FMS-like tyrosine kinase 3 (FLT3) internal tandem duplication (FLT3-ITD) is a well-characterized genetic alteration associated with poor prognosis in acute myeloid leukemia (AML). Although FLT3 inhibitors, such as gilteritinib and quizartinib, initially show clinical efficacy, resistance frequently emerges because of secondary mutations. Through a kinase inhibitor library screen, we identified XL999, which exhibits potent antileukemic activity across diverse FLT3-mutant AML cell lines and primary patient samples. Mechanistically, biochemical and molecular docking analyses revealed that XL999 directly binds to FLT3-ITD, potentially independent of the common F691 resistance site, thereby suppressing downstream signaling pathways to induce cell cycle arrest and apoptosis. In preclinical mouse models harboring refractory FLT3-ITD-D835Y or ITD-F691L mutations, oral administration of XL999 effectively reduced the leukemic burden across the peripheral blood, spleen, and bone marrow, significantly prolonging survival compared with gilteritinib and quizartinib, with no overt systemic toxicity observed in short-term mouse tolerability studies. Furthermore, XL999 demonstrated robust efficacy in patient-derived xenograft models, complemented by a favorable oral bioavailability profile. Together, these findings suggest that XL999 may represent an orally bioavailable FLT3 inhibitor with the potential to overcome clinically relevant secondary resistance mutations in FLT3-ITD-positive AML, supporting its further investigation as a therapeutic candidate.
Tertiary lymphoid structures (TLS) are ectopic lymphoid aggregates formed under chronic inflammatory tumor microenvironments. Clinical cohorts across solid cancers demonstrate that mature TLS with intact germinal centers serve as independent favorable prognostic biomarkers and predict superior responses to immune checkpoint inhibitors. However, the differences in maturity, spatial localization and cellular composition of TLSs result in heterogeneity in their immunostimulation or immunosuppression, thereby exhibiting dual functions of promoting or anti-tumor. Furthermore, there are significant differences in the maturity assessment methods and definitions of TLS, which jointly hinder the clinical evaluation and targeted treatment of TLSs. In this review, we integrate current understanding of TLS development, molecular drivers, and stromal licensing, and dissect the key mechanisms by which TLS contribute to anti-tumor immunity within tumors. We consolidate standardized TLS detection methods encompassing pathology, multi-omics, and radiogenomics, and further explore the translational value of TLS maturation status, germinal center-like responses, and intratumoral versus peritumoral localization in prognosis and immune checkpoint blockade. We systematically elaborate therapeutic strategies for inducing functional TLS, dissect the intrinsic mechanisms by which TLS synergistically boost the efficacy of adoptive cell therapy and tumor vaccines, and outline major bottlenecks hindering clinical translation. Collectively, this work constructs a comprehensive multi-dimensional framework for TLS research and provides actionable insights to develop TLS-targeted immunotherapies for human cancers.
Recent years have witnessed considerable progress in computer-aided drug discovery, driven by the incorporation of computational technologies within both academic and pharmaceutical environments. This evolution is marked by a significant accumulation of data pertaining to detailed three-dimensional structural information, ligand properties, and their interactions with therapeutic targets. The augmentation of computational capabilities and the accessibility of extensive chemical libraries containing billions of drug-like small molecules have further facilitated this transition. To effectively utilize these resources, it is imperative to employ rapid computing methods for virtual screening, which encompass structure-driven in silico screening across vast molecular spaces, supported by efficient recurrent profiling techniques. Furthermore, advancements in deep learning methodologies are required to improve the accuracy of prediction concerning target functionalities and ligand characteristics, even when complete receptor structures are not available. Here, this review discusses the expansion of chemical space, advanced virtual screening, deep learning, molecular dynamics (MD) simulations, Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) modelling, and challenges for drug discovery. It examines how experimental validation integrates computational predictions with laboratory testing for effective candidate selection. Finally, it outlines future research directions for artificial intelligence-driven, ultra-high performance computing (UHPC) in drug discovery, offering new prospects for the economical creation of safer and more efficacious molecule-level therapies.
