Bone health is critically dependent on the precise regulation of bone-forming cells, which are significantly impacted by glucocorticoids. Although DNA damage-regulated autophagy modulator 1 (DRAM1) plays a key role in autophagy, its involvement in glucocorticoid-induced osteoporosis has not been fully explored. Here, we showed that alterations in N⁶-methyladenosine (m⁶A) levels contributed to the regulation of DRAM1 under dexamethasone treatment. In vitro, our research revealed that high doses of dexamethasone impaired the m⁶A-dependent regulation of DRAM1 by YTH m⁶A RNA binding protein F1 (YTHDF1), leading to decreased DRAM1 levels and subsequent disruption of autophagy-associated osteogenic differentiation in human bone marrow mesenchymal stem cells (hBMSCs) and MC3T3-E1 cells. Additionally, the classic Wnt/β-catenin pathway, which plays a critical role in bone formation, was shown to be modulated by DRAM1 during osteogenic differentiation. In vivo experiments showed that DRAM1 protein expression in the femurs of Ythdf1 knockout (KO) mice was significantly lower than that in the wild-type (WT) group, and that resveratrol not only mitigated dexamethasone-induced bone damage but was also associated with increased DRAM1 expression. These findings advance our understanding of how glucocorticoids hinder bone formation and suggest that targeting the m⁶A-dependent YTHDF1/DRAM1 regulatory axis may offer novel strategies for osteoporosis treatment.

Background: Endoplasmic reticulum oxidoreductase 1α (ERO1A) is the core engine of oxidative protein folding in the endoplasmic reticulum (ER), playing a central role in maintaining ER redox homeostasis and modulating the unfolded protein response (UPR). Its aberrant overexpression in multiple solid tumors has established ERO1A as a critical regulator of tumor progression, microenvironment remodeling, and therapy resistance, positioning it as an emerging therapeutic target in precision oncology.

Main body: This review systematically synthesizes the structural biology, catalytic mechanisms, and regulatory networks of ERO1A, encompassing transcriptional control, post-translational modifications, and compensatory alternative oxidases. We detail the multidimensional oncogenic functions of ERO1A, including intracellular promotion of proliferation, apoptosis resistance, migration, invasion, and epithelial-mesenchymal transition, as well as extracellular remodeling of the tumor microenvironment via VEGF-driven angiogenesis, PD-L1-mediated immune evasion, metabolic reprogramming, and induction of CD8⁺ T cell exhaustion and recruitment of immunosuppressive cells. The clinical relevance of ERO1A as an independent prognostic biomarker and its association with chemotherapy and immune checkpoint inhibitor resistance are critically evaluated. Current therapeutic strategies targeting ERO1A are classified into FAD-competitive inhibitors, non-competitive inhibitors, allosteric inhibitors, and emerging PROTAC degraders, with discussion of their mechanisms, selectivity, and translational hurdles.

Conclusion: ERO1A represents a promising yet challenging therapeutic node that connects ER stress adaptation with tumor pathogenesis. Overcoming the limitations of current inhibitors—particularly poor isoform selectivity, off-target effects, and pharmacokinetic deficiencies-through structure-guided optimization, allosteric modulation, or protein degradation technologies will be essential for clinical translation. Future efforts should focus on biomarker-driven patient stratification and rational combination with immunotherapies or conventional chemotherapies to maximize therapeutic benefit.

Background: Acute lung injury (ALI) is a common inflammatory pulmonary disorder, with increasing evidence implicating ferroptosis as a critical type of cell death in its pathogenesis. Ubiquitin-specific protease 43 (USP43) is an important deubiquitinating enzyme that plays a significant role in both inflammation and ferroptosis regulation. In this study, we mainly analysed whether USP43 could participate in the process of ALI by regulating the ferroptosis process, and clarified its molecular mechanism.

