Background: Gastrointestinal cancers remain a leading cause of cancer-related morbidity and mortality worldwide, with surgery being central to curative treatment. Tumour-targeted fluorescence-guided surgery (tFGS) has emerged as a promising approach to improve intraoperative visualisation and oncological precision.
Methods: We conducted a systematic review of preclinical and clinical studies on tFGS in gastrointestinal oncology, registered in PROSPERO (ID: 558994) and following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Searches of PubMed and Embase identified 133 eligible studies.
Results: Nine tracers have currently been evaluated in clinical trials, targeting the following biomarkers: carcinoembryonic antigen, vascular endothelial growth factor, epidermal growth factor receptor, folate receptor α and/or β, integrin αvβ3, 5-aminolevulinic acid and acidic tumour microenvironment.Clinical trials demonstrated that tracers like SGM-101 and panitumumab–IRDye800CW can achieve high tumour-to-background ratios (TBRs) up to 6.1 ex vivo and alter surgical strategy in up to 35% of cases. Preclinical research identified additional promising targets, including mucins, epithelial cell adhesion molecules, urokinase receptors, tumour-associated glycoproteins, gamma-glutamyl transferase, organic anion transporting polypeptides, human epidermal growth factors 1/2 and Lewis antibodies, with TBRs frequently exceeding three.Despite encouraging feasibility and safety data, translation into routine practice is hampered. Confirmation from early health technology assessments is needed to advance tFGS. Besides, standardisation of protocols and phase III confirmatory trials are required to establish clinical benefit and support regulatory approval.
Conclusion: tFGS holds considerable potential to transform surgical oncology in GI cancers by enabling more precise resections and guiding organ-preserving strategies. Future innovations in multimodal tracers, NIR-II fluorophores and theranostics may further enhance the precision and therapeutic potential of tFGS.
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Background: Ageing is a significant risk factor for pathophysiological alterations in the heart, but the intrinsic mechanisms by which these occur have yet to be fully elucidated. Purinergic 2×7 receptor (P2X7R) is important for the pathogenesis of numerous cardiovascular diseases; nevertheless, its function in the process of cardiac ageing remains uncertain.
Methods: This study utilised P2X7R knockout (P2X7R−/−) mice. An ageing model was established by either maintaining mice until they reached 20 months of age or performing chronic subcutaneous injection of D-galactose (D-gal). Recombinant adeno-associated virus serotype 9 (AAV9) was employed to achieve cardiac-specific overexpression of P2X7R and nuclear receptor subfamily 4 group A member 1 (NR4A1). Cardiac function and histopathological changes in cardiac tissues were evaluated. Transcriptome sequencing was further applied to elucidate the potential mechanisms of P2X7R in cardiac senescence.
Result: Our result show that serum levels of P2X7R increase with advancing age in humans and that P2X7R expression is upregulated during cardiac senescence in mice. P2X7R deficiency alleviates ageing-related cardiac dysfunction, senescence phenotypes and impaired mitophagy. Cardiomyocyte-specific overexpression of P2X7R with AAV9 exacerbates the myocardial dysfunction, senescence phenotype and mitophagy disruption induced by D-gal. Mechanistically, P2X7R promotes human antigen R (HuR) nucleocytoplasmic shuttling in ageing hearts, thereby increasing the mRNA stability of tripartite motif containing 26 (TRIM26) and the expression of the E3 ubiquitin ligase TRIM26. TRIM26 subsequently mediates NR4A1 ubiquitination, leading to its proteasomal degradation, which subsequently suppresses mitophagy in cardiomyocytes and ultimately accelerates cardiac ageing.
Conclusions: Our findings provide valuable insights into the role of P2X7R in cardiac ageing and identify the HuR/TRIM26/NR4A1 axis as a key signalling pathway through which P2X7R regulates cardiac ageing.
