutic target.

Circular RNAs (circRNAs) play crucial roles in cancer progression and metastasis. However, the expression profiles and biological roles of circRNAs in non-small cell lung cancer (NSCLC) remain unclear. In this study, we found a novel circRNA, hsa_circ_0001936(circBRWD3), in NSCLC and investigated the biological role of circBRWD3 in NSCLC progression in vitro and in vivo. qRT-PCR was used to quantify the expression of circBRWD3 in NSCLC tissues, serum and cells. Cell growth curves, colony formation, Edu assay and transwell assays were conducted to study the function of circBRWD3. Finally, the molecular mechanism of circBRWD3 was revealed by Bioinformatic analyses, RNA pull-down analysis, RNA Fluorescence in situ hybridization (FISH), RNA-Binding Protein Immunoprecipitation (RIP), western blot, dual-luciferase reporter gene assays and rescue experiments. Our results showed that circBRWD3 was upregulated in NSCLC tumor tissues, patient serum and cell lines. CircBRWD3 knockdown inhibited NSCLC cells' growth and metastasis both in vitro and in vivo, while circBRWD3 overexpression had the opposite effect. Mechanistically, circBRWD3 binds to the TRIM25 protein to enhance its stability and mediate ubiquitin degradation of the p53 protein. In addition, circBRWD3 acted as a sponge for miR-135a-5p and miR-135b-5p to regulate the expression of SPTBN2, which promoted NSCLC progression. Our findings uncovered a tumor oncogenic function of circBRWD3 in NSCLC through the regulation of TRIM25 protein stability and miR-135a-5p/miR-135b-5p/SPTBN2 axis. CircBRWD3 may act as a promising oncogenic biomarker and therapeutic target for NSCLC.

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

Tumour-infiltrating FOXP3+ regulatory T cells (Tregs) exert suppression of anti-tumour immunity in non-small cell lung cancer (NSCLC), contributing to poor prognosis and immunotherapy resistance. The BCL-2 family pro-survival protein, MCL-1, is a critical controller of lymphoid Treg viability, yet its role in tumour-infiltrating Tregs remains poorly defined. Here we find that tumour-infiltrating effector Tregs in human NSCLC exhibit an activation-associated shift in BCL-2 family pro-survival protein expression typified by elevated MCL-1 expression. Pharmacological inhibition of MCL-1 with the BH3 mimetic S63845 induced moderate apoptotic cell death in both human and murine tumour-infiltrating Tregs, coincident with transient enhancement of CD8⁺ T cell activity. Combined MCL-1 inhibition and anti-PD1 immunotherapy further reduced tumour-infiltrating effector Treg abundance and influenced CD8⁺ T cell dynamics, although these effects were not sufficient to extend long-term survival. Mechanistically, we found that IL-33 upregulated MCL-1 expression and was required to support activated tumour-infiltrating Tregs. These results establish MCL-1 as an important regulator for tumour-infiltrating Treg survival and highlight the potential of repurposing BH3 mimetics to modulate immune suppression in NSCLC.

Despite the promise of immune checkpoint blockade (ICB), only a minority of non-small-cell lung cancer (NSCLC) patients achieve long-term benefits. In this study, we present a single-cell spatial transcriptomic landscape of NSCLC, revealing a previously uncharacterized link between elevated glutathione peroxidase 8 (Gpx8) and resistance to PD-1 blockade. Through CyTOF, CODEX, and ATAC-sequencing analyses, we demonstrate that Gpx8 knockout in both immunocompetent and humanized mouse models suppress tumor growth. This suppression is accompanied by increased infiltration of antitumor T lymphocytes, reduced enrichment of pro-tumorigenic myeloid cells, and the formation of tertiary lymphoid structures (TLS). Mechanistically, Gpx8 inhibits the activity of the RNA-binding protein Celf1(CUGBP Elav-like family member 1) through disulfide bonding between cysteine 79 of Gpx8 and cysteine 177 of Celf1. This interaction stabilizes CCAAT-enhancer-binding protein β (C/EBPβ) mRNA, promotes CSF1 secretion, and drives the recruitment of myeloid-derived suppressor cells (MDSCs) into the tumor microenvironment. Notably, resistance to anti-PD-1 treatment in Gpx8-expressing NSCLCs can be overcome through enforced expression of Celf1, CSF1R blockade, or a mimic peptide designed to disrupt the Gpx8-Celf1 interaction. Furthermore, anti-PD-1 or rCSF1 treatment activates C/EBPβ and upregulates Gpx8 transcription, establishing a Gpx8-C/EBPβ-CSF1 feedback loop that contributes to immune evasion. These findings provide new insights into the role of Gpx8 in modulating the tumor microenvironment and offer a potential framework for enhancing the sensitivity of NSCLC to PD-1 blockade therapy.

Metabolic alterations and neutrophils define the pre-metastatic niche, but the mechanisms connecting them remain incompletely understood. In this issue of Immunity, Qian et al. show that endothelial-derived palmitate induces neutrophils to release lipocalin-2, disrupting vascular integrity and promoting lung metastasis.

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related mortality worldwide, and the search for novel therapeutic strategies is ongoing. Chalcones, a class of naturally occurring and synthetically derived open-chain flavonoid precursors, have attracted attention because of their chemical versatility and broad biological activity. This review synthesizes current findings on the molecular mechanisms through which chalcones exert their effects in NSCLC, including modulation of oxidative stress, epithelial-mesenchymal transition, DNA damage responses, and diverse cell death pathways. Chalcones have been shown to interfere with key signaling cascades, such as TGF-β, PI3K/AKT/mTOR, JAK/STAT, and Wnt/β-catenin, with reported effects on tumor proliferation, migration, invasion, and therapy resistance. Their structural flexibility enables chemical optimization aimed at improve potency, selectivity, and pharmacokinetic profiles, supporting further preclinical investigation. Despite promising preclinical results, challenges remain regarding pharmacokinetics, bioavailability, and context-dependent off-target effects. Future research should prioritize integrated mechanistic studies, structure-activity relationship analyses, selective cytotoxicity assessment, and advanced delivery strategies to improve tumor specificity. Chalcones represent chemically versatile multitarget scaffolds for further optimization in NSCLC, particularly in strategies aimed at exploiting epithelial-mesenchymal plasticity, redox adaptation, and regulated cell death.

Fiber morphology (length > 5 μm; respirable diameter < 3 μm) and biopersistence have been linked to their potential to cause fibrosis, lung cancer, and malignant pleural mesothelioma. Among the mechanisms involved, frustrated phagocytosis occurs when macrophages attempt but fail to fully internalize and clear long rigid fibers. Although nanofibers could meet these criteria and trigger frustrated phagocytosis, their small diameters may enable them to entangle, causing them to lose their fiber-like morphology and affecting their toxicological potential. The toxicological assessment of (nano)fibers relies on animal studies; therefore, there is an urgent need to establish in vitro alternatives. Carbon nanotubes, the most commercially prevalent class of nanofibers, have been extensively investigated, and some have demonstrated pathogenic potential, by causing inflammation initiated by cathepsin B translocation from the lysosomes into the cytosol. Independent studies have indicated that only long and rigid carbon nanofibers lead to a decrease of lysosomal enzymes (including multiple cathepsins) inside macrophages and increased levels in the extracellular environment. Thus, different roles for cathepsin B have been proposed in response to nanofiber exposure. To reconcile these observations, this review examines the underlying mechanisms by assessing in vitro studies, particularly how in vitro macrophages respond to carbon-based nanomaterials of distinct morphologies, discusses the limitations of current in vitro models, and evaluates potential approaches for assessing nanofiber toxicity.

ASCOLT compared adjuvant aspirin versus (vs) placebo in 1587 patients with colorectal cancer (CRC) and reported no significant improvement in disease-free survival (DFS). Subsequently, two randomised controlled trials (RCTs) reported benefit of adjuvant aspirin in CRC patients with somatic PI3K-related mutations. The ASCOLT Translational Research (TR) study was a preplanned subgroup analysis by PIK3CA mutation status and COX-2 overexpression, and an exploratory analysis including PTEN mutation status. Among 778 participants who commenced study medication, tissue was evaluable for targeted next generation sequencing of PIK3CA and PTEN in 289 tumours and by Sanger sequencing for PIK3CA exon 9 or 20 (9/20) mutations in a further 108. PTGS2/COX-2 expression was assessed by immunohistochemistry in 450. Hazard ratio (HR) and 95% confidence intervals (CI) for DFS of aspirin vs placebo in subgroups were assessed in Cox models. A systematic review of completed RCTs of adjuvant aspirin in CRC patients with PIK3CA mutations was also undertaken. NCT00565708 and ACTRN12614000513617. Among 397 patients with tumour assessable for PIK3CA, there were 80 recurrences, 40 deaths and 86 DFS events after 5 years follow-up. Among 69 (17%) with PIK3CA mutations (any exon), there were 8 vs 8 DFS events on placebo vs aspirin (HR 0.93 (95% CI 0.35-2.47); in 45 (11%) with exon 9/20 mutations, 7 vs 4 events (HR 0.72 (95% CI 0.21-2.46) and in 84 (21%) with PIK3CA or PTEN mutations, 8 vs 9 events (HR 1.23 (95% CI 0.47-3.19). Tumours were positive for COX-2 overexpression in 307 (69%) with 28 vs 34 events (HR 0.99 (95% CI 0.60-1.63). A meta-analysis of trials in patients with PIK3CA exon 9/20 mutations showed reduced DFS events with HR 0.61 (95% CI 0.39-0.96). In ASCOLT TR, adjuvant aspirin was not associated with significantly improved DFS in CRC with PI3K-related mutations or COX-2 overexpression, albeit with wide confidence intervals. Combined results of the three published trials showed improved DFS among patients with PIK3CA exon 9/20 mutations. Moderate, but important, effects of aspirin in other patient groups have not been excluded. National Health and Medical Research Council, Australia; Cancer Australia; National Cancer Centre Singapore Cancer Fund; Rising Tide Foundation; SingHealth Duke-NUS Academic Clinical Programme; Lee Foundation; Lee Kim Tah Foundation; Silent Foundation; Australasian Gastro-Intestinal Trials Group.

Identifying high-risk patients for recurrence after endoscopic resection (ER) 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 ER alone across 27 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 clinicopathological 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 <1,000μm indicated low risk, whereas invasion ≥1,000μm required morphological assessment, with polypoid lesions classified as high risk and flat lesions further stratified using a 2,000μm threshold. Three clinically distinct recurrence risk subtypes were identified in T1 CRC following ER, suggesting that morphological 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.

The therapeutic landscape of colorectal cancer (CRC) has evolved with the identification of molecular subtypes, including mismatch repair-deficient/microsatellite instability-high, POLE mutations, RAS/BRAF alterations, and HER2 amplification, enabling use of precision therapies and immune checkpoint inhibitors for selected populations. However, most microsatellite-stable (MSS) tumors remain resistant due to tumor heterogeneity, adaptive resistance, and an immunosuppressive tumor microenvironment (TME). Advances in molecular profiling, spatial biology, and immune characterization have revealed vulnerabilities beyond canonical signaling, facilitating novel strategies such as antibody-drug conjugates (ADCs), bispecific antibodies, DNA damage response (DDR) targeting, TME-directed therapies, cellular therapies, and epigenetic modulation. HER2-directed ADCs, notably trastuzumab deruxtecan, have shown clinically meaningful activity in HER2-positive metastatic CRC, providing proof of concept for ADC-based therapy. Additional targets under investigation include CEACAM5, LGR5, EGFR, HER3, MET, B7-H3, and CDH17. Bispecific antibodies and co-stimulatory agonists are being developed to overcome antigen heterogeneity and pathway redundancy, with the EGFR-MET bispecific antibody amivantamab showing initial clinical efficacy. Moreover, next-generation immune checkpoint inhibitors and multitarget combinations aim to reinvigorate T-cell responses in MSS tumors. Early chimeric antigen receptor T-cell studies targeting CEA and GUCY2C demonstrate feasibility and manageable toxicity, although tumor-intrinsic and TME barriers persist. Emerging strategies increasingly focus on modulating the TME to enhance immune infiltration and effector function, targeting CCR8, TGF-β, adenosine, CSF1R, CXCR1/2, STING, and CD47. DDR and epigenetic therapies offer additional opportunities to sensitize resistant tumors. Integrated, multidimensional biomarker approaches and artificial intelligence-driven interpretation of tumor and TME features are expected to guide personalized therapy, anticipate resistance, and broaden the benefit of targeted and immune-based interventions in CRC.

Accurate profiling of microRNAs (miRNAs) is often limited by the severe background leakage of conventional enzyme-free DNA amplification. Herein, we develop a 1D DNA nanowire-mediated dispersion-to-localization catalytic hairpin assembly (DL-CHA) platform for the highly sensitive detection of miRNAs. Methodologically, responsive hairpins are physically isolated on spatially dispersed nanowire scaffolds to rigorously suppress target-independent interactions. Upon specific recognition of miR-21, a pseudo-intramolecular cascade is triggered, dynamically cross-linking the scaffolds into a highly fluorescent DNA ladder. As a result, this DL-CHA platform yields an exceptional signal-to-background ratio of 12.4, a broad linear range of 0.05 to 250 nM, and an ultra-sensitive limit of detection of 50 pM without the need for exogenous enzymes. Furthermore, the rigid nanowire architecture ensures robust nuclease resistance and enables highly efficient, transfection-free internalization for the in situ imaging of endogenous miRNAs in living cells. Clinically, the platform accurately discriminated colorectal cancer patients from healthy controls using non-invasive liquid biopsies of human serum. This paradigm offers a robust, low-background molecular tool for precision cancer diagnostics.

The gut microbiota is integral to host physiology, contributing to metabolic homeostasis, epithelial barrier integrity, immune balance, and bidirectional communication along gut-organ axes. Disruption of this ecosystem, commonly referred to as dysbiosis, is increasingly implicated in a wide range of gastrointestinal and extra-intestinal diseases. Rather than reflecting isolated compositional changes, microbiota-related pathology often involves interconnected disturbances in barrier function, microbial metabolism, immune regulation, genotoxicity, inflammatory and oncogenic signaling, and long-range communication with distal organs. However, key challenges remain, particularly in resolving causality, accounting for interindividual heterogeneity, and translating complex microbiome data into robust clinical tools. In this review, we summarize the role of the gut microbiota in maintaining host homeostasis and outline the concept, drivers, and consequences of dysbiosis. We then discuss the major mechanisms through which the gut microbiota contributes to disease development and progression, using colorectal cancer as a representative gastrointestinal example and gut-organ axes as a framework for extra-intestinal disorders. We further highlight current translational advances in microbiota-based biomarkers, dietary modulation, biotic and postbiotic strategies, fecal microbiota transplantation, and emerging precision microbiota therapies. By integrating mechanistic insights with translational perspectives, this review offers an updated framework for interpreting the gut microbiota in health and disease and may help inform the future development of more precise, mechanism-informed diagnostic and therapeutic strategies.

Multidrug resistance and invasive metastasis constitute pivotal clinical bottlenecks that severely compromise curative outcomes of malignant tumors. Conventional chemotherapy and immunotherapy frequently fail to achieve satisfactory efficacy due to drug resistance barriers and tumor immune escape. Herein, a hyaluronic acid‑cinnamaldehyde Schiff base micelle nanoplatform loading quaternary ammonium‑modified carbon dots (HACA@QASCDs) is rationally constructed, which achieves targeted killing of drug‑resistant tumor cells, remodeling of immunosuppressive microenvironments, and inhibition of distant metastasis via a sequential cascade of irreversible membrane perforation, mitochondria‑dependent apoptosis, and immunogenic cell death (ICD). HACA@QASCDs actively accumulate in drug‑resistant CT26 (DR‑CT26) cells through HA‑CD44 recognition and enable pH‑triggered QASCDs release in acidic tumor microenvironments. The liberated QASCDs elicit irreversible membrane perforation, leading to lactate dehydrogenase leakage, disrupted calcium homeostasis, mitochondrial depolarization, and subsequent intrinsic apoptosis. Such membrane damage simultaneously ignites ICD, and the released damage‑associated molecular patterns effectively drive dendritic cell maturation and M2‑to‑M1 macrophage polarization. In vivo evaluations in bilateral syngeneic tumor models reveal that HACA@QASCDs alone yields 54.2% primary tumor inhibition and 28.6% distant tumor inhibition. Upon combination with αPD‑L1, the distant tumor inhibition rate is markedly elevated to 68.7%. By integrating membrane perforation‑mediated direct cytotoxicity and ICD‑evoked immune activation, HACA@QASCDs offers a highly potent and clinically translatable synergistic strategy to surmount tumor multidrug resistance and block invasive metastasis.

Cancer cells acquire malignant traits through epigenetic remodeling driven by inflammatory secretomes in the tumor microenvironment. However, how immune-derived inflammatory factors regulate cancer cell epigenetics remains poorly understood. Here, we show that IL-26 produced by tumor-specific type 17 T cells acts as a key mediator of epigenetic reprogramming associated with cancer malignancy. We identify selective accumulation of IL-26-expressing T cells in colorectal tumors resistant to immune checkpoint blockade therapy. IL-26 functions as a noncanonical cytokine by translocating into the nucleus of tumor cells, where it directly binds STAT1 and forms transcriptional complexes with NF-κB and AP-1. Nuclear IL-26 induces a transcriptionally active chromatin state characterized by BRD4 and H3K27ac enrichment, resulting in upregulation of CXCL chemokines. This process promotes neutrophil infiltration and enhances immune evasion and tumor progression by suppressing CD8 + T cell responses. Together, these findings support a model in which tumor-specific type 17 T cells directly reprogram cancer cell epigenetics through IL-26, reshaping the tumor immune microenvironment to promote immune evasion.

Targeted drug delivery in the gastrointestinal tract remains challenging because therapeutics must overcome multiple hierarchical barriers before reaching diseased tissue. Here, we present a multistage delivery platform that integrates magnetic microrobots, a pH-responsive protective coating, and platelet membrane-coated nanoparticles (PNPs) in one platform. A fillable design enables the formation of an internal magnetic layer for microrobot actuation, while the pH-responsive coating protects the cargo during transit and selectively degrades upon pH change, releasing cancer cell-targeting PNPs. In an in vitro colon cancer model that reproduces key gastrointestinal features, including flow, pH variation, and villi-like structures, this strategy increased nanoparticle retention and enhanced cancer cell cytotoxicity compared to nanoparticles administered alone. Ex vivo studies in porcine stomach and intestine further demonstrated robust locomotion on compliant and folded tissue surfaces. These results establish an environment-responsive hierarchical delivery strategy for more precise oral delivery in complex gastrointestinal settings.

Colorectal cancers (CRCs) with deficient DNA mismatch repair (dMMR) and microsatellite instability (MSI) are believed to be intrinsically immunogenic and respond more favorably to immune checkpoint inhibitor (ICI) therapy. However, a significant fraction of dMMR/MSI CRCs do not respond or eventually develop resistance to ICIs. The mechanisms underlying dMMR/MSI-associated immunogenicity is unclear. The aim of this study was to investigate the role and mechanism of MSI-induced antitumor immunity in CRCs. We used transplant syngeneic tumor models, immune cell co-culture assays, and air-liquid interface (ALI) culture of tumor-derived organoids to investigate the mechanism by which dMMR elicits an antitumor immune response. We also analyzed the gene expression databases of CRC cancer patients receiving ICI treatment. We found that inactivating Mlh1 causes endoplasmic reticulum (ER) stress and death receptor 5 (Dr5)-mediated apoptosis in syngeneic colorectal tumors. Sustained immune response against Mlh1-deficient tumors requires nuclear Ligase 3 (Lig3)-mediated release of extrachromosomal circular DNAs (eccDNAs) from apoptotic cells. A feedback Dr5/Lig3 amplification loop perpetuates apoptosis and immune cell activation in Mlh1-deficient syngeneic tumors. This feedback mechanism is critical for the response to ICI therapy in mice, which is supported by a significant association between DR5 or LIG3 expression and the efficacy of ICI therapy in cancer patients. Our results reveal a functional link between dMMR and antitumor immunity, which may be useful for improving ICI therapy in tumors with different MMR statuses.

Colorectal cancer (CRC) with microsatellite instability (MSI) is often treated with immune checkpoint inhibitors (ICIs), such as anti-PD-1 antibodies. However, a substantial fraction of MSI CRCs do not respond to ICIs. Recent studies have identified the DNA helicase WRN as a synthetic lethal target in MSI cancer cells, leading to the development of several small-molecule WRN inhibitors that are currently in clinical trials. In this study, we found that targeting WRN in MSI CRC cells triggered a robust antitumor immune response. Cell death induced by WRN inhibition was selective in MSI CRC cells and led to the release of extrachromosomal circular DNA (eccDNA), which directly stimulated immune cell activation and cytokine production. The deletion of nuclear ligase LIG3, a key mediator of eccDNA biogenesis, abolished the antitumor and immunogenic effects of WRN inhibition in MSI CRC cells and tumors. Furthermore, WRN inhibition potentiated anti-PD-1 therapy in MSI CRC models, including syngeneic mouse tumors and patient-derived tumor organoids. Together, these results reveal eccDNA-mediated immunogenic effects of WRN inhibition in MSI CRC, further strengthening the rationale for combining WRN inhibitors with ICIs.