Methods: To investigate the functional role of USP43 in ALI, Usp43 knockout mice, USP43 knockdown and overexpressed human bronchial epithelial BEAS-2B cells and mouse alveolar type II epithelial MLE12 cells were treated with lipopolysaccharide (LPS) in vivo and in vitro. Subsequently, ferroptosis inhibitor and activator, Ferrostatin-1 and Erastin, were taken to explore the effect of ferroptosis on the regulation function of USP43 in ALI. Finally, co-immunoprecipitation (Co-IP), ubiquitination and rescue assays were conducted to determine the regulation mechanism of USP43.

Results: The expression of USP43 was up-regulated during ALI. Usp43-KO mice exhibited aggravated lung injury, inflammation and ferroptosis. Consistently, USP43 knockdown exacerbated LPS-stimulated cellular damage, inflammation and ferroptosis, while its overexpression exerted the opposite effect in vitro. Furthermore, the regulation effects of USP43 on ALI mainly depended on ferroptosis by administering a ferroptosis inducer and inhibitor, respectively. Mechanistically, USP43 was found to inhibit K48-linked polyubiquitination of solute carrier family 7 member 11 (SLC7A11), thereby stabilising SLC7A11. Overexpression of SLC7A11 rescued the ferroptosis, cellular and lung tissue injury and inflammation aggravated by USP43 knockdown or deficiency.

Conclusion: USP43 prevents the progression of ALI by mediating K48-linked deubiquitination of SLC7A11, thereby inhibiting ferroptosis. Targeting USP43 may represent a potential therapeutic strategy for ALI treatment by enhancing SLC7A11 stability and inhibiting ferroptosis.

Background: Sarcomas are rare, diverse malignancies with limited therapeutic options and poor clinical outcomes. Preclinical models that preserve tumour biology are urgently needed to advance mechanistic understanding and functional precision oncology. We established and comprehensively characterized 29 early-passage patient-derived sarcoma cell (PDC) cultures from 19 patients, representing 11 sarcoma subtypes. Multi-region and multi-site sampling enabled generation of PDCs from spatially distinct areas of individual tumours and from matched primary, recurrent and metastatic lesions.

Methods: PDCs underwent genomic, transcriptomic and proteomic profiling alongside extracellular vesicle (EV) biomarker evaluation and phenotypic and drug-response assays.

Results: Copy-number variant (CNV) analysis revealed recurrent alterations affecting key regulators of cell cycle control and growth signalling. Bulk RNA-sequencing captured substantial inter- and intra-subtype heterogeneity. Proteomic and EV analyses recapitulated subtype- and site-specific differences. A functional drug screen of 38 clinically relevant and investigational agents identified both shared and divergent therapeutic vulnerabilities in the different PDCs.

Discussion: These results demonstrate that early-passage sarcoma PDCs can be regarded as biologically faithful and experimentally tractable models that capture lineage identity, tumour evolution and functional heterogeneity. Integrated multi-omic and functional profiling reveals therapeutic vulnerabilities not evident from genomic data only, supporting PDCs as valuable platforms for translational sarcoma research.

Background: Oral squamous cell carcinoma (OSCC) remains a major clinical challenge, with delayed diagnosis, frequent resistance to therapy, and poor long-term survival.

Methods: This review systematically evaluates the methodological framework for applying AI to oral microbiome data in OSCC. Emerging paradigms, including self-supervised learning for leveraging unlabelled data and explainable AI (XAI) techniques for model interpretability, are also discussed. Model evaluation relies on cross-validation, hyperparameter optimisation, and performance metrics such as AUC, accuracy, sensitivity, specificity, and F1-score.

Results: Multiple studies demonstrate that AI-based classifiers, especially random forest models built on salivary or tissue-derived microbial features, achieve outstanding discrimination between OSCC patients and healthy controls in retrospective, single-centre cohorts, with reported AUC values exceeding 0.99 and accuracy >95%. However, these exceptional metrics should be interpreted with caution, as they are susceptible to cohort size, sampling site heterogeneity, batch effects, feature-selection bias, and the absence of independent external validation. Beyond binary diagnosis, AI has been successfully applied to predict lymph node metastasis, explore tumour metabolic reprogramming, and assess environmental interactions. Integrated multi-omics approaches further enhance the specificity and clinical relevance of microbial biomarkers.

