Atherosclerotic plaque progression is shaped by coordinated inflammatory and remodeling programs involving immune cells and vascular wall cells. Inflammatory macrophage activation and vascular smooth muscle cell (VSMC) phenotypic remodeling are central features of human atherosclerosis, but their transcriptomic relationships during plaque progression remain incompletely characterized. This study integrated single-cell and bulk transcriptomic datasets to examine highly inflammatory macrophage states, VSMC remodeling-related transcriptional programs, and candidate ligand-receptor expression patterns in human atherosclerotic plaques. Human atherosclerotic plaque single-cell RNA sequencing data from GSE260657 and bulk transcriptomic data from GSE28829 were analyzed. After quality control, 7628 cells were retained for single-cell analysis. Major cell types were annotated using canonical markers, followed by reclustering of macrophages and VSMC-related cells. Functional module scoring, differential expression analysis, Gene Ontology biological process enrichment, and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed to characterize macrophage transcriptional states. Slingshot was applied to infer VSMC pseudotime ordering. CellChat and NicheNet were used to prioritize candidate ligand-receptor expression patterns and ligand-associated VSMC target gene programs. External bulk transcriptomic analysis was performed to examine whether single-cell-derived inflammatory and remodeling signatures were represented at the tissue-transcriptome level during plaque progression. Macrophage reclustering identified a highly inflammatory macrophage state characterized by prominent inflammatory activation, cytokine-response, and stress-response features. Genes upregulated in this population were enriched in pathways related to tumor necrosis factor (TNF) response, nuclear factor kappa B signaling, leukocyte activation, cytokine signaling, lipid and atherosclerosis, toll-like receptor signaling, and inflammasome-associated inflammation. VSMC reclustering revealed contractile VSMCs, PTHLH+ synthetic VSMCs, KRT7+ VSMC-like cells, interferon-responsive VSMCs, pericyte-like mural cells, and osteogenic/modulated VSMCs. Pseudotime analysis showed a broad contractile-to-osteogenic/modulated transcriptional continuum accompanied by increased expression of remodeling-associated genes and selected inflammatory or remodeling-associated receptor genes. CellChat and NicheNet analyses prioritized candidate ligand-receptor and ligand-associated target gene expression patterns involving SPP1-CD44, TNF-TNFRSF1A, IL1B-IL1R1/IL1RAP, MIF-ACKR3, PDGFB-PDGFRB, and FN1-SDC1/ITGB1. In GSE28829, inflammatory macrophage-, osteogenic/modulated VSMC-, candidate ligand-receptor expression-, SPP1-CD44 candidate axis-, and NicheNet-prioritized target program-related signatures were more prominent in advanced plaques and were positively correlated with each other. This integrative transcriptomic analysis identified a highly inflammatory macrophage state and a VSMC remodeling continuum in human atherosclerotic plaques. Candidate ligand-receptor and ligand-associated target gene expression patterns linked inflammatory macrophage activation with osteogenic/modulated VSMC remodeling at the computational level. External bulk data further showed coordinated enrichment of inflammatory and remodeling signatures in advanced plaques. These findings provide a descriptive and hypothesis-generating transcriptomic framework for understanding inflammatory macrophage activation and VSMC remodeling in human atherosclerosis.
Multiorgan fibrosis is a complex, maladaptive response to chronic injury, where the extracellular matrix (ECM) shifts from serving merely as a structural framework to becoming an active, sustained contributor to disease progression. We propose a unifying mechanobiological framework in which chronic injury promotes fibroblast activation and ECM remodeling, whereas the stiffened and remodeled ECM feeds back through integrins, mechanosensitive ion channels, collagen receptors, and Hippo-YAP/TAZ signaling to sustain fibrogenesis. Regulation of this process is governed by bidirectional feedback between cellular signaling and the matrix's physical properties across organs such as the lung, heart, liver, and kidney. This narrative review article presents recent advances in understanding mechanisms of cell-cell and ECM communication, paratensile signaling, mechanical feedback loops, cellular crosstalk and secretomes, quantitative therapeutic thresholds, and translational strategies for tissue remodeling. These developments collectively offer promising insights for the development of novel pharmacological agents targeting mechanotransduction pathways, ECM normalization, and cell-specific sensitization. We further define quantitative thresholds as diagnostic, action, and response thresholds that may support clinical decisions such as referral, treatment initiation, therapeutic monitoring, and clinical-trial enrichment. Finally, we provide specific details regarding the treatment of liver fibrosis before concluding with research agenda.