Diet composition shapes tissue function and disease risk by modulating nutrient availability, metabolic state and cellular dynamics1. In the gastrointestinal tract, obesogenic high-fat diets enhance small-intestinal stem cell activity and tumorigenesis2. However, the impact of ketogenic diets (KDs), which contain even higher lipid content but reduce circulating insulin and induce ketogenesis, remains poorly understood3. This is particularly relevant for patients with familial adenomatous polyposis who face a high risk of small-intestinal tumours4. Here we combine dietary, genetic and metabolic manipulations in mouse models of spontaneous intestinal adenoma formation to dissect the role of systemic and epithelial ketogenesis in intestinal cancer. We show that KD accelerates tumour burden and shortens survival, independent of ketone metabolites. Through genetic manipulation of the ketogenic pathway, we modulate the production of local and systemic ketone metabolites; however, neither inhibition nor augmentation of the ketogenic enzyme 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 nor disruption of ketolysis altered tumorigenesis. Combined intestinal loss of PPARα/δ/γ attenuates KD-driven intestinal stem cell expansion, proliferation and clonogenicity, whereas inhibition of downstream fatty acid oxidation through CPT1A loss limits adenoma formation specifically under KD, linking tumour initiation to fatty acid oxidation of dietary lipids rather than lipid accumulation. These findings reveal that dietary lipid content, through fatty acid oxidation rather than ketone metabolism, influences intestinal tumorigenesis and highlight the need for nuanced consideration of dietary strategies for cancer prevention in genetically susceptible populations.

Colorectal cancer stem cells (CSCs) drive tumor progression through poorly understood metabolic-epigenetic crosstalk. Here, we identify mitochondrial RNA polymerase POLRMT as a key link connecting mitochondrial transcription to CSC maintenance. Clinically, POLRMT is overexpressed in colorectal cancer (CRC) tissues and correlates with poor prognosis. Genetic ablation or pharmacological inhibition of POLRMT suppresses CSC self-renewal and tumorigenicity across cell line-derived CSCs, CRC organoids, and xenograft models. Mechanistically, POLRMT deficiency triggers mitochondrial dysfunction, which unexpectedly elevates the demethylase KDM6B expression, α-ketoglutarate (α-KG) levels, and Dickkopf-1 (DKK1) expression, thereby transcriptionally silencing Wnt/β-catenin signaling and collapsing the CSC niche. Restoration of β-catenin rescues tumorigenicity in POLRMT-knockout (KO) cells, confirming the hierarchy of this signaling cascade. Crucially, POLRMT catalytic activity and mitochondrial localization are indispensable for sustaining this axis. Our work unveils POLRMT as a metabolic gatekeeper that licenses CSC plasticity through KDM6B-α-KG/H3K27me3-mediated chromatin remodeling, proposing the mitochondrial transcription machinery as a therapeutic target for dismantling the CSC hierarchy in CRC.

Cellular turnover and death result in shedding of DNA into the extracellular space. In the context of cancer, the constant turnover of cancer cells leads to the release of tumor-derived DNA (tDNA) into the circulation. Circulating tDNA (ctDNA) provides molecular insights into the genetic alterations present within cancer cells, which has been exploited to noninvasively detect and monitor tumors. In addition to its utility in cancer monitoring, tDNA has also been implicated in the formation of neutrophil extracellular traps that can promote oncogenesis and metastasis. Despite the appreciation that tDNA/ctDNA can affect the tumor microenvironment (TME) to shape cancer progression and its promise in liquid biopsy applications, the mechanisms that control tDNA accumulation in the TME and the functional impacts of tDNA remain lacking. In this issue of Cancer Research, Cao and colleagues identify a role for mutant KRAS in reducing colorectal tDNA through upregulation of CD9, which remodels membrane tension to promote endocytosis. Knockdown of CD9 increases extracellular tDNA and reprograms tumor-associated macrophages to sensitize tumors to PD-1 blockade. Together, these studies identify novel roles for mutant KRAS in tDNA turnover and tDNA in immune evasion. See related article by Cao et al., p. 3394.

T-cell exhaustion in the tumor microenvironment undermines antitumor immunity and limits immunotherapy efficacy. Further defining the metabolic triggers of this dysfunctional state could provide therapeutic targets for circumventing immunosuppression. In this study, we identified soluble uric acid (UA)-an abundant purine metabolite frequently elevated in patients with cancer-as a metabolic checkpoint that drives the exhaustion of CD8+ T cells and immune evasion in colorectal cancer. In hyperuricemic mouse models, elevated UA accelerated tumor progression in immunocompetent hosts, but not in T cell-deficient ones, by functionally exhausting tumor-infiltrating CD8+ T cells. Mechanistically, UA directly bound the kinase scaffold kinase suppressor of Ras 1 (KSR1) and hyperactivated MEK-ERK signaling, leading to chronic MAPK stimulation that upregulated inhibitory receptors, including PD-1 and Tim-3, on CD8+ T cells and blunted their cytotoxic function. Genetic disruption of this UA-KSR1-MAPK axis via Tim-3 knockout or Ksr1 knockdown restored T-cell effector activity and tumor control. Notably, pharmacologic UA depletion with the clinical xanthine oxidase inhibitor febuxostat reinvigorated CD8+ T cells, slowing tumor growth and markedly enhancing the efficacy of both chemotherapy and adoptive T-cell therapy in vivo. These findings establish soluble UA as a metabolic immune checkpoint that subverts antitumor T-cell immunity. Targeting UA metabolism may offer a strategy to overcome immune resistance and improve the efficacy of cancer immunotherapies. A common metabolic byproduct, soluble uric acid, can act as an immune checkpoint that drives T-cell exhaustion, redefining how systemic metabolism shapes cancer progression.

Peritoneal metastasis (PM) of colorectal cancer (CRC) remains a major therapeutic challenge due to the limited efficacy of current systemic and intraperitoneal treatments. To overcome these limitations, we developed an injectable, self-healing hydrogel constructed from a dual dynamic cross-linked network formed by borate ester and Schiff base bonds between phenylboronic acid-modified carboxymethyl chitosan (CMCS-PBA) and oxidized dextran (ODEX). This hydrogel was engineered for sustained intraperitoneal co-delivery of oxaliplatin (OXA) and resveratrol-loaded mesoporous silica nanoparticles (MSNs@RES). The system exhibited excellent biocompatibility and significantly inhibited cancer cell proliferation, migration, and invasion while inducing apoptosis and immunogenic cell death (ICD) in vitro. In a murine model of CRC peritoneal metastasis, the dual-drug-loaded hydrogel markedly suppressed tumor progression, reduced ascites formation, and prolonged survival. Proteomic analysis further revealed that treatment significantly modulated key signalling pathways, including HIF-1α-mediated angiogenesis, apoptosis-related cascades, and TGF-β-driven epithelial-mesenchymal transition (EMT). Collectively, this work presents a rationally designed localized delivery platform with significant potential for the treatment of colorectal cancer peritoneal metastasis.

Therapy-resistant cancers remain largely incurable because malignant cells acquire metabolic adaptations that sustain survival under chronic cytotoxic and oxidative stress. One such adaptation is constitutively enhanced macropinocytosis, enabling aggressive cancer cells to scavenge extracellular nutrients and maintain redox homeostasis. Here, we introduce a macropinocytosis-exploiting polymer-metal strategy that converts this metabolic dependency into a lethal vulnerability. We developed PPS02, a polyaspartic acid sodium salt-based metal complex that is preferentially internalized by cancer cells via macropinocytosis, while remaining largely excluded from normal epithelial cells. This cancer-preferential uptake enables intracellular delivery of selenomethionine and ferrous iron, resulting in intracellular H2O2 accumulation, mitochondrial reactive oxygen species overload, and activation of necroptotic cell death. Macropinocytic activity was significantly elevated in patient-derived metastatic colorectal cancer cells but remained minimal in normal colonic epithelial cells, demonstrating pronounced cancer selectivity. Accordingly, PPS02 exhibited negligible cytotoxicity toward normal colonic epithelial cells while effectively suppressing the viability of both nonmetastatic and platinum-resistant metastatic colorectal cancer cells. In patient-derived xenograft models, PPS02 induced sustained tumor regression without overt systemic toxicity under the experimental conditions, whereas cisplatin failed to control metastatic tumors and caused significant adverse effects. Collectively, these findings support macropinocytosis-driven redox imbalance as a therapeutically exploitable vulnerability and demonstrate a polymer-metal platform that preferentially induces necroptosis in drug-resistant cancer while sparing normal tissues.

Glioblastoma (GBM) is an aggressive and lethal brain tumor marked by profound local and systemic immune dysfunction. Despite evidence of peripheral immune impairment, the clinical relevance of these alterations for diagnostic or therapeutic purposes remains poorly defined. We performed multimodal single-cell profiling of peripheral blood mononuclear cells from a single-center cohort of treatment-naïve GBM patients and healthy donors, integrating mass and flow cytometry with single-cell RNA-sequencing. Unsupervised clustering, pseudo-temporal trajectory analyses and cell-cell communication inference were applied to map immune states and their interactions. GBM blood profiles were characterized by heterogeneous changes in classical monocytes, encompassing expanded, reduced and unchanged subsets with distinct functional states, including antigen-presenting, interferon and metabolic programs. Additional myeloid adaptations included myeloid-derived suppressor cell (MDSC) expansion and loss of non-classical monocytes. Trajectory analyses identified a differentiation continuum, evolving from antigen-presenting to metabolic monocyte subsets, and positioning MDSCs as an intermediate state. Antigen‑presenting monocytes displayed tumor‑migratory, precursor‑like profiles that corresponded to tumor‑associated macrophages in public GBM datasets. Across subsets, circulating monocytes shared a "GBM-classical monocytic signature" characterized by low MHC class II expression, altered cell-cell communication and upregulation of anti-inflammatory mediators, including IL1R2 and CD163. Notably, complementary myeloid expression signatures were identified across patients, indicating distinct systemic immune phenotypes. In parallel, lymphocyte alterations included decreased proportions of CD4+ T, natural killer (NK) and CD56+ T cells, retaining relatively conserved activation profiles, exemplified by up-regulation of alarmins S100A8/S100A9. These findings delineate systemic immune reprogramming in primary GBM, characterized by coordinated myeloid and lymphocyte alterations. The identification of circulating monocyte states with transcriptional continuity to the tumour microenvironment, alongside distinct patient-level systemic myeloid signatures, provides a framework for exploring peripheral blood as a source of immune biomarkers in GBM.

Alterations in the BRCA1 DNA repair associated (BRCA1) and BRCA2 DNA repair associated (BRCA2), two key DNA repair genes, are strongly linked to adult-onset malignancies and play a significant role in tumor development. Although BRCA1 and BRCA2 are firmly recognized as hereditary cancer risk genes in adult populations, their role in central nervous system (CNS) tumor susceptibility in younger patients is still debated. We analyzed 367 pediatric, adolescent, and young adult (AYA) patients with primary CNS tumors. Tumors were classified according to the WHO Classification of Tumors of the CNS, fifth edition. All patients underwent clinical exome sequencing, including BRCA1 and BRCA2. Rare germline BRCA1/2 variants were identified in 17 of 367 patients (4.6%). Among them, 5 patients (1.4% of the overall cohort) carried pathogenic or likely pathogenic variants, whereas 12 patients (3.3%) carried variants of uncertain significance (VUSs). Our findings are consistent with emerging evidence suggesting that monoallelic pathogenic BRCA1/2 variants may act as low-penetrance susceptibility factors in pediatric and AYA CNS tumors rather than as primary oncogenic drivers. Interpretation is limited by cohort heterogeneity, lack of a noncancer control group, and absence of systematic somatic analyses to assess biallelic inactivation or homologous recombination deficiency. Nevertheless, these data support the inclusion of BRCA genes in germline testing panels for pediatric and AYA CNS tumors, with relevant implications for genetic counseling and long-term surveillance.

Meningiomas in pediatric and adolescent/young adult patients are poorly characterized biologically and clinically, and risk stratification is largely extrapolated from adult tumors. We analyze 293 tumors from patients aged 0-39 years using integrated histopathological and molecular profiling. Youth-onset meningiomas are enriched for NF2 and SMARCE1 alterations and exhibit a gain-dominated copy-number landscape, including recurrent chr17q gain, whereas canonical adult high-risk features, such as chr1p loss, lack prognostic significance. Adult-derived prognostic frameworks, including WHO grade, methylation-based stratification and integrated risk scores, fail to predict progression in patients ≤21 years of age. Tumors segregate into age-enriched epigenetic clusters defined by SMARCE1, NF2 and BAP1 alterations. Among NF2-altered tumors, patterns of Merlin inactivation, shaped by germline status and co-occurring copy-number variations, delineate biologically divergent subsets. In patients ≤21 years, extent of resection is the dominant predictor of outcome, while molecular features further refine risk assessment. These findings define pediatric and young adult meningiomas as a distinct molecular entity and support age-adapted risk refinement that integrates molecular features with strong clinical determinants.

Focused ultrasound-mediated blood-brain barrier opening (BBBO) is hypothesized to enhance liquid biopsy by facilitating the release of tumor material into the circulation. This study aimed to evaluate the impact of MR-guided FUS-BBBO on plasma extracellular vesicle (EV) concentrations and size distributions, alongside cell-free DNA (cfDNA) profiles, to determine the clinical efficacy of this approach in glioblastoma (GB) patients. We performed 14 MR-guided focused ultrasound (FUS)-BBBO procedures in patients with GB with blood sampling 1 h before and 1 h after sonication. Plasma EVs were isolated by size-exclusion chromatography and quantified by tunable resistive pulse sensing technology. Cell-free DNA (cfDNA) concentration and fragment profiles were assessed using fluorometric and electrophoretic methods. EV concentration showed no consistent change after FUS-BBBO, with marked inter-patient and inter-procedure heterogeneity. In contrast, EV size displayed a reproducible diameter reduction of ∼8-10 nm, driven predominantly by contraction of the upper tail of the size distribution, indicating selective modulation of larger vesicle subpopulations. cfDNA concentrations and fragmentation showed a heterogeneous pattern, with occasional patient-specific increases. Exploratory analyses revealed no significant associations between biomarker changes and procedural parameters except for treated volume. Our findings challenge the assumption that clinical BBBO uniformly increases biomarker abundance. The consistent shift toward smaller EVs suggests a selective modulation rather than an increased release. While BBBO may enhance the detectability of analytes that poorly cross the BBB, this may not apply to EVs, which are able to traverse the BBB under physiological conditions, further supporting their value as liquid-biopsy substrates in neuro-oncology. Future studies integrating clinical data with mechanistic models are necessary to refine FUS-enhanced diagnostic strategies.

Molecular profiling has become an integral part of glioma classification. The extent to which incidence of molecularly-defined adult-type gliomas vary by demographics is unknown. We describe national-level incidence and overall survival patterns of selected glioma subtypes by race/ethnicity. We generated standardized average annual age-adjusted incidence rates of molecularly-defined glioma subtypes by race/ethnicity from the Central Brain Tumor Registry of the United States (CBTRUS) using newly-diagnosed cases from January 1, 2018 to December 31, 2022. Survival data from the National Cancer Database (NCDB) were used from newly-diagnosed cases from January 1, 2018 to December 31, 2021 (with follow up through December 31, 2022) to evaluate four-year overall survival, median survival, and multivariable Cox Proportional hazards ratios. CBTRUS identified 68,172 glioma cases, IDH-wildtype glioblastoma was most common (n = 51,548). Non-Hispanic White individuals had significantly higher incidence of all gliomas compared to other groups (p > 0.001). Non-Hispanic Black individuals had the lowest incidence for IDH-mutant astrocytoma and oligodendroglioma, and shared lowest incidence of IDH-mutant glioblastoma and IDH-wildtype astrocytoma with non-Hispanic other individuals, who had the lowest incidence of IDH-wildtype glioblastoma (p < 0.001). The odds of having an IDH-wildtype (versus IDH-mutant) astrocytoma or glioblastoma were significantly lower for males, those of older age, and non-Hispanic Black individuals (p < 0.001). Non-Hispanic other individuals also had increased adjusted overall survival for most glioma subtypes compared to other racial/ethnic groups (p < 0.001). Four-year overall survival was lowest in all racial/ethnic groups for IDH-wildtype glioblastoma (p < 0.001). Our findings reveal significant disparities in incidence and survival by race/ethnicity with notable variations present based on glioma biomarkers.

UFMylation, a recently identified ubiquitin-like modification, is essential for cellular stress homeostasis, particularly endoplasmic reticulum (ER) stress regulation. However, its biological and therapeutic exploration has been hindered by the absence of potent small-molecule inhibitors. Here, we report the first discovery of two compounds targeting the UFMylation E3 ligase complex core protein DDRGK1: Osimertinib, originally designed as an EGFR T790M selective inhibitor, acting through a previously unrecognized covalent mechanism, and CP-24, a novel non-covalent inhibitor. Both compounds disrupt the DDRGK1-UFL1 interaction, globally suppress UFMylation, inhibit ER-phagy, and induce ER stress. In glioblastoma (GBM), pharmacological UFMylation inhibition markedly reduces tumor cell viability and sensitizes cells to Temozolomide and radiotherapy. Both compounds also exert strong immunomodulatory activity, promoting macrophage polarization toward an anti-tumor M1 state. In vivo, Osimertinib, benefiting from superior pharmacokinetics, significantly suppresses tumor growth in immunodeficient models and achieves tumor-free outcomes in 65% of immunocompetent mice. These tumor-free mice develop durable anti-GBM immune memory, rapidly clearing tumors upon rechallenge, an outcome unattainable by previous GBM treatments. Mechanistically, Osimertinib enhances anti-tumor immunity by promoting macrophage M1 polarization, T cell expansion, and reducing PD-1 protein levels. Collectively, our study introduces Osimertinib and CP-24 as valuable chemical probes for dissecting UFMylation biology and highlights Osimertinib's potential for off-label use in EGFR-wildtype GBM. More broadly, we establish UFMylation inhibition as a dual-targeting therapeutic strategy that disrupts tumor survival pathways and reprograms the immune microenvironment, offering a promising avenue for durable GBM control.

Human glioma is a devastating primary brain tumor with a dismal prognosis, necessitating the identification of novel therapeutic targets. The small nuclear ribonucleoprotein C (SNRPC), a core spliceosome component, is implicated in cancer, but its role in glioma remains unexplored. This study aimed to delineate the expression, function, and underlying mechanisms of SNRPC in human glioma. The bioinformatic analysis revealed that SNRPC is significantly upregulated in glioma tissues, with expression levels correlating strongly with higher tumor grade, aggressive molecular subtypes (IDH wild-type, 1p/19q non-codeleted), and poor patient prognosis, establishing it as a robust independent biomarker. Single-cell analysis pinpointed SNRPC enrichment in malignant glioma cells, and co-expression studies linked it to mitochondrial metabolism and oxidative phosphorylation. SNRPC expression is also upregulated in locally-resected glioma tissues and various glioma cell types. Functionally, silencing (by targeted shRNA) or knocking out (via CRISPR/Cas9 method) of SNRPC profoundly attenuated glioma cell proliferation, migration, and invasion, while inducing G1-S arrest and apoptosis. These effects were specifically observed in malignant glioma cells, sparing normal astrocytes. Mechanistically, loss of SNRPC led to severe mitochondrial dysfunction, characterized by impaired mitochondrial respiration, ATP depletion, membrane depolarization, and excessive ROS (reactive oxygen species) production. Conversely, SNRPC overexpression enhanced mitochondrial bioenergetics and promoted malignant phenotypes. SNRPC promoted glioma malignancy by regulating the expression of a key oncogene tumor necrosis factor alpha-induced protein 2 (TNFAIP2), which acts as a key downstream effector to drive increased cell proliferation and migration. Critically, SNRPC knockdown suppressed subcutaneous glioma xenograft growth and disrupted mitochondrial bioenergetics. Its knockout also impeded the intracranial glioma growth in an orthotopic mouse model. Our findings establish SNRPC as a pivotal driver of glioma malignancy by sustaining mitochondrial hyperfunction and TNFAIP2 expression essential for tumor cell proliferation.

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

Fluorescence lifetime imaging microscopy (FLIM) enables quantitative measurement of molecular environments, interactions and protein conformations. However, a large number of photons are required for accurate lifetime determination, restricting its practical applications in fast deep-tissue imaging. Here, we present event-based first-photon FLIM (EFLIM), a self-supervised denoising method that infers fluorescence lifetime at extremely low light. By representing each excitation event as a binary process instead of histogram accumulation, EFLIM reduces photon requirement by over two orders of magnitude compared to state-of-the-art algorithms, leading to an apparent mean lifetime measurement below one photon per pixel with strong robustness to intensity artifacts. To demonstrate EFLIM's applicability, we observed transient intracellular dynamics of ligand-dependent molecular states, captured putative vesicle-mediated contacts between different lymphocytes through multiplexed imaging in a single spectral channel and achieved rapid label-free visualization of tumor heterogeneity in human glioma tissue. These results illustrate EFLIM's strong potential in neuroscience, cell biology, immunology and pathology by probing dynamic molecular processes in vivo.