Conclusions: The convergence of AI and oral microbiome analysis is reshaping the diagnostic and therapeutic landscape of OSCC, and explore microbiome-targeted combination therapies. Addressing these challenges will be pivotal to realising truly intelligent, personalised management and ultimately improving outcomes for OSCC patients.

Background: Diabetes accelerates atherosclerotic plaque expansion and loss of stability, but the plaque-resident mechanisms through which the diabetic milieu promotes macrophage lipid-oxidative injury are not fully understood.

Methods: We used apolipoprotein E-deficient (ApoE^−/−) mice with Mex-3 RNA-binding family member A (MEX3A) deficiency and primary bone marrow-derived macrophage (BMDM) experiments under diabetic conditions to investigate the role of MEX3A in diabetic atherosclerosis and macrophage lipid-peroxidation injury.

Results: MEX3A loss worsened diabetic atherosclerosis while leaving body weight, glycaemia and lipid measurements largely unchanged relative to diabetes alone. MEX3A deficiency enlarged aortic lesions, enhanced lipid deposition, increased macrophage content, expanded necrotic cores and thinned fibrous caps, while reducing collagen content. In lesional macrophages, loss of MEX3A coincided with increased lipid reactive oxygen species, malondialdehyde, increased labile iron, 4-hydroxynonenal, a disturbed glutathione redox state, mitochondrial abnormalities compatible with ferroptotic stress, lower GPX4/SLC7A11 and higher ACSL4. CRISPR/Cas9-mediated Mex3a knockout in BMDMs recapitulated this phenotype, whereas restoring MEX3A blunted it. RNA immunoprecipitation-qPCR together with actinomycin D decay analyses demonstrated preferential recovery of Slc7a11 and Gpx4 transcripts with MEX3A and decreased stability of Slc7a11 and Gpx4 mRNAs after Mex3a knockout. GPX4 or SLC7A11 overexpression reduced lipid-peroxidation injury in Mex3a-knockout macrophages, whereas Gpx4 or Slc7a11 knockdown weakened the protection conferred by MEX3A re-expression. Ferrostatin-1 partially attenuated macrophage lipid peroxidation and plaque injury.

Conclusions: Together, these results place MEX3A among the protective regulators of diabetic plaque stability and support a MEX3ASLC7A11/GPX4-linked ferroptosis-associated mechanism in plaque macrophages.

Background: Sepsis-induced acute kidney injury (S-AKI) is a major global public health concern, yet effective therapeutic strategies remain limited. Mitochondrial dysfunction in renal tissues is a key pathogenic mechanism underlying S-AKI. GTS-21, a selective α7 nicotinic acetylcholine receptor (α7nAChR) agonist, exhibits anti-inflammatory and renoprotective effects in S-AKI.

Methods: We investigated the role of α7nAChR in S-AKI using both in vitro (lipopolysaccharide (LPS)-induced renal tubular cell injury) and in vivo (caecal ligation and puncture (CLP)-induced septic mice) models, with GTS-21 treatment.

Results: GTS-21 significantly attenuated mitochondrial dysfunction, suppressed apoptosis, and alleviated inflammation, thereby protecting renal tubular cells and renal tissues against LPS- and CLP-induced injury. Mechanistically, GTS-21 activated α7nAChR and upregulated myocyte enhancer factor 2 (MEF2), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), and heme oxygenase-1 (HO-1), which collectively mediate its anti-oxidative, anti-apoptotic and anti-inflammatory effects.

Conclusion: These findings suggest that GTS-21 may represent a potential therapeutic strategy for sepsis-induced kidney injury.

Background: The interplay between tumour cells and tumour-associated macrophages (TAMs) within the tumour microenvironment is crucial for the progression of non-small cell lung cancer (NSCLC). The underlying mechanisms involving RNA modification and exosomal communication remain incompletely understood.

