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.

Interleukin-22 (IL-22) is a cytokine that regulates tissue inflammation and repair, primarily through activation of STAT3 signaling. Although its protective effects in acute pancreatitis (AP) are well established, the specific contribution of its receptor, IL-22Ra1, and the underlying signaling mechanisms remain incompletely defined. Notably, the C-terminal region of IL-22Ra1, although devoid of tyrosine residues, has been reported to promote STAT3 activation through unconventional mechanisms. Here, we investigated the role of IL-22Ra1 and its C-terminal (STAT3-associated) domain in a mouse model of caerulein-induced AP. We show that deletion of the C-terminal region of IL-22Ra1 markedly reduces STAT3 activation in the pancreas, confirming its role in amplifying canonical signaling. Unexpectedly, this reduction does not worsen pancreatic injury. In contrast, both IL-22- and IL-22Ra1-deficient mice develop more severe pancreatitis, characterized by increased serum amylase levels, exacerbated inflammatory responses, and enhanced tissue damage. Notably, IL-22Ra1 deficiency results in a more severe phenotype than IL-22 deficiency alone, indicating that IL-22Ra1 mediates protective effects beyond IL-22 signaling. Consistently, acinar-to-ductal metaplasia during the regenerative phase is impaired in both IL-22- and IL-22Ra1-deficient mice but preserved upon C-terminal truncation. Together, these findings identify IL-22Ra1 as a central regulator of pancreatic protection through mechanisms that are independent of maximal STAT3 activation and extend beyond IL-22 itself, suggesting the involvement of additional IL-22Ra1 ligands. These results support the exploration of receptor-activating strategies, such as agonistic antibodies, to promote tissue protection and regeneration.

IgA nephropathy (IgAN) is a prevalent glomerular disease characterized by IgA deposition, chronic inflammation, and fibrosis, leading to end-stage renal disease. This study explored B-cell translocation gene 2 (BTG2) as a biomarker and treatment target for IgAN progression. Through integrated expression datasets and functional analyses, BTG2 emerged as a key gene with downregulation in renal tissues of IgAN patients, showing high diagnostic accuracy. BTG2 influenced the immune microenvironment by modulating T follicular helper cells and M0 macrophages. Moreover, BTG2 overexpression mitigated IgA deposition and fibrosis, whereas its silencing exacerbated these pathological changes. The study highlights BTG2 as a promising diagnostic tool and therapeutic target, shedding light on potential treatment strategies for IgAN.

[This corrects the article DOI: 10.1002/ccs3.70047.].

Breast cancer represents the most common malignant tumor in women globally. Isoleucyl-tRNA synthetase 2 (IARS2), a member of the aminoacyl-tRNA synthetase family, promotes tumorigenesis across various cancers. Herein, the effects and mechanisms of IARS2 in breast cancer were investigated. IARS2 is upregulated in breast cancer tissues compared to normal tissues, correlating with the poor prognosis of patients with breast cancer. In order to knock down and overexpress IARS2, plasmids carrying short hairpin RNA targeting IARS2 and IARS2-overexpression plasmids were transfected into breast cancer cells, respectively. Our results demonstrated that silencing IARS2 repressed breast cancer cell progression. IARS2 deficiency attenuated tumor growth in MDA-MB-231 cell xenograft mouse models. Conversely, overexpression of IARS2 aggravated malignant cell behaviors in vitro and accelerated tumor growth in vivo. Mechanistically, IARS2 depletion stimulated ubiquitination-mediated degradation of β-catenin in breast cancer cells. Notably, the β-catenin inhibitor XAV-939 reversed IARS2-driven breast cancer malignant progression. Furthermore, differentially expressed genes were identified in IARS2-overexpressed breast cancer cells with or without XAV-939 in the light of mRNA-seq results. Overall, our findings uncovered that silencing IARS2 blocked the β-catenin signaling axis, thereby impeding breast cancer progression. IARS2 might become a promising novel therapeutic target for breast cancer intervention.

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.

[This corrects the article DOI: 10.1002/ccs3.70055.].

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.

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.

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.

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.

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.