Defective neutrophil-derived exosomes induce macrophage activation through miR-122-5p in Behçet’s Disease
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PROVIDER: PRJNA1062868 | ENA |
REPOSITORIES: ENA
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Project description:Background Behçet’s disease (BD) is a life-threatening systemic vasculitis characterized by polymorphonuclear neutrophil (PMN) and macrophage activation. This study aimed to elucidate the potential dysregulation of BD PMN exosomes on macrophage activation. Methods BD PMN exosomes were isolated, quantified, incubated with macrophages, and analyzed by miRNA sequencing (n=4). RNA sequencing of miRNA-transfected macrophages (n=6) was analyzed using transcription factor co-regulation network and miRNA target prediction to identify key regulatory factors, which were validated by dual luciferase reporter assay and miRNA/siRNA transfection. Results BD PMN exosomes were decreased and negatively correlated with CRP. PMN exosomes suppressed IL-6, TNF-α, CD80 and CD86 expressions on macrophages, which were attenuated in BD PMN exosomes. miRNA sequencing showed decreased miR-122-5p in BD PMN exosomes, which targeted IRF5, suppressed TLR4 signaling and IFN-β autocrine, and downregulated macrophage activation. Conclusions BD PMN exosomes were decreased in both quantity and miR-122-5p, which impaired the immunoregulatory potential on macrophages through degrading IRF5 and suppressing IFN-β autocrine, indicating a new interaction mechanism between PMNs and macrophages.
2025-07-23 | GSE252885 | GEO
Project description:Renal interstitial fibrosis (RIF) is a common pathological feature and final manifestation of chronic progressive kidney disease. Although exosome-mediated intercellular communication is known to play a crucial role in RIF, the mechanism by which injured tubular epithelial cells (TECs) contribute to fibrogenesis remains incompletely understood. In this study, we investigated the role of exosomal miR-122-5p derived from TECs in modulating fibroblast activation and renal fibrosis. Exosomes were isolated from kidneys of unilateral ureteral obstruction (UUO) mice and from TGF-β1-stimulated HK-2 cells. These exosomes were either co-cultured with fibroblasts (NRK-49F cells) or injected into UUO mice via the tail vein. High-throughput miRNA profiling was used to identify differentially expressed miRNAs in exosomes derived from HK-2 cells, and miR-122-5p was selected for further investigation based on expression level and bioinformatic prediction. Functional analyses were performed using miRNA mimics, inhibitors, and target validation assays. The results showed that exosomal miR-122-5p was significantly downregulated in both fibrotic kidneys and TGF-β1-induced HK-2 cells. In vivo and in vitro, restoration of miR-122-5p levels markedly attenuated renal fibrosis, as evidenced by the reduction of fibrotic markers including α-smooth muscle actin, fibronectin, and collagen I. Mechanistically, miR-122-5p was found to directly target hypoxia-inducible factor 1-alpha (HIF-1α), thereby inhibiting activation of the TGF-β1/Smad signaling pathway. These findings suggest that decreased expression of miR-122-5p in TEC-derived exosomes promotes renal fibrosis through the HIF-1α/TGF-β1/Smad axis, while reintroduction of miR-122-5p can effectively reverse these effects. Taken together, our study provides new insights into the intercellular communication between TECs and fibroblasts via exosomal miRNAs, and identifies miR-122-5p as a potential therapeutic target for the treatment of renal interstitial fibrosis.
2025-09-19 | GSE305699 | GEO
Project description:To further explore the mechanism of miR-122-5p in mice with NEC.
2025-01-17 | GSE286479 | GEO
Project description:Emerging evidence shows that small extracellular vesicles (S-EVs) play a critical role in cancer biology. However, the role of S-EVs in pediatric anaplastic large cell lymphoma (ALCL) is still largely unknown. Small RNA sequencing of plasma S-EVs revealed a peculiar microRNA profile in pediatric ALCL patients compared to healthy donors (HD). In particular, the liver-specific miR-122-5p was more abundant in ALCL plasma S-EVs compared to HD. Elevated levels of miR-122-5p correlated with advanced stage disease and impaired hepatic function in ALCL patients. In vitro experiments demonstrated that miR-122-5p inhibits glucose consumption in stromal cells, and the increased glucose availability enhances ALCL cell aggressiveness. Moreover, miR-122-5p promoted tumor dissemination in vivo. This study identified a new molecular factor promoting ALCL cell dissemination. This finding sets the ground for investigating the clinical use of miR-122-5p antagonists in ALCL patients to potentially improve the clinical outcome.