Methods: Multiplex immunofluorescence and flow cytometry were performed to evaluate M2 macrophage polarization. Exosomes were isolated by ultracentrifugation and validated by transmission electron microscopy, nanoparticle tracking analysis, and exosomal marker blots. To investigate the molecular mechanism, methylated RNA immunoprecipitation (MeRIP)-qPCR and dual-luciferase reporter assays were used to validate m⁶A modification sites on NFIC and miR-194-5p; RNA immunoprecipitation (RIP) confirmed the interaction between ZNF106 and interleukin-6 (IL-6) mRNA; chromatin immunoprecipitation (ChIP) was employed to detect STAT3 binding to the METTL3 promoter. The in vivo function of the identified feedback loop was assessed using an orthotopic xenograft mouse model of NSCLC.

Results: A negative feedback loop between METTL3 and NFIC was demonstrated in NSCLC cells. METTL3 suppressed miR-194-5p expression and its loading into exosomes through m⁶A methylation. NSCLC-derived exosomal miR-194-5p was internalized by macrophages and directly targeted ZNF106, thereby inhibiting M2 polarization. In macrophages, ZNF106 stabilized IL-6 mRNA and promoted exosomal IL-6 secretion, thereby activating the JAK2/STAT3 pathway and upregulating METTL3. This IL-6-driven METTL3 upregulation formed a positive feedback loop that sustains M2 polarization and tumour progression. In vivo disruption of this loop reduced tumour growth and metastasis.

Conclusions: These findings establish a closed regulatory circuit initiated by an NFIC/METTL3 negative feedback loop. In this circuit, METTL3-mediated m⁶A modification of exosomal miR-194-5p in NSCLC cells derepresses ZNF106 expression in macrophages, leading to IL-6 production that activates the JAK2/STAT3 pathway and upregulates METTL3 in tumour cells, thereby perpetuating M2 polarization and malignant progression. This circuitry offers potential nodes for therapeutic intervention in NSCLC.

Objective: To determine whether macrophage gasdermin D (GSDMD)-dependent pyroptosis drives acute cardiac allograft rejection and whether targeting GSDMD improves graft survival.

Methods: GSDMD activation was assessed by immunoblotting. Graft survival wasanalysed in global and macrophage-specific GSDMD-deficient recipients. Single-cell RNA-seq identified GSDMD-expressing populations, while immune infiltration and cytokines were measured by flow cytometry and immunohistochemistry; IL1R1 knockout and cytokine challenge were used for validation.

Results: GSDMD deficiency markedly prolonged graft survival. Single-cell profiling showed increased GSDMD expression and pyroptosis primarily in recipient-derived M1 macrophages during acute rejection. Macrophage-specific GSDMD deficiency reduced CD8⁺T cell/macrophage infiltration and decreased TNF-α and IL-1β. TNFα/IL6 activated NF-κB and JAK–STAT3 to upregulate GSDMD, and IL-1β was verified as a key effector for CD8⁺T cells mediated-rejection. GSDMD pyroptosis inhibitors mitigated rejection and extended graft survival.

Conclusion: Recipient M1 macrophages promote acute rejection via TNFα/IL6-driven NF-κB/STAT3 activation of GSDMD-mediated pyroptosis. Targeting GSDMD represents a promising strategy to enhance cardiac allograft survival.

Background: Transferrin receptor 1 (TfR-1), a key mediator of cellular iron uptake, is significantly upregulated across a broad spectrum of malignancies, and its overexpression highly correlated with poor clinical outcomes, such as in breast cancer. However, its precise role in breast cancer progression remains unclear.

Methods: We integrated clinical breast cancer tissue specimens and public transcriptomic datasets to analyse the clinical correlation between TfR-1 expression and tumour metastasis. A series of in vitro cell functional assays and in vivo xenograft tumour metastasis models were performed to validate the regulatory effect of TfR-1 on breast cancer proliferation and metastasis. Co-immunoprecipitation, phosphorylation detection and protein stability assays were further applied to dissect the intermolecular regulatory network among TfR-1, Hematopoietic Cell Kinase (HCK) and Ubiquitin Specific Peptidase 32 (USP32).