High mobility group box 1 (HMGB1) is a nuclear protein that functions as a damage-associated molecular pattern molecule upon extracellular release. It has an established role in sterile inflammation, yet its involvement in UV-induced human skin inflammation remains poorly characterized. This study investigated whether UV irradiation and chronic photoaging trigger HMGB1 release in human skin and whether this release drives proinflammatory cytokine expression and photoaging. Both acutely UV-irradiated and chronically photoaged human skin exhibited HMGB1 release, which temporally correlated with the upregulation of photoaging-related critical proinflammatory cytokines, IL-1β, IL-6, IL-8, and TNF-α. In cultured cells, UV irradiation reduced intracellular HMGB1 protein without altering mRNA levels while simultaneously increasing extracellular HMGB1 in supernatants, confirming active secretion rather than passive leakage. HMGB1 knockdown and blockade of TLR4, its principal receptor, significantly attenuated UV-induced proinflammatory cytokine expression in both cell types. In mouse skin, disruption of the HMGB1-TLR4 axis reduced neutrophil infiltration, suppressed cytokine expression, and mitigated dermal damage (photoaging). Collectively, these findings demonstrate that UV irradiation promotes active HMGB1 release from skin cells, amplifying cutaneous inflammation and exacerbating dermal damage via TLR4 signaling. Therapeutic targeting of the HMGB1-TLR4 axis therefore represents a promising strategy for mitigating UV-induced skin inflammation and its downstream pathologies, including photoaging.
Renal ciliopathies encompass a spectrum of genetic disorders arising from structural or functional impairments of primary cilia, specialized organelles critical for mechanosensation and signal transduction within renal epithelial cells. These disorders are characterized by cystogenesis, driven by dysregulated ciliary signaling, leading to uncontrolled epithelial proliferation, aberrant growth, and loss of cellular polarity. The clinical trajectory evolves from initial cyst formation to advanced tubulointerstitial fibrosis and progressive renal failure. This progression is governed by pathogenic variants in genes encoding ciliary proteins. While advancements in genetic testing have established prenatal diagnosis as a pivotal tool for early identification, definitive diagnosis and therapeutic intervention remain challenging. These difficulties stem from several factors: incomplete understanding of the molecular mechanisms underlying cyst formation and fibrosis; limitations in prenatal diagnostic accuracy owing to phenotypic overlap and incomplete penetrance; and the marked genetic heterogeneity and diverse clinical trajectories of renal ciliopathies. Existing studies have primarily focused on unidirectional modulation of individual pathways, whereas the systematic integration of signaling network cascades remains largely unaddressed. This review systematically elucidates the molecular mechanisms and aberrant signaling pathways in renal ciliopathies, links genetic heterogeneity to clinical phenotypes, and lays a theoretical basis for prenatal diagnosis and novel therapies.
Fast-spiking parvalbumin-positive (PV+) interneurons, a specialized class of inhibitory neurons, possess unique morphological and functional properties that govern spatiotemporal precision in local microcircuits, large-scale network synchronization, and memory-related computations. Since their initial identification in the late 19th century, technological innovations in cellular neuroscience have progressively elucidated the multifaceted roles of these neurons. In this review, we first delineate the embryonic origins and developmental trajectory of PV+ interneurons, emphasizing their unique properties for high-frequency firing with remarkable temporal precision. These specialized features enable PV+ interneurons to orchestrate network oscillations and critically modulate memory encoding, consolidation, and retrieval, despite their substantial metabolic demands. We subsequently integrate multiple lines of evidence implicating region- and subtype-specific PV+ interneuron impairment as a pivotal pathological hallmark in Alzheimer's disease (AD). Furthermore, we dissect molecular and cellular mechanisms driving PV+ interneuron dysfunction in AD, including numerical alterations, morphological remodeling, and electrophysiological disruptions. Critically, we propose that the pathological transformation of PV+ interneuron physiology emerges as a key driver in AD progression, bridging cellular dysfunction to system-level cognitive failure.