The glioblastoma tumor-immune microenvironment (TIME) is an immunosuppressive barrier to therapy that encumbers glioblastoma responses to immune checkpoint inhibition (ICI). Immunosuppressive cytokines, protumor macrophages and myeloid cells, and exhausted T cells are all hallmarks of the glioblastoma TIME. In this study, we integrated spatial and single-cell analyses of patient-matched human glioblastoma samples before and after ICI treatment with genetic, immunologic, single-cell, and pharmacologic studies in preclinical models to show that interleukin 6 (IL6) neutralization reprograms the glioblastoma TIME to sensitize mouse glioblastoma allografts to ICI and radiotherapy. Rare human glioblastomas that achieved clinical responses to ICI had lower pretreatment IL6 levels compared with glioblastomas that did not respond to ICI. Diverse immunostimulatory gene therapies suppressed local IL6 levels in mouse glioblastoma allografts, and IL6 from glioblastoma cells and the tumor microenvironment was associated with reduced survival in preclinical models and in patients. IL6 blockade with a neutralizing antibody transiently sensitized mouse glioblastoma allografts to ICI by decreasing immunosuppressive regulatory T cells and increasing MHCII+ monocytes, CD103+ migratory dendritic cells (DC), CD11b+ conventional DCs, and effector CD8+ T cells. IL6 blockade plus ICI sensitized mouse glioblastoma allografts to immunostimulatory ablative radiotherapy. Together, these data suggest that IL6 signaling contributes to ICI resistance in glioblastoma and provides a combination treatment strategy that could be used for patients. Integration of spatial and single-cell analyses of glioblastoma samples with immunologic, single-cell, and pharmacologic studies showed that IL6 neutralization reprograms the immune microenvironment to sensitize glioblastomas to ICI and radiotherapy.

Medulloblastoma (MB) is the most common malignant pediatric brain tumor. The non-WNT/non-SHH subtypes (Groups 3 and 4) exhibit high genetic heterogeneity and limited therapeutic options. Kinesin family member 14 (KIF14) is selectively expressed during nervous system development. However, its role in non-WNT/non-SHH MB remains poorly understood. In this study, analysis of the GEO database revealed that high KIF14 expression correlates with poor prognosis in patients with non-WNT/non-SHH MB. Silencing KIF14 in MB cells impaired cytokinesis, leading to p53-mediated cell cycle arrest and apoptosis. Mechanistically, KIF14 knockdown disrupted the nucleocytoplasmic shuttling of the p53-binding protein 53BP1, resulting in its nuclear accumulation. This shift inhibited homologous recombination (HR)-mediated repair of DNA double-stranded breaks (DSBs), thereby enhancing cisplatin-induced DNA damage. In vitro experiments and intracranial orthotopic xenograft models confirmed that KIF14 silencing potentiates the antitumor efficacy of cisplatin treatment. Overall, these findings highlight the critical role of KIF14 in promoting cell proliferation and DNA repair in non-WNT/non-SHH MB and suggest that targeting KIF14 may represent a novel therapeutic strategy to enhance the efficacy of DNA-damaging agents in this challenging MB subtype.

Glioblastoma (GBM) is the most common type of primary malignant brain tumor, characterized by a poor prognosis, high recurrence rate, and elevated mortality. In recent years, gene-targeted therapies leveraging small molecule compounds have gained momentum as a promising avenue for GBM intervention. Myoferlin (MYOF), a type II membrane protein of the Ferlin family, has emerged as a key regulator of membrane dynamics-governing processes such as vesicular trafficking, endocytosis, and membrane repair. In this study, we explore the previously uncharted role of MYOF in GBM progression and its potential as a diagnostic and therapeutic target. Our data reveal that silencing MYOF markedly suppresses glioma growth both in vitro and in vivo. Mechanistically, MYOF knockdown disrupts the nuclear translocation of phosphorylated STAT3 (P-STAT3), a critical oncogenic signaling event. Notably, we identified Entacapone (ENT), a small molecule capable of targeting MYOF, which significantly impedes glioma development across experimental models. These findings position MYOF as a novel molecular lever in GBM pathogenesis and highlight ENT as a potential therapeutic agent that exerts anti-glioma effects by blocking MYOF-mediated P-STAT3 nuclear import.The diagram illustrates a novel regulatory mechanism of IL-6/STAT3 signaling involving MYOF. Upon IL-6 stimulation, STAT3 undergoes phosphorylation. MYOF then binds to phosphorylated STAT3 and facilitates its translocation into the nucleus to regulate target gene expression. The compound ENT acts as a targeted inhibitor of this process by binding to MYOF, thereby blocking the nuclear transport of phosphorylated STAT3 and suppressing downstream signaling.

In cancer cells, pyruvate generated through aerobic glycolysis is preferentially converted into lactate. Here, we examined the impact of aerobic glycolysis on innate immune regulation within the tumor microenvironment. We identified lactate as a potent suppressor of stimulator of interferon genes (STING)-mediated innate immune signaling. Lactate directly bound the cyclic GMP-AMP (cGAMP)-binding domain of STING, thereby inhibiting cGAMP binding, STING activation, and interferon regulatory factor 3 (IRF3)-dependent cytokine expression. Mechanistically, activation of epidermal growth factor receptor (EGFR) promoted phosphorylation of lactate dehydrogenase A (LDHA) by pyruvate kinase M2 (PKM2) at serine 161 (S161). This increased LDHA activity and lactate production, which suppressed immune cell infiltration and promoted tumor growth. Pharmacological PKM2 inhibition relieved lactate-mediated STING suppression, reduced tumor immune evasion, and synergized with anti-PD-1 therapy. In human glioblastoma, elevated LDHA S161 phosphorylation correlated with reduced STING activation, diminished cytotoxic immune cell infiltration, and poor survival. Thus, oncogenic signaling directs PKM2-generated pyruvate toward LDHA-mediated lactate production, which directly inhibits STING to silence innate immune activation.

IMC-C103C is an ImmTAC bispecific (MAGE-A4×CD3) T cell engager targeting a peptide from the cancer-testis antigen MAGE-A4 presented by HLA-A*02:01. This phase 1/2 study in advanced solid tumors (IMC-C103C-101; NCT03973333) investigated the safety and preliminary anti-tumor activity of IMC-C103C. HLA-A*02:01+ patients with previously treated advanced solid tumors received IMC-C103C by weekly intravenous infusion, with step-up dosing introduced at higher dose levels. Dose-escalation decisions were guided by the mTPI-2 method. Key objectives included evaluating safety and preliminary anti-tumor activity; pharmacokinetics, pharmacodynamics, and biomarkers were also assessed. 68 patients were enrolled in 10 dose-escalation cohorts (n=56; 64% ovarian carcinoma (OC)) and one OC expansion cohort (n=12). In dose escalation, patients received doses ranging from 0.5 to 240 µg. The dose-limiting toxicity rate at the two highest dose levels (1/6 and 2/10) did not exceed the maximum tolerated dose. A lower-dose regimen of 15-45-140 µg, with clinical activity, and a more favorable tolerability profile, was selected as the expansion dose. There were no treatment-related discontinuations or deaths. The most common treatment-related adverse events were cytokine-mediated, including grade 1/2 cytokine release syndrome and associated symptoms. While MAGE-A4 expression was observed in most patient's tumors (71% positive), the level of expression was low (median H-score=16). Initial clinical and pharmacodynamic activity was observed at doses of 15 µg, with more consistent activity observed at doses ≥90 µg. At doses ≥90 µg, patients with MAGE-A4+ OC had deeper reductions in tumor size and circulating tumor DNA and trended to have longer overall survival than patients with MAGE-A4- OC. IMC-C103C was well tolerated at doses up to 140 µg, induced dose-dependent pharmacodynamic changes, and demonstrated clinical activity in multiple patients with MAGE-A4+ solid tumors. Clinical activity was dependent on MAGE-A4 expression; however, expression was especially low in the higher dosing cohorts, limiting adequate efficacy assessment. NCT03973333.

Long-term monitoring of female reproductive hormones, such as anti-Müllerian hormone (AMH) and luteinizing hormone (LH), is essential for fertility preservation. However, their concentrations in biofluids fluctuate over several orders of magnitude (sub-pg to ng/mL), posing a challenge for conventional assays in longitudinal studies. Here, we present an ultrasensitive electrochemical immunosensor that leverages dual-enhanced 3D gold nanostructure array (USE-IDEA) for quantifying reproductive hormones in undiluted saliva and plasma. The protein-covered high-curvature nanostructure imparts the sensor with abundant binding sites, not only boosting target-probe interactions, but facilitating enzyme-catalyzed proximity labeling at the immunointerface. This dual-enhancing assay achieves ultrahigh sensitivity up to 0.29 fg/mL (105-fold better than enzyme-linked immunosorbent assay), and a broad dynamic range (10-3 - 104 pg/mL) for AMH and LH. This sensitive assay enabled accurate discrimination between healthy individuals and patients with ovarian diseases (AUC = 1.0). Clinical monitoring revealed cyclic salivary LH fluctuations across menstrual phases and strong correlations with plasma levels for both AMH and LH (Pearson's r = 0.973 and 0.847, respectively). This ultrasensitive assay provides a promising tool for precise clinical monitoring and fertility counseling.

Inter-patient heterogeneity complicates predicting treatment response in ovarian cancer (OC). We developed OMICS-FUSE, an early-fusion multi-omics predictive model integrating proteomic, transcriptomic, and methylomic data from OC patients, evaluated across five machine learning algorithms with SHapley Additive exPlanations (SHAP) and experimental validation. The early-fusion Random Forest model achieved excellent predictive accuracy (AUC = 0.939, accuracy = 0.896, F1 = 0.939), with performance comparable to or surpassing that of the best-performing single-omics models. Nevertheless, the multi-omics framework yielded superior balance across accuracy and F1 score. SHAP analysis identified key determinants of treatment response, including CLEC2A, MYH4, and methylation of SYT12_1, with functional enrichment implicating immune regulation, metabolic pathways, and drug resistance signaling. Experimental validation confirmed six hub genes (CASP8, AQP8, CAV1, FN1, CREB1, KDR), exhibiting expression patterns associated with drug resistance, immune regulation, and prognosis. This multi-omics machine learning model enables robust, interpretable prediction, uncovering molecular signatures for therapeutic stratification and precision oncology in OC.

PARP inhibitors (PARPi) selectively target cancers with homologous recombination deficiency (HRD), yet emerging evidence suggests that additional determinants beyond BRCA/HRD status modulate PARPi response. Here, by integrating unbiased proteomic profiling with systematic drug-response phenotyping across 11 ovarian cancer cell lines, we identify MAP2K6 as a previously unrecognized regulator of PARP1 activation and an important determinant of PARP inhibitor sensitivity in BRCA-wild-type ovarian cancer. MAP2K6 promotes global PARP1-mediated PARylation and enhances cellular responsiveness to the PARP inhibitor olaparib both in vitro and in vivo. Mechanistically, MAP2K6 can phosphorylate and activate HMGA1, which in turn transcriptionally upregulates NAMPT, resulting in increased intracellular NAD+ levels, and sustained PARP1 activation. Genetic depletion of HMGA1 or NAMPT abolishes MAP2K6-driven PARylation and PARPi sensitization, whereas loss of MAP2K6 reduces NAD+ levels, attenuates PARP1 activation, and confers resistance to PARPi. We propose that MAP2K6 promotes PARP inhibitor sensitivity by coupling metabolic control of NAD+ supply to PARP1 activation, thereby potentially helping extend the therapeutic scope of PARP inhibition beyond homologous recombination-deficient ovarian cancer.

Polyploid giant cancer cells (PGCCs) are chemoresistant tumor cells associated with poor patient outcomes. PGCCs in tissue are identified by their extremely large sizes and the presence of massive or several nuclei. Although nuclear dysmorphia is also a common characteristic of tumor cells, little is known about the shape of nuclei in PGCCs. We show that the nuclear lamina in giant nucleated PGCCs is highly wrinkled across diverse cancer patient tissues. We investigated the cause of this hyperwrinkling in ovarian PGCCs in vitro. Laminar hyperwrinkling in PGCCs was independent of cell shape or cytoskeletal forces. Instead, measurements combined with computational modeling show that laminar hyperwrinkling is an intrinsic property of PGCCs, arising from a disproportionate amount of laminar excess area. PGCCs also displayed attenuated mechanosensitivity of cell spreading and YAP nuclear localization compared with control cells. Thus, laminar hyperwrinkling may disrupt PGCC mechanobiological pathways, slowing their proliferation.

Poly(ADP-ribose) polymerase inhibitors (PARPi) are a first-line treatment for epithelial ovarian cancer (EOC) patients, but the development of resistance limits long-term therapeutic efficacy. Tumor acidosis is a hallmark of the tumor microenvironment that has been shown to promote resistance to cancer therapies, suggesting that it may impact PARPi response. Here, we demonstrated that the acidic tumor microenvironment drives a p300-dependent mechanism of PARPi resistance in EOC. Pathologically acidic pH enhanced DNA damage repair, reduced PARPi-induced PARP1 trapping, and attenuated the anti-tumor efficacy of PARPi. A CRISPR-Cas9 screen identified p300 as a druggable mediator of acidosis-induced PARPi resistance. Mechanistically, acidic pH activated an ERK-p300-PARP1 signaling axis that acetylated PARP1 at lysine 505 (PARP1 K505Ac), thereby alleviating PARPi-mediated PARP1 trapping and DNA damage. Elevated PARP1 K505Ac was associated with clinical resistance to PARPi and poor overall survival. In patient-derived and syngeneic EOC models, pharmacologic inhibition of p300 synergized with PARPi to suppress tumor growth. Together, these findings identify p300 as a key mediator of acidosis-induced PARPi resistance and a promising therapeutic target to enhance PARPi efficacy.

High-grade serous (tubo-ovarian) carcinoma (HGSC) is the most lethal gynecologic malignancy and the most common form of ovarian cancer. Most patients (75%) are diagnosed at advanced disease stages, with widespread peritoneal dissemination leading to a poor prognosis. In this study, we characterized the interactions between fibroblasts and immune cells that facilitate the formation of a hospitable niche in metastatic peritoneal ovarian cancer. Analysis of the dynamic changes in cancer-associated fibroblasts (CAF) and immune cells isolated from patients with HGSC and mouse models revealed transcriptional reprogramming during HGSC progression. Functionally, peritoneal fibroblasts promoted myeloid cell recruitment and T-cell dysfunction. Distinct peritoneal metastatic niches featured reciprocal communication between fibroblasts and T cells. In human HGSC peritoneal metastases, the TIGIT-CD155 immune checkpoint signaling axis was upregulated in T cells and CAFs, respectively, and correlated with worse outcomes. This study implicates CAFs as key modulators of the immune microenvironment in intraperitoneal HGSC metastasis and provides potential avenues for therapeutic intervention of stromal-immune interactions to inhibit the metastatic spread of ovarian carcinoma. Fibroblast-T cell interactions shape a permissive metastatic niche to support intraperitoneal metastasis of ovarian cancer and offer a promising immunotherapeutic target to interrupt disease progression.

Lipid phosphates serve as signaling molecules involved in diverse cellular processes such as cell proliferation, migration, angiogenesis, inflammation, immunity and cancer progression. Phospholipid phosphatases (PLPPs) modulate these signals by catalyzing the dephosphorylation of lipid phosphates. Here, we report the cryo-EM structure of PLPP3, revealing a tetrameric assembly. PLPP3 contains six transmembrane helices (TMs) and an extracellular domain that contains two extracellular loops. TMs 1-4 create a hydrophobic cleft that holds the tails of a phospholipid while the extracellular domain forms a positively charged pocket to accommodate the polar head group. Two conserved catalytic histidine residues in this pocket coordinate a putative zinc ion previously identified as a PLPP3 inhibitor. Structural mapping of somatic mutations with functional analysis reveals that PLPP3 acts as a tumor suppressor in melanoma. Together, our findings provide critical insights into the structure, substrate engagement, inhibitory mechanism, and cancer-related function of PLPP3.

Tumor evolution is driven by various mutational processes, ranging from single-nucleotide variants (SNVs) to large structural variants (SVs) to dynamic shifts in DNA methylation. Current short-read sequencing methods struggle to accurately capture the full spectrum of these genomic and epigenomic alterations due to inherent technical limitations. To overcome that, here we introduce an approach to identify and analyze the genomic and epigenetic events in different stages of tumoral evolution from long-read sequencing of single-cell derived sublines. We then use it to profile 23 sublines of a mouse cutaneous melanoma cell line, characterized with distinct growth phenotypes and treatment responses. We develop a computational framework for harmonization and joint analysis of different variant types in the evolutionary context. Uniquely, our framework enables detection of recurrent amplifications of putative driver genes, generated by independent SVs across different lineages, suggesting parallel evolution. In addition, our approach revealed gradual and lineage-specific methylation changes associated with aggressive clonal phenotypes. We also show our set of phylogeny-constrained variant calls along with openly released sequencing data can be a valuable resource for the development and benchmarking of computational methods.

Gut microbiota can modulate cancer immunotherapy and enhance the efficacy of programmed cell death protein 1 (PD-1) blockade in tumors, yet the responsible microbes and underlying mechanisms remain incompletely understood. Publicly available anti-PD-1-treated melanoma microbiome cohorts were reanalyzed. Causal validation was performed using oral Blautia coccoides (B. coccoides) supplementation in B16-F10 melanoma-bearing mice. Untargeted and targeted liquid chromatography-tandem mass spectrometry-based metabolomics, CD8+ T-cell depletion, receptor identification and binding analyses, and downstream transcriptomic and biochemical assays were used to identify the key metabolite and investigate its mechanism of action. We identified Blautia as enriched in melanoma patients responding to immune checkpoint inhibitors, with higher abundance associated with non-progression. In melanoma-bearing mice, oral B. coccoides suppressed tumor growth and increased intratumoral effector CD8+ T cells. Metabolomic profiling identified acetate as a prominent B. coccoides-associated metabolite. Acetate enhanced CD8+ T-cell effector function. CD8+ T-cell depletion largely abrogated the antitumor effects of both B. coccoides and acetate, supporting a central role for CD8+ T cells. Mechanistically, these findings support a model in which acetate is associated with a TLR3-linked signaling pathway in CD8+ T cells, accompanied by PI3K/Akt activation and enhanced effector function. Notably, B. coccoides or acetate potentiated anti-PD-1 therapy in mouse melanoma models, supporting their potential as adjunctive strategies for melanoma immunotherapy. These findings support a model of an acetate-TLR3-linked PI3K/Akt signaling axis linking microbiota-associated metabolites to CD8+ T cell-mediated antitumor immunity. Importantly, our study highlights both B. coccoides and its derivative, acetate, as preclinical adjunctive candidates to potentiate anti-PD-1 efficacy in melanoma.

The tumour microenvironment comprising tumour cells, immune infiltrates, stromal components, and the extracellular matrix (ECM) plays a central role in cancer biology, modulating key hallmarks, such as proliferation, immune evasion, and metastasis. The ECM is a dynamic network of collagens, elastin, glycoproteins, and proteoglycans and emerging evidence highlights its role in tumour progression, with loss of matrix organisation altering tumour cell behaviour through both pro- and anti-tumourigenic mechanisms. Uveal melanoma (UM), the most common primary intraocular malignancy in adults, carries a poor prognosis due to its propensity for hepatic metastasis and limited therapeutic options. The uveal tract, comprising the choroid, ciliary body, and iris, relies on highly specialised ECM architectures, and we describe in this review how they support nutrient exchange, lens accommodation, and light regulation while mediating essential biochemical and biomechanical signalling. Understanding this baseline ECM is essential for interpreting the pathological matrix remodelling that occurs in UM. We describe marked compositional differences in ECM between low- and high-metastatic risk primary UM, revealing key mechanisms of tumour progression, including upregulation of structural collagens, fibronectin, laminins, and ECM-remodelling enzymes that activate oncogenic signalling pathways enhancing proliferation, invasion, and metastatic competence. We also explore the hepatic metastatic niche, where stellate cell-driven ECM remodelling promotes a fibrotic, immune-restrictive microenvironment that limits therapeutic efficacy. Finally, we discuss experimental constraints limiting ECM research and highlight advances in decellularised scaffolds, three-dimensional culture systems, and multi-omics technologies to dissect tumour-matrix interactions and uncover ECM-targeted therapeutic vulnerabilities with the potential to improve UM's poor clinical outcomes.

Oncogene-directed therapies can induce profound tumor regression in oncogene-addicted cancers, but their long-term benefit is often limited by resistance and relapse. Here we show that oncogene inactivation rapidly induces senescence and a pro-inflammatory senescence-associated secretory phenotype (SASP). In vivo, oncogene inactivation-induced senescence (OIIS) predisposes tumors to relapse, accompanied by polyploidy, chromosomal instability, acquisition of alternative oncogenic pathways including mouse double minute 2 homolog (Mdm2) upregulation, and tumor microenvironmental remodeling toward neovascularization and immunosuppression. Spectral flow cytometry reveals a shift from an immune-activated to an immunosuppressive milieu during relapse. OIIS features are also observed in human BRAFV600E melanoma cells treated with vemurafenib, supporting clinical relevance. Together, our findings establish OIIS as a double-edged process: it initially restrains tumor growth but simultaneously creates conditions that favor recurrence. By defining the genetic, metabolic and microenvironmental hallmarks of OIIS, our study highlights adaptations to oncogene deprivation that limit the durability of targeted therapies.

Therapeutic monoclonal antibodies (mAbs) are central to cancer treatment but often show incomplete efficacy. We show that transient pharmacologic inhibition of complex N-glycans in host cells ("glycoengineering") enhances the in vivo activity of multiple depleting mAbs, including mAbs already engineered for heightened potency. In preclinical models, glycoengineering improved α-CD20-mediated tumor clearance and survival through FcγRIIIa- and natural killer (NK) cell-dependent pathways. In B16-F10 melanoma, glycoengineering similarly enhanced anti-CD25 depletion of intratumoral regulatory T cells (Tregs). Notably, glycoengineering produced minimal changes in equilibrium binding affinity but markedly increased the mechanical durability of IgG-FcγRIIIa interactions under physiological shear stress. These results establish antibody effector function as a mechano-immunological process in which IgG-FcγR interactions can be tuned for resilience to physiological forces, thereby moving beyond the current affinity-centric paradigm in mAb engineering. Integrating mechanobiology into therapeutic development may enable mAbs optimized for the dynamic forces of human physiology, which provides a route to enhance next-generation immunotherapies.