The continued antigenic evolution of SARS-CoV-2 Omicron subvariants has progressively eroded vaccine-elicited protective immunity, driving demand for next-generation candidates that confer broad-spectrum protection against phylogenetically divergent strains. Here we report the design and preclinical evaluation of SV, an mRNA vaccine encoding a heterodimeric receptor-binding domain (RBD) antigen. In this construct, a previously optimized monomeric RBD (BSCOV06) is tandemly linked to the KP.3 RBD, presenting two antigenically distinct RBDs within a single immunogen. A two-dose SV regimen in BALB/c mice elicited high-titer neutralizing antibodies with potent cross-reactivity against BA.1, XBB.1.5, JN.1, KP.3, and the phylogenetically distant XDV variant. Integrated B cell receptor (BCR) and T cell receptor (TCR) repertoire profiling revealed that SV drives qualitatively distinct adaptive immune remodeling relative to BSCOV06. Key features included elevated class-switched somatic hypermutation, sustained naive B cell engagement, broad polyclonal T cell expansion, and extensive VJ gene-usage reprogramming across both lymphocyte compartments. In BALB/c and K18-hACE2 transgenic mice, SV conferred robust protection against JN.1 and XDV challenge, substantially reducing pulmonary viral loads and attenuating histopathological injury. Notably, SV achieved immunogenicity and cross-protective efficacy comparable to or exceeding those of the three-dose BSCOV06 schedule, supporting the potential of heterodimeric antigen design. These findings support SV as a promising broad-spectrum COVID-19 vaccine candidate. More broadly, they suggest that heterodimeric RBD architectures incorporating antigenically divergent variants may represent a generalizable platform for countering viral immune evasion, with implications for future SARS-CoV-2 variants and other rapidly evolving viral pathogens.
Abstract Severe pneumonia caused by respiratory virus infection is one of the main threats to the lifespan of the aged population and physiological aging fundamentally drives poor disease outcomes in respiratory viral infections, necessitating investigation of the underlying mechanisms and the development of effective countermeasures. Pulmonary transcriptomic profiling reveals that diffuse cell death, inadequate antiviral responses, myeloid-driven excessive inflammation and immune-thrombosis dictate the pulmonary microenvironment are typical molecular pathology characteristics of the SARS-CoV-2-infected aged hamsters rather than the adult controls. The elevated pathological baseline and dysregulated immune responses are demonstrated as the key host factors of lethal viral pneumonia in the aged hamsters. Meanwhile, SARS-CoV-2 infection in the adult hamsters usually resulted in an aged-like pulmonary transcriptomic signature, suggesting a potential link among aging, dysregulated immune responses and severe illness. To reverse the progression of lethal severe pneumonia in the aged hamsters, we initiated a multidimensional combination therapy of dexamethasone, heparin and the broad-spectrum viral decoy CoVR-MV at three days after infection. Although single- and dual-drug therapies were insufficient to achieve functional cure, the three-drug combination therapy resulted in a potent reduction of high mortality, body weight loss, viral load, lung pathology and cytokine storm through the synergism of immunoregulatory, anticoagulant and antiviral effects, effectively intercepting the multifaceted pathogenic network. This study highlights physiological aging as a core driver of critical COVID-19 pathogenesis and provides valuable clues for the rational design of multi-drug and multi-target therapies against respiratory viral infections and lethal severe pneumonia in the elderly population.
