Vascular cell adhesion molecule-1 (VCAM-1) has been implicated in tumor progression, but its role and molecular mechanisms in colorectal cancer (CRC) remain incompletely defined. This study investigated the clinical association and functional mechanisms of VCAM-1 in CRC. Immunohistochemistry using a tissue microarray containing 90 CRC samples showed that VCAM-1 expression was higher in CRC tissues than in adjacent non-tumorous tissues. Reanalysis of the original survival dataset showed that VCAM-1 positivity was associated with better overall survival in this cohort, whereas its associations with most clinicopathological variables were not statistically significant. This survival association should be interpreted cautiously and requires validation in larger independent cohorts. In vitro, VCAM-1 knockdown in HCT116 and SW480 cells suppressed proliferation, migration, and invasion and promoted apoptosis. Mechanistically, VCAM-1 depletion inhibited RhoA/LIMK/Cofilin signaling, reduced F-actin organization and fluorescence intensity, and reversed epithelial-mesenchymal transition (EMT)-related changes; RhoA-Q63L expression partially restored downstream signaling, cytoskeletal organization, migration, and invasion. VCAM-1 also contributed to CRC cell adhesion to endothelial cells and migration in an M2 macrophage co-culture setting. In vivo, stable VCAM-1 knockdown in HCT116 cells reduced tumor growth in a subcutaneous model. In a tail-vein BLI model, the same knockdown decreased whole-body experimental colonization burden. Together, these findings suggest that VCAM-1 may promote CRC cell progression through RhoA/LIMK/Cofilin-related cytoskeletal remodeling and tumor microenvironment-associated interactions. However, the clinical prognostic direction observed in the current cohort differs from the tumor-promoting functional findings, and the prognostic and therapeutic relevance of VCAM-1 should be further evaluated in larger studies.
Excessive inflammation contributes to adverse cardiac remodeling after myocardial infarction (MI), but the mechanisms linking cardiomyocyte injury to immune activation remain incompletely understood. Here, we investigated the role of the endoplasmic reticulum chaperone HSP90B1 in cardiomyocyte injury and inflammatory crosstalk after MI. We examined HSP90B1 expression in human myocardial tissues, mouse MI hearts, and oxygen-glucose deprivation (OGD)-treated AC16 cardiomyocytes. Hsp90b1+/− mice were used to evaluate post-MI survival, cardiac function, and myocardial remodeling. HSP90B1 was selectively silenced or overexpressed in AC16 cardiomyocytes, and transcriptomic and pharmacological analyses were performed to investigate the signaling pathways involved. A non-contact Transwell co-culture system was also established to assess the effects of cardiomyocyte HSP90B1 on macrophage inflammatory activation. HSP90B1 was increased in human and mouse myocardial tissues after MI and in OGD-treated AC16 cardiomyocytes. Hsp90b1+/− mice showed improved post-MI survival, preserved cardiac function, and reduced adverse remodeling. In AC16 cardiomyocytes, HSP90B1 promoted OGD-induced pyroptosis and was associated with activation of NF-κB/NLRP3 signaling, whereas pharmacological inhibition of these pathways reduced pyroptosis-related responses. In a non-contact cardiomyocyte–macrophage co-culture system, cardiomyocyte HSP90B1 expression influenced the pro-inflammatory phenotype of macrophages. Our findings identify HSP90B1 as a key regulator of cardiomyocyte pyroptotic signaling and post-MI inflammatory remodeling, with cardiomyocyte HSP90B1 expression associated with M1-like macrophage polarization.