Background: Myocardial ischemia/reperfusion (I/R) injury induces an intense inflammatory response and involves multiple cell death pathways. PANoptosis, an integrated cell death process involving pyroptosis, apoptosis and necroptosis, is a major driver of cardiomyocyte loss during I/R injury. However, the epitranscriptomic control of PANoptosis is poorly understood.
Methods: We investigated the role of ALKBH3, an mRNA N1-methyladenosine (m1A) demethylase, in the regulation of cardiomyocyte PANoptosis using hypoxia/reoxygenation models in vitro and murine I/R models in vivo. Integrated transcriptomic and m1A epitranscriptomic profiling identified downstream targets. Loss- and gain-of-function studies of ALKBH3, AIM2, ZBED6 and STAT1 (siRNA or plasmid overexpression) were coupled with assessments of cell death phenotypes, inflammasome activity and gene expression. Molecular interactions and transcriptional/translational regulation were examined using co-immunoprecipitation, chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays.
Results: Cardiomyocyte-restricted ALKBH3 overexpression mitigates I/R injury in vivo. Mechanistically, ALKBH3 acts as a key suppressor of PANoptosis by inhibiting AIM2. ALKBH3 demethylates m1A onZBED6 mRNA, enhancing ZBED6 translation and limiting cardiomyocyte PANoptosis. Although ZBED6 does not bind directly to the AIM2 promoter, it physically interacts with STAT1, a transcriptional activator of AIM2, and represses STAT1-driven AIM2 expression. ZBED6 overexpression reduces AIM2 levels and PANoptosis, whereas AIM2 knockout attenuates the exacerbation of cardiac injury and PANoptosis induced by ALKBH3 silencing.
Conclusions: These findings identify the ALKBH3/ZBED6/STAT1/AIM2 signalling axis that epitranscriptomically breaks cardiomyocyte PANoptosis, highlighting a tractable therapeutic target that limits cell death and improves myocardial outcomes after I/R.
Background: Clonal haematopoiesis (CH) is an age-related condition increasingly recognised for its relevance in haematologic malignancies. In chronic lymphocytic leukaemia (CLL), its prevalence and clinical implications are gaining attention, particularly in the context of prolonged patient survival and the widespread adoption of targeted therapies. A comprehensive understanding of the biological and clinical significance of CH in CLL is therefore essential.
Methods: This review synthesises current evidence on the biological basis, epidemiology and clinical impact of CH in CLL. Data from prospective clinical trials, real-world cohorts and translational studies were analysed to explore the associations between CH, genomic instability, immune dysregulation and inflammaging. Particular attention was given to the interaction between CH and contemporary therapeutic strategies, including Bruton tyrosine kinase (BTK) inhibitors and BCL2 inhibitors, and their potential influence on long-term outcomes.
Results: Available evidence indicates that CH is relatively frequent in patients with CLL and may contribute to disease biology through mechanisms involving genomic instability, chronic inflammation and immune system alterations. Emerging data suggest that CH can influence prognosis, treatment-related toxicities and cardiovascular risk, as well as predispose to therapy-related myeloid neoplasms. The interplay between CH and targeted agents may further modulate long-term outcomes, although the impact of CH on Richter transformation remains incompletely defined.
Conclusions: CH represents a clinically relevant factor in the management of CLL in the era of targeted therapies. Its detection may have important implications for risk stratification, toxicity monitoring and survivorship care. Further prospective studies are needed to clarify its prognostic value and to integrate CH assessment into routine clinical practice and personalised treatment algorithms.
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Background: Medulloblastoma (MB) represents the most prevalent malignant paediatric brain tumour, characterised by the sonic Hedgehog molecular subtype (SHH-MB), which is driven by aberrant activation of the SHH signalling cascade. Suppressor of fused (SUFU), a core member of SHH signal transduction, functions as a tumour suppressor by inhibiting the activity of transcription factors glioma-associated oncogene homologue (GLI)-triggered SHH signalling pathway. While ubiquitin-mediated proteasomal degradation of SUFU has been shown to modulate SHH signalling, the regulatory factors involved in SUFU deubiquitination and their specific roles in MB pathogenesis remain largely undefined.