Results: Clinical data analysis revealed that elevated TfR-1 expression was tightly linked to metastatic phenotype in breast cancer. Functional experiments confirmed that upregulated TfR-1 robustly boosted the proliferative and metastatic capacity of breast cancer cells in vitro and in vivo. Mechanistically, TfR-1 dual-regulates HCK, it directly induces HCK phosphorylation, while simultaneously activating USP32 to block HCK protein degradation and sustain HCK abundance. Activated HCK further triggers STAT3 transcription factor signalling, which drives the upregulation and secretion of matrix metalloproteinase 9 (MMP9). In addition, HCK reciprocally phosphorylates TfR-1 to establish a positive feedback circuit amplifying the prometastatic signalling.

Conclusions: This work identifies a previously uncharacterized prometastatic function of TfR-1 independent of its canonical iron transport activity in breast cancer. We uncover a reciprocal TfR-1/HCK positive feedback axis that activates the USP32-HCK-STAT3-MMP9 signalling cascade to facilitate breast cancer metastasis, which provides novel candidate therapeutic targets for metastatic breast cancer intervention.

Background: Atherosclerotic cardiovascular disease (ASCVD) is a leading cause of global morbidity and mortality, but its genetic architecture remains incompletely understood. This study aims to uncover novel genetic insights into ASCVD through multivariate genomic analysis and molecular structure predictions.

Methods: We analysed genomic data from over 3.8 million individuals across various ASCVD phenotypes, including coronary heart disease, stroke, transient ischemic attack, peripheral artery disease and abdominal aortic aneurysm. Advanced tools such as Genomic Structural Equation Modeling, fine-mapping, FUSION, FOCUS and other functional annotation methods were applied to identify causal single nucleotide polymorphisms associated with ASCVD. Protein structural analysis was performed using AlphaFold3, and AI-driven thermodynamic analysis (ThermoMPNN) assessed the stability and functional consequences of mutations.

Results: Our analysis revealed 347 genome-wide significant variants linked to ASCVD, distributed across 213 loci. Ninety of these variants were not identified in any of the five input GWAS datasets. Upon cross-referencing with large-scale external GWAS, 19 of the 90 variants showed no prior association with any cardiovascular or metabolic trait, 15 were previously reported only in risk factor GWAS, and 56 had been reported in direct ASCVD endpoint GWAS. The latter group includes the DCLRE1B rs11552449 missense mutation. Nevertheless, AI-based structural and thermodynamic analyses revealed that this mutation (H61Y) disrupts DCLRE1B protein stability, increases conformational flexibility, and alters solvent-accessible surface area—mechanistic insights that have not been previously described.

Conclusions: This study provides a hypothesis-generating genetic landscape of ASCVD, unveiling novel variants and their molecular impacts. These findings enhance our understanding of ASCVD mechanisms and may offer potential avenues pending experimental validation and targeted therapies in cardiovascular disease.

Background: Chemotherapy resistance remains a critical hurdle in advanced prostate cancer (PCa). Succinylation, an essential post-translational modification linking cellular metabolism with epigenetic regulation, has been implicated in tumour progression; however, its contribution to PCa chemoresistance remains poorly defined.

Objective: This study aimed to evaluate the prognostic significance of succinylation in PCa, develop a succinylation-based biomarker, and elucidate the mechanisms driving chemotherapy resistance.

Methods: We generated a succinylation score (SS) by applying single-sample gene set enrichment analysis (ssGSEA) to transcriptomic profiles from the TCGA-PRAD cohort. Its relationships with survival, the tumour microenvironment (TME), and treatment susceptibility were examined using CIBERSORT, GSEA, TIDE, oncoPredict, and single-cell RNA sequencing (scRNA-seq). Findings were functionally validated in patient-derived organoids, PCa cell lines, and xenograft models through genetic manipulation, chemosensitivity assays, and mechanistic studies.