Periodontitis is a common disorder that causes tooth loss. Osteogenic differentiation of periodontal ligament stem cells (PDLSCs) is crucial for repairing periodontal defects. This study aimed to investigate the underlying mechanism by which Shikonin promotes the osteogenic differentiation of PDLSCs. PDLSCs were isolated from the periodontal ligaments of orthodontically extracted premolars. Flow cytometry was used to characterize the cell surface markers of PDLSCs. PDLSCs were cultured in osteogenic induction medium and treated with Shikonin. Alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining were used to evaluate ALP activity and assess calcium nodule formation, respectively. Protein levels were determined by Western blot analysis. A dual-luciferase reporter assay and chromatin immunoprecipitation analysis confirmed the interaction between the ETS transcription factor 1 (ELK1) and cementum protein 1 (CEMP1) promoter. Shikonin promoted the osteogenic differentiation of PDLSCs (about 7.6 folds for ALP staining and about 6.9 folds for ARS staining with 1 μm Shikonin, p < 0.01). Shikonin-mediated enhancement of osteogenic differentiation was achieved by promoting the phosphorylation of ELK1 through the activation of the p38 MAPK signaling pathway (p < 0.05). Moreover, ELK1 accelerated osteogenic differentiation and transcriptionally activated CEMP1 expression (p < 0.05). Activation of the ELK1/CEMP1 axis contributed to Shikonin-induced osteogenic differentiation of PDLSCs (p < 0.05). Shikonin promotes the osteogenic differentiation of PDLSCs in vitro by modulating the p38 MAPK/ELK1/CEMP1 axis.
Alcohol consumption has been a risk factor for more than 200 diseases, including esophageal cancer. Cell communication network 1 (CCN1), a matricellular protein, is highly expressed in esophageal squamous cell carcinoma (ESCC) but is barely detectable in esophageal adenocarcinoma (EAC). Alcohol consumption has been identified as a major contributor to ESCC development, but its role in EAC is uncertain. This study examines the impact of acute (30 min) or prolonged (12 h) alcohol exposure (5, 100, and 500 mM) on CCN1 expression and function in esophageal epithelial cells, including normal (HEEC), ESCC (KYSE150 and KYSE410), and EAC (OE19 and OE33), in association with the activity of MMP2, MMP9, and MMP14. It was found that alcohol exposure promoted CCN1 expression in both normal and tumor cells but induced CCN1 cleavage exclusively in EAC cells, generating an 18-kDa fragment that promotes tumor growth. MMP9, which was active only in EAC cells, was found to mediate this cleavage. Forced activation of MMP9 in either normal or ESCC cells improved cell viability, whereas inhibition of MMP9 in EAC cells attenuated cell survival. Taken together, alcohol exposure promotes MMP9-mediated CCN1 cleavage in EAC, converting CCN1 from a pro-death to a pro-survival factor for EAC. This makes alcohol consumption a risk factor not only for ESCC development but also for EAC progression. A preprint can be found at d197for5662m48.cloudfront.net.
The 13th International Workshop on the CCN Family of Genes, held in Nice as part of the inaugural ARBIOCOM World Conference, brought together investigators with diverse research backgrounds, disciplines, and expertise. Building on the long-standing mission of the International CCN Society and the expanding ARBIOCOM initiative, the meeting highlighted the growing importance of interdisciplinary exchange in accelerating discovery in normal and pathological signaling networks. Scientific sessions featured presentations by renowned speakers and early-career investigators alike, creating a highly collaborative environment that fostered discussion of emerging concepts and translational opportunities. Among the major themes, this joint conference-meeting highlighted "hot" research topics such as the nuclear and extracellular roles of CCN proteins in regulating gene expression, tissue development and remodeling, stem cell differentiation and therapies, and disease initiation and progression. Presentations on skeletal biology emphasized how extracellular matrix proteins and matricellular factors shape bone, cartilage, and connective tissue development and pathology, whereas vascular sessions revealed new mechanistic links between extracellular cues, endothelial remodeling, macrophage-driven vessel regression and vascular smooth muscle homeostasis. Other talks extended the relevance of new signaling molecules in intestinal regeneration, hepatic inflammation, metabolic control, and cancer progression, illustrating the broad biological reach of these pathways. The meeting also emphasized the value of integrating new model systems, advanced single-cell approaches, and functional in vivo studies to uncover mechanisms that were previously unknown. Collectively, the scientific sessions and discussions identified and filled key knowledge gaps and generated new hypotheses with strong potential for future discoveries. By connecting established expertise with complementary scientific perspectives, this meeting provided a fertile platform for cross-pollination of ideas and highlighted several avenues for future research aimed at developing novel diagnostic and therapeutic strategies and tools.