Patient-reported outcome measures (PROMs) complement oncological endpoints by capturing what matters most to patients. The TrueNTH surgical collaboration presented 1-yr PROMs following robotic prostatectomy using accessible visual formats. We present 5-yr PROMs from the phase 3, international PACE-B trial, which randomised men with localised prostate cancer to stereotactic body radiotherapy (SBRT) or conventionally fractionated radiotherapy (CRT). PROMs are reported from baseline to 5 yr and presented using waffle charts to enable visual alignment with surgical outcomes. At 5 yr, urinary incontinence outcomes were favourable and comparable between SBRT and CRT. Leak-free rates were 64% (164/258) for SBRT and 69% (172/249) for CRT, pad-free rates were 91% (233/257) for SBRT and 90% (225/250) for CRT, and moderate or big urinary leakage problems were reported by only 6% (15/250) for SBRT and 4% (9/244) for CRT. Intercourse-adequate erections declined in both groups from baseline to 5 yr: from 35% (133/374) to 17% (43/250) for SBRT, and from 40% (157/391) to 20% (47/240) for CRT. Moderate or big sexual problems increased in both groups, from 24% (87/367) to 31% (74/242) for SBRT and from 22% (85/382) to 32% (77/238) for CRT. Bowel effects were low and comparable between groups, with moderate or big bowel problems reported by 2% (7/397) at baseline and 5% (12/265) for SBRT at 5 yr, and 2% (9/415) at baseline and 5% (13/253) for CRT at 5 yr. Stool incontinence as a moderate or big problem rose from <1% (1/374) to 2% (4/240) in the SBRT group and from <1% (1/395) to 3% (7/238) in the CRT group.

To present a summary of the 2026 version of the European Association of Urology (EAU)-European Association of Nuclear Medicine (EANM)-European Society for Radiotherapy & Oncology (ESTRO)-European Society of Urogenital Radiology (ESUR)-International Society of Urological Pathology (ISUP)-International Society of Geriatric Oncology (SIOG) Guidelines on the treatment of relapsing, metastatic hormone-sensitive and castration-resistant prostate cancer (PCa). The Panel performed a literature review of new data, covering the time frame between 2023 and 2025. The Guidelines were updated and a strength rating for each recommendation was added based on a systematic review of the evidence. Risk stratification for relapsing PCa after primary therapy may guide salvage therapy decisions. The range of treatment options for metastatic PCa has broadened, including androgen receptor pathway inhibitors (ARPI), metastasis-directed therapy, PARP inhibitors and their combinations, as well as PSMA-based therapy. The recommendations in the EAU Guidelines are based on clinical evidence and do not account for variations in cost, reimbursement structures, or resource availability across healthcare systems. The evidence in the field of relapsing, metastatic, and castration-resistant PCa is evolving rapidly. These PCa Guidelines reflect the multidisciplinary nature of PCa management. A full version is available from the EAU Guidelines Office or online (http://uroweb.org/guideline/ prostate-cancer/).

Cohesin organizes three-dimensional genome architecture by extruding DNA loops, but the mechanisms specifying its chromatin entry sites remain unclear. Here we show that the cohesin loader NIPBL is prepositioned at highly tissue-specific sites, largely distinct from the broadly conserved binding profiles of CTCF and cohesin. Motif and chromatin immunoprecipitation-sequencing analyses identify pioneer transcription factors, particularly FOXA1, as key mediators of the tissue-specific NIPBL recruitment. FOXA1 directs NIPBL to intratopologically associating domain (TAD) regions, enabling symmetric loop extrusion, whereas ETS factors guide NIPBL to TAD boundaries, supporting asymmetric extrusion. FOXA1 depletion disrupts NIPBL binding and impairs intra-TAD loops selectively, whereas NIPBL loss reduces both intra-TAD and boundary-to-boundary interactions. In prostate cancer, the recurrent FOXA1 R219S mutation redirects FOXA1-NIPBL to TAD boundaries, fostering a more insulated and aggressive genome. These findings reveal how pioneer factors orchestrate cohesin loading to shape tissue-specific three-dimensional genome organization and how this mechanism is co-opted in cancer.

Prostate specific membrane antigen (PSMA)-positive exosomes hold significant potential for the diagnosis and risk assessment of prostate cancer. However, accurate detection is severely hindered by their low abundance in blood and interference from similarly sized particles. In this study, we have developed a protospacer adjacent motif (PAM)-assembled clustered regularly interspaced short palindromic repeat (CRISPR)-Cas12a activation-based fluorescent and colorimetric dual-modal biosensor for the highly sensitive detection of PSMA-positive exosomes. In this work, two split strands respectively containing CD63 and PSMA aptamers are utilized to bind CD63 and PSMA on the exosome surface, forming a template that induces the opening of a hairpin DNA (HP DNA). A PAM site forms via hairpin-to-double-stranded structure transition. CRISPR-Cas12a recognizes PAM, activates to cleave FAM-labeled probes for fluorescence, while cerium dioxide nanozyme (CeO2 NZ) (with phosphatase-mimicking activity) hydrolyzes cleavage products. Hydroxyl radicals from hydrolysis oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to induce colorimetry. CRISPR-Cas12a-nanozyme dual recognition significantly improves prostate cancer exosome detection selectivity and sensitivity. Under optimized conditions, the limits of detection for the fluorescence and colorimetric modes reach 49 particles/μL and 63 particles/μL, respectively. By mutually validating dual detection modes, this biosensing technology accurately distinguishes prostate cancer patients from healthy individuals, holding great promise for early diagnosis.

Spatial transcriptomics is powerful but costly; hematoxylin and eosin (H&E) images are routine. We present Coladan-human3K, the largest human spatial transcriptomics resource (~ 3,000 profiles), and Coladan, a trimodal (image, language, spatial-gene) whole-slide framework predicting genome-wide genes per spot with calibrated uncertainty while preserving foundation-model representations. Across 32 Visium datasets, Coladan improves Pearson correlation from 0.230 to 0.431 (~ 1.9 ×), shows pathway-level enrichment consistency, and transfers zero-shot to VisiumHD and spot-level Xenium. Classification token (CLS) embedding-only perturbation performs on par with expression-based baselines, enabling image-only virtual perturbation without measured expression, illustrated on normal and cancer prostate sections for in-situ hypothesis generation.

Homeobox protein Hox-B13 (HOXB13) is an oncogenic transcription factor that is associated with prostate cancer risk. Unlike most transcription factors (TFs), HOXB13 is a methyl-plus TF and has been shown to be preferentially recruited to DNA prostate cancer risk loci. Here, we miniaturized HOXB13 to create metal-stapled mimetics that can target its cognate primary DNA-binding site in a sequence-specific, methyl-sensitive manner. Targeted profiling of 5-methylcytosine (5mC) recognition using V269X mimetic mutants revealed that leucine can further enhance both methyl sensitivity and methyl specificity through methyl-methyl contact with 5mC and decreased methyl-pi interactions with cytosine. Collectively, our study revealed novel insights into the molecular interactions of HOXB13 with methylated DNA and new tools that will provide a structural basis for future development of a new class of sequence-specific methylated DNA binders.

Endoscopic screening for gastric cancer reduces mortality, but the optimal upper age limit and modifying role of comorbidity remain unclear. This nationwide retrospective cohort study used the Korean National Health Insurance Service database linked to national mortality records. Individuals aged ≥40 years who underwent screening esophagogastroduodenoscopy between 2005 and 2010 were included and propensity score-matched to non-screened controls. The primary outcome was gastric cancer-specific mortality. Comorbidity was assessed using the Charlson Comorbidity Index (CCI). Hazard ratios (HRs) were estimated in the matched cohort. Among 555 904 propensity score-matched individuals (277 952 per group), gastric cancer-specific mortality HRs showed an age-dependent gradient: <1.0 at 50-59 years (HR 0.56, 95%CI 0.34-0.92), 60-64 (0.65, 0.40-1.07), and 65-69 (0.82, 0.58-1.17); close to 1.0 at 70-74 (0.99, 0.75-1.30) and 75-79 (0.92, 0.68-1.24); and elevated at ≥80 years (2.21, 1.49-3.27). Comorbidity further modified this association: HRs were <1.0 in individuals with CCI <3 (0.86, 0.74-1.01) but elevated in those with CCI ≥3 (1.88, 1.42-2.49). In age-CCI analyses, point estimates favored screening across age groups up to age 75-79 years among individuals with CCI 0, but HRs exceeded 1.0 in most age groups with CCI ≥3. Post-endoscopy respiratory or cerebrocardiovascular events increased with age and comorbidity. The mortality benefit of gastric cancer screening endoscopy diminished with advancing age and comorbidity burden, becoming questionable beyond age 70. At age ≥80 or in individuals with substantial comorbidity, potential harms may outweigh benefits.

Perioperative immunotherapy improves outcomes in locally advanced gastric or gastroesophageal junction cancer (GC/GEJC), but reliable predictive biomarkers remain elusive. In this biomarker-stratified, randomized, multicenter phase 2 Mountain-02 trail (NCT06374901), 136 patients with operable cT3-4aN + M0 GC/GEJC were enrolled and stratified by tumor-specific MHC class II (tsMHC-II) expression status and then randomized (1:1) to receive perioperative tislelizumab plus chemotherapy or chemotherapy alone. Among tsMHC-II-positive patients, adding tislelizumab to chemotherapy significantly increased the major pathological response (mPR) rate compared with chemotherapy alone (61.8% vs. 26.5%, p = 0.003), meeting the pre-specified primary endpoint. In contrast, no significant benefit was observed in the tsMHC-II-negative subgroup. Within the combination therapy arm, tsMHC-II-positive patients also achieved numerically higher mPR (61.8% vs. 23.5%, p = 0.001) and higher pathological complete response (pCR) rate (32.4% vs. 5.9%, p = 0.006) than tsMHC-II-negative patients. These findings support tsMHC-II as a promising predictive biomarker for perioperative immunotherapy and warrant further validation in larger studies.

Gastric cancer primarily originates from gastric stem/progenitor cells and is driven by somatic mutations. Although Helicobacter pylori is the main risk factor, how it drives malignancy is still not well understood. Analysis of single-cell RNA sequencing data reveals that human gastric cancer correlates with suppressed BMP signalling, a crucial niche signal for gastric stem cells, in stromal cells rather than epithelial cells. Genetic disruption of BMP signalling in Col1a2+ stromal cells or Acta2+ myocytes/pericytes alone, but not in gastric stem cells themselves, triggers mutations in gastric stem cells and initiates cancer development. Mechanistically, loss of BMP signalling increases stromal production of Wnt ligands, which dose-dependently drive transcription-replication collisions, R-loops, and DNA damage in gastric stem cells. The resulting DNA damage and carcinogenesis can be prevented by small-molecule inhibitors targeting Wnt pathway. Importantly, we identify inflammation as a key disruptor of stromal BMP signalling, as seen in patient samples with chronic atrophic gastritis and H. pylori-infected mouse gastric samples, which is associated with DNA damage. Together, these findings show how chronic inflammation derails niche signalling to drive stem cell mutations and gastric cancer and highlight promising avenues for early prevention.

MicroRNAs (miRNAs) are promising biomarkers for cancer diagnosis due to their stability in body fluids and disease-specific expression profiles. However, current detection methods suffer from limitations including cumbersome workflows, heavy instrumentation for signal readout, or vulnerability in minimizing instrumentation. To address these challenges, we describe a novel point-of-care miRNA detection platform executable with "off-the-shelf", personal glucose meter (PGM), termed 'KEY-FACT (Kinases Ensemble-driven glucose phosphorYlation upon Fuel-Aided CRISPR acTivation)'. Upon recognition of target miRNA, a fuel-assisted toehold-mediated strand displacement reactions liberate guide RNAs (gRNAs) to activate Cas13a to cleave a chimeric reporter probe, producing 2',3'-cyclic adenosine monophosphates (cAMP). Subsequent dephosphorylation and kinases ensemble-mediated phosphorylation/dephosphorylation cycles lead cAMP to consume a large amount of glucose. A user can immediately measure resulting glucose level change with PGM on the spot. This strategy allows sensitive, prompt detection of miR-135b, a gastric cancer (GC) biomarker, with a limit of detection (LOD) of 1.4 pM within 2 h. KEY-FACT is specific to the target miRNA and is applicable to body fluids such as human serum with dilution (95.2% < recovery rates <104.3%, coefficients of variation ≤13%). Owing to its simple probe design, KEY-FACT was readily expanded to detect another GC biomarker, miR-21, with comparable sensitivity (LOD = 1.5 pM). The proposed platform fulfills minimal instrumentation and thus enables cost-effective, field-deployable analysis, paving the way for practical, on-demand miRNA diagnostics.

Metabolites sculpt the immunosuppressive tumor microenvironment (TME) that facilitates immune evasion. As a crucial signaling lysophospholipid, lysophosphatidylserine (LysoPS) correlates with advanced disease stages in multiple tumor types. However, the mechanisms by which LysoPS drives gastric cancer peritoneal metastasis remain undefined. Single-cell transcriptomic profiling of primary tumors, normal peritoneum, and metastatic lesions delineated mechanisms underlying LysoPS-mediated tumor-associated macrophages (TAMs) reprogramming. Immunohistochemistry and multiplex immunofluorescence validated GPR34-high TAMs infiltration in peritoneal metastases. Functional validation was performed using molecular assays and in vivo models. LysoPS accumulated in ascites from patients with gastric cancer peritoneal metastasis, establishing an immunosuppressive TME that drove malignant progression. Using single-cell transcriptome sequencing, we identified a distinct subset of TAMs highly expressing GPR34 enriched in gastric cancer peritoneal metastases, which correlates with tumor progression and immune evasion. Mechanistically, LysoPS engagement of GPR34 activated ERK/c-Jun signaling, transcriptionally upregulating AXL and CD36 to enhance efferocytosis. This effect drove TAMs toward an immunosuppressive phenotype, characterized by enhanced interleukin-10 and transforming growth factor-β secretion. GPR34 inhibitor attenuated M2-like TAMs infiltration while bolstering cytotoxic T cells recruitment and curtailing programmed cell death protein 1 (PD-1)+T cells accumulation. Furthermore, combination with anti-PD-1 therapy synergistically suppressed tumor growth beyond monotherapy efficacy. LysoPS-GPR34 axis synergized with efferocytosis to amplify TAMs immunosuppressive properties, fostering an immune-evasive microenvironment; GPR34 inhibitor thus represents a promising strategy to potentiate PD-1 blockade efficacy in gastric cancer peritoneal metastasis.

Gastric cancer develops through a sequential carcinogenic process beginning with pyloric metaplasia, a key feature of which is the transdifferentiation of gastric chief cells into spasmolytic polypeptide-expressing metaplastic (SPEM) cells. We found that SOX9, a transcription factor, is highly expressed in SPEM cells in both human and murine metaplasia. We therefore investigated the impact of SOX9-mediated SPEM cell plasticity and function on metaplasia development and progression during gastric carcinogenesis. We utilized two Sox9 knock-out mouse models, GIFrtTA/+;TetO-CreTg/+; Sox9flox/flox (GCS) and GIFrtTA/+;TetO-CreTg/+;LSL-KrasG12DTg/+;Sox9flox/flox (GCKS) to examine the effects of SOX9 loss on metaplasia development following mucosal injury and carcinogenesis by immunofluorescence and single-cell RNA-sequencing. We also confirmed our findings in human patient tissue microarrays and utilized SPEM organoids to identify genes regulated by SOX9. Sox9 knock-out in chief cells resulted in the failure of metaplasia development in both acute and chronic injury as chief cells were unable to transdifferentiate into SPEM cells. The GCKS mouse failed to progress through carcinogenesis due to lack of a SPEM cell subpopulation responsible for metaplasia progression and fibroblast recruitment to gland bases. Instead, homeostatic gastric cell lineages were repopulated in the glands. We also identified a TOP2A-expressing SPEM cell subpopulation that contributes to metaplasia progression, and putative SOX9 downstream genes associated with lineage plasticity. These results demonstrate that SOX9 is a master regulator of SPEM cell lineage evolution and metaplasia progression. Therefore, SOX9-mediated SPEM cell plasticity is a central mechanism in carcinogenesis.

This trial shows that excellent outcomes can be achieved without routine renorrhaphy in selected patients undergoing purely off-clamp robotic partial nephrectomy. Beyond demonstrating noninferiority, it highlights the challenges of evaluating surgical innovation within conventional randomized trial frameworks.

This study was carried out to screen PANoptosis-related biomarker genes and the involved underlying mechanism in clear cell renal cell carcinoma (ccRCC). Differentially expressed (DE) PANoptosis-related genes were identified. Then prognostic biomarker genes were screened, and used to construct a prognostic model. The predictive ability of the model was explored by receiver operating characteristic (ROC) curves. Then the correlation between risk score with clinical features and immune cell infiltration were evaluated. The expression level and prognosis of gelsolin (GSN) were examined in datasets. Furthermore, in vivo and in vitro experiments were employed to investigate underlying mechanisms. ScRNA-seq analysis uncovered that PANoptosis scores varied significantly across 9 cell types in the tumor microenvironment. A total of 26 DE-PANoptosis-related genes were identified by intersecting DEGs and PANoptosis-related genes. Then 3 prognosis biomarker genes were screened and used to build the prognostic model, namely TLR3, GSN and TNFRSF1A. The risk score was associated with tumor grade, stage (I, II, III, IV), pathologic T-Stage, etc, and also significantly correlated with immune cells and expression of immune checkpoint proteins. In addition, GSN was upregulated in ccRCC, and high expression of GSN indicated a favorable OS. In vivo and in vitro experiments revealed that GSN liquid-liquid phase separation (LLPS) directly induces PANoptosis in ccRCC by inhibiting YAP-dependent glycolysis. In conclusion, using TLR3, GSN and TNFRSF1A, the established PANoptosis-related prognostic model was established and showed good predictive performance for predicting prognosis of ccRCC patients. GSN LLPS directly induces PANoptosis in ccRCC by inhibiting YAP-dependent glycolysis.

The global incidence and prevalence of kidney diseases continue to rise, posing a serious public health challenge. SIRT6 is an NAD⁺-dependent histone deacetylase with broad essential regulatory roles across various pathophysiological processes, including DNA repair, chromatin accessibility, telomere stability, and glycolipid metabolism. As an epigenetic regulator specifically expressed in kidney tissues, SIRT6 serves as a central mediator of kidney homeostasis. Notably, accumulating evidence has implicated aberrant SIRT6 expression in the onset and development of various kidney diseases, such as acute kidney injury, diabetic kidney disease, hypertensive nephropathy, renal fibrosis, and renal cell carcinoma. In the present review, we provide an overview of the sirtuin family, systematically characterize the enzymatic activities and critical biological functions of SIRT6, and discuss its molecular mechanisms across various kidney diseases, focusing on its cell type-specific functions. We further summarize the latest research advances in SIRT6-targeted modulators for improving kidney diseases and analyze the challenges associated with their clinical application. Overall, we highlight SIRT6 as a highly promising novel target in the treatment and prevention of kidney diseases, with strong potential for clinical translation.

Chromosomal instability (CIN), a hallmark of malignancy, remains poorly understood in clear cell renal cell carcinoma (ccRCC). Here, we identify spindle and kinetochore-associated protein 3 (SKA3) as a critical regulator of CIN in ccRCC. BAP1 loss-of-function mutations, prevalent in ccRCC, drive aberrant SKA3 overexpression. Mechanistically, BAP1 deubiquitinates nuclear receptor co-repressor 1 (NCOR1) to enhance its recruitment to the SKA3 promoter, thereby repressing SKA3 transcription. Additionally, BAP1-mediated histone H2AK119 deubiquitination at the TRIM25 promoter activates the expression of TRIM25, facilitating ubiquitin-proteasome-mediated degradation of SKA3. BAP1 deficiency disrupts both regulatory pathways, leading to aberrant accumulation of SKA3, which fuels CIN and correlates with metastasis progression and poor prognosis. In conclusion, our findings establish dysfunction of the BAP1-SKA3 axis as a molecular driver of CIN in ccRCC and suggest SKA3 as a potential therapeutic target for BAP1-mutant ccRCC.

Clear cell renal cell carcinoma (ccRCC) exhibits glycolytic addiction due to VHL mutation, making it vulnerable to metabolic intervention. While conventional inhibitors targeting core housekeeping glycolytic enzymes show robust antitumor efficacy, their clinical use is limited by on-target systemic toxicity. Here, we identify enolase 2 (ENO2) as an isoform-specific glycolytic vulnerability specific to VHL-deficient ccRCC. Although ENO2 is physiologically redundant (compensated by ENO1) in normal glycolysis, it becomes essential for sustaining ccRCC malignancy via VHL loss-HIF2α-driven glycolytic flux. Targeting ENO2 specifically suppressed glycolysis in VHL-deficient ccRCC cells, reducing their malignancy and enhancing their response to axitinib, doxorubicin, and gemcitabine. Importantly, pharmacological ENO2 blockade displayed superior tumor selectivity compared to traditional glycolysis inhibitors, effectively eliminating VHL-deficient ccRCC cells while sparing VHL-intact normal and malignant cells, despite causing slightly weaker glycolytic suppression. Mechanistic investigations revealed that ENO2 ablation inactivated up to ~78% of VHL loss-induced oncogenic effectors, including a pleiotropic oncoprotein MDK, through dual modulation of lactate metabolism and interferon signaling. Collectively, this work reveals ENO2 as a genotype-specific metabolic dependency in ccRCC, thereby enabling precision glycolytic therapy. Our work also suggests that targeting physiologically redundant metabolic isoenzymes may offer a precision medicine strategy for cancers with defined genetic alterations.