The liver possesses an extraordinary capacity to regenerate after injury or surgical resection, a process highly dependent on the coordinated orchestration of the immune microenvironment. Although macrophages are recognized as pivotal coordinators of hepatic tissue repair, the precise checkpoints governing their functional transitions during regeneration remain elusive. Here, we identify the glutamine transporter SLC1A5 (Solute Carrier Family 1 Member 5) as a critical metabolic gatekeeper of macrophage function during liver regeneration. Using a mouse model of partial hepatectomy, we show that SLC1A5 is markedly upregulated in monocyte-derived macrophages at the peak of regeneration. Myeloid specific deletion of Slc1a5 (Slc1a5fl/flLyz2cre) severely impairs hepatocyte proliferation and diminishes the expression of macrophage derived regenerative factors. Mechanistically, Slc1a5 deficiency depletes intracellular glutamine, which triggers macrophage senescence and drives a pro-inflammatory phenotype. This senescent state selectively downregulates Gas6 (Growth Arrest Specific 6), a crucial bridging ligand for efferocytosis, thereby impairing apoptotic cell clearance and exacerbating local inflammation. Strikingly, exogenous Gas6, senolytic Quercetin therapy, or in vivo L-glutamine supplementation successfully alleviates macrophage senescence, reinstates Gas6 mediated efferocytosis, and rescues defective liver regeneration. Collectively, our findings reveal a novel ‘Slc1a5-glutamine-senescence-efferocytosis’ axis that dictates macrophage driven tissue repair. This study not only uncovers a fundamental immunometabolic mechanism but also highlights glutamine supplementation and senolytics therapy as promising clinically strategies to accelerate liver regeneration.
Pathological deposition of endogenous material within tissues represents a central, unifying mechanism across a broad spectrum of prevalent human diseases. Atherosclerosis and Alzheimer's disease (AD) exemplify this paradigm: atherosclerosis is driven by subendothelial accumulation of apolipoprotein B–containing lipoproteins, cholesterol crystals, and hydroxyapatite, while AD is characterized by cerebral deposition of misfolded amyloid-β (Aβ) and/or hyperphosphorylated tau. Emerging evidence implicates substantial lipid dyshomeostasis in AD pathogenesis, including lipid-droplet-accumulating microglia and widespread lipidomic disruption, blurring the categorical boundary between "lipid" and "protein" deposition diseases. Apolipoprotein A-I (apoA-I) dysfunction bridges both domains: post-translational modifications and point mutations impair reverse cholesterol transport in atherosclerosis and simultaneously promote apoA-I amyloid fibril formation in systemic amyloidosis. Across atherosclerosis, AD, systemic amyloidoses, immune complex–mediated nephropathies, crystal arthropathies, and lysosomal storage disorders, shared pathophysiologic processes emerge: production–clearance imbalance, conformational transitions favoring aggregation or crystallization, microenvironmental modulation by extracellular matrix components, and chronic sterile inflammatory responses driven by NLRP3 inflammasome activation. In this review, we synthesize deposition biology using atherosclerosis and AD as primary paradigms, integrate mechanistic insights from related disorders, highlight convergent molecular pathways including NLRP3, proteostasis, and glymphatic clearance, and discuss diagnostic and therapeutic strategies. A conceptual framework unifying these conditions as variations on a common deposition theme is proposed to guide broadly applicable precision therapies.
RNA splicing expands the functional output of eukaryotic genomes by enabling individual precursor messenger RNA (pre-mRNA) to generate multiple mature transcripts with protein‑coding and regulatory properties. Its fidelity and plasticity depend on coordinated interactions among the spliceosome, trans-acting splicing factors, cis-regulatory elements, and chromatin- and RNA-associated regulatory mechanisms. However, how these components collectively generate cell- and tissue-specific splicing programs, and how their disruption drives disease, remain incompletely understood. In this review, we integrate the molecular regulation of RNA splicing with its physiological, pathological and therapeutic consequences. We describe how spliceosome assembly, splicing regulatory elements, splicing factors, epigenetic modifications, and post-transcriptional processes determine splice-site selection. We then examine how regulated isoform programs support development, tissue specialization, homeostasis, circadian timing, and stress adaptation, and how their failure contributes to cancer and diverse non-neoplastic diseases. In cancer, we highlight the bidirectional interplay between splicing dysregulation and the tumor microenvironment, through which metabolic reprogramming and immune suppression reinforce aberrant splicing. Finally, we assess strategies that modulate the spliceosome, splicing-factor activity, or disease-associated transcripts, and present a perspective on how multi-omics, artificial intelligence, targeted delivery, and combination with immunotherapy could collectively advance the discovery and precision of splicing-directed therapies. We suggest that safe clinical translation will require greater selectivity, reduced off-target toxicity, and preservation of essential physiological splicing.