This study aims to elucidate the role of suppressor of cytokine signaling 3 (SOCS3) in glioma stem cells (GSCs) via single-cell RNA sequencing (scRNA-seq), focusing on its regulation of STAT3-mediated self-renewal, apoptosis resistance, and tumor microenvironment (TME) remodeling. ScRNA-seq data from 19 high-grade glioma patients were analyzed using Seurat, Harmony, and SingleR for clustering, annotation, and SOCS3 stratification (SOCS3-High: n = 4; SOCS3-Low: n = 15). Differential gene analysis, pathway enrichment, and CellChat were employed for TME characterization. In vitro, SOCS3-overexpressing/silenced GSC11 models were tested via MTT, TUNEL, neurosphere assays, and STAT3 pathway modulation (IL-6). In vivo, intracranial xenografts in nude mice evaluated tumor growth and survival. SOCS3 was downregulated in GSCs and neurons. SOCS3-Low GSCs exhibited 777 differentially expressed genes enriched in T-cell receptor, p53, and JAK-STAT axis, suppressed T-cell/microglia infiltration, and promoted oligodendrocyte precursor cell/astrocyte survival. SOCS3 overexpression reduced GSC proliferation, induced apoptosis, inhibited neurosphere formation, and suppressed STAT3 phosphorylation and stemness markers (OCT4/SOX2/NANOG). IL-6 reactivated STAT3, reversing SOCS3-mediated tumor suppression. In vivo, SOCS3 overexpression attenuated tumor growth and prolonged survival, counteracted by IL-6. Low SOCS3 expression contributes to glioma progression by promoting STAT3 activation and an immunosuppressive TME. Targeting the SOCS3-STAT3 axis may offer therapeutic potential.
To explore the mechanism by which plasma exosome miR-339-3p regulates myocardial remodeling in heart failure (HF). Plasma exosomes were isolated from 5 patients with HF and 5 controls, and cell uptake was determined by transmission electron microscopy, western blot and PKH26 labeling. Angiotensin II treated AC16 cells to induce cell hypertrophy. Cell counting kit-8 assay and flow cytometry were used to detect cell viability and apoptosis to evaluate the effect of HF-exo. High-throughput sequencing was performed on exosomal microRNAs to identify key differential mirnas (miR-339-3p), and database screening was used to verify downstream target proteins ubiquitin-specific protease 25 (USP25) by dual-luciferase reporter gene assay and reverse transcription-quantitative polymerase chain reaction/western blot. Searchtool for the retrieval of interacting genes, the Cancer Genome Atlas and co-immunoprecipitation were used to screen USP25-interacting proteins (DDX58). Analysis revealed that inhibition of miR-339-3p completely reversed cardiomyocyte injury induced by HF-exo. miR-339-3p directly targets USP25, down-regulates its expression, and impairs its K48-related DDX58 deubiquitination. Overexpression of USP25 can reverse myocardial injury induced by mir-339-3p, and DDX58 silencing can eliminate the protective effect of USP25. Plasma exosome miR-339-3p promotes myocardial remodeling in HF through the miR-339-3p-USP25-DDX58 axis and is a potential target for the diagnosis and treatment of HF.