Methods: Mass spectrometry and co-immunoprecipitation were employed to identify the interaction between USP9X and SUFU. Clinical correlation analyses were conducted using MB tissue microarrays and publicly available datasets. Ubiquitination assays, functional cell-based experiments, and orthotopic xenograft models were performed to evaluate the biological role of USP9X in SHH-MB. Chaperone-mediated autophagy (CMA) inhibitors were utilised to investigate their regulatory effects on USP9X expression and SHH-MB progression.
Results: Mass spectrometry identified the deubiquitinase ubiquitin-specific protease 9X (USP9X) as a previously unrecognised SUFU-binding partner. Notably, USP9X and SUFU exhibit a positive correlation in a MB tissue array, with both exhibiting low expression levels that are associated with adverse prognostic outcomes. Loss of USP9X in SHH-MB enhances cell proliferation in vitro as well as orthotopic MB xenograft tumourgenicity in vivo. Mechanically, USP9X deubiquitinates and stabilises SUFU, thereby negatively regulating SHH signal transduction. Interestingly, SHH signalling promotes SUFU ubiquitination through CMA-dependent degradation of USP9X protein levels, facilitating pathway activation. Combined inhibition of CMA and SHH pathway had a synergically therapeutic effect on SHH-MB.
Conclusions: These results establish CMA and USP9X as pivotal regulators of SHH medulloblastoma MB progression, emphasising their potential as novel therapeutic targets for medulloblastoma, and combinatorial inhibition of CMA and Smoothened (SMO) may provide a strategy to overcome intrinsic or acquired resistance to SMO monotherapy in SHH-MB.
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Background: Pyroptosis, apoptosis and necroptosis (PANoptosis) simultaneously occur and are extensively cross-linked in infectious and inflammatory diseases. However, the co-existence and regulation of macrophage pyroptosis, apoptosis and necroptosis in atherosclerosis have not yet been investigated.
Methods: Atherosclerotic specimens from human lower extremity amputation and carotid endarterectomy were analysed. Ox-LDL-induced macrophages and high-fat diet (HFD)-fed ApoE−/− mice were employed as in vitro and in vivo models, respectively. Galectin-3 was recognised as a key differentially expressed protein and gene related to PANoptosis by integrative proteomic and transcriptomic analysis of atherosclerotic murine aortas. Single-cell transcriptomic analysis of human carotid endarterectomy specimens investigated the cellular distribution of galectin-3. Galectin-3 is a potent driver of macrophage activation and elicits inflammation through NLRP3 inflammasome activation. To elucidate the role of galectin-3/NLRP3 in atherosclerosis, galectin-3 siRNA transfection in macrophages was conducted, galectin-3 and ApoE dual-deficiency mice were produced, AAV-F4/80-shGalectin-3 was injected, and NLRP3 agonist nigericin was administered.
Results: A substantial content of inflammatory factors, the activation of NLRP3/GSDMD/CASP3/CASP8/RIPK3/pMLKL, and the upregulation of galectin-3 were detected in advanced human and mouse atherosclerotic lesions. Galectin-3 was predominantly expressed in atherosclerotic macrophages, and Galectin-3-positive macrophages were mainly distributed in the atherosclerotic core in comparison with the proximal adjacent artery. Ox-LDL induced apoptosis, pyroptosis and necroptosis in macrophages, as evidenced by the activation of NLRP3/GSDMD/CASP3/CASP8/RIPK3/pMLKL and the secretion of proinflammatory cytokines. Galectin-3 interacted with NLRP3. Genetic knockdown of galectin-3 alleviated ox-LDL-induced activation of inflammatory cell death, which was pronouncedly abrogated by NLRP3 agonist nigericin. Genetic galectin-3 deficiency attenuated, and conversely nigericin exacerbated macrophage death, vascular inflammation and atherosclerosis in HFD-fed ApoE−/− mice. Mechanistically, galectin-3 activated the TLR4/MyD88/NF-κB/NLRP3 axis and induced pyroptosis, apoptosis and necroptosis in macrophages.