Abstract Obesity is a global health problem linked to the development of metabolic syndrome and comorbidities such as metabolic dysfunction-associated steatotic liver disease (MASLD) and metabolic dysfunction-associated steatohepatitis (MASH). Pleiotrophin (PTN) is a cytokine known for its role in tissue regeneration and energy metabolism. However, its function in hepatic metabolism and its role in MASLD progression remain to be elucidated. Here, we investigated the contribution of PTN to hepatic metabolism using male and female wild-type ( Ptn +/+ ) and PTN-deficient ( Ptn −/− ) mice fed a standard or high-fat diet (HFD) for 6 months, as well as primary hepatocytes and Huh7 cells. Ptn -deletion protected both sexes against HFD-induced body weight gain, fasting hyperglycemia, hyperinsulinemia, insulin resistance and metainflammation. Furthermore, PTN deficiency prevented hepatic steatosis, reduced triacylglyceride accumulation and protected against liver fibrosis. Mechanistically, PTN deficiency was associated with increased AMPK activation, reduced ACC abundance and phosphorylation, constitutively low DGAT2 expression and altered AKT signalling. In vitro, PTN directly promoted lipid accumulation and triacylglyceride synthesis in primary hepatocytes and Huh7 cells, supporting a direct role for PTN in regulating hepatocyte lipid metabolism. Our results highlight PTN as a key modulator of hepatic lipid metabolism, systemic inflammation and extracellular matrix remodelling during obesity. These findings identify PTN as a promising therapeutic target for MASLD, MASH and related metabolic disorders, and point to a sexual dimorphism in adaptive metabolic strategies, with females demonstrating a greater degree of protection against the liver-damaging effects of diet-induced obesity.
The Piezo1 channel is a mechanosensitive, non-selective cation channel that converts mechanical forces into electrochemical signals, playing pivotal roles in vertebrate physiology. Structurally, Piezo1 features a distinctive trimeric propeller structure that undergoes conformational changes in response to membrane tension, enabling mechanogating. Accordingly, Piezo1 is involved in a broad spectrum of physiological processes, including vascular development and homeostasis, bone and cartilage formation, skeletal muscle growth, neural development, sensory perception, immune regulation, and cellular volume regulation. Accumulating evidence indicates that mutations or dysregulation of Piezo1 are closely associated with a variety of human diseases, including genetic diseases, cardiovascular diseases, infectious diseases, autoimmune diseases, and cancer. Therefore, Piezo1 has emerged as a potential therapeutic target. Currently, the exploration of pharmacological modulators targeting Piezo1, as well as emerging approaches such as gene therapy, artificial intelligence (AI)-driven drug discovery, and advanced drug delivery systems, offer potential avenues for the development of Piezo1-targeted therapeutic strategies. However, these approaches still face significant challenges regarding specificity, in vivo targeting, and context-dependent effects. This review systematically summarizes the structure, mechanogating mechanisms, physiological and cellular functions of Piezo1, as well as its associations with human diseases. Based on this, the limitations of current Piezo1-targeted therapeutic strategies and their future developmental directions are highlighted, while the therapeutic potential of targeting Piezo1 is emphasized.
Abstract Metabolic syndrome (MetS)-associated cognitive impairment is a growing global health crisis. The hippocampus is particularly vulnerable to metabolic stress, yet the precise molecular mechanisms linking systemic metabolic dysfunction to cognitive decline remain unclear. High-mobility group box 1 (HMGB1), a damage-associated molecule, is elevated in MetS. However, its specific role in simultaneously regulating hippocampal neuronal ferroptosis and microglial activation is poorly defined. Here we show that HMGB1 exacerbates MetS-associated cognitive impairment by promoting both neuronal ferroptosis and microglial inflammation. Using a high-fat high-glucose diet mouse model and primary co-cultures, we found that HMGB1 neutralization alleviated cognitive deficits, reduced neuronal ferroptosis, and suppressed microglial activation. Mechanistically, extracellular HMGB1 acts through neuronal toll-like receptor 4 (TLR4) to trigger the ubiquitination and degradation of nuclear factor erythroid 2-related factor 2 (NRF2). This HMGB1-induced loss of NRF2 impairs autophagic flux, thereby aggravating ferroptosis and neuroinflammation. Furthermore, in vitro conditioned medium experiments revealed a reciprocal pathological aggravation, where ferroptotic neurons and activated microglia both secrete HMGB1 to exacerbate each other's pathological state. These findings position HMGB1 as a pivotal mediator linking metabolic stress specifically to neuronal ferroptosis and neuroimmune activation. Targeting the HMGB1-NRF2-autophagic flux axis therefore offers a promising therapeutic strategy for mitigating cognitive impairment in patients with MetS.