Hexokinase 2 (HK2), a key enzyme in the Warburg effect, is associated with poor prognosis in colorectal cancer (CRC). HK2 is known to exert glycolysis-independent functions, yet the underlying targets and mechanisms remain unclear. This study aims to investigate the functional role of ZNF366 as a direct target of HK2 in colorectal cancer and reveal a non-metabolic function of HK2 in CRC. HK2-binding proteins were identified by immunoprecipitation and mass spectrometry. ZNF366 expression was assessed by western blotting and qRT-PCR. Cell proliferation, invasion, and migration were evaluated by functional assays. In vivo tumor growth was examined using a xenograft model. Molecular mechanisms were dissected by co-immunoprecipitation and ubiquitination assays. ZNF366 is downregulated in CRC tissues and cells. Overexpression of ZNF366 suppresses cell proliferation, invasion, and migration in vitro, and inhibits tumor growth in vivo, accompanied by decreased levels of pERK1/2, Cyclin D1, and MMP9, whereas ZNF366 knockdown increases their expression, indicating a negative regulatory role of ZNF366 on malignant phenotypes. HK2 and ZNF366 protein levels are inversely correlated in CRC specimens. HK2 reduces ZNF366 protein levels without affecting its mRNA expression. Mechanistically, UBE3A is identified as the E3 ubiquitin ligase for ZNF366. HK2 enhances the interaction between ZNF366 and UBE3A, leading to increased polyubiquitylation and degradation of ZNF366, which in turn correlates with enhanced malignancy of CRC cells. Our findings suggest that HK2 facilitates UBE3A-dependent ZNF366 degradation, which may partially contribute to the tumor-promoting effect of HK2, revealing a novel HK2-UBE3A-ZNF366 regulatory axis that provides insights into the non-metabolic functions of HK2 in CRC.
Intervertebral disc degeneration (IVDD) is a progressive, multifactorial disorder in which nucleus pulposus (NP) cells operate within chronic hypoxia, nutrient limitation, acid stress and mechanical loading. Metabolites in this niche are not only substrates for energy production but also signaling molecules and donors or regulators of covalent protein modifications. Metabolism-related post-translational modifications (mrPTMs), including lactylation, O-GlcNAcylation and succinylation, provide a molecular interface between metabolic state and protein activity, stability, localization and turnover. Recent studies in IVDD have linked these modifications to ferroptosis, oxidative stress, mitochondrial dysfunction, endoplasmic-reticulum and autophagic quality control, senescence and extracellular-matrix disruption, while crotonylation, glutarylation, β-hydroxybutyrylation and itaconate-related modifications remain emerging areas. However, donor abundance, modifying-enzyme activity, global modification signals, site occupancy and protein abundance are often assessed separately, and direct PTM crosstalk in NP cells remains incompletely resolved. Here, we synthesize current evidence using a framework that connects the metabolic and mechanical microenvironment with metabolic inputs, modifying enzymes, substrate sites, cellular processes, NP cell dysfunction and IVDD. We distinguish site-resolved evidence from candidate mechanisms, integrate established interventions with substrate-selective and local-delivery strategies, and outline priorities for flux-resolved, site-resolved and clinically translatable research.
The filamentous cyanobacterial genus Microcoleus is globally distributed and plays an important role in biological soil crusts. CRISPR–Cas immunity is a prokaryotic defense mechanism that mediates interactions with mobile genetic elements, and may thereby contribute to genome evolution. Yet, its diversity and evolutionary significance in cyanobacteria remain poorly understood. Here, we integrate CRISPR–Cas locus identification, phylogenetic analyses, genome and environmental association analyses and spacer target search to examine CRISPR–Cas diversity and evolution across 14 Microcoleus species. We found that CRISPR–Cas systems are widespread in Microcoleus genus, but their distribution does not follow species-level phylogeny. Instead, locus heterogeneity and module-specific evolutionary histories indicate repeated gain, loss and exchange of CRISPR–Cas components, suggesting that gene flow has contributed to their diversification. CRISPR spacer composition and sharing patterns further indicate dynamic exposure to mobile genetic elements, potentially linked to environmental context. Although a large proportion of spacer targets remain unidentified, expanded protospacer searches revealed plasmid-like elements as prominent targets among the identifiable sequences. Together, our results suggest that CRISPR–Cas systems in Microcoleus are highly dynamic and reflect interactions between gene flow, mobile genetic elements and environmental context, highlighting their potential role as variable components of cyanobacterial genome evolution.