Neoadjuvant immunochemotherapy (nICT) has emerged as a promising neoadjuvant strategy for esophageal squamous cell carcinoma (ESCC). Identification of the factors affecting the responsiveness to nICT could help further improve treatment efficacy. Here, we performed single-cell analysis on 14 ESCC patients undergoing nICT and revealed tumor microenvironment (TME) features associated with differential treatment responses. Nonnegative matrix factorization (NMF) identified five coordinated cellular programs with distinct response associations. Specifically, the NMF3 program mainly comprising immunosuppressive cell subsets was enriched in the minimal or no pathological tumor regression (TRS3) group, in which regulatory T cells (Tregs) and dendritic cells (DCs) exhibited close correlation. In addition, elevated expression of mitochondrial transcription factor A (TFAM) in DCs was associated with increased Treg infiltration in the TRS3 group. A myeloid-specific Tfam knockout mouse model showed that TFAM deficiency reversed the immunosuppressive TME, inhibited tumor growth, and enhanced response to anti-PD-1 therapy in ESCC. Mechanistically, TFAM deficiency in DCs activated the STING-TBK1-IRF3 pathway, thereby promoting DC maturation to enhance anti-tumor immunity. Overall, this study characterized the TME in residual ESCC after nICT and revealed an association between elevated TFAM expression in DCs and poor responsiveness to nICT. These findings indicate the critical role of TFAM deficiency in DCs in activating anti-tumor immunity, highlighting the potential of targeting TFAM to improve the efficacy of immunotherapy and optimize therapeutic strategies.

Neoadjuvant therapy (NAT) followed by radical surgery is the standard treatment for locally advanced esophageal squamous cell carcinoma (LA-ESCC), yet many patients fail to achieve pathologic complete response (pCR). Predictive biomarkers and biological mechanisms underlying NAT resistance remain elusive. We aimed to address this challenge through longitudinal multi-omics profiling of patient plasma during NAT. We performed longitudinal plasma proteomic and metabolomic analyses in a discovery cohort of patients with LA-ESCC (n = 40) receiving neoadjuvant chemotherapy (nCT) or neoadjuvant immunochemotherapy (nICT) to reveal molecular features relevant to treatment response. Predictive biomarkers for pCR were assessed in plasma samples from an independent validation cohort (n = 93) by ELISA. Mechanistic studies were conducted through in vitro cell co-culture, in vivo mouse experiments, and analysis of ESCC tissue samples from another validation cohort (n = 33). Multi-omics analysis revealed distinct molecular profiles between treatment responders and non-responders. Metabolomic profiling revealed dynamic treatment-associated changes in amino acid and lipid metabolism during NAT, and proteomic profiling highlighted baseline differences related to neutrophil activity and inflammatory responses. DEFA1 was validated as an unfavorable predictive biomarker for achieving pCR after nICT. Mechanistic studies demonstrated that DEFA1 fosters an immunosuppressive microenvironment by promoting myeloid-derived suppressor cell (MDSC) differentiation and impairing CD8+ T cell cytotoxicity. This study delineates the longitudinal plasma multi-omic landscape associated with NAT response in LA-ESCC and identifies DEFA1 as a predictive biomarker and potential therapeutic target for overcoming immunotherapy resistance.

Pathological complete response (pCR) following neoadjuvant therapy is associated with favorable prognosis in esophageal adenocarcinoma. However, whether long-term outcomes differ by neoadjuvant modality among patients achieving pCR remains uncertain. To compare survival outcomes and recurrence patterns in patients with esophageal or gastroesophageal junction adenocarcinoma achieving pCR after perioperative 5-fluorouracil, leucovorin, oxaliplatin, and docetaxel (FLOT) chemotherapy vs the Chemoradiotherapy for Oesophageal Cancer Followed by Surgery Study protocol (CROSS). This cohort study was conducted at 14 high-volume European esophagogastric cancer centers between January 2018 and December 2024 with follow-up from surgery to death or last contact among patients with esophageal or junctional adenocarcinoma treated with neoadjuvant therapy followed by curative esophagectomy. Patients with squamous carcinoma, noncurative resection, incomplete pathological response, or 90-day mortality were excluded. Neoadjuvant perioperative FLOT chemotherapy or CROSS chemoradiotherapy followed by curative-intent esophagectomy. The primary outcome was overall survival. Secondary outcomes included disease-free survival, recurrence patterns, and nodal stage-stratified survival. Survival was estimated using Kaplan-Meier methods and multivariable Cox proportional hazards models. Among 2717 eligible patients, 297 achieved pCR and were analyzed (mean [SD] age, 64.4 [9.7] years; 256 [79.5%] men). Of these, 150 received CROSS and 147 FLOT. FLOT patients were younger and more frequently clinically node-positive at baseline, while postoperative outcomes were similar. On unadjusted analysis, FLOT was associated with better overall survival (hazard ratio [HR], 0.32; 95% CI, 0.17-0.60; P < .001) and disease-free survival (HR, 0.32; 95% CI, 0.19-0.55; P < .001). After adjustment, FLOT remained independently associated with better overall survival (HR, 0.34; 95% CI, 0.16-0.69; P = .003) and disease-free survival (HR, 0.35; 95% CI, 0.19-0.62; P < .001). Recurrence occurred in 13 of 147 patients receiving FLOT (8.8%) compared with 37 of 150 receiving CROSS (24.7%). Distant metastases predominated in the CROSS group (27 of 37 recurrences [70.3%]), whereas recurrence patterns were more heterogeneous after FLOT (locoregional, 2 of 13 recurrences [15.4%]; distant, 2 of 13 recurrences [38.5%]; both, 6 of 13 recurrences [30.8%]). The median (IQR) time to recurrence was longer after FLOT than CROSS (16.8 [12.3-21.5] vs 12.8 [7.1-26.7] months). In subgroup analyses, FLOT was associated with better survival in cN0 or 1 disease (HR, 0.32; 95% CI, 0.15-0.67; P = .002). In this cohort study, perioperative FLOT in patients achieving pCR for esophageal adenocarcinoma was associated with superior survival, lower recurrence risk, and later recurrence compared with CROSS. These findings suggest the prognostic significance of pCR may vary by neoadjuvant modality and should be interpreted in the context of treatment strategy.

Radioresistance remains a major barrier in esophageal squamous cell carcinoma (ESCC). This study demonstrates small nucleolar RNA SNORA23 as a pathogenic epigenetic driver of intrinsic radioresistance, with its overexpression tightly correlating with advanced T-stage, lymph node metastasis, and adverse clinical outcomes in a chemoradiotherapy cohort. Mechanistically, SNORA23 directly binds the ARID domain of histone demethylase KDM5C via a structurally defined G11-Ser169 interface-obstructing KDM5C chromatin binding to derepress DNA repair scaffolding gene SFPQ. This chromatin reprogramming enables SFPQ-facilitated recruitment of RAD51 and Ku80 to DNA double-strand breaks, accelerating homologous recombination (HR) and non-homologous end joining (NHEJ). SFPQ high expression recapitulates SNORA23-mediated repair enhancement and reduced radiosensitivity. Therapeutically, SNORA23-targeting antisense oligonucleotides (ASOs) disrupt this axis, impair DNA repair, and synergize with radiotherapy to suppress tumor growth and prolong survival in vivo without evident systemic toxicity These findings define a snoRNA-chromatin-repair axis driving therapeutic resistance and support SNORA23 inhibition as a promising strategy to overcome radioresistance in ESCC.

Emerging evidence implicates human endogenous retroviruses (HERV) in cellular senescence and stemness suppression, suggesting that they may also play a role in tumor cell senescence. In this study, we identified the histone methyltransferase disruptor of telomeric silencing 1-like (DOT1L) as a key epigenetic repressor of HERV-K in adenocarcinoma of the esophagogastric junction (AEG). Comparison of the expression of HERV and epigenetic regulators in AEG using two independent datasets revealed an inverse correlation between DOT1L and HERVs, and DOT1L was also overexpressed in AEG and correlated with poor clinical prognosis. Pharmacologic inhibition of DOT1L in vitro and in vivo diminished H3K79 methylation, reactivated HERV-K expression, and triggered STING-dependent innate immune signaling, thereby inducing tumor cell senescence and conferring potent antitumor effects. By promoting the assembly and secretion of HERV-K-derived retrovirus-like particles (RVLP), DOT1L inhibition propagated senescence to neighboring tumor cells via STING pathway activation. Together, this study not only establishes DOT1L as a druggable epigenetic target in AEG but also proposes a therapeutic strategy that leverages HERV-K and RVLPs to drive tumor cell senescence and intercellular senescence transmission for antitumor therapy. Pharmacological DOT1L inhibition not only reactivates HERV-K to trigger autonomous senescence through a STING-mediated immune response but also promotes formation of HERV-K-derived retrovirus-like particles that propagate senescence, suppressing gastrointestinal cancer progression.

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

The initiation and progression of cancer are driven by the dynamic interplay between somatic mutations and the tumor microenvironment. Identifying core cell populations driving malignant transformation and understanding the intercellular interactions within the tumor microenvironment are, therefore, crucial for early diagnosis and effective treatment. Here, we construct a spatiotemporal atlas of oral squamous cell carcinoma (OSCC) progression by integrating multi-omics approaches. Our analysis reveals that the critical driver gene PLAU activates the TGF-β pathway via ITGB1, thereby promoting the conversion of fibroblasts into the COL11A1⁺ fibroblast (COL11A1⁺ Fib). These specialized fibroblasts remodel the extracellular matrix (ECM), collectively establishing an immunosuppressive niche composed of malignant epithelial cells, COL11A1+ Fib, and regulatory T cells (Tregs). Importantly, targeting COL11A1+ Fib alleviates this immunosuppressive niche and curbs OSCC progression. Our findings underscore the potential of COL11A1+ Fib as a predictive biomarker, especially in combination with immunotherapy. Collectively, this work demonstrates that the PLAU-ITGB1-TGF-β axis drives the formation of an Epi-COL11A1+ Fib-Treg immunosuppressive niche that fuels OSCC malignancy.

Head and neck squamous cell carcinoma (HNSCC) is often diagnosed at advanced stages, resulting in poor clinical outcomes. Ferroptosis resistance presents a major challenge in the treatment of HNSCC, highlighting the need to elucidate the mechanisms that enable HNSCC cells to evade ferroptosis. In this study, we conducted a genome-wide CRISPR-Cas9 knockout screen and identified trafficking protein particle complex subunit 4 (TRAPPC4) as a key regulator of ferroptosis resistance in HNSCC. Across a comprehensive set of experimental models, including HNSCC cell lines, patient-derived organoids, cell-derived xenografts, patient-derived xenografts, Trappc4 conditional knockout mice, and lymph node and lung metastasis models, TRAPPC4 promoted tumor progression by inhibiting ferroptosis. Mechanistically, TRAPPC4 decreased chromatin accessibility at a distal regulatory element upstream of TRIM55, thereby limiting FOS-dependent transcription. This repression reduced TRIM55-mediated GPX4 ubiquitination and degradation, resulting in GPX4 stabilization and ferroptosis resistance. Structure-based high-throughput virtual screening identified pitavastatin (PTV) calcium as a TRAPPC4-binding compound that promoted TRAPPC4 degradation. Notably, PTV calcium synergized with the ferroptosis inducer RSL3 to enhance ferroptotic activity and suppress HNSCC progression. These findings delineate a TRAPPC4-FOS-TRIM55-GPX4 signaling axis that drives ferroptosis resistance and tumor progression and highlight TRAPPC4 as a promising therapeutic target for ferroptosis-based intervention in HNSCC. TRAPPC4 enables head and neck squamous cell carcinoma to resist ferroptosis by regulating TRIM55-mediated GPX4 degradation, providing a potential therapeutic target to inhibit cancer progression.

Head and neck squamous cell carcinomas (HNSCC) are preceded by potentially malignant precursor lesions, highlighting the need for strategies to discern which precancerous lesions harbor a high risk for malignant transformation. Here, we developed a unique genetically engineered mouse model (GEMM) by inducing oral epithelial cell-specific amplification of Sox2, which recapitulates the malignant transformation of epithelial dysplasia to HNSCC with high histologic and phenotypic resemblance to human tumors. SOX2 promoted the release of IL-1α and CCL2, expanding inflammatory monocytes. The recruited myeloid cells exhibited a distinct, early, high-risk signature of high levels of IL-1α, SLC2A1, SPP1, and low levels of type-I interferon (IFN-I) targets, and intralesional SLC2A1high myeloid cells were more suppressive than their SLC2A1low counterparts. Brief priming of myeloid cells with IL-1α desensitized them to STING agonists and enhanced the suppressive effects of myeloid-derived suppressor cells on T-cell activation. Mechanistically, IL-1 activation repressed the expression of DHHC3/7 enzymes that palmitoylate STING. Early blockade of IL-1 signaling using pharmacologic and genetic approaches similarly reduced the most suppressive subsets of myeloid cells, promoted immune surveillance against the progression of epithelial dysplasia, and extended survival. This work establishes a central high-risk myeloid cell transcriptomic program, characterized by elevated levels of IL-1α, SLC2A1, and SPP1, that leads to irreversible immune escape upon transformation of precancerous epithelial lesions.

Neoadjuvant immunochemotherapy (nICT), defined in this study as PD-L1 blockade combined with cisplatin-based chemotherapy, provides substantial clinical benefits in muscle-invasive bladder cancer (MIBC). However, the tumor immune microenvironment (TIME) is highly heterogeneous, resulting in variable patient responses and persistent therapy resistance. This study investigates key immune cell subsets, intercellular communication networks, and spatial distributions within the TIME of MIBC, with a focus on cellular and spatial features associated with nICT response. Patients were classified into responder and non-responder groups according to RECIST 1.1-based radiological evaluation. Responders were defined as patients achieving complete or partial response, whereas non-responders were defined as patients with stable or progressive disease after adequate nICT exposure and evaluable imaging. Representative computed tomography images and fresh tissue samples were collected from four no-treatment controls (NT), three nICT responders (R), and three nICT non-responders (NR). Samples underwent single-cell RNA sequencing and spatial transcriptomics sequencing. Cell populations were annotated to assess infiltration abundance, cell-cell communication networks, and pseudotime trajectories. Resistance-associated cell subsets and their spatial niches were identified. Following quality control, the tumor microenvironment (TME) was classified into ten major cell subsets based on canonical markers. Cross-sectional group comparative analyses showed significant remodeling of the TME in the NR and R groups. B-cell infiltration was elevated in the NR group, whereas macrophages, fibroblasts, and mast cells were enriched in the R group. Functional alterations in activated CD4 + T cells and impaired differentiation of naive CD8 + T cells were identified as key drivers of acquired therapy resistance. Accumulation of macrophage-derived CXCL8 was also observed as a resistance driver. Cancer-associated fibroblasts (CAFs) served as a physical barrier, contributing to the establishment of an immunosuppressive TME. Notably, strong spatial co-localization between macrophage-derived CXCL8 and CAFs was observed in NR tumors, suggesting a cooperative role in mediating resistance to nICT. Further cell-cell communication analysis using spatial transcriptomics indicated that SPP1 signaling from macrophages (CXCL8) to CAFs contributes to TME reprogramming and resistance to nICT. The integrated analysis identifies distinct immunological features that underlie differential nICT responses in MIBC. These findings provide a theoretical basis for optimizing personalized neoadjuvant therapy strategies.

Bladder cancer is frequent and highly recurrent. Despite recent advances, knowledge gaps remain in molecular mechanisms underlying disease progression. In this study, we apply integrated multi-omic analyses to a cohort of 48 bladder cancer patients to comprehensively profile genetic, epigenetic, transcriptomic, and spatial features. Combining cell-free DNA sequencing, long-read tumor DNA-sequencing, RNA-sequencing, and spatial transcriptomics, we explore molecular alterations driving bladder cancer. We find frequent somatic LINE-1 (L1) insertions, and show that these L1 insertions are active and occur early in bladder cancer development. We link somatic L1 insertion with downstream genomic rearrangements and chromosomal instability, including increased structural variant and extrachromosomal DNA (ecDNA) counts in patients with high L1 counts. We identify variable ecDNA enrichment across tissue architecture, with highest enrichment overlapping differential expression of APOBEC3B and immune response pathways. In summary, our results support a model whereby L1 retrotransposition triggers downstream genomic instability and viral mimicry response.

This publication represents a summary of the updated 2026 European Association of Urology (EAU) Guidelines for nonmuscle-invasive bladder cancer (NMIBC), TaT1 and carcinoma in situ (CIS). The information presented herein is limited to urothelial carcinoma, unless specified otherwise. The aim is to provide practical recommendations on the clinical management of NMIBC, with a focus on clinical presentation. For the 2026 guidelines on NMIBC, new and relevant evidence was identified, collated, and appraised via a structured assessment of the literature. Databases searched included MEDLINE, EMBASE, and the Cochrane Library. Recommendations within the guidelines were developed by the panel to prioritise clinically important care decisions. The strength of each recommendation was determined according to a balance between desirable and undesirable consequences of alternative management strategies, the quality of the evidence (including the certainty of estimates), and the nature and variability of patient values and preferences. Key recommendations emphasise the importance of thorough diagnosis, treatment, and follow-up for patients with NMIBC. The guidelines stress the importance of defining patients' risk stratification and treating them appropriately. Key updates in the 2026 NMIBC Guidelines summary include: the addition of two new tables addressing risk factors for bladder cancer and prognostic factors for progression in subtypes; the addition of two new tables summarising the treatment options for Bacillus Calmette-Guérin (BCG)-unresponsive tumours; inclusion of a new section addressing the addition of immune checkpoint inhibitors to BCG in selected high- and very high-risk NMIBC BCG-NAÏVE patients; a new table addressing the role of urinary markers in follow-up; and the addition of a new section on pragmatic de-intensification strategy for NMIBC. This overview of the 2026 EAU guidelines offers valuable insights into risk factors, diagnosis, classification, prognostic factors, treatment, and follow-up of NMIBC. They are designed for effective integration into clinical practice. The EAU has issued updated clinical practice guidelines on NMIBC. The guidelines provide recommendations for diagnosis, treatment, and follow-up, with a particular focus on quality of life for patients.

Cuproptosis has emerged as a promising anti-tumor strategy. However, the efficient accumulation of copper ions inside tumors and the copper homeostasis regulatory mechanisms in tumor cells remain challenges for cuproptosis-based cancer therapy. To address these obstacles, we developed a covalent organic framework (COF)-based engineered nanomotor CP-Motor@LF3, which achieved amplified cuproptosis through potent tumor penetration and efficient delivery of the Wnt/β-catenin inhibitor LF3. The engineered nanomotor consists of an Au hemispherical shell and spherical COF carrier co-loaded with copper peroxide (CuO2, CP) and LF3. The ordered hydrophobic porous structure endows the COF carrier with exceptional drug-loading capacity, achieving an LF3 loading amount 3.4-fold higher than mesoporous SiO2. Under 808 nm near-infrared irradiation, the Janus self-thermophoresis nanomotor achieves rapid and deep tumor penetration. CP decomposes in an acidic tumor microenvironment to release Cu2+ for cuproptosis and synergistic chemodynamic therapy. Concurrently, the controlled release of LF3 further amplified the cuproptosis and chemodynamic therapy through inhibiting the Wnt/β-catenin signaling pathway. Consequently, CP-Motor@LF3 exhibits potent anti-tumor activity against bladder cancer both in vitro and in vivo, with post-treatment tumor volumes being 8.67-fold smaller than the control group, demonstrating superior therapeutic efficacy and promising clinical potential.

G protein-coupled receptors (GPCRs) are targets for almost a third of all FDA-approved drugs. GPCRs are known to signal through both heterotrimeric G proteins and β-arrestins. Traditionally these pathways were viewed as largely separable, with G proteins primarily initiating downstream signaling while β-arrestins modulate receptor trafficking and desensitization in addition to regulating their own signaling events. Recent studies suggest an integrated role of G proteins and β-arrestins in GPCR signaling, however the cellular and biochemical requirements for G protein:β-arrestin interactions remain unclear. Here, we show that G proteins and β-arrestins can directly interact. Through utilization of β-arrestin-biased receptors and artificially enforced β-arrestin relocalization, we demonstrate that recruitment of β-arrestin to the plasma membrane is sufficient to interact with the G protein Gαi. Using purified proteins, we show that Gαi directly interacts with β-arrestin. In addition, we find that Gαi family members differ in their degree of association with β-arrestin, and that a large degree of this selectivity resides within the alpha helical domain of Gαi. These findings delineate the cellular and biochemical conditions that drive direct interactions between G proteins and β-arrestins and illuminate the molecular basis for how they interact.