Cancer remains a major global health burden and the second leading cause of mortality worldwide. Recent advances in cancer immunotherapy have emphasized the critical role of the tumor microenvironment (TME) in determining therapeutic outcomes, leading to the classification of tumors into immunologically “hot” and “cold” phenotypes. Cold tumors are characterized by low immunogenicity, limited immune cell infiltration, and a highly immunosuppressive microenvironment, resulting in poor prognosis and resistance to immune checkpoint inhibitors. Despite the development of multiple immunotherapeutic strategies, effective activation of antitumor immunity in cold tumors remains a major clinical challenge. Current approaches aim to initiate immune responses through priming strategies such as cancer vaccines and adoptive T-cell transfer, while simultaneously overcoming immunosuppressive signaling via immune checkpoint blockade. Additional strategies include depletion of myeloid-derived suppressor cells and enhancement of co-stimulatory pathways. However, these approaches are often limited by inefficient delivery, poor tumor penetration, and systemic toxicity. Nanotechnology has emerged as a promising platform for tumor microenvironment reprogramming. Nanocarriers enable targeted delivery of immunomodulatory agents, enhance antigen presentation, and improve immune activation while overcoming biological barriers such as dense stroma and abnormal vasculature. By integrating nanotechnology with immunotherapy, new opportunities arise to convert cold tumors into hot, immune-responsive phenotypes, thereby improving therapeutic efficacy and clinical outcomes.
Inflammatory bowel disease (IBD) is a chronic inflammatory disorder characterized by epithelial barrier disruption, persistent mucosal inflammation, and progressive intestinal fibrosis, for which effective therapeutic options remain limited. Thymosin β4 (Tβ4) is a highly conserved endogenous peptide with established roles in tissue repair and immune regulation, but its contribution to IBD pathogenesis has not been fully elucidated. Here, we found that TMSB4X, the gene encoding Tβ4, was downregulated in colonic tissues of IBD patients. To investigate its functional significance, we generated Tmsb4x-deficient mice and demonstrated that loss of endogenous Tβ4 markedly increased susceptibility to dextran sulfate sodium (DSS)-induced colitis. Conversely, oral administration of recombinant human Tβ4 (rhTβ4) significantly improved survival, alleviated body weight loss, reduced epithelial injury, and suppressed inflammatory cytokine production in both prophylactic and therapeutic colitis models. Furthermore, rhTβ4 attenuated intestinal fibrosis, as evidenced by reduced expression of fibrosis-associated markers and decreased collagen I deposition. Transcriptomic analysis revealed that rhTβ4 partially restored DSS-induced gene dysregulation and suppressed mineralocorticoid receptor (MR, NR3C2) signaling, a pathway further supported by reporter assays and downstream target gene analyses. Collectively, these findings identify Tβ4 as an endogenous protective factor against intestinal inflammation and fibrosis and suggest that pharmacological restoration of Tβ4 activity may represent a promising therapeutic strategy for IBD through modulation of mineralocorticoid receptor signaling.