Conclusions: Macrophage-derived galectin-3 contributed to pyroptosis, apoptosis and necroptosis in concert, promoted vascular inflammation and atherosclerosis through the upregulation of TLR4/MyD88/NF-κB/NLRP3 pathway.
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Background: Urothelial carcinoma (UC) is a biologically heterogeneous disease, and current molecular classifications have limited integration into clinical decision-making. To further pursue precision oncology efforts in UC, we developed a molecular classification framework applicable to transcriptomic and proteomic data from non–muscle-invasive bladder cancer (NMIBC), muscle-invasive bladder cancer (MIBC) and urothelial cancer cell lines.
Methods: Using a whole-transcriptome self-organised map and regularised semi-supervised clustering of 4439 bulk NMIBC and MIBC transcriptomes and proteomes, and 33 UC cell lines, we identified three molecular UC clusters. Making use of both in silico and in vitro approaches, we selected promising treatment approaches for each cluster.
Results: The three developed clusters displayed distinct signatures of mRNA, proteins, biological processes, metabolism and essential driver genes. They also differed in prognosis and machine learning-predicted treatment vulnerabilities and resistance. High-risk, stroma-rich Cluster #1 cancers were predicted to respond to selected cytotoxic drugs, ferroptosis inducers and PARP inhibitors. For the aggressive, fast-proliferating, immune-infiltrated Cluster #2 tumours with basal/squamous differentiation, cytotoxic agents and EGFR/ERBB- and MEK/ERK-targeting therapies were proposed. Cluster #3 cancers of predominantly luminal papillary phenotype with scarce stroma and immune infiltration were enriched with NMIBC and low-risk malignancies. For patients with Cluster #3 tumours, selected epigenetic drugs or EGFR/FGFR inhibitors may represent attractive treatment options.
Conclusions: Our novel molecular taxonomy holds promise as a practical framework for patient risk stratification and clinical trials in UC. Our molecular classification scheme may facilitate personalised transcriptome- and proteome-based risk assessment and clinical trial design for the development of various therapeutics.
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Background: Throughout oral squamous cell carcinoma (OSCC) progression, tumor-associated macrophages (TAMs) lose their antigen-presenting capacity and anti-tumor function. The mechanisms that cause this dysfunction are not fully understood. CD74 is essential for antigen-presenting process, while little direct evidence describes its role in TAMs' immune function.
Methods: We integrated single-cell transcriptomic analysis, clinical cohort validation and CD74 conditional knockout mouse model to investigate the role of CD74 in TAMs during OSCC progression. Metabolomic analysis and mechanistic studies were performed to dissect how lactate-mediated metabolic reprogramming regulates CD74 expression.
Results: We demonstrate that lactate accumulation in TME induces metabolic reprogramming of TAMs, which drives the acetylation of C/EBPα, and consequently suppresses CD74 expression. This downregulation of CD74 impairs the antigen-presenting capacity of TAMs, suppresses T cell activation, and ultimately promotes OSCC progression and recurrence.
Conclusions: Our findings reveal the critical role of the lactate-C/EBPα-CD74 axis in shaping TAMs function, and provide potential therapeutic target for OSCC immunotherapy.
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Introduction: Patients undergoing multiple wound repair surgeries often develop moderate-to-severe anxiety and depression. However, there is a lack of effective rapid emotional intervention strategies during the perioperative period.