This study investigated the pathological relevance of the serine palmitoyltransferase long chain base subunit 2 (Sptlc2)-ceramide axis in nonobese nonalcoholic fatty liver disease (NAFLD), focusing on hepatic steatosis, inflammation, oxidative stress, and mitochondrial dysfunction. A nonobese NAFLD rat model was established using a high-temperature dry-fried soybean diet. Integrated liquid chromatography-mass spectrometry-based proteomic and metabolomic analyses were used to identify candidate pathways. Sptlc2 function was validated by AAV2/8-mediated liver-directed knockdown in vivo, lentiviral knockdown in primary hepatocytes, and C2-ceramide rescue experiments. Multi-omics profiling identified Sptlc2 as a sphingolipid metabolism-related candidate in the model. Sptlc2 knockdown reduced long-chain ceramide accumulation, hepatic lipid deposition, inflammatory cytokine expression, oxidative stress, and hepatocyte injury. In primary hepatocytes, Sptlc2 silencing improved mitochondrial respiration, membrane potential, calcium and reactive oxygen species homeostasis, and mitochondrial ultrastructure. These protective effects were partially reversed by C2-ceramide in vitro and in vivo. The Sptlc2-ceramide axis contributes to ceramide accumulation, hepatic lipotoxicity, inflammatory activation, and mitochondrial dysfunction in this nonobese NAFLD model, suggesting its potential relevance as a therapeutic target for further investigation.
Over the past 2 decades, major advances have contributed to the elucidation of the structural and biochemical bases underlying the biological activities attributed to CCN proteins. The concept that CCN proteins exhibit bifunctional "moonlighting" properties in both the extracellular matrix (ECM) and the cell nucleus has recently emerged. CCN proteins participate in dual signaling processes, integrating combinatorial interactions with regulatory ligands, cell surface receptors or associated co-receptors such as heparan sulfate proteoglycans (HSPGs), LRPs, TrkA, Notch, integrins, BMP-4, TGF-β, and FGFR2, as well as transcription factors in the nuclear compartment. In this manuscript, we propose an exploratory integrative model that brings together previously unassociated observations into a coherent framework. In this model, the C-terminal module, present in all CCN proteins except CCN5, is proposed to direct the formation of homo- and heterodimers, which constitute a fundamental level of transcriptional regulation.
Extracellular matrix (ECM) remodeling is a critical component of colorectal cancer (CRC) progression and tumor microenvironment organization. Members of the A Disintegrin and Metalloproteinase with Thrombospondin Motifs (ADAMTS) metalloproteinase family are known regulators of ECM structure; however, the transcriptional regulation of ADAMTS14 and its potential role in inflammation-associated ECM remodeling remain poorly understood. In this study, we investigated whether inflammatory signaling regulates ADAMTS14 expression and explored its association with ECM organization in CRC. Interleukin-6 (IL-6) stimulation significantly increased ADAMTS14 expression at both mRNA and protein levels in CRC cells. Promoter deletion analyses identified a critical IL-6-responsive region between -145 and -43 bp upstream of the transcription start site, suggesting transcriptional responsiveness of ADAMTS14 to inflammatory signaling. Inhibition experiments demonstrated that Extracellular Signal-Regulated Kinase, c-Jun N-terminal Kinase, Phosphatidylinositol 3-Kinase, and Nuclear Factor Kappa B pathways were associated with IL-6-induced ADAMTS14 expression. Transcriptomic analyses of The Cancer Genome Atlas CRC datasets revealed that ADAMTS14 expression is elevated in tumors and is associated with inflammatory signaling, stromal activation, fibroblast-related gene expression, and ECM organization pathways. Functional enrichment analyses indicated that ADAMTS14-correlated genes are primarily involved in ECM organization, collagen fibril organization, and connective tissue development. Together, these findings identify ADAMTS14 as an inflammation-responsive ECM-associated metalloproteinase and suggest that IL-6 signaling may be associated with ECM-related transcriptional programs through regulation of ADAMTS14 expression. Our findings further suggest that ADAMTS14 expression may be associated with inflammatory and stromal-related transcriptional programs in CRC.