A central challenge in nanomedicine is the scalable and precise engineering of nanocarriers that are simultaneously tunable, immune-compatible, and capable of encapsulating diverse bioactive cargos. Extracellular vesicles (EVs), including native red blood cell-derived EVs (RBCEVs), offer biocompatibility but remain limited by heterogeneous composition, low yield, poor scalability, and limited control in cargo loading. Here, we introduce a bottom-up strategy for developing engineered red blood cell extracellular vesicles (eRBCEVs) through microfluidic diffusional mixing of purified RBC lipids and components. Parametric optimization through multi-physics simulation enabled predictive control over flow rates, lipid concentration, and channel geometry. eRBCEVs exhibited comparable encapsulation efficiency across molecular cargos of distinct sizes and properties, including oligonucleotides, gold nanoparticles, hemoglobin, erythrocruorin (∼3.6 MDa), and full adeno-associated virus (AAV ∼25 nm) particles, validated through cryo-EM, chemical mapping, and high-resolution TIRFM. Surface conjugation with CD47 peptide was established for comparison with RBCEVs; the bio-conjugations with α-PD-L1 antibodies yielded functionalized vesicles with uptake in PD-L1-positive human tumor organoids. In vivo biodistribution and pharmacokinetic analysis in mice demonstrated sustained circulation and tissue distribution in the liver, spleen, and lungs. Neutrophil activation and macrophage uptake assays confirmed minimal immunogenicity compared to conventional particles and free-protein controls. Collectively, our platform enables the scalable synthesis of customizable eRBCEVs that preserve RBC lipid features, support cargo flexibility, and potential for scale-up. This work establishes eRBCEVs as a next-generation therapeutic platform for programmable, patient-specific delivery in gene therapy, immunomodulation, and translational nanomedicine.

Extracellular ATP (eATP) accumulates in the tumor microenvironment (TME), where its metabolite adenosine is implicated in promoting immunosuppressive pathways. We found that immunotherapy further increased eATP concentrations in the TME. eATP directly signaled through the purinergic receptor P2RY2 in tumor cells to suppress T cell function, independently of its metabolites. Mechanistically, eATP-P2RY2 signaling drove expression of cyclooxygenase (COX)-1 and COX-2 and downstream accumulation of prostaglandin E2 (PGE₂). Genetic deletion or pharmacologic inhibition of P2RY2 reduced baseline intratumoral PGE₂ and prevented therapy-induced PGE₂ accumulation, changed the TME immune infiltrate, restored antitumor T cell responses, and overcame resistance to immune checkpoint blockade, CAR-T, and TCR-T therapies. A P2RY2 antagonist monoclonal antibody recapitulated the in vivo therapeutic benefit and enhanced human autologous tumor-infiltrating lymphocyte responses against matched tumor cells ex vivo. Thus, persistent eATP-P2RY2 signaling in the TME drives a PGE₂-mediated immunosuppressive circuit that operates at baseline and is further amplified during immunotherapy, driving adaptive resistance.

Programmed death receptor 1 (PD-1) blockade produces high response rates in resectable and locally advanced cutaneous squamous cell carcinoma (CSCC), but how many doses are needed and whether surgery or radiation after immunotherapy (consolidation) adds benefit in deep responders (patients with substantial clinical responses) remain unclear. Observational inference is challenging because treatment decisions are response-guided, doses accumulate over time, and treatment selection depends on patient characteristics that also affect outcomes. We conducted a retrospective cohort study of 189 patients with resectable, borderline-resectable, locally advanced, or limited metastatic CSCC treated with immune checkpoint inhibitors as part of management (2019-2025). We summarized responses, treatment discontinuation patterns, and time-to-event outcomes. To evaluate dose-response relationships, we fit Bayesian regression models adjusting for prespecified baseline confounders. Directed acyclic graphs were used to formalize causal assumptions and identify minimally sufficient adjustment sets. For event-free survival (EFS), we used a prespecified landmark analysis among patients receiving ≥2 doses to reduce guarantee-time bias. Objective response occurred in 119/189 (63%), including 52/189 (27.5%) complete responses. Nearly half of patients received ≤2 doses (92/189, 48.7%). Among 50 complete responders managed without surgery, 1 recurrence was observed over a median follow-up of 25.4 months. Dose-response models showed a consistent but modest positive association between additional doses and response: in a linear model, each additional dose was associated with ~1.09 fold higher odds of response (posterior probability >95%). Flexible threshold models suggested early concentration of benefit, strongest at 2 versus 1 dose (93.6% posterior probability of benefit), with persistent uncertainty in effect magnitude (66% probability of ≥6 percentage-point absolute increase). In the EFS landmark modeling cohort (n=177), receipt of ≥3 versus 2 doses showed a 93% posterior probability of reduced event risk, but the 89% credible interval spanned the null (HR 0.61-1.01), indicating substantial uncertainty regarding incremental downstream benefit. In real-world CSCC, durable disease control frequently occurred after limited immunotherapy exposure, and clinical responders often did well without routine surgical consolidation. Although additional doses may modestly improve outcomes, observed gains appear concentrated early with uncertain incremental value beyond two doses. These patterns support conceptualizing treatment as frontline immunotherapy with response-guided subsequent treatment rather than fixed-duration neoadjuvant therapy.

T cell targeting of cancer cells alters the tumor antigen landscape in preclinical models. Here, we examined the impact of immunoediting on the antigenic landscape of primary, treatment-naive human tumors. Cutaneous squamous cell carcinoma tumors from immunocompetent and immunosuppressed patients revealed consistent tumor mutational signatures; however, high-immune-infiltrate tumors from immunocompetent patients had lower overall mutational burdens and lower clonal mutational burdens compared with low-infiltrate tumors from immunocompetent patients and tumors from immunosuppressed patients. The lower clonal mutational burden in high-immune-infiltrate tumors from immunocompetent patients persisted after accounting for tumor purity and growth rate. Predicted neoantigen: major histocompatibility complex (MHC) class I binding affinity decreased with increasing variant allele frequency, demonstrating restriction of mutations encoding MHC-binding neoantigens. Neoantigens with features shared with validated immunogenic neoantigens were decreased in clonal relative to subclonal cancer cell populations in high-immune-infiltrate tumors from immunocompetent patients. Thus, the immune system restricts cancer cells expressing immunogenic antigens from clonal populations in primary, treatment-naive human tumors.

Progression from differentiated thyroid cancer to anaplastic thyroid cancer (ATC) involves profound epithelial plasticity and remodeling of the tumor microenvironment (TME), but how BRAFV600E and RAS driver mutations shape these processes remains unclear. Here, we integrated single-nucleus RNA-seq, spatial transcriptomics, and bulk RNA-seq across BRAFV600E- and RAS-driven thyroid tumors to delineate mutation-specific progression trajectories. BRAFV600E-driven tumors exhibited a gradual dedifferentiation trajectory with immune pathway activation, whereas RAS-driven tumors displayed abrupt transitions characterized by aneuploidy, epithelial-mesenchymal transition, hypoxia, and extracellular matrix remodeling. Cancer-associated fibroblasts (CAFs) emerged as key regulators, with mutation-specific ligand-receptor interactions: integrin-based signaling predominated in BRAFV600E-mutant ATCs, while PLAU-PLAUR, TNFSF10-TNFRSF10B, and AREG-EGFR were additionally enriched in RAS-driven ATCs. These CAF-epithelial circuits were spatially validated and associated with poor prognosis. Together, our findings reveal mutation-dependent epithelial and TME dynamics associated with thyroid cancer dedifferentiation and highlight the potential importance of molecular-tailored approaches in the management of advanced thyroid cancer.

Dysregulation of Polycomb Repressive Complex 2 (PRC2) contributes to cancer. Of its catalytic subunits, Enhancer of Zeste (EZH) 1 and EZH2, EZH2 mutations are extensively studied, but the role of EZH1 in cancer remains largely unexplored. Here, we investigate the thyroid cancer-associated EZH1Q571R mutation and uncover a mechanism that extends beyond catalytic gain of function. Using biochemical, single-molecule, epigenomic, and transcriptomic analyses, we demonstrated that EZH1Q571R significantly enhances chromatin compaction and stimulates PRC2-EZH1 catalytic activity, thereby rewiring PRC2-chromatin interactions. This altered engagement enables PRC2 activity within H3K36me2-marked chromatin typically refractory to H3K27 methylation, leading to widespread epigenetic and transcriptional reprogramming. Notably, enhanced chromatin compaction is observed with EZH1Q571R but not with the corresponding EZH2Q570R mutation, indicating an EZH1-specific mechanism. Functionally, EZH1Q571R accelerated tumor growth in vivo and induced pronounced micronuclei formation ex vivo, reflecting aberrant chromatin compaction and genome instability. Together, our study demonstrates that EZH1Q571R promotes a permissive epigenomic landscape for the progression of follicular thyroid cancer.

Skin cancer remains a major global health concern, necessitating the development of innovative therapeutic strategies that enhance treatment efficacy while minimizing adverse effects. This review aims to explore the emerging role of herbal nanogels as a promising nanotechnology-based drug delivery system for skin cancer therapy. A comprehensive review of recent literature was conducted focusing on nanogel-based drug delivery systems, herbal bioactive compounds, and stimulus-responsive nanotechnology approaches for cancer treatment, particularly skin cancer. Nanogels, defined as crosslinked nanoscale polymeric networks, have gained significant attention due to their high drug-loading capacity, tunable physicochemical properties, and ability to respond to various internal and external stimuli. They can be synthesized using techniques such as emulsion methods, micro-molding, and photolithography, allowing precise control over size and morphology. Advanced stimulus-responsive nanogels, including tumor microenvironment-sensitive and ligand-targeted systems (e.g., LHRH-targeted nanogels), enable controlled and site-specific drug release. Recent developments in herbal nanogels integrate plant-derived bioactive compounds within nanogel matrices, enhancing solubility, stability, and therapeutic efficiency against skin cancer. Herbal nanogels represent a promising interdisciplinary approach combining nanotechnology and phytomedicine, offering targeted, less invasive, and potentially more effective treatment options for skin cancer. Despite significant progress, challenges such as clinical translation, large-scale production, and long-term safety evaluation remain. Continued research in smart and multifunctional nanogel systems may significantly advance future skin cancer therapies.

Neuroblastoma, a pediatric cancer derived from sympathetic ganglia of the peripheral nervous system, frequently metastasizes, driving poor outcomes. Primary neuroblastomas are well-characterized, but the mechanisms underlying metastasis remain poorly understood. Here, by using single-cell and spatial multiomics, we identified that lymph node metastases in high-risk neuroblastomas display distinctive cellular heterogeneity and plasticity, marked by mesenchymal-like and stem-like states and heightened epithelial-to-mesenchymal transition activity compared to primary adrenal tumors. In addition, compared to primary adrenal masses, the metastatic niche display increased immunosuppressive myeloid programs, heightened immune checkpoint signaling, and lymphocyte exhaustion, which are indicative of immune evasion and dysfunction. Notably, metastatic neuroblastomas show elevated eIF4F translation machinery and XPO1 levels. Dual inhibition of eIF4A and XPO1 synergistically halted tumor growth and prolonged survival in xenograft models. Together, our multiomics studies reveal the molecular and cellular plasticity that contributes to therapy resistance and highlight exploitable therapeutic vulnerabilities in high-risk metastatic neuroblastomas.

The nasopharynx constitutes a critical niche in the upper respiratory tract, harboring a diverse microbiota linked to nasopharyngeal carcinoma (NPC), the mechanistic roles of which remain poorly understood. Here, we established the Nasopharyngeal Mucosal and Tumor-resident Bacterial Catalog (NMTBC) that comprises 5311 bacterial isolates representing 127 species, with 1006 of them being fully sequenced and annotated, providing a comprehensive culturable resource facilitating mechanistic dissection of the microbiome-tumor interactions. With NMTBC, we uncovered a Fusobacterium-Prevotella mutualism and revealed heterotypic bacterium-bacterium interactions involving transcriptional reprogramming and metabolic cross-talk. Using single-bacterial transcriptomics, we mapped a high-resolution transcriptomic trajectory, showing the ability of a single strain to differentiate into functionally distinct subpopulations that cooperate to sustain mutualism. By analyzing a multicenter NPC cohort, we showed that Fusobacterium and Prevotella co-colonization in NPC tumors correlated with unfavorable clinical outcomes after conventional radiochemotherapy. Analysis of RNA-seq data from two previous phase 3 clinical trials showed that coenrichment of Fusobacterium-Prevotella predicted better response to anti-PD-1 immunotherapy, highlighting their important role in microbiota-mediated immunomodulation. Overall, this study establishes a comprehensive nasopharyngeal bacterial catalog through culturomics, which offers valuable insights into microbiome-derived biomarker discovery and immunotherapy patient stratification in clinical practice.

Photodynamic therapy (PDT) is effective for localized cancers, but its reliance on external light limits treatment depth and uniformity. Herein, we report a programmable supramolecular chemiluminescence (CL)-induced PDT platform that generates reactive oxygen species (ROS) in the absence of light irradiation and displays intrinsic cancer selectivity. This platform is constructed through the supramolecular interaction between enzyme-responsive chemiluminescent spiroadamantyl phenoxy-1,2-dioxetane probes (ADOP, ADOE, and ADOG) and luminescent iridium(III) complexes bearing a trimethyl-β-cyclodextrin (TMCD) host [Ir(N∧C)2(bpy-TMCD)](Cl) (HN∧C = 2-phenylpyridine (Hppy) (1a), 2-phenylquinoline (Hpq) (2a), and 2-(1-naphthyl)benzothiazole (Hbsn) (3a)). The chemiluminescent probes exhibited CL upon reaction with alkaline phosphatase (ALP), porcine liver esterase (PLE), and β-galactosidase (β-gal); while the iridium(III) complexes showed intense phosphorescence with efficient singlet oxygen production, distinct intracellular localization, and tunable (photo)cytotoxicity. In aqueous solutions, mixing the chemiluminescent probes with TMCD-tagged iridium(III) complexes afforded supramolecular adducts that, upon enzyme activation, underwent chemiluminescence resonance energy transfer (CRET) from the dioxetane donors to the iridium(III) acceptors. Remarkably, the ALP-responsive conjugate formed from probe ADOP and complex 3a (adduct ADOP-3a) selectively generated ROS and elicited potent cytotoxicity in cancerous HeLa cells and spheroids under light-free conditions, with apoptosis as the predominant cell death pathway, while remaining noncytotoxic in normal HEK293 cells. This modular platform couples supramolecular host-guest assembly, enzyme-specific activation, and CRET-induced ROS generation to overcome the limitations of conventional PDT, enabling cancer-selective, light-free therapy.

Olorofim, a novel dihydroorotate dehydrogenase inhibitor, may be efficacious in patients with disseminated coccidioidomycosis (DCM) who lack alternative treatment options. To evaluate olorofim effectiveness and adverse events in patients with DCM. Single-group, open-label, phase 2b study. (ClinicalTrials.gov: NCT03583164). Ten U.S. sites. Forty-one patients with DCM and limited or no treatment options. Patients received olorofim alone or in combination with ongoing standard of care during an 84-day main treatment phase. Extended treatment was offered to patients. Mycoses Study Group-European Organization for Research and Treatment of Cancer (MSG-EORTC) criteria for global response based on subcategories of clinical, radiologic, and mycologic response were adjudicated by an independent data review committee (DRC) at days 42 and 84 (main treatment phase). Because the slow pace of serologic improvement in DCM limits global response to stable at best, this article focuses on patient clinical responses. Treatment-emergent adverse events (TEAEs) were compiled for both treatment phases. Forty-one patients with DCM were enrolled from May 2019 to August 2022. Thirty-nine (95.1%) did not have immunosuppression. Central nervous system disease was present in 30 (73.2%) patients, and 13 (43.3%) had a ventriculoperitoneal shunt with or without an Ommaya reservoir. Clinical success as adjudicated by the DRC occurred in 31 of 41 patients (75.6% [95% CI, 59.7% to 87.6%]) at day 42 and 30 of 41 patients (73.2% [CI, 57.1% to 85.8%]) at day 84. The main TEAE was hepatic biochemistry elevation in 9 of 41 patients (21.9%), which was managed by liver enzyme monitoring and dose reduction or pause in 8 patients (19.5%) and drug discontinuation in 1 patient (2.4%). This was a single-group, open-label trial, but a randomized controlled trial would be preferable. Olorofim showed effectiveness in patients with DCM with limited or no therapeutic options. F2G, Ltd.

The prevalence and long-term prognostic significance of coronary atherosclerosis detected by coronary computed tomography angiography (CCTA) in young adults remain insufficiently defined. We aimed to evaluate the prevalence of coronary atherosclerosis and its association with long-term cardiovascular outcomes among young adults undergoing clinically indicated CCTA. We performed a retrospective cohort study of consecutive patients undergoing CCTA at two academic centres between 2006 and 2021. Patients with prior coronary artery disease, end-stage renal disease, or malignancy were excluded. Young adults were defined as men ≤50 years and women ≤60 years. Coronary artery disease (CAD) was classified as no CAD, non-obstructive plaque (1-49% stenosis), or obstructive CAD (≥50% stenosis). The primary outcome was a composite of cardiovascular death, non-fatal myocardial infarction, or ischaemic stroke. Associations between CAD severity and outcomes were assessed using cause-specific proportional hazards models with multivariable adjustment. Among 10,247 young adults (median age 47 years; 56% women), 29.0% had non-obstructive plaque and 8.7% had obstructive CAD. Over a median follow-up of 6.7 years (interquartile range 4.2-10.4), the risk of the primary outcome increased stepwise with increasing CAD severity. Compared with no CAD, obstructive CAD was associated with a more than two-fold higher risk of events [adjusted hazard ratio (HR) 2.6, 95% confidence interval (CI) 1.9-3.6], while non-obstructive CAD was associated with a modest, non-significant increase in risk (adjHR 1.2, 95% CI 0.9-1.7). Among patients with non-obstructive CAD, event rates increased with greater plaque extent. In routine clinical practice, approximately 40% of young adults undergoing CCTA demonstrate coronary atherosclerosis, which is independently associated with adverse long-term cardiovascular outcomes. These findings support the role of CCTA in early risk stratification and targeted preventive strategies among younger individuals.

Merkel cell carcinoma (MCC) recurs in 17-30% of patients with stage I-II disease, with local recurrence rates between 2-27%. Per established guidelines, conventional postoperative radiotherapy (cPORT; ∼50 Gray across 25 fractions) should be considered for localized MCC when risk factor(s) are present. cPORT can be morbid and logistically challenging. Single fraction 8 Gray radiotherapy (SFRT) has utility and low toxicity for palliation but remains underexplored postoperatively. Evaluate postoperative SFRT for stage I-II MCC, compared to observation and cPORT. Local recurrences were assessed for observation (n=108 patients), cPORT (n=156), or SFRT (n=43) groups. Observation patients had fewer per-patient risk factors (mean: 1.2 versus 1.6 for SFRT [p=0.001] and 1.6 for cPORT [p=0.020]). Despite this, 3-year local control was better for cPORT (97.4%; p<0.001) and SFRT (100%; p=0.012) groups than observation (91.0%), particularly in patients with narrow surgical margins (≤1 cm). Adverse events occurred in 21% of SFRT patients (all Grade 1). Single-center retrospective study. The SFRT group experienced improved local control relative to observation and had comparable local control to the cPORT group. For early-stage narrowly excised MCC, SFRT is feasible, well tolerated, and warrants exploration in a prospective multi-center study.

Behçet's disease (BD) is a multisystem inflammatory disorder with diverse phenotypes and incompletely defined immune mechanisms. This study aimed to map immune dysregulation in BD at high resolution, comparing active vs remission states and identifying pathways linked to clinical phenotypes. We performed single-cell RNA sequencing on 247,028 peripheral blood mononuclear cells from 34 patients with BD and 12 healthy controls. Transcriptomic profiling, differential gene expression, pathway enrichment analyses, and phenotype-stratified comparisons were used to delineate immune cell alterations associated with disease activity and clinical subtypes. All 3 monocyte subsets were markedly expanded in BD and demonstrated dominant interferon (IFN)-γ-associated activation, robust heat-shock responses, and enhanced antigen-presentation programmes. In active disease, monocytes exhibited pronounced type II IFN signatures, which reversed in remission alongside restoration of regulatory and metabolic pathways. Remission was instead characterised by increased expression of type I IFN-regulated genes and activation of serine protease inhibitor (SERPIN)-associated programmes linked to tissue stabilisation. Clinical phenotype stratification revealed distinct transcriptional signatures in peripheral blood monocytes, including enrichment of heat-shock and stress-response pathways in monocytes from patients with vascular BD and tumour necrosis factor/NF-κB-associated programmes in monocytes from patients with ocular BD. Patients without organ involvement demonstrated an increased type I IFN gene signature. This study provides a high-resolution immune atlas of BD, identifying monocyte-driven dysregulation as a central feature. Our findings map the immune heterogeneity of BD, identify activity- and phenotype-linked peripheral blood monocyte states, and suggest immune pathways suitable for targeted intervention.

Metabolic dysfunction-associated steatotic liver disease (MASLD) includes a spectrum of progressive liver conditions ranging from isolated steatosis to metabolic dysfunction-associated steatohepatitis (MASH), advanced fibrosis and cirrhosis. Currently, MASLD is the leading cause of chronic liver disease worldwide. MASLD is strongly associated with type 2 diabetes, cardiovascular disease, chronic kidney disease and certain extrahepatic cancers. MASLD shares a common pathogenesis with cardiometabolic diseases, especially type 2 diabetes, primarily driven by unhealthy dietary habits, dysfunctional adipose tissue, insulin resistance and low-grade inflammation. Substantial heterogeneity in the pathophysiology of MASLD may influence its rate of progression, its relationship with cardiometabolic diseases and its treatment response. In addition to lifestyle interventions, including a healthy low-energy diet and increased physical activity levels, pharmacological treatment of MASLD/MASH is recommended. For individuals with type 2 diabetes and MASLD/MASH, treatment should preferably include glucagon-like peptide-1 (GLP-1) receptor agonist-based therapies and sodium-glucose cotransporter 2 (SGLT2) inhibitors, which have been shown to improve MASLD/MASH and provide established cardiorenal benefits. In this narrative review, we assess the efficacy of these pharmacotherapies and discuss other treatment approaches for MASLD/MASH, with a focus on their metabolic benefits.