Methods: This multi-centre, randomized, double-blind, placebo-controlled trial involved 130 adult patients (65 in the esketamine group and 65 in the placebo group). Participants were randomly assigned to receive either esketamine (0.2–0.3 mg/kg) or saline intravenously during surgery. The primary outcome was the response rate (proportion of patients with ≥50% reduction in MADRS total score from baseline) on postoperative days (PODs) 1–3, evaluated using the Montgomery–Åsberg depression rating scale (MADRS). The secondary outcome was the remission rate (proportion of patients with MADRS total score ≤10) on postoperative days (PODs) 1–3; scores on the Patient Health Questionnaire-9 (PHQ-9), the Hospital Anxiety and Depression Scale-Anxiety subscale (HADS-A); and esketamine-related neuropsychiatric adverse events assessed using the Young Mania Rating Scale (YMRS), Clinician-Administered Dissociative States Scale (CADSS), and Brief Psychiatric Rating Scale (BPRS) within 30 days after surgery.
Results: The esketamine group showed a significantly higher response rate than the placebo group on POD 1–3. (POD 1: 53.8% vs. 26.2%, p = 0.001; POD 2: 60.0% vs. 40.0%, p = 0.009; POD 3: 73.8% vs. 53.8%, p = 0.018). The esketamine group also showed a higher remission rate and lower MADRS scores (POD 1: 33.8% vs. 10.8%, p = 0.002; POD 2: 40.0% vs. 23.1%, p = 0.038; POD 3: 56.9% vs. 23.1%, p < 0.001). Esketamine improved HADS-A and PHQ-9 scores by POD 3 without increasing neuropsychiatric adverse events within 30 days postoperatively.
Conclusions: The results demonstrate that the intraoperative use of low-dose esketamine can rapidly and effectively alleviate moderate-to-severe anxiety and depressive symptoms in the early postoperative period (POD 1–3) among patients requiring repeated debridement surgeries without increasing neuropsychiatric or systemic adverse events within 30 days after surgery.
Background: Metabolic reprogramming is a key cancer hallmark, with dysregulated fatty acid metabolism critical for tumorigenesis and progression. The acyl-CoA synthetase long-chain (ACSL) family (ACSL1–ACSL6) catalyzes ATP-dependent activation of long-chain fatty acids into acyl-CoA, a bioactive intermediate in lipid synthesis, β-oxidation, membrane biogenesis, and signal transduction. Dysregulated ACSL expression is widespread in malignancies, but their non-metabolic functions (ferroptosis regulation, tumor immune microenvironment remodeling) and translational potential of targeted therapies remain to be systematically summarized.
Methods: This narrative review comprehensively synthesizes existing literature on the biological functions of ACSL family members in cancer. We retrieved and analyzed studies focusing on ACSL-mediated lipid metabolic reprogramming, ferroptosis modulation, and immunomodulatory effects, with particular emphasis on isoform-specific mechanisms and the context-dependent roles (pro-tumorigenic or tumor-suppressive) of the ACSL family across different cancer types. Additionally, we summarized emerging therapeutic strategies targeting ACSL isoforms and their translational potential.
Results: ACSL isoforms exert distinct context-dependent effects:ACSL1 promotes immunosuppressive TIME via M2 macrophage polarization;ACSL3/4 have antagonistic roles in ferroptosis;ACSL5 exerts dual effects via lipid metabolism, apoptosis, and immunity;ACSL6 involves autophagy and hematological malignancies. Dysregulation correlates with tumor progression, drug resistance, and immunotherapy response, while emerging ACSL-targeted drugs show substantial translational potential.
Conclusions: The ACSL family serves as a key regulatory node integrating lipid metabolism, ferroptosis, and tumor immunity. Its isoform-specific mechanisms and context-dependent characteristics highlight its potential as a precise therapeutic target. Future research should focus on optimizing isoform-selective inhibitors, clarifying their synergistic effects with existing therapies (e.g., immune checkpoint inhibitors, radiotherapy), and validating their translational efficacy through clinical trials to advance the development of innovative cancer treatment strategies.