Results: High SS correlated with favourable prognosis, lower Gleason scores, absent lymph node metastasis, and an immune-active TME enriched in CD8⁺ precursor exhausted T cells. High-SS tumours showed enhanced sensitivity to docetaxel and cisplatin. scRNA-seq identified KAT2A as a key driver in low-SS malignant clusters with chemoresistance features. KAT2A was elevated in chemoresistant tissues, cell lines, and organoids. KAT2A knockout sensitised cells to chemotherapy, while ectopic expression promoted resistance in vitro and in vivo. Mechanistically, KAT2A-mediated succinylation of PIK3R2 at K477 and K564 inhibited its ubiquitination and proteasomal degradation, stabilising PIK3R2 to drive chemoresistance. The KAT2A inhibitor Butyrolactone 3 synergised with standard chemotherapy to suppress tumour growth.

Conclusion: The succinylation score serves as a robust prognostic biomarker integrating metabolic and immunological features in PCa. The KAT2A-PIK3R2 succinylation pathway represents a newly defined driver of chemoresistance and points to MB-3-based combination therapy as a potential strategy for resistant advanced disease.

Background: Atherosclerotic cardiovascular disease (ASCVD) is the leading cause of morbidity and mortality worldwide. Despite effective lipid-lowering treatments, substantial residual risks remain. In atherosclerosis, vascular smooth muscle cells (SMCs) undergo dedifferentiation, promoting disease progression. Membrane-type I matrix metalloproteinase (MT1-MMP/MMP14) promotes SMC dedifferentiation. However, the effect of inhibiting MMP14 in adults, particularly those with existing atherosclerotic plaques, is unclear.

Methods: We developed an inducible conditional SMC-specific MMP14 knockout mouse model. Cardiac and vascular function were assessed using echocardiography and wire myography, respectively. Atherosclerosis progression and regression were evaluated in Ldlr^−/− mice with or without MMP14 deficiency. snRNA-seq of the aortas from Ldlr^−/− mice was performed to determine the effect on SMC populations.

Results: MMP14 expression was elevated in SMCs within fibroatheroma compared with the pathological intima thickening in coronary aortas from patients with ASCVD. Conditional knockout of SMC MMP14 in adult mice did not change plasma cholesterol levels or basic cardiac and vascular function. However, atherosclerosis development was reduced, and the regression of existing plaques was enhanced in Ldlr^−/− mice lacking SMC MMP14. snRNA-seq revealed increased fibroblast-like SMCs and reduced foam cell-like SMCs in MMP14-deficient Ldlr^−/− mice compared to Ldlr^−/− mice. Furthermore, SMC MMP14 deficiency decreased SMC proliferation and migration, accompanied by reduced platelet-derived growth factor receptor (PDGFR) β levels and attenuated PDGF signalling.

Conclusion: SMC MMP14 promotes atherosclerosis in adult mice, likely through reducing PDGF signalling and inhibiting SMC migration and proliferation.

Introduction: Metabolic dysfunction-associated fatty liver disease (MAFLD) is a common chronic liver condition marked by abnormal lipid metabolism.

Objectives: Small nucleolar RNA host gene 5 (SNHG5) is involved in the regulation of cell proliferation and apoptosis and was previously identified as a profibrotic factor in liver fibrosis. However, its role in MAFLD remains unclear. This study aimed to elucidate the contribution of SNHG5 to MAFLD progression.

Methods and results: SNHG5 expression was markedly elevated during MAFLD progression, whereas SNHG5 inhibition suppressed lipid accumulation. Transcriptomic sequencing of primary hepatocytes overexpressing SNHG5 demonstrated significant downregulation of monocarboxylate transporter 1 (MCT1), a key lactate transporter. Integrated lactyl-proteomic and proteomic analyses further revealed that SNHG5 overexpression promoted lactylation of the non-histone protein AT-rich interaction domain 1A (Arid1a) at K391, thereby driving excessive lipid accumulation. Acyltransferase assays indicated that both P300 and HBO1 participated in SNHG5-mediated Arid1a lactylation. In vivo, hepatocyte-specific SNHG5 knockout markedly attenuated hepatic lipid accumulation and MAFLD progression, whereas simultaneous hepatocyte-specific deletion of SNHG5 and MCT1 restored lipid accumulation compared with SNHG5-deficient mice.