Evaluating soft tissue elasticity provides critical biomechanical insights essential for the precise characterization of various physiological and pathological conditions. Here, we present the clinical validation of a wireless, compact wearable system designed for direct, location-specific monitoring of the elastic modulus of the skin and underlying tissues via vibro-rotational biomechanical dynamics. Validated by computational models and experiments, the device uses a tunable skin interface to enable depth-controlled measurements across diverse anatomical sites. Two human subject studies, one involving patients with cancer-related lymphedema and the other involving patients with systemic scleroderma, yield data that correlate with standard clinical metrics. This technology offers the potential for longitudinal assessments in these and other contexts, in both clinical and home settings.

Steatotic liver disease (SLD) is a leading cause of chronic liver disease worldwide impacting more than 30% of the adult population. With the rise of obesity, metabolic syndrome and alcohol use disorder, the prevalence of the two main subtypes of SLD- metabolic dysfunction-associated steatotic liver disease (MASLD) and alcohol-associated liver disease (ALD)- continues to increase. There has concurrently been an acknowledgement of a third subtype of SLD- MetALD- wherein an individual has both components of metabolic dysfunction and regular alcohol intake. The public health significance of the SLD epidemic is substantial given risk for progression to cirrhosis, end-stage liver disease and development of hepatocellular carcinoma (HCC). Individuals with stage two fibrosis or above and those with active inflammation with hepatocyte injury are at highest risk for adverse liver-related outcomes and overall mortality. Current screening and risk stratification recommendations highlight the importance of identifying individuals at highest risk of clinical outcomes using non-invasive testing (NIT) as these individuals would benefit from liver-directed pharmacotherapy. Importantly, the performance of NITs can vary substantially based on NIT selected, cut-points applied and patient population evaluated. The evidence base for NIT performance is strongest in MASLD with comparatively limited data in ALD and emerging data in MetALD. This review discusses NIT diagnostic performance in SLD and their role in chronic disease management.

Severe cutaneous toxic effects from chemotherapy and radiation often require treatment interruptions. High-dose oral vitamin D (hdVD) has shown potential as a rapid immunomodulator in experimental models and small human studies, but robust clinical data in oncology populations remain limited. To evaluate the clinical response, safety profile, and time to improvement for hdVD in patients with toxic erythema of chemotherapy (TEC) and acute radiation dermatitis (ARD). This retrospective multicenter case series included 33 patients treated across 3 academic medical centers between December 2021 and January 2024. Eligible participants were those receiving hdVD (100 000 international units) for TEC or ARD with at least 10 days of follow-up. One or 2 oral doses of 100 000 international units of cholecalciferol or ergocalciferol. The primary outcomes were time to patient-reported symptomatic relief and clinician-assessed objective improvement in erythema (using a 5-point Likert scale). Secondary outcomes included changes in serum calcium and the ability to continue anticancer therapy. Among 33 patients (mean [SD] age, 60.9 [14.6] years; 19 [58%] female), 28 (85%) had TEC and 5 (15%) had ARD. Subjective symptom relief was reported by 26 of 30 patients (87%) within 10 days of treatment. The median (range) time to improvement was 5 (1-28) days overall and 3 (1-16) days for the inpatient subgroup. The mean (SD) Likert erythema score decreased from 4.36 (0.60) at baseline to 2.21 (1.36) by day 10. Patients with neutrophilic eccrine hidradenitis and Stevens-Johnson syndrome/toxic epidermal necrolysis-like subtypes showed the most rapid responses. No meaningful changes in serum calcium levels were observed, and 24 of the 33 patients (73%) continued anticancer therapy without interruption. No treatment-related adverse events were reported during the study. In this case series, hdVD was associated with rapid symptomatic and objective improvement in chemotherapy- and radiation-induced toxic effects of the skin without treatment-related adverse events. These findings support further study of hdVD as a supportive care approach in oncodermatology.

Biologic agents are commonly used with conventional immunosuppressants for treatment of immune-mediated dermatologic diseases, raising concerns about tuberculosis (TB) risk, particularly in high-burden geographic regions. Previous studies have shown inconsistent results across biologic classes, with most focusing on active TB and limited data on latent TB infection (LTBI) conversion. To evaluate the incidence of active TB and LTBI conversion in dermatologic patients receiving systemic therapy and to explore differences by biologic class and regional TB burden. PubMed/MEDLINE, Embase, Web of Science Core Collection, and the Cochrane Library were searched from database inception to October 1, 2025. Controlled vocabulary and free-text terms were used for dermatologic diseases, systemic therapies, and tuberculosis-related outcomes. Clinical trial registries and reference lists were also screened. Randomized clinical trials, cohort studies, case-control studies, and cross-sectional studies reporting active TB or LTBI conversion in dermatologic patients receiving systemic therapy were included. Two reviewers independently screened records and performed full-text assessment. Data extraction and risk-of-bias assessment (Newcastle-Ottawa Scale and Cochrane Risk of Bias tool) were conducted independently by 2 reviewers following PRISMA guidelines. Single-arm incidence rates were pooled using random-effects models with logit transformation. Prespecified subgroup analyses were conducted by biologic class and regional TB burden. Primary outcomes were incidence of LTBI conversion (among patients with negative baseline results of tuberculin skin test or interferon-gamma release assay) and active TB during follow-up. Of 4726 records identified, 31 studies comprising a total of 15 005 patients met inclusion criteria and were included in the analysis. The pooled incidence of LTBI conversion was 4.3%, highest with tumor necrosis factor inhibitors, followed by interleukin (IL)-17 and ustekinumab (IL-12/23 p40 inhibitor). The overall incidence of active TB was 1.0% and it was more frequent in high-burden regions. In this systematic review and meta-analysis, TB-related risk varied by biologic mechanism and epidemiologic context. Risk assessment and monitoring should integrate dermatologic treatment class and regional TB burden to guide clinical decision-making.

Cancer cells counteract oxidative stress through upregulation of antioxidant networks. Peroxiredoxin 3 (PRX3), a mitochondrial antioxidant enzyme, regulates reactive oxygen species homeostasis and promotes tumor cell survival. The natural compound thiostrepton (TS) covalently inhibits PRX3, disrupting redox balance and selectively induces tumor cell death. Mesothelioma, an aggressive malignancy, has limited therapeutic options, particularly in relapsed or refractory settings. Here, we demonstrate genetic deletion of PRX3 impairs mitochondrial bioenergetics and suppresses mesothelioma growth, while pharmacological inhibition of PRX3 with TS induces apoptosis in patient-derived mesothelioma explants. In a phase 1 trial treating patients with relapsed pleural mesothelioma and malignant pleural effusion (NCT05278975), weekly local intrapleural treatment with the TS formulated drug product RSO-021 at 90 mg is well tolerated leading to disease control in 67% of patients at 12 weeks and is associated with tumor reductions. Primary endpoints of safety, tolerability and dose finding were met, and secondary endpoints of pharmacokinetics, objective response rate, disease control rate, and progression free survival are explored. Genomic screening identified Solute Carrier Family 7 member 11 (SLC7A11) as a mediator of TS resistance, suggesting combined targeting may further enhance the pro-oxidant activity of RSO-021.

Micro- and nanoplastics (MNPs) are emerging risk factors for cardiovascular diseases. The present study aimed to evaluate the burden of MNPs in coronary blood across the spectrum of coronary artery disease (CAD), and their association with air pollution exposure and inflammation. Cross-sectional study, including 61 consecutive patients undergoing coronary angiography for suspected CAD, stratified into: ST-segment elevation myocardial infarction (STEMI, n = 19), chronic coronary syndromes (CCS, n = 20), and controls with normal coronary arteries (n = 22). MNPs were quantified in coronary and peripheral blood using pyrolysis-gas chromatography-mass spectrometry and laser direct infrared spectroscopy. Air pollution exposure data were collected on the day of the invasive procedure (acute exposure) and over the preceding 2 years (chronic exposure). MNPs were detected significantly more frequently in STEMI patients (84.2%) than in CCS (40%) and controls (31.8%) (P = .002), with higher concentration and greater polymer diversity [median of 3 polymers (interquartile range: 2-4), P < .001]. Polyethylene was the predominant polymer (97%). The same polymers were consistently identified in peripheral and coronary blood samples from individual patients, with the highest concentrations in coronary blood (P < .001). STEMI patients showed higher levels of interleukin-6 and tumour necrosis factor-α (P≤.006) and were exposed to higher levels of PM2.5 (P≤.012). MNP detection was more frequent among smokers and patients exposed to PM2.5 > 15 µg/m3 (P = .006), with all patients presenting both factors showing detectable MNPs (P < .001). In multivariable analysis, smoking history emerged as the only independent predictor of MNP presence (odds ratio 5.69, 95% confidence interval 1.33-26.63, P = .023). STEMI patients exhibited a greater burden of MNPs in coronary blood than CCS and controls. MNP detection frequently co-occurred with elevated inflammatory biomarkers, greater PM2.5 exposure, and smoking, suggesting a potential association between environmental exposure and CAD.

Cancer-associated thrombosis (CAT) is a major cause of morbidity and mortality in patients with gastrointestinal (GI) malignancies and often negatively impact anti-cancer treatment delivery. A prothrombotic risk markedly varies across primary sites, from pancreatic, gastric to colorectal cancers and is influenced by tumor-driven inflammation, disease stage, and exposure to contemporary systemic therapies, including newly, immune checkpoint inhibitors, targeted agents, and antibody-drug conjugates. Despite its clinical relevance, current risk assessment models, including the pivotal Khorana score (KS), show limited accuracy in GI tumors and do not adequately account for treatment-related or biology-driven variability. This review synthesizes current evidence on the incidence, mechanisms, and clinical determinants of CAT in GI cancers and discusses approaches to primary thromboprophylaxis and secondary prevention, with attention to bleeding risk and cardiovascular considerations. Emerging strategies such as molecular profiling, inflammatory biomarkers, and ctDNA-based assessment of minimal residual disease (MRD) show promise for improving individualized risk prediction and guiding more precise anticoagulation strategies. A precision-medicine framework integrating tumor biology, dynamic biomarkers, and treatment-specific cardiovascular risk is needed to optimize CAT management and to inform future trials designed to refine primary and secondary prevention across the spectrum of GI malignancies.

Elevated lipoprotein(a) [Lp(a)] levels are an established risk factor for atherosclerotic cardiovascular disease, but the association between Lp(a) and venous thromboembolism (VTE) remains unclear. Sex and hormonal status may modify the relationship between Lp(a) and VTE. The present study included participants from the UK Biobank with available baseline Lp(a) data. Individuals with a history of VTE or cancer, as well as those using anticoagulants, were excluded. Multivariable-adjusted Cox models were used to assess the association between Lp(a) levels ≥125 nmol/L and incident VTE in premenopausal women, postmenopausal women, and men. Subgroup analyses stratified premenopausal women by oral contraceptive (OCP) use and postmenopausal women by menopausal hormone therapy (MHT) use. Among 55 302 premenopausal women, 129 045 postmenopausal women, and 189 013 men, the proportions with Lp(a) ≥ 125 nmol/L were 14.0%, 19.0%, and 15.0%, respectively. Over a median (interquartile range) follow-up of 13.6 (12.9-14.4) years, 8186 VTE events occurred (cumulative incidence 2.2%). Lp(a) ≥ 125 nmol/L was associated with incident VTE in premenopausal women [adjusted hazard ratio (aHR) 1.32; 95% confidence interval (CI) 1.04-1.66; P = .02] but not in postmenopausal women (aHR 1.03; 95% CI 0.94-1.13; P = .47; Pinteraction = .03) or men (aHR 1.00; 95% CI 0.92-1.08; P = .94). OCP use did not modify the Lp(a)-VTE association among premenopausal women (Pinteraction = .61). However, among postmenopausal MHT users, Lp(a) ≥ 125 nmol/L was associated with higher VTE risk (aHR 1.48; 95% CI 1.03-2.12; P = .03; Pinteraction = .04). Elevated Lp(a) was associated with VTE in premenopausal women and in postmenopausal MHT users, suggesting that hormonal context may influence Lp(a)-associated thrombotic risk.

Aging is a biologically tractable process. Telomerase reverse transcriptase (TERT) has emerged as an upstream regulator coordinating several hallmarks of aging across preclinical models. Beyond maintaining telomeres, TERT influences mitochondrial health, epigenetic regulation, inflammation and stem cell function. Multiple translational strategies are being explored to modulate TERT. In mice and human cell models, restoration of physiological-range TERT expression characteristic of younger cells, or related telomere-focused interventions, has been associated with improvements in selected age-related phenotypes without a detectable increase in cancer. Simultaneously, human genetics links common variation in the TERT locus to increased risk of several cancers, underscoring the need for careful mechanistic and long-term safety evaluations. Together, mounting evidence indicates that TERT occupies an important position in aging biology with the potential to affect healthspan. This Perspective reviews current evidence for TERT's canonical and noncanonical roles and outlines a cautious therapeutic framework for evaluating TERT-directed geroprotective strategies.

Transcription blockage frequently occurs in tumor cells, and aberrant R-loop formation during this process drives genome instability. However, the regulation of R-loop homeostasis and its contribution to tumorigenesis remain to be investigated. Here, we report that the RNA exonuclease REXO4 resolves R-loops by 3'-5' exonucleolytic cleavage of the RNA strand within RNA-DNA hybrids. Accessible RNA ends, generated by endonucleases, are required for this process, and N6-methyladenosine (m6A) modification on RNA moieties promotes REXO4 localization and R-loop removal in human cells. REXO4 ablation-induced DNA damage stimulates an interferon response and tumor immune infiltration, suppressing mouse squamous cell carcinoma (SCC) progression. Importantly, inhibition of REXO4 potentiates the anti-tumor efficacy of PD-1 blockade against SCC by recruiting and activating CD8+ T cells. Thus, our study provides mechanistic insight into how m6A couples with an exonuclease in R-loop clearance and genome maintenance and uncovers a druggable epitranscriptional machinery that constrains the innate immune response and enables SCC immune evasion.

Adenomyosis (AM) is frequently associated with compromised embryo implantation. The leukemia inhibitory factor (LIF)/signal transducer and activator of transcription 3 (STAT3) signaling pathway is crucial for endometrial receptivity; however, its mechanistic role in AM-related implantation failure remains insufficiently elucidated. This study aimed to investigate the inactivation mechanism of the LIF/STAT3 pathway in AM and to evaluate a novel nanotherapeutic strategy for restoring implantation capacity. Clinical endometrial samples from AM patients and controls were analyzed. A three-dimensional (3D) co-culture system simulating the endometrial microenvironment was established. Interventions were performed using a tofacitinib-loaded biomimetic nanogel (Tofa-NG). Comprehensive analyses included bulk RNA Sequencing (RNA-seq), immunohistochemistry (IHC), RT-qPCR, proteomics, inflammatory cytokine profiling, and metabolomics to dissect the pathological links. The LIF/STAT3 signaling pathway was significantly downregulated in the endometrium of AM patients, correlating directly with implantation failure. Pathologically elevated inflammatory cytokines suppressed LIF expression via NF-κB pathway activation, exacerbating the inflammatory microenvironment. Metabolomic profiling revealed a strong association between LIF/STAT3 pathway inactivation and aberrant cellular energy metabolism. The engineered Tofa-NG facilitated targeted drug delivery, effectively mitigated local inflammation, and successfully reactivated the LIF/STAT3 pathway. Consequently, this intervention significantly improved the embryo implantation success rate in the experimental model. This study identifies the inactivation of the LIF/STAT3 pathway as a central mechanism underlying embryo implantation defects in AM, intricately linked to chronic inflammation and metabolic dysregulation. The Tofa-NG strategy demonstrates promising therapeutic potential by rectifying the signaling deficit and ameliorating the endometrial microenvironment. These findings provide a novel theoretical foundation for developing targeted, personalized treatments for infertility associated with AM.

Senile osteoporosis (SOP) is a chronic age-related skeletal disorder characterized by progressive bone loss and high fracture risk, with pathogenesis linked to excessive reactive oxygen species (ROS), osteoprogenitor cell senescence, and macrophage inflammation. Prussian Blue (PB) nanozymes show antioxidative potential for SOP but are limited by poor targeting, short circulation time, and unclear mechanisms, restricting clinical translation. Herein, we fabricated macrophage membrane-camouflaged citrate-modified PB (M@CPB) nanozymes for targeted SOP therapy and elucidated the underlying mechanism. Macrophage membrane coating prolonged the blood half-life of M@CPB by 1.47-fold and enhanced its bone tissue accumulation by 3.09-fold relative to CPB. In vitro, M@CPB inhibited O₂⁻ by 42.17% and scavenged H2O2 by 22.38%, reduced osteoprogenitor cell senescence, promoted M1-to-M2 macrophage polarization, and decreased the secretion of receptor activator of nuclear factor-κB ligand (RANKL) and tumor necrosis factor-α (TNF-α) to 47.41% and 53.10% of the model group, respectively. In vivo, M@CPB increased bone volume/tissue volume ratio by 3.83-fold relative to SOP mice, effectively ameliorating bone loss, motor dysfunction, and depressive-like behaviors. Collectively, M@CPB mitigated SOP by intervening in the ROS-senescence-inflammation axis and balancing bone remodeling, providing an innovative therapeutic strategy for SOP and potentially other age-related disorders.

Glycan-mediated interactions are vital to development, microbial colonisation, immune signalling, and cancer progression. Glycan microarrays have revolutionised glycobiology by enabling high-throughput analysis of these complex interactions, supported by techniques that reveal kinetics and dynamics in solution or at the cellular level. We introduce multifunctional glycan probes based on a tri-functional Fmoc-Amino-Azido (FAA) linker, enabling multi-platform investigation of glycan-mediated interactions. These FAA probes support glycan presentation on both covalent and non-covalent array platforms, allowing direct comparison of glycan recognition by diverse proteins. Notably, certain viral adhesins and immune lectins show a preference for the non-covalent platform. The azido group allows further functionalisation via 'click chemistry', enabling biotinylation for immobilisation on bio-layer interferometry biosensors for influenza virus binding, or fluorescent tagging for flow cytometry analysis of glycan-lectin interactions on cells. These versatile probes offer a unified platform for in-depth interrogation of glycan interactions using complementary approaches, with strong potential to advance glycan-based diagnostics and therapeutics.

The Nav1.5 channel, a major isoform of voltage-gated sodium ion channel, is mainly found in ventricular cardiomyocytes, playing a key role in generating essential cardiac action potentials for normal heart rhythms. Mutations in Nav1.5 have been associated with severe heart conditions such as long QT syndrome, Brugada syndrome, cardiac conduction disorders, atrial fibrillation, and dilated cardiomyopathy. Recent research has linked Nav1.5 to cardiac fibrosis and proposed its role in non-cardiac illnesses, including specific neurological disorders and cancers, subjects that will be reviewed in this paper. On the other hand, sodium-glucose cotransporter 2 inhibitors (SGLT2i), initially designed to manage diabetes by facilitating glucose excretion through urine, have demonstrated unexpected and encouraging cardioprotective benefits in clinical trials. This review compares the important SGLT2 inhibitors empagliflozin, dapagliflozin, and canagliflozin in terms of their interactions with Nav1.5 and their therapeutic effects on the heart. We also investigate new medications and compounds being developed to regulate Nav1.5 function, providing a preview of potential future treatments. Past attempts to develop late INa inhibitors and difficulties in transitioning from the research phase to clinical trials have raised doubts about the optimal design of such trials. In addition, we cover the applications of molecular dynamics simulations in understanding the mechanism of action of these drugs within the Nav1.5 channel computationally. We hope that this review identifies new opportunities to generate more effective inhibitors using novel methods and advanced multiscale molecular modelling techniques.

In people living with HIV (PLWH), persistent systemic inflammation drives comorbidities, including cardiovascular and cerebrovascular diseases. Monocyte-mediated inflammation, rather than T-cell activation, is a major contributor to chronic inflammation. However, the underlying mechanisms remain elusive. Here, we investigated the role of Dll4 (delta-like 4)-Notch signaling in monocyte-mediated inflammation. In clinical specimens, we quantified monocyte mDll4 (membrane-bound Dll4) expression and plasma levels of exDll4 (extracellular Dll4). We also analyzed the association between circulating exDll4 and monocyte subsets in PLWH. In vitro, human primary monocytes, endothelial cells, and T cells were stimulated with lipopolysaccharide or TNF-α (tumor necrosis factor-α) to assess DLL4 mRNA and Dll4 protein expression and exDll4 secretion. RNA sequencing was performed after DLL4 silencing to determine downstream signaling pathways. In addition, molecular and functional studies examined the effects of Dll4-Notch1 signaling on interferon-related gene expression, monocyte adhesion, and transmigration. Both male and female PLWH exhibited significantly increased mDll4 expression, activation of Dll4-Notch1 signaling, and increased Notch-targeted inflammatory factors. Although monocyte mDll4 did not differ by sex in PLWH, plasma exDll4 was significantly elevated in males, but not females, compared with HIV-uninfected individuals. In male PLWH, plasma exDll4 levels paralleled monocyte mDll4 expression. Circulating exDll4 was positively associated with proinflammatory monocytes and negatively associated with classical monocytes in male PLWH. Mechanistically, lipopolysaccharide or TNF-α robustly increased DLL4 mRNA and protein expression in human primary monocytes, along with exDll4 secretion from monocytes, but not from endothelial cells or T cells. RNA sequencing after DLL4 silencing showed that Dll4-Notch signaling suppresses a broad anti-pathogen program regulated by interferon and NF-κB, as well as cell-mobility-related genes. At the molecular level, Dll4 activated Notch1, which upregulated Hes1, forming a heterotrimer with STATs (signal transducers and activators of transcription), and repressed interferon pathway-related gene expression. Functionally, loss of Dll4 significantly reduced monocyte adhesion and transmigration. Proinflammatory cues upregulate Dll4 in monocytes and activate Dll4-Notch signaling, thereby suppressing antiviral responses and enhancing monocyte transmigration. Together, these effects sustain systemic inflammation in PLWH of both sexes. Our data identify monocyte-derived Dll4 as a promising biomarker and therapeutic target for systemic inflammation.