Background: Ninjurin1 (NINJ1) is a transmembrane protein originally identified as a nerve injury-induced adhesion molecule. Recent discoveries have revealed its essential role in plasma membrane rupture (PMR) during lytic cell death, positioning NINJ1 as a critical mediator at the intersection of vascular biology, inflammation, and programmed cell death. Its complex and context-dependent biology makes it a compelling target for cardiovascular research.
Methods: This review comprehensively synthesizes evidence from structural, molecular, cellular, and in vivo studies on NINJ1. We integrated data on NINJ1's structural biology, its cell-type-specific roles in endothelial cells, macrophages, smooth muscle cells, and pericytes, and its contributions to major cardiovascular diseases, including atherosclerosis, myocardial infarction, aortic aneurysm, and ischemia-reperfusion injury. Emerging therapeutic strategies targeting NINJ1 oligomerization were also evaluated.
Results: NINJ1 exhibits a fundamental biological paradox in cardiovascular pathophysiology. In its membrane-bound form, NINJ1 transitions from an autoinhibited homodimer to an active polymeric filament upon cell death stimulation, executing PMR and releasing damage-associated molecular patterns (DAMPs) that amplify vascular inflammation. In contrast, its soluble MMP-9-cleaved ectodomain (sNINJ1) suppresses macrophage activation, attenuates monocyte-endothelial interactions, and exerts potent atheroprotective effects. NINJ1 is dynamically regulated across multiple cardiovascular pathologies and contributes to endothelial dysfunction, plaque instability, myocardial injury, and pericyte-mediated vascular remodeling.
Conclusions: NINJ1 is a pivotal and therapeutically tractable mediator of cardiovascular inflammation. Its dual roles in promoting PMR-driven DAMP release and in limiting inflammation through sNINJ1 signaling provide complementary avenues for therapeutic intervention. Strategies targeting NINJ1 oligomerization or exploiting sNINJ1-mimetic peptides hold promise for the treatment of inflammatory cardiovascular diseases and warrant further translational investigation.
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Background: Short stature (SS) is a common growth disorder with multiple aetiologies. Variants in the MMP13 gene can result in varying degrees of SS, typically accompanied by pronounced skeletal abnormalities. This study aimed to investigate the genetic basis of SS in a family lacking significant imaging abnormalities and elucidate the underlying pathogenic mechanism.
Methods: Trio whole-exome sequencing was performed in a Chinese pedigree with SS to identify pathogenic variants, followed by Sanger sequencing validation. Patient-derived induced pluripotent stem cell model and CRISPR/Cas9-generated Mmp13R459fs homologous mutant mouse model were established to verify the pathogenicity of the variant. Western blotting, immunofluorescence staining, co-immunoprecipitation coupled with mass spectrometry (Co-IP/MS), histological staining and transmission electron microscopy were used to evaluate the effects of the variant on MMP13 protein function and chondrocyte development.
Results: A heterozygous frameshift variant, NM_002427.4:c.1372del(p.Arg458Valfs*31), was identified in the MMP13 gene. Mmp13R459fs mutant mice recapitulated the SS phenotype in patients, with growth plate abnormalities that were present only during the growth phase and resolved earlier than those in Mmp13 knockout mice. Co-IP/MS in HEK293T cells revealed significantly increased HSPA5 expression in the mutant, and enhanced interaction between MMP13 mutant and HSPA5 was confirmed, leading to their retention within the endoplasmic reticulum (ER). In patient-derived chondrocytes, misfolded MMP13 protein upregulated HSPA5 expression, induced significant ER dilation, activated unfolded protein response and increased chondrocyte apoptosis, ultimately contributing to MMP13-related SS.
Conclusion: This study for the first time reports the MMP13 c.1372del (p.Arg458Valfs*31) variant causes autosomal dominant SS without obvious skeletal abnormalities. The variant is associated with defective MMP13 protein secretion and ER stress. These findings expand the mutational spectrum and genotype‒phenotype correlations of the MMP13 gene, providing a novel pathogenic mechanism of SS that is important for the precise diagnosis and treatment.
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