To investigate the molecular mechanism by which gemcitabine (GEM) inhibits ovarian cancer (OC) progression, focusing on the hypoxia-inducible factor 1-alpha (HIF1A)/UBR5/LATS2 axis and Hippo pathway. Bioinformatics analysis of gene expression omnibus datasets identified HIF1A-associated modules. Transcriptional regulation of UBR5 by HIF1A was validated via ChIP-PCR and dual-luciferase assays. Anti-tumor effects of GEM were assessed in OC cell lines and an orthotopic mouse model using functional assays (CCK-8, transwell), Western blot, Co-IP, and ubiquitination analysis. HIF1A transcriptionally upregulates UBR5 in OC. GEM downregulated the HIF1A/UBR5 axis, suppressing OC cell proliferation, migration, and invasion. Mechanistically, UBR5 promoted LATS2 ubiquitination and degradation. GEM inhibited this interaction, stabilizing LATS2, activating the Hippo pathway, and suppressing downstream YAP1/FGFR1 signaling. In vivo, GEM inhibited tumor growth and downregulated HIF1A, UBR5, and FGFR1. GEM suppresses OC progression by targeting the HIF1A/UBR5 axis, which subsequently stabilizes LATS2, activates the Hippo pathway, and inhibits YAP1/FGFR1 signaling, revealing a novel therapeutic mechanism.
Atrial fibrillation (AF) is a common cardiac arrhythmia often accompanied by structural remodeling of the atria, particularly fibrosis, and disruption of normal mitochondrial function. N(1)-methyladenosine (m1A), a methylation of RNA, is gaining attention for its role in diverse biological processes. This study aimed to explore the role of the m1A methyltransferase tRNA methyltransferase 10C (TRMT10C) in AF pathogenesis. In the study, TRMT10C and m1A methylation levels were upregulated in AF rats, accompanied by excessive mitochondrial fission and myocardial fibrosis. Knockdown of TRMT10C inhibited the expression levels of mitochondrial fission-related proteins Drp1 and Fis1, reduced collagen deposition (collagen I, Postn, collagen III, and fibronectin), and AF progression. In vitro results showed that TRMT10C knockdown inhibited TGF-β1-induced cardiac fibroblasts proliferation and migration, whereas overexpression of transferrin receptor (TFRC) reversed this effect. Mechanistically, TRMT10C enhanced the stability of TFRC mRNA by promoting m1A methylation, driving mitochondrial fission and fibrosis. Collectively, our findings elucidate a novel TRMT10C-TFRC m1A axis driving pathological mitochondrial dynamics and fibrosis in AF, offering new insight into cell-signaling pathways underlying atrial disease and potential therapeutic targets.
Cellular communication network (CCN) proteins are key matricellular regulators of cartilage development, yet their species-specific roles and network-level context remain unclear. This study integrated bulk RNA sequencing from chicken and mouse embryonic limb bud micromass cultures and human mesenchymal stem cell chondrogenesis with co-expression, protein-protein interaction, and ortholog analyses to construct CCN-centered regulatory networks across models. CCN1 and CCN2 emerged as dominant, conserved hubs enriched in collagen-containing extracellular matrix, cartilage development, and growth factor signaling modules, whereas CCN3-CCN6 showed lower context-dependent expression and connectivity. Functional and ortholog analyses revealed moderate pathway conservation, with high conservation of IGF, EGFR, and HIF-1 signaling, but reduced overlap in hypoxia and mechanosensing/Hippo categories, indicating species-specific tuning of environmental sensing. A focused ortholog screen identified multifunctional conserved hubs, including COL2A1, TGFBR1, SMAD3, RUNX2, HIF1A, IGF1, SPP1, and CD44. Single-cell RNA-seq meta-analysis of human iPSC-derived chondrogenesis and embryonic limb datasets showed CCN1/2 expression and homologous network activity peaking in mesenchymal and early chondrocyte populations, consistent with model-dependent persistence into hypertrophic and ossification stages in vivo. Overall, this work defines a conserved CCN1/2-centered axis integrating extracellular matrix formation with growth factor and mechanical cues, providing a framework for model selection and CCN-targeted cartilage regeneration strategies.
