Investigation of differential microRNAs in the exosomes of alcohol-induced H9c2 myocardial cells
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ABSTRACT:
ORGANISM(S): Rattus norvegicus
PROVIDER: GSE193545 | GEO | 2022/01/15
REPOSITORIES: GEO
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Project description:To investigate the function of exosomes from cardiopulmonary progenitors in the regulation of gene expression in endothelial cells.We performed gene expression profiling analysis using data obtained from RNA-seq of H9C2 treated with or without CPPs exosomes.
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Project description:Recent evidence suggests that hypoxia caused by acute myocardial infarction (AMI) can induce evident cardiomyocyte apoptosis. Exosomes, as paracrine signaling mediators by delivering cytosolic components (including miRNA, mRNA, protein), play crucial roles in intercellular communication. However, the systemic regulation of exosome-mediated miRNA delivery and subsequent regulation effect underlying AMI are not well-understood to date. Here, using small RNA-seq, we investigated the miRNAome of H9C2 cells and corresponding paracrine exosmes after hypoxia stress, and identified total of 92 and 62 differentially expressed (DE) miRNAs in cells and exosomes from hypoxia vs. normoxia condition. We found that hypoxia strongly induced the expression of hypoxamiRs in H9C2 cells and altered cardiomyocyte-derived exosomal miRNAome. Functional enrichment analysis revealed extensive roles of DE exosomal miRNAs in HIF-1 signaling pathway and cell apoptosis process, such as TNF signaling pathway, MAPK signaling pathway, mTOR signaling pathway. Furthermore, the gain- and loss-of-function analysis demonstrated the anti-apoptotic effects of hypoxia-induced exosomal miRNAs, i.e. miR-21-5p, miR-378-3p, miR-152-3p, let-7i-5p, and then the luciferase reporter assay confirmed that miR-152-3p and let-7i-5p implemented their anti-apoptotic function by directly targeting Atg12 and Faslg, respectively. In short, this study revealed that hypoxia-exosomes derived from cardiomyocyte, loaded abundant cardio-protective miRNAs, mediate crosstalk among cardiomyocytes and prevent cardiomyocytes apoptosis after hypoxia. Methods: we established the anoxia model using H9C2 cells, an immortal cardiac muscle cell line, to imitate hypoxia condition caused by AMI in vitro, and then small RNA-seq were employed to investigate the miRNA transcriptome of H9C2 cells and its exosomes collected from hypoxia and normoxia culture medium. Results: we identified the miRNAome of H9C2 cells and its exosmes under both hypoxia and normoxia, including 331, 338, 144, 74 unique mature miRNAs from hypoxia-cells, normoxia-cells, hypoxia-exosomes, normoxia-exosomes, respectively. Total of 92 and 62 DE miRNAs were identified from cells and exosomes. Among of which, the DE exosomal miRNAs were mainly involved in HIF-1 signaling pathway and cell apoptosis process, such as TNF signaling pathway, MAPK signaling pathway, mTOR signaling pathway. Interestingly, we found that some DE exosomal miRNAs, including miR-21-5p, miR-378-3p, miR-152-3p, let-7i-5p, have an anti-apoptotic and pro-viability effects in H9C2 cells under hypoxic stress. In addition, Atg12 and Faslg to be respective target of miR-152-3p and let-7i-5p were partly elucidated the anti-apoptosis mechanism of hypoxia-exosomes. Conclusions:In brief, this study illustrated a potential mechanism that, under hypoxic stress, hypoxia pre-perceived cardiomyocytes can spontaneously secrete the hypoxamiRs enriched exosomes, loaded a large amount of cardio-protective miRNA, which mediate crosstalk among cardiomyocytes and prevent apoptosis to heighten the hypoxia adaptability of cardiomyocytes. In more detail, we first discovered that miR-152-3p and let-7i-5p, enriched in hypoxia-exosomes derived by cardiomyocytes, played an anti-apoptosis role via mitochondrial pathway(intrinsic pathway) and death receptor pathway (extrinsic pathway), respectively.
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Project description:The human HEK293 / 293T and rat cardiomyoblast H9c2 cell lines are commonly employed for microRNA-mRNA interaction studies. Here, I provide microRNA sequencing data obtained from each of these lines to better document which microRNAs are endogenously expressed at high or low levels. Small RNA sequencing profiles were generated from cultured HEK293 and H9c2 cells on Illumina HiSeq 2000 instruments.