2023-08-08 | GSE209838 | GEO
Project description:Objective: The objective was to study the expression profile of serum exosomal ncRNAs in patients with PBC and to explore their potential molecular mechanisms. Methods: The expression profile of ncRNAs in serum exosomes of PBC patients was analyzed by RNA-sequence. Bioinformatics analysis predicted target genes and related signaling pathways. Spearson correlation analysis and receiver operating characteristic (ROC) curve were used to explore the relationship between ncRNAs and PBC clinical indicators and application value. Primary human intrahepatic biliary epithelial cells (HIBECs) were cultured in vitro and lipopolysaccharide (LPS) induced HIBECs inflammatory injury model. The expression of miR-122-5p is regulated by miR-122-5p mimics and inhibitors. Enzyme linked immunosorbent assay (Elisa) measured levels of inflammatory cytokines. HIBECs cycle and apoptosis were detected by flow cytometry and HIBECs activity was detected by CCK8. The biomarkers of epithelium-mesenchymal transition (EMT), fibrogenesis, and p38 mitogen-activated protein kinase (MAPK) pathway were detected by real time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). Dual luciferase reporter assay to confirm the binding of miR-122-5p. Results: 8531 differential expressed (DE) lncRNAs and 263 DE miRNAs were identified in serum exosomes of PBC patients. RNA-sequence and verification results showed that miR-122-5p was increased in serum exosomes of PBC patients, and was positively correlated with liver injury and cholestasis indexes. Exosomal miR-122-5p combined with gp210 and sp100 could improve the sensitivity of PBC diagnosis. In vitro experiments showed that the high expression of miR-122-5p promoted the proliferation of HIBECs, and inhibited the apoptosis, EMT and fibrosis of HIBEC. TNFRSF19, p38, p-p38, and inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8, IL-12, IL-17, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and interferon (IFN)-γ were down-regulated and the effect was largely reversed after blocking p38 MAPK pathway. In addition, TNFRSF19 has binding site with miR-122-5p, which is the target gene of miR-122-5p, and could regulate the level of apoptosis signal-regulated kinase 1 (ASK1). Then ASK1 further regulate the downstream p38 MAPK signaling pathway. Conclusion: MiR-122-5p regulates ASK1 by targeting TNFRSF19 and plays an important role in EMT and fibrosis of PBC through p38 MAPK signaling pathway. MiR-122-5p may be a new marker for the diagnosis of PBC and a potential therapeutic target.
2025-10-29 | GSE231432 | GEO
Project description:Hepatocellular carcinoma (HCC) remains the third leading cause of cancer deaths; however, its therapeutic options are limited. Understanding the molecular mechanisms of HCC could shed light on new therapies. Emerging studies indicate the important role of long-noncoding RNAs (lncRNAs) in the pathogenesis of HCC. One of the early discovered lncRNAs, taurine-upregulated gene 1 (TUG1), has been up-regulated in HCC tissues, but its impact on HCC cells’ transcriptomics remains unexplored. We established TUG1-knockdown HCC cells and performed RNA-seq analysis. KEGG analysis revealed the significant enrichment of downregulated genes involved in glycolysis. Accordingly, TUG1-depleted HCC cells showed impairments in glucose uptake, ATP synthesis, and lactate production. Clinical HCC tissue data revealed positive gene expression correlations between TUG1 and glycolysis genes identified by RNA-seq. To identify a molecular function of TUG1 in glycolysis, we explored the competing endogenous (ceRNA) model and identified five microRNAs (miRNAs) with the highest number of TUG1 binding sites using bioinformatic tools. Amongst these miRNAs, microRNA-122-5p (miR-122-5p) displayed an inverse relationship for gene expression with most TUG1-regulated glycolysis genes, including PKM, ALDOA, ENO2, and PFKM. Dual luciferase assays confirmed the direct interaction between TUG1 and miR-122-5p and between miR-122-5p and the 3’ untranslated regions (3’UTR) of PKM and ALDOA. We further showed that inhibition of miR-122-5p alleviated the suppression of glycolysis induced by TUG1 depletion. Together, our RNA-seq analysis of TUG1-depleted HCC cells in combination with clinical data reveals the crucial function of TUG1 in promoting glycolysis via sponging miR-122-5p, which is a negative regulator of multiple glycolytic enzymes.