Spatial profiling of proteins and protein interactions facilitates understanding of cell functions within tissues and is essential for studies in signaling, immunity, and cancer. We present spatial proximity sequencing (Sprox-seq) for simultaneous profiling of surface proteins, protein complexes, and mRNAs, recording the tissue location of each molecule. Sprox-seq profiled 32 proteins, 528 pairwise interactions, and thousands of mRNAs with spatial resolution across human tonsils and germinal centers. Mapping tissue-wide protein interactions recapitulated RNA-defined tissue architecture but also revealed higher interaction complexity in the light zone. Protein-interaction trajectories uncovered a B cell state transition distinct from that inferred by RNA. Integrated protein-complex and mRNA analysis related spatially enriched complexes with mitotic pathways. Sprox-seq captured cell-cell interactions, such as B cell-follicular dendritic cell interactions mediated by the receptor complex VLA-4-VCAM1. Sprox-seq provides a spatially resolved multi-modal view of cell states and an integrated study of protein and cellular interactions across tissues.

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

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

Alternative lengthening of telomeres (ALT) is a telomerase-independent telomere maintenance mechanism active in a subset of aggressive cancers. Targeting ALT remains a therapeutic challenge due to its complexity and lack of clinically specific inhibitors. Here, we report that the natural compound Withaferin A (Wi-A) selectively impairs ALT cancer cell viability by disrupting Rad51-dependent homologous recombination (HR), a critical pathway for resolving telomeric replication stress and DNA damage in ALT cells. Comparative analysis using isogenic ALT and telomerase-positive cells revealed that Wi-A selectively downregulates PI3K/AKT signaling and induces replication stress, telomeric DNA damage, and DNA repair deficiency in ALT cells. Mechanistically, Wi-A destabilizes the Rad51 nucleoprotein filament and promotes Rad51 degradation via a RFWD3-dependent mechanism localized within ALT-associated PML bodies (APBs). Depletion of either PML or RFWD3 rescues Rad51 stability and reduces sensitivity of ALT cells to Wi-A. Together, these findings uncover a novel mechanism by which Wi-A selectively targets ALT cells through Rad51 destabilization within APBs, highlighting its potential to exploit ALT-specific vulnerabilities in telomere maintenance and DNA repair for therapeutic development.

Extrachromosomal circular DNA (eccDNA), characterized by abundant amplicons and high chromatin accessibility, contributes to tumor heterogeneity and transcriptional activity; however, its single-cell dynamics in cutaneous squamous cell carcinoma (CSCC) remain poorly understood. Here, we developed a single-cell framework eccDNAscope that reconstructs eccDNA structures and infers clonal trajectories from scATAC-seq data. Integrative analysis of 154,624 single cells revealed that eccDNAs were broadly distributed across diverse cell types but exhibited striking enrichment at promoter regions (79.94%), where they were associated with elevated expression of cell type-specific eccDNA-associated genes. The framework identified both known oncogenic eccDNAs (MYC) and previously unrecognized eccDNAs (PTMA, NRP2, and KLF7), which were independently validated by DNA fluorescence in situ hybridization (FISH), super-resolution microscopy, Immuno-FISH, and Circle-seq. These eccDNAs displayed enhanced transcriptional activity in the epithelial cell populations in which they were detected. We further identified CSCC-specific eccDNAs that were enriched in cell cycle-related pathways, including CACYBP, which was exclusively detected in CSCC, but was not observed in actinic keratosis (AK) or para-tumor tissues. Finally, eccDNA-based phylogenetic reconstruction identified conserved and tissue-specific eccDNA programs during disease evolution, and highlighted recurrent eccDNAs, particularly PTMA and CACYBP, as potential drivers of CSCC progression. Collectively, our single-cell analysis provides a framework for dissecting eccDNA-mediated oncogenesis and offers insights into therapeutic strategies targeting eccDNA-positive clones in cancer.

Critical periods are developmental windows that allow experience-dependent refinement of neuronal circuitry and function. While the consequences of sensory deprivation or alteration during the visual critical period have been well characterized, the impact of ethological experiences requiring active interaction with the sensory world is largely unexplored. Here, we use prey capture learning to assess structural and functional plasticity associated with visual learning in the primary visual cortex of critical-period mice. We show that prey capture learning supports improved temporal frequency discrimination and profoundly remodels visual circuitry by increasing spine turnover and moving spine density across the dendritic arbors of pyramidal neurons to new stable values. This widespread and persistent rewiring is absent in adults and supported by tumor necrosis factor α (TNF-α)-dependent homeostatic plasticity that contributes to behavioral improvement. Ethological critical period learning can thus co-opt homeostatic mechanisms to reset synaptic weight bounds to enable widespread synaptic remodeling to support improved visual function and behavior.

Neuromyelitis optica spectrum disorder (NMOSD) is a chronic inflammatory autoimmune disease affecting the central nervous system (CNS), characterized by anti-aquaporin 4 (AQP4) antibody-mediated damage to astrocytes, resulting in subsequent demyelination. Our prior work identified the protective effects of the apolipoprotein E130-149 (ApoE130-149) peptide in NMOSD mice by promoting astrocyte-microglia intercellular communication. However, its therapeutic potential is restricted due to the limited penetration of the blood-brain barrier (BBB) with systemic administration. Here, we designed a heavy-chain ferritin (HFn)-based nanocarrier containing the ApoE130-149 peptide (HFn-ApoE130-149), specifically engineered for CNS delivery. HFn-ApoE130-149 was constructed through genetic engineering by fusing the coding sequence of HFn with that of the ApoE130-149 peptide in a recombinant plasmid. An acute NMOSD mouse model was induced by transcranial co-injection of AQP4-IgG and human complement (hC) into the brain. The distribution of Cy5.5-labeled HFn-ApoE130-149 post intravenous injection was tracked using in vivo fluorescence imaging to confirm its presence in the brain and peripheral organs. Lesions in the brain were quantified using T2-weighted 7 Tesla magnetic resonance imaging (7T-MRI). Neuropathological features of NMOSD were evaluated by immunostaining of brain sections. Neuroinflammation and immune cell infiltration were analyzed via flow cytometry. The key signaling pathways regulated by HFn-ApoE130-149 were investigated through Western blot (WB) analysis. The interaction between HFn-ApoE130-149 and its receptors was validated through co-immunoprecipitation and visualized on microglia using proximity ligation assay (PLA). Finally, the therapeutic effect on spatial learning and memory was evaluated using the Morris water maze (MWM) test. The HFn-ApoE130-149 effectively crossed the BBB, attenuated lesion progression and demyelination, as well as preserved AQP4 expression and astrocytic integrity in NMOSD mice. The treatment induced a spatial and phenotypic restructuring of the astrocytic response, notably reducing excessive astrocyte accumulation around lesions while encouraging a proliferative and reparative phenotype. Furthermore, HFn-ApoE130-149 influenced microglial polarization towards an anti-inflammatory state, reducing infiltration of peripheral immune cells. Mechanistically, HFn-ApoE130-149 exerted its anti-inflammatory effects through the low-density lipoprotein receptor-related protein 1 (LRP1) -nuclear factor kappa B (NF-κB) signaling axis in microglia. Functional binding of HFn-ApoE130-149 to LRP1 suppressed inhibitor of NF-κB (IκBα) phosphorylation, thereby inhibiting NF-κB nuclear translocation and the subsequent release of pro-inflammatory cytokines, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Knocking down LRP1 reversed these effects, highlighting the importance of the LRP1-NF-κB signaling axis in the nanotherapeutic's efficacy. Treatment with HFn-ApoE130-149 improved spatial learning and rescued memory deficits in NMOSD mice. This study demonstrates that the engineered nanodrug HFn-ApoE130-149 is a promising targeted therapy for alleviating NMOSD pathology by enhancing BBB penetration and suppressing neuroinflammation through the LRP1-NF-κB signaling axis.

Aromatic camptothecins (CPTs) are clinically potent TOP1 inhibitors constrained by an "aromaticity-solubility" paradox, which leads to poor bioavailability and efflux-mediated resistance. Here, we report a modular strategy to reprogram the planar CPT scaffold into multifunctional topoisomerase I (TOP1) degraders by integrating hydrophobic tagging (HyT) with supramolecular self-assembly. Adamantane HyT introduces an orthogonal aliphatic module, converting classical inhibitors into proteasome-dependent degraders with enhanced membrane permeability while preserving TOP1-DNA binding affinity. This tag synergistically acts as a supramolecular anchor, enabling host-guest assembly with poly(β-cyclodextrin) to form stable, pH-responsive nanoparticles without pharmacophore modification. Our lead candidate, SN-38-A2, demonstrates potent TOP1 degradation and achieves superior tumor regression in xenograft models compared to clinical irinotecan. This synergistic HyT-supramolecular approach rebalances aromaticity for optimized drug-like properties, establishing a paradigm to transform solubility-limited warheads into high-performance degraders with integrated delivery.

Molecular motors, known for their ability to undergo controlled unidirectional rotation under external stimuli, have gained growing interest for their potential applications in biological systems and smart materials. Meanwhile, glycosylated macromolecules are utilized in lectin-binding, making them potential candidates to combat pathogenesis, cancer, and biofilms. However, glycopolymers are limited by their current inability to adapt in response to external stimuli, restricting their potential for future precision therapies. By integrating multivalent glycopolymeric structures with first-generation molecular rotary motors, it becomes possible to control carbohydrate-lectin interactions using intrinsic motor functions. Herein, we present photoadaptive glycopolymer-based first-generation molecular motors functionalized with immunologically important monosaccharides, β-d-glucopyranoside and β-d-mannopyranoside. The lectin-binding properties were subsequently investigated using surface plasmon resonance with DC-SIGN and Langerin, key players within the human innate immune system. Additionally, a competition assay with the envelope glycoprotein of HIV, namely gp120, was conducted to ascertain the inhibitory potential of these photoresponsive glycopolymer-based molecular motors. It was established that the light-controlled conformation of the molecular motor impacted both the overall binding affinity and half-maximal inhibitory concentration values. These findings create opportunities to control the binding affinity of glycopolymers and emphasize the potential of light-driven glycopolymers to modulate key interactions in cell-specific targeted delivery of therapeutics.

Uniform predominance of lower- and mid-chlorinated polychlorinated biphenyls (PCBs), including PCB-8, -18, -28, -44, -52, -66, -101, -105, -118, -138, and -153, across urban and suburban/rural indoor dust from a central Indian city indicates strong influence of open, partially open and closed application technical formulations used in built environments that remain largely unaccounted in India's National Implementation Plan. Mean of Σ28 PCBs (103.3 ng/g) and Σ12 DL-PCBs (30 ng/g) were strongly influenced by urban outliers. Homolog profiles were dominated by lower to mid-chlorinated congeners (di - hexa), accounting for more than 94% of total PCBs, with uniform distribution across urban and suburban/rural sites. No statistically significant differences were observed in household characteristics or PCB levels between urban and suburban/rural sites. However, residences above 20 years exhibited higher PCB loading in urban hotspots. Despite this trend, lack of statistical significance indicates that building age alone may not be the primary influencing factor. Strong correlations with open, partially open, closed application formulations (Aroclor 1248, Kanechlor 400, Clophen A50, Kanechlor 500, and Pyralene DP-3) indicated source profiles consistent with congener fingerprints. Congener-specific fingerprints suggested legacy building materials, localized combustion, vehicular and workshop operations, domestic emissions and coal-powered industrial facilities as possible sources. Site-specific exceedances of hazard indices (HI > 1) at urban hotspot locations for adults and children are predominantly driven by dermal exposure. Incremental lifetime cancer risk exceeded regulatory benchmarks for adults at multiple urban hotspots, suggesting indoor PCB exposure in India may be underestimated due to undocumented sources within the built environment.

The indoor use of household insecticides introduces complex mixtures of volatile organic compounds (VOCs) into residential environments, raising potential health concerns. This study characterized the emission profiles of total VOCs (TVOCs) and targeted aromatic compounds (BTEXS) from 19 household insecticides, comprising aerosol insect sprays (IS), liquid vaporizers (EV), mat-type fumigators (MA), and mosquito coils (MC), using environmental test chambers and TD-GC-MS analysis. Emission rates (ERs) and emission factors (EFs) were calculated, and health risks were assessed using non-carcinogenic hazard quotients (HQ) and incremental lifetime cancer risks (ILCR). Solvent-based IS and EV products exhibited low ERs for aromatic compounds but contributed to substantial chemical burdens due to aliphatic carrier solvents, as reflected by high EFs. Trace benzene emissions from MA devices were attributed to thermal degradation (pyrolysis) of the paper-pulp pad under continuous heating. MC acted as continuous combustion sources, releasing high levels of BTEXS. Non-carcinogenic risks were acceptable across all product types (HQ < 1.0); however, chronic inhalation of pyrolytic coil smoke may pose a carcinogenic risk from benzene (ILCR > 1.0 × 10⁻⁶) in unventilated rooms. Exposure modeling identified the indoor air exchange rate as the primary determinant of human exposure; active ventilation reduced pollutant concentrations by up to 84%. These findings highlight the importance of adequate indoor ventilation during household insecticide use.

Somatostatin (SRIF) and its receptors (SSTR) are key players in the regulation of neuroendocrine neoplasms (NEN), with significant implications for diagnosis, prognosis, and treatment. The field of SSTR-targeted drugs/ligands (SRLs) is continuously evolving, with emerging novel therapeutic approaches aiming to address the numerous unmet needs and limitations of the standard-of-care (SOC) SRLs. In the present manuscript, we aim to provide a concise review of the field, focusing on the new and ongoing developments and potential novel approaches to be considered in the future. The paper (1) provides insights into the pathophysiology of SSTR in relation to unmet needs in the SRL efficacy and limitations, (2) presents a state-of-the-art assessment of knowledge on the SOC SRLs, and (3) depicts current ongoing and future research, highlighting areas of insufficient knowledge for SRLs in NEN routine practice.

The industrial production of monoclonal antibodies (mAbs) has been suffering from the lack of efficient technology and equipment for quickly acquiring high-yield specific hybridoma cells (HSHCs). This work presented an efficient microfluidic device that integrated single-cell pairing and electrofusion of myeloma and lymphocyte cells, microfluidic chemiluminescence immunoimaging (μCLII) screening of HSHCs, selective retrieval and proliferation of HSHCs to secrete mAb through microfluidic control and programmable signal generator modules. The designed pump-equipped lantern-shaped array chip achieved single-cell screening and undamaged retrieval of HSHCs through a pump-controlled surfing force. Using PCSK9-mAb as a model, this device demonstrated a cell fusion efficiency of 38% with an asymmetric cage-shaped structure, the precise sorting of HSHCs within 2 h, and a high PCSK9-mAb yield of ∼3.0 pg per cell during 18-h culture. The whole procedure including cell fusion and selection of HSHCs could be completed within 8 days, indicating a greatly shortened period of monoclonal antibody production compared to 3-4 months of commercial technology, and thus showing its promising applications in multiple scenarios, such as single-cell analysis, mAbs screening and drug discovery.

Epigenetic regulation, including DNA methylation, histone modifications, non-coding RNAs, and higher-order chromatin remodeling, plays a central role in the biology of neuroendocrine neoplasms (NENs). Advances in high-throughput profiling have uncovered epigenetic alterations across pheochromocytomas/paragangliomas (PPGLs), gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs), lung neuroendocrine neoplasms (LNENs), medullary thyroid carcinoma (MTC), and pituitary NETs (PitNETs). These alterations converge on pathways governing cell cycle control, telomere maintenance, hypoxia signaling, epithelial-mesenchymal transition, and chromatin architecture. Importantly, epigenetic signatures not only mirror genetic backgrounds (eg, SDHx, MEN1, ATRX, RET) but also provide independent layers of prognostic and predictive information. Distinct methylation profiles, histone modification patterns, and deregulated microRNA/long non-coding RNA networks have been consistently linked to tumor aggressiveness, metastatic potential, and therapeutic resistance. The clinical translation of these insights is rapidly evolving. Methylation-based classifiers and circulating epigenetic markers are emerging as promising tools for early diagnosis, risk stratification, and longitudinal monitoring. Moreover, epigenetic pathways represent attractive therapeutic targets, with DNA methyltransferase inhibitors, histone deacetylase inhibitors, and RNA-modifying enzyme modulators under active investigation. However, significant challenges remain, including methodological heterogeneity, small or retrospective cohorts, and the limited functional validation of candidate biomarkers. Future priorities include prospective multi-institutional validation, integration of epigenomics with other molecular layers in multidimensional classifiers, and the application of spatial and single cell approaches to resolve intra-tumoral heterogeneity. Ultimately, epigenetic research has redefined our understanding of NEN pathogenesis and progression, positioning the epigenome as a promising frontier in precision oncology. Through robust validation, epigenetic biomarkers and therapies may transform the clinical management of NENs.

Macrophages are central regulators of immunity, tissue homeostasis, and repair, whose plasticity enables them to integrate cues from their local microenvironment. Dysregulated macrophages contribute to chronic inflammation, fibrosis, impaired wound healing, and tumor progression. Biomaterials enable the modulation of macrophages through localized delivery of immunomodulatory signals and by creating instructive microenvironments. In this review, we examine macrophage-biomaterial interactions, emphasizing how surface chemistry, topography, mechanics, porosity, and degradation profiles shape macrophage recruitment and phenotype. We discuss emerging strategies for macrophage modulation, including programmable systems that provide spatiotemporal control over immune signaling. We further highlight how these applications can improve disease outcomes. Finally, we discuss design considerations, characterization, and translational challenges that must be overcome to advance macrophage-targeted biomaterials toward clinical implementation.

Similar to CD8+ T cells, stem-like CD4+ T cells sustain responses to tumors, chronic infections, and inflammation. In this issue of Immunity, Wen et al. and Agesta et al. reveal EOMES as a critical regulator of stem-like CD4+ T cell maintenance and differentiation.

Alveolar rhabdomyosarcoma (aRMS) is a fusion-driven pediatric cancer with poor survival and limited therapeutic options. To uncover novel vulnerabilities, we employed complex-based analysis of the DepMap functional genomic data, identifying CDK8 as a dependency in aRMS. Both CDK8 knockout and pharmacologic inhibition impaired tumor cell growth and induced myogenic differentiation in vitro and in vivo. Compared to genetic loss, CDK8 pharmacologic inhibition induced more dynamic transcriptional changes. With a genome-scale CRISPR-Cas9 drug modifier screen, we determined that the maximal anti-tumor activity of the CDK8 inhibitor requires the presence of the Mediator kinase module, including CDK8, and transcriptional cooperation with the SAGA complex. We further identified SIX4 as a key transcription factor mediating CDK8 inhibitor-induced transcriptional activation of myogenic differentiation genes and impaired tumor proliferation. These findings suggest a distinct gain-of-function mechanism of the CDK8 inhibitor and establish a strong rationale for CDK8 inhibition as a differentiation-inducing therapeutic strategy in aRMS.

Ewing sarcoma is characterized by a chromosomal translocation resulting in the fusion protein EWSR1::FLI1. We utilize endogenous EWSR1::FLI1 target gene reporters in patient-derived cell lines to perform a high-throughput phenotypic screen to identify small molecules that impair the EWSR1::FLI1 transcriptional program. We discovered that inhibitors of cyclin-dependent kinase 8 (CDK8), including a novel pyridyl imidazole, altered transcription of EWSR1::FLI1 target genes and CDK8 co-localized with EWSR1::FLI1 preferentially on single GGAA DNA binding motifs. Using pooled CRISPR screening, biochemical studies, and chromatin profiling, we discovered that CDK8 inhibitors suppressed proliferation of Ewing sarcoma cells through a gain-of-function mechanism by inducing molecular trapping of the CDK8 kinase module and Mediator complex on chromatin. This mechanism has implications for the development of small molecules to target transcription and suggests that impairment of transcriptional regulatory complex dynamics might serve as a vulnerability in cancers in which the dominant oncogenes act as dosage-sensitive transcription factors.

Somatic mutations often predict survival in childhood cancers, as exemplified in rhabdomyosarcoma, where FOXO1 gene fusion status is a key prognostic feature. Here, we examined single-cell transcriptomics and discovered that cancer cells of lethal disease converge on a common cell state with a shared transcriptional landscape, irrespective of fusion status. Nuclear transcriptomics, chromatin accessibility, spatial transcriptomics, perturbation studies, and previously published data sets validated the overarching high-risk cell state. The convergent cell state only partially overlapped with transcriptional effects of the FOXO1 fusion and unexpectedly exhibited neural features. Overall, these findings delineate a cell state of high-risk rhabdomyosarcoma cells that transcends conventional molecular and histological boundaries, suggesting an overarching disease phenotype that could transform target discovery and inform clinical practice.