Major depressive disorder (MDD) and schizophrenia (SCZ) are severe psychiatric disorders, the molecular mechanisms of which remain incompletely understood. Increasing evidence implicates neuroinflammatory signaling, autophagy dysregulation, and unfolded protein response (UPR) alterations in their pathophysiology. Here, we analyzed dorsolateral prefrontal cortex (DLPFC) samples from postmortem human brains of 28 MDD subjects, 28 SCZ subjects, and 28 matched controls. Gene expression levels of key inflammatory, autophagy, and UPR-related markers were assessed by RT-qPCR, while selected proteins were quantified by Western blot and ELISA. Logistic and linear regression models were applied to evaluate disease-associated alterations and the influence of sex, age, and cause of death. Transcriptional analyses revealed pathway-specific alterations in both disorders, with IRE1α emerging as the most consistently upregulated marker across MDD and SCZ. Sex-stratified analyses indicated that risk-associated transcriptional changes were more prominent in men with MDD, whereas women with SCZ showed broader transcriptional alterations. Age-related effects were mainly detected at the mRNA level, particularly in autophagy-related genes. In contrast, protein analyses showed a generalized downregulation of several inflammatory (AIM2, NLRP3), autophagy (ATG7, mTOR, RAB5A), and UPR-related (IRE1α) proteins in both disorders. In SCZ subjects who died by suicide, increased IL18, CASPASE-5, and IRE1α transcription, together with increased CASPASE-8 protein levels, suggested enhanced inflammatory and stress-related signaling. Overall, these findings reveal a marked transcription-protein dissociation in key cellular stress pathways in the DLPFC of MDD and SCZ subjects, supporting multilayer regulation of inflammatory, autophagy-related, and UPR responses in the psychiatric brain.
Abstract Liver sinusoidal endothelial cells (LSECs) form a specialized discontinuous microvascular interface between sinusoidal blood and the hepatic parenchyma. Functioning as a central regulator of hepatic homeostasis, LSECs integrate metabolic, inflammatory, and hemodynamic signals to maintain sinusoidal permeability, immune tolerance, vascular tone, and lipid exchange. During acute injury or sustained hepatic stress, LSECs may lose their differentiated sinusoidal phenotype and acquire features of capillarization, including fenestrae loss, subendothelial matrix accumulation, impaired scavenging activity, and attenuation of KLF2-eNOS-NO signaling. Capillarized LSECs are not merely a byproduct of hepatic damage; instead, they may function as early endothelial sentinels and important contributors to the amplification of parenchymal injury, inflammation, fibrogenesis, and tumor immune evasion. This review summarizes the physiological functions of LSECs and examines mechanisms through which LSEC dysfunction contributes to acute liver injury, metabolic dysfunction-associated steatotic liver disease, viral hepatitis, cirrhosis, and hepatocellular carcinoma. By comparing conserved mechanisms with disease-specific spatial and molecular triggers, this review highlights how LSEC phenotypic switching disrupts the angiocrine–immune–fibrotic axis and contributes to disease progression. Recent advances in single-cell and spatial omics, LSEC subpopulation heterogeneity, and capillarization-aware therapeutic delivery are further discussed. Finally, therapeutic reprogramming of dysfunctional LSECs toward a quiescent, fenestrated, and tolerogenic phenotype is proposed as a strategy to restore sinusoidal homeostasis and improve treatment of liver diseases.
The pathogenesis of prevalent metabolic diseases such as obesity, atherosclerosis, metabolic dysfunction-associated steatotic liver disease, and diabetes is intricately linked to dysregulated lipid metabolism. Peroxisome proliferator-activated receptor gamma (PPARγ) is a key transcriptional regulator of lipid homeostasis and a well‑researched therapeutic target. Although biochemical signaling pathways have been the traditional focus, recent studies now highlight the mechanical microenvironment (matrix stiffness, fluid shear stress, and tensile strain) as a pivotal physical metabolism regulator. However, how mechanical signals integrate with PPARγ to control lipid metabolism across tissues and diseases remains poorly defined. This review details the molecular mechanisms by which mechanical cues influence PPARγ expression, activity, and post‑translational modifications, focusing on Yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ), neural precursor cell expressed developmentally down-regulated protein 4 (NEDD4)-mediated ubiquitination, and protein kinase Cα (PKCα)-extracellular signal-regulated kinase (ERK) pathways. We further explore the critical role of PPARγ in mechano‑metabolic coupling in adipose tissue, liver, and vasculature. Under normal physiological conditions, mechanical loading suppresses PPARγ to promote osteogenesis and vascular homeostasis; under pathological conditions, aberrant signals and PPARγ dysfunction establish a vicious cycle of “mechanical imbalance–metabolic disorder–tissue remodeling.” This review suggests that PPARγ may function as a mechano‑metabolic downstream transcriptional transducer linking the mechanical microenvironment to metabolic reprogramming, thereby offering a novel theoretical framework and translational perspective for the physical intervention and targeted therapy of common metabolic diseases.