Conclusion: SNHG5 promotes lipid accumulation and MAFLD progression through induction of MCT1-mediated Arid1a K391 lactylation.

Background: The systematic link between cellular spatial organization and its biomechanical consequences remains a critical knowledge gap in human oncology.

Objective: To quantitatively map the cellular architecture of solid tumours and elucidate its mechanistic impact on mechanical signalling, immune infiltration, and patient survival.

Methods: We developed a scalable digital pathology framework, processing 7910 H&E whole-slide images across 21 solid tumours. A deep learning pipeline was employed to segment over 4.7 billion nuclei, enabling the calculation of cell density and nearest neighbour distance (NND) as key spatial metrics. To bridge morphology and function, we integrated these metrics with bulk RNA-seq data from 19 TCGA cohorts (n = 7401) using rigorous histological matching. Furthermore, to resolve microenvironmental heterogeneity, we performed unsupervised clustering of over 60 000 T-cell transcriptomes from independent cohorts. These findings were validated through high-resolution Visium-HD spatial transcriptomics to correlate physical proximity with localized gene expression.

Results: While tumours exhibited significant heterogeneity, NND, but not cell density, emerged as a primary determinant of biomechanical and immune signatures. A trend-level association was observed between lower NND and higher Hippo/YAP/TAZ pathway activity across nine matched cancer types (Spearman's ρ = -.65, p = .058, n = 9). In addition, lower NND was significantly correlated with increased CD4⁺ memory T-cell (CD4⁺ T_Mem cell) abundance (Spearman's ρ = -.86, p < .01). Single-cell analyses confirmed that CD4⁺ T_Mem cells intrinsically express mechanical stress signalling markers, which spatial transcriptomics localized to CD4⁺ T_Mem cell aggregation zones characterized by high pathway activity. Clinically, this spatial-mechanical-CD4⁺ T_Mem cell axis was associated with prognosis in breast, oesophageal, liver, and lung adenocarcinomas, where high mechanical signalling generally predicted poor outcomes but could be modulated by CD4⁺ T_Mem infiltration levels.

Conclusion: Our study identifies low NND as a spatial correlate of biomechanical crowding that is associated with CD4⁺ T_Mem cell programming and adverse clinical outcomes. By integrating deep learning-based spatial metrics with multi-omics, we highlight spatial mechanics as a critical, potentially targetable dimension of the tumour microenvironment for future immunotherapies.

Background: Septic cardiomyopathy (SCM) is the leading cause of mortality among patients diagnosed with sepsis. Nevertheless, the precise mechanisms underlying its pathogenesis remain poorly understood. Deubiquitinating enzymes (DUBs) play a vital role in various cardiovascular diseases.

Methods: This study investigated deubiquitinating enzymes in septic myocardial injury via public RNA-Seq. Pyroptosis was modelled in primary neonatal rat cardiomyocytes using lipopolysaccharide (LPS) and nigericin, with levels assessed by IL-1β ELISA, Western blot, PI staining, CCK-8 and LDH assays. Potential substrates of OTUD5 were screened by Co-immunoprecipitation. Cardiomyocyte-specific OTUD5-knockout mice generated by CRISPR/Cas9 were subjected to LPS- or Caecal ligation and puncture (CLP)-induced sepsis models for cardiac function and pyroptosis evaluation. AAV9-mediated cardiomyocyte-specific OTUD5 overexpression in NLRP3-knockout mice was used to validate OTUD5-NLRP3 functional interaction.

Results: This study identifies the deubiquitinating enzyme OTUD5 as being significantly upregulated in myocardial tissue subjected to sepsis induced by LPS and CLP. Cardiomyocyte-specific knockout of OTUD5 leads to exacerbated septic myocardial injury and pyroptosis. Mechanistically, OTUD5 directly interacted with NLRP3, with its C224 site facilitating deubiquitination to inhibit NLRP3 activation. Notably, the protective effects associated with OTUD5 overexpression were lost in NLRP3 knockout mice, underscoring its dependence on NLRP3 for function.