2025-07-29 | GSE273253 | GEO
Project description:DEmiRNA-mRNA regulatorynetwork reveals miR-122-5p as aregulatory factor of argininemetabolism in necrotizingenterocolitis
| PRJNA1209570 | ENA
Project description:Hepatitis C virus uniquely requires the liver specific microRNA-122 for replication, yet global effects on endogenous miRNA targets during infection are unexplored. Here, high-throughput sequencing and crosslinking immunoprecipitation (HITS-CLIP) experiments of human Argonaute (Ago) during HCV infection showed robust Ago binding on the HCV 5’UTR, at known and predicted miR-122 sites. On the human transcriptome, we observed reduced Ago binding and functional mRNA de-repression of miR-122 targets during virus infection. This miR-122 “sponge” effect could be relieved and redirected to miR-15 targets by swapping the miRNA tropism of the virus. Single-cell expression data from reporters containing miR-122 sites showed significant de-repression during HCV infection depending on expression level and number of sites. We describe a quantitative mathematical model of HCV induced miR-122 sequestration and propose that such miR-122 inhibition by HCV RNA may result in global de-repression of host miR-122 targets, providing an environment fertile for the long-term oncogenic potential of HCV.
2015-01-06 | GSE64679 | GEO
Project description:The liver-specific microRNA, miR-122, is an essential host factor for replication of hepatitis C virus (HCV), an important infectious cause of chronic liver disease and hepatocellular carcinoma. miR-122 stabilizes the positive-strand HCV RNA genome and promotes viral RNA synthesis by binding two closely spaced sites (S1 and S2) near the 5’ end of the genome in association with Ago2. Ago2 is essential for both host factor activities, but whether other host proteins are involved is unknown. Using a quantitative proteomics approach, we identified TNRC6A (GW182) and its paralogs (TNRC6B and TNRC6C), as functionally important components of the miR-122/Ago2 host factor complex binding HCV RNA. Depletion of any two TNRC6 proteins reduced HCV replication in Huh-7.5 cells,but did not reduce viral RNA stability or translational activity, but rather dampened miR-122 stimulation of viral RNA synthesis. However, TNRC6 depletion had no effect on replication of HCV in which S2 was mutated so that miR-122 binds only S1, whereas it significantly enhanced replication when S1 was mutated and only S2 bound by miR-122. Consistent with this, we found that TNRC6 proteins preferentially associate with the S1 site, and that the association of Ago2 with S2 is increased in TNRC6-depleted cells. Collectively, these data suggest a model in which TNRC6 proteins, which are known to interact with Ago2, preferentially direct the miR-122/Ago2 complex to S1 while restricting its association with S2, thereby fine tuning the spatial organization of miR-122/Ago2 complexes bound to the viral RNA.
2019-04-24 | PXD013054 | Pride
Project description:Hepatitis C virus uniquely requires the liver specific microRNA-122 for replication, yet global effects on endogenous miRNA targets during infection are unexplored. Here, high-throughput sequencing and crosslinking immunoprecipitation (HITS-CLIP) experiments of human Argonaute (Ago) during HCV infection showed robust Ago binding on the HCV 5’UTR, at known and predicted miR-122 sites. On the human transcriptome, we observed reduced Ago binding and functional mRNA de-repression of miR-122 targets during virus infection. This miR-122 “sponge” effect could be relieved and redirected to miR-15 targets by swapping the miRNA tropism of the virus. Single-cell expression data from reporters containing miR-122 sites showed significant de-repression during HCV infection depending on expression level and number of sites. We describe a quantitative mathematical model of HCV induced miR-122 sequestration and propose that such miR-122 inhibition by HCV RNA may result in global de-repression of host miR-122 targets, providing an environment fertile for the long-term oncogenic potential of HCV. AGO HITS-CLIP libraries were generated from single cell clones of miR-122 deleted Huh7.5 cells using CRISPR (KO), unedited controls (WT), or cells transfected with GFP instead of CRISPR. Libraries were generated with a 4nt index read, a common priming sequence, followed by a 5nt degenerate barcode terminiating in a G. Files have been demultiplexed such that the 5nt degenerate barcode has been appended as the first 5 nucleotides of the read.
2015-01-06 | E-GEOD-64679 | biostudies-arrayexpress