Neonatal necrotizing enterocolitis (NEC) continues to be the most severe gastrointestinal emergency affecting preterm infants, with reported mortality rates ranging from 20% to 30%. The absence of distinct early biomarkers results in delayed intervention and unfavorable outcomes, and the molecular mechanisms underlying the dysregulated immune response in NEC remain incompletely defined. We integrated five gene expression omnibus datasets (36 NEC cases and 34 controls) using weighted gene co-expression network analysis and systematically evaluated 113 machine learning algorithm combinations. The best-performing model (RF + XGBoost) identified candidate diagnostic genes, which were validated through independent bulk RNA-sequencing cohorts, single-cell RNA sequencing (11,308 intestinal cells), single-cell virtual gene knockout (scTenifoldKnk), and experimental models including mice and the intestinal epithelial cells cell line. Combinatorial in silico perturbation was further performed using Geneformer, and a ferroptosis gene panel was validated by qRT-polymerase chain reaction in the mouse NEC model. The RF + XGBoost model achieved high diagnostic accuracy (area under the curve = 0.979, 95% CI: 0.940-1.000). Three key biomarkers were identified: SLC26A3, CCL20, and CXCL5. Multi-platform validation showed consistent downregulation of SLC26A3 and upregulation of CCL20 and CXCL5 in NEC (all p < 0.001). Single-cell analyses revealed cell-type-specific dysregulation, with CCL20 markedly elevated in macrophages (log2FC = 3.85) and CXCL5 broadly upregulated across enterocytes, macrophages, and fibroblasts. Immune profiling demonstrated elevated proportions of M0 macrophages and activated mast cells, alongside reduced naive B cells and naive CD4 T cells, and CXCL5 expression was strongly correlated with neutrophil infiltration (r = 0.88, p < 0.001). Virtual knockout analysis revealed that perturbation of CCL20 and CXCL5 produced overlapping downstream networks converging on major histocompatibility complex class II-related antigen presentation genes, whereas SLC26A3 knockout perturbed a distinct set of epithelial barrier and innate immune genes. FTH1 was the sole gene perturbed across all three knockouts, implicating ferroptosis as a potential convergence point in NEC pathogenesis. Geneformer-based combinatorial perturbation indicated subadditive rather than synergistic interactions among the three biomarkers, with SLC26A3 appearing as the dominant node. Extension to a 9-gene ferroptosis panel showed coordinated directional shifts across the tested genes, and qPCR measurements in the NEC mouse ileum were consistent with the predicted expression changes for the five measured genes. SLC26A3, CCL20, and CXCL5 constitute a candidate molecular signature for early NEC diagnosis. Combinatorial perturbation analysis suggests subadditive rather than synergistic interactions among the three biomarkers, with SLC26A3 appearing as the dominant node. Virtual knockout network analyses suggest that CCL20 and CXCL5 share downstream regulatory circuits linked to antigen presentation, while SLC26A3 primarily perturbs the epithelial barrier and innate immune genes. Ferroptosis emerged as a coordinated multi-node convergence point, with computational predictions corroborated by qPCR measurements in a mouse NEC model. These findings provide a framework for a mechanistic study and potential targeted intervention in NEC.
Silent mating type information regulators (sirtuins and SIRTs) have emerged as central nodes linking cellular metabolism to stress adaptation and diseases. SIRT1 stands out as a ubiquitously expressed, NAD+-dependent deacetylase with remarkably broad and sometimes paradoxical functions. Beyond its canonical roles in chromatin regulation and DNA repair, SIRT1 regulates key pathways controlling metabolism, inflammation, circadian rhythms, mitochondrial biogenesis, mitophagy, immune response, and cellular survival. Growing evidence, including several studies published in JCCS, positions SIRT1 at the intersection of cardiovascular, metabolic, neurovascular, and oncogenic processes, highlighting both its biological complexity and therapeutic potential. Mechanistically, SIRT1 modulates through deacetylation, a diverse network of substrates including histones H3/H4, tubulin, BMAL1, p53, NF-κB, FOXO, PGC-1α, and eNOS, yet, its functional outcomes are highly context dependent. Although SIRT1 activation is often protective in metabolic and cardiovascular and neurovascular settings, its role in cancer remains dualistic, acting as both tumor suppressor and promoter. Similarly, its contribution to longevity remains unresolved, with conflicting results across experimental models. Pharmacological targeting of SIRT1, through compounds such as resveratrol and synthetic modulators, has generated considerable enthusiasm, but challenges related to specificity, bioavailability, and off-target effects persist. Here, I discuss SIRT1 not simply as a therapeutic target but as a molecular "shapeshifter," whose context-dependent actions demand more precise biomarker-guided strategies for clinical translation.