Conclusions: This study has confirmed that OTUD5 inhibits pyroptosis by suppressing the activity of NLRP3, thereby ameliorating septic cardiomyopathy. OTUD5 is worthy of further exploration as a potential therapeutic target for septic cardiomyopathy.

Background: Tumour-node-metastasis staging does not fully explain prognostic heterogeneity in non-small cell lung cancer. We evaluated whether haematoxylin-and-eosin whole-slide images could estimate histological subtype, pathological stage probabilities, survival risk and spatially grounded biological associations.

Methods: SparseAGE-MTL, a weakly supervised multi-task multiple-instance learning model with a shared projection-topology encoder and endpoint-specific heads, was trained and benchmarked in 954 The Cancer Genome Atlas cases using seven pathology feature spaces and 18 comparator models. External evaluation used 948 tissue-microarray and 324 whole-slide cases. Attention maps were co-registered with 10x Visium spatial transcriptomics and integrated with bulk transcriptomics, immune-infiltration estimates and ESTIMATE scores. Analyses included paired model comparisons, false-discovery-rate correction, Cox models, calibration assessment and decision curve analysis.

Results: In the CONCH feature space, SparseAGE-MTL achieved 93.73% accuracy, 98.19% area under the receiver-operating-characteristic curve and 93.08% F1-score for adenocarcinoma/squamous cell carcinoma classification in internal benchmarking; external area-under-the-curve values were approximately .91 and .82. Stage estimation had lower discrimination, with external overall area under the curve approximately .70 and cohort-dependent calibration. Risk-score-defined groups differed in overall survival in both histological subtypes and showed similar external trends. High-attention regions were enriched at tumour–stroma or tumour–immune interfaces and were associated with B-cell, fibroblast, C1QC, COL1A1, epithelial–mesenchymal transition, metastasis and hypoxia signals. Higher risk cases showed malignant pathway activation, lower immune/stromal scores, higher tumour purity and subtype-specific immune/stromal differences. Adding the risk score to the clinical model increased external pooled concordance index from approximately .620 to .672.

Conclusions: In retrospective cohorts, SparseAGE-MTL generated subtype-classification, stage-probability and survival-risk outputs from routine pathology images. Subtype classification had higher numerical performance than stage estimation. Survival-risk and attention outputs were associated with outcome and spatial/transcriptomic features, but prospective, treatment-annotated validation is required before clinical use.

Background: The resolution of inflammation is actively driven by omega-3 polyunsaturated fatty acids (PUFAs) via their specialized pro-resolving mediator (SPM) derivatives, including resolvin E1 (RvE1), whose role has been well established. However, clinical application of these mediators is hampered by inherent instability and elevated production costs.

Methods: To surmount these obstacles, we have engineered a biosynthetic platform based on the probiotic Escherichia coli Nissle 1917 (EcN) that enables controlled, sustained RvE1 production through inducible expression of COX2 and 5-LOX, designated EcN-RvE1. The catalytic capacity, intestinal persistence, and therapeutic efficacy of the platform were evaluated in vitro and in LPS-induced acute inflammation and DSS-induced colitis murine models.

Results: In this study, we validate the capacity of EcN-RvE1 to catalyse the conversion of eicosapentaenoic acid (EPA) to RvE1 and confirm its ability to achieve long-term intestinal persistence. In murine models of acute inflammation and colitis, EcN-RvE1 exerts marked anti-inflammatory and tissue-protective effects, which are mediated by the regulation of inflammatory cytokine expression and the amelioration of gut microbiota dysbiosis. Moreover, EcN-RvE1 using Euglena gracilis as a photosynthetic protist-based source of PUFAs also exhibits protective anti-inflammatory activity.

Conclusion: Collectively, we report a probiotic engineering platform for the biosynthesis of RvE1, offering a novel strategy for harnessing the anti-inflammatory potential of PUFAs derivatives in clinical settings.