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Project description:Exosomes have recently been shown to play a key role in cell-to-cell communication through delivery of various functional content, including microRNAs (miRNAs). We investigated the potential roles of exosomal miRNA derived intrafollicular cells in polycystic ovary syndrome (PCOS). Using microarray profiling, a total of 492 miRNAs and 220 miRNAs were found in follicular fluid-derived exosomes and serum-derived exosomes, respectively, in PCOS and non-PCOS females. By excluding miRNAs existing in serum-derived exosomes, we found 179 miRNAs which were specifically expressed in follicular fluid-derived exosomes both in PCOS and non-PCOS females. Using microarray profiling, a total of 492 miRNAs and 220 miRNAs were found in follicular fluid-derived exosomes and serum-derived exosomes, respectively, in PCOS and non-PCOS females. By excluding miRNAs existing in serum-derived exosomes, we found 179 miRNAs which were specifically expressed in follicular fluid-derived exosomes both in PCOS and non-PCOS females.
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Project description:The human HEK293 / 293T and rat cardiomyoblast H9c2 cell lines are commonly employed for microRNA-mRNA interaction studies. Here, I provide microRNA sequencing data obtained from each of these lines to better document which microRNAs are endogenously expressed at high or low levels.
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Project description:Dental pulp cells obtained from several donors proliferated actively in a serum-free medium STK2. The growth rate of dental pulp cells from most donors was higher in the serum-free medium than that in a medium containing 10% serum. DNA microarray analyses showed that gene expression profile of dental pulp cells grown in the serum-free medium was similar to that of cells grown in a medium containing 10% serum. However, several genes related to cell proliferation were up-regulated in dental pulp cells grown in the serum-free medium.
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Project description:Most cancer-related deaths are caused by distant metastases, which are tumour cells that have escaped from a primary tumour and passed into the bloodstream to colonize a new organ. In this context, communication between tumour and stromal cells is essential. Indeed, tumor cells interact with cells in the tumor microenvironment and are able to modify them to their advantage. Both extracellular vesicles (EVs) and exosomes are heterogeneous populations of small vesicles present in the tumor microenvironment and in body fluids that have recently emerged as powerful mediators involved in this communication and their transport in fluids. Tumor cells release large quantities of exosomes containing tumor markers, which can then spread to distant locations.
The exosomes are of endosomal origin. They are composed of proteins, lipids, RNA and DNA, and they circulate in the bloodstream. They can be internalized by specific distant cells and thus deliver a functional message. It has recently been shown that tumor exosomes containing pro-metastatic factors form pre-metastatic niches, before the tumor cells actually arrive, while determining the metastatic organotropism of tumors. These properties are now opening up new avenues of research in tumor biomarkers. In recent years, several studies have highlighted different markers contained specifically in exosomes derived from cancer cells. Consequently, exosomes are considered as potential reservoirs of tumor biomarkers that could be clinically useful for the non-invasive diagnosis of cancer, with the advantage of being performed by liquid biopsy. The study of microRNA (miRNA) is of particular interest. Indeed, miRNAs are small non-coding RNAs (between 21 and 25 nucleotides) involved in the regulation of gene expression and which are frequently deregulated in cancer. Several studies underline that the variation of free miRNAs in the blood is correlated with the progression of the disease, particularly in colon cancer. However, the stability of free miRNAs is controversial. Therefore, exosomes represent a very advantageous means of transporting miRNAs in the blood, as they are able to protect miRNAs from degradation by RNAase.
The hypothesis of the project is that circulating exosomes derived from tumours contain markers including specific miRNAs that could be used as biomarkers of early prognosis (survival and progression), easily measured in blood samples from patients with colon cancer. But other molecules contained in exosomes could also be of interest.
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Project description:Kashin-Beck disease (KBD) is an endemic, chronic degenerative joint disease in China. Exosomes miRNAs, as signaling molecules in intercellular communication, can transfer specific biological martials into target cell to regulate the their function and might participate in the pathogenesis of KBD. We isolated serum and chondrocytes-derived exosomes, miRNA sequencing revealed exosomes miRNA profiles and differentially expressed miRNAs (DE-miRNAs) were identified. The target genes were predicted of known and novel DE-miRNAs with TargetScan 5.0 and miRanda 3.3a database. Single-cell RNA sequencing (scRNA-seq) was performed to identify chondrocyte clusters and their gene signatures in KBD. And we performed comparative analysis between the serum and chondrocytes-derived exosomes DE-miRNA target genes and differentially expressed genes of each cell clusters.A total of 20 DE-miRNAs were identified in serum-derived exosomes. In the miRNA expression of chondrocytes-derived exosomes, 53 DE-miRNAs were identified. 16063 predicted targets were identified as the target genes in the serum-derived exosomes, 57316 predicted targets were identified