Ultra-low-pass whole-genome sequencing (ULP-WGS) of cell-free DNA (cfDNA) offers a cost-efficient strategy for cancer detection, but its clinical application is limited by extreme data sparsity and poor model generalization. We developed Fragmentia-AI™ WGS, a mutation-calling-independent framework that uses a transformer-based multiple-instance learning architecture with sequential fine-tuning across tumor fraction (TF) strata to extract latent cancer-associated signals from ULP-WGS data. Model performance was evaluated in multiple independent cohorts, including a pan-cancer test set covering 17 cancer types, an external public dataset generated on a different sequencing platform, and a technical variability cohort with heterogeneous pre-analytical and experimental conditions. Clinical relevance was assessed by correlating model predictions with progression-free survival (PFS) in patients with advanced non-small cell lung cancer receiving chemoimmunotherapy. Sequential fine-tuning across TF strata significantly improved performance in low-TF samples, achieving a 35.6
miR-210-3p is a well-established hypoxia-induced microRNA that is commonly upregulated in a wide range of solid tumors, traditionally linked to mitochondrial repression and hypoxia-inducible factor (HIF) signaling. However, its functional role in cancer remains complex and highly context dependent. Here we perform a comprehensive pan-cancer transcriptomic analysis together with functional assays, revealing that miR-210-3p not only mediates classical hypoxic responses but also amplifies mitotic gene expression through activation of FOXM1. Mechanistically, this effect is shown to be dependent on HIF1α but not on HIF2α and, importantly, it has not been recapitulated by hypoxia alone. Notably, activation of the mitotic program is observed in breast cancer cells but not in head and neck squamous carcinoma models, highlighting a strong degree of context dependency across tumor types. In breast cancer cells, miR-210-3p overexpression enhances FOXM1 phosphorylation, upregulates kinetochore regulators, and induces mitotic defects, correlating with poor prognosis in aggressive tumors. Together, these findings position miR-210-3p as a molecular integrator linking pseudohypoxia to mitotic dysregulation, contributing to tumor aggressiveness by sustaining HIF activity and promoting mitotic stress. This dual functionality reconciles its paradoxical effects on proliferation and highlights its potential as a therapeutic target in cancers characterized by pseudohypoxia and mitotic abnormalities.
Iron overload-related liver injury is a major clinical problem in hereditary hemochromatosis, transfusional siderosis, and chronic liver diseases, yet current treatments such as iron chelation therapy and phlebotomy are limited by safety and efficacy issues. Cannabinoid receptor type 2 (CB2R) has been implicated in the regulation of iron metabolism, but its role and underlying mechanism in iron overload remain unclear. Here, we investigated whether CB2R can prevent liver damage caused by iron overload and explored its potential mechanism. Male C57BL/6J mice received 100 mg/kg iron-dextran intraperitoneally to establish an iron overload model. Liver damage and iron deposition were evaluated via histology, serum biochemistry, and molecular analyses. Further studies using primary hepatocytes and Kupffer cells exposed to ferric ammonium citrate (FAC) were performed to explore cell type-specific mechanisms in vitro. Pharmacological CB2R activation with the selective agonist JWH133 reduced liver iron deposition, oxidative stress, and inflammation, whereas CB2R-knockout (KO) exacerbated these phenotypes. Mechanistically, CB2R activation suppressed signal transducer and activator of transcription 3 (STAT3)-dependent hepcidin transcription in hepatocytes, thereby enhancing ferroportin 1 (FPN1)-mediated iron efflux. Concurrently, CB2R promoted nuclear factor erythroid 2-related factor 2 (Nrf2)/FPN1 signaling in Kupffer cells, decreasing the labile iron pool. These findings identify CB2R as a key regulator of hepatic iron homeostasis and suggest that targeting CB2R may offer a novel therapeutic strategy for iron overload-related diseases.