Sepsis often has a dysregulated inflammatory response and is accompanied by cardiac dysfunction. This study aimed to explore the mechanism of long non-coding RNA MIR503HG (MIR503HG) in regulating sepsis and the inflammatory responses, and sepsis-induced cardiac dysfunction (SICD). 102 sepsis patients were divided into an SICD group (n = 31) and a non-SICD group (n = 71). A cecal ligation and puncture (CLP) sepsis rat model was constructed. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to evaluate the target gene expression. Enzyme-linked immunosorbent assay was used to measure the levels of myocardial damage markers and inflammatory factors. RNA immunoprecipitation and Dual luciferase assay were used to determine the targeting relationship. Kyoto Encyclopedia of Genes and Genomes analysis was used to predict signaling pathways of target genes. MIR503HG expression was significantly down-regulated in sepsis patients and SICD patients, and it has good diagnostic value for these two types of diseases. Its expression was significantly negatively correlated with myocardial injury markers (cardiac troponin I [cTnI], creatine kinase-MB [CK-MB]), cardiac function indicators (left ventricular ejection fraction), inflammatory factors (interleukin-6, tumor necrosis factor-alpha), and disease severity scores (Sequential Organ Failure Assessment, Acute Physiology and Chronic Health Evaluation II). Furthermore, it exerted a protective effect in sepsis: MIR503HG overexpression could effectively alleviate SICD and mitigate the inflammatory response, as evidenced by decreased left ventricular end-diastolic pressure, increased left ventricular systolic pressure, reduced levels of CK-MB and cTnI, as well as restored myocardial systolic/diastolic capacity (maximal rate of left ventricular pressure rise/fall [±dP/dt]). Mechanistically, MIR503HG functions as a molecular sponge that sequesters microRNA-497-5p (miR-497-5p), thereby lifting brain-derived neurotrophic factor (BDNF). Delivery of miR-497-5p-agomiR partially offset the cardioprotective and anti-inflammatory effects of MIR503HG, whereas overexpression of BDNF partially restored them. MIR503HG relieved sepsis and reduced cardiac dysfunction and inflammatory response by regulating the miR-497-5p/BDNF axis.
Endoplasmic reticulum stress (ERS) is a major event associated with myocardial ischemia-reperfusion injury (MIRI). Imperatorin (IMP) has cardioprotective effects, but its role in MIRI has not been reported. H9c2 cells were injured by the hypoxia/reoxygenation (H/R) method. The MIRI mouse model was prepared by coronary artery ligation and by using IMP, and the PI3K/AKT agonist 740 Y-P and inhibitor LY294002 were used for intervention. Cardiomyocyte apoptosis was determined through flow cytometry and TUNEL staining. Western blot, transmission electron microscopy, calcium assay kits, and Fluo-4 AM fluorescent probe were used to evaluate ERS and Ca2+ levels. In addition, the PI3K/AKT/Nrf2 pathway protein expressions were detected. IMP could significantly reduce cardiomyocyte apoptosis in H9c2 cells and myocardial tissue of MIRI mice, reduce ER swelling and damage, restore ER membrane structure, and reduce Ca2+ content and Caspase 12 and C/EBP homologous protein (CHOP) protein levels. IMP also increased p-PI3K and p-AKT proteins and the nuclear translocation of Nrf2 in H9c2 cells and myocardial tissue of MIRI mice. After IMP treatment combined with 740 Y-P intervention, the effect of IMP on improving cardiomyocyte apoptosis and ERS was further promoted, and the apoptosis rate, Ca2+ content, and caspase 12 and CHOP protein levels were significantly reduced. LY294002 weakened the improvement of IMP, and apoptosis and ERS levels increased significantly. IMP attenuated MIRI by inhibiting ERS and myocardial apoptosis through activating the PI3K/AKT/Nrf2 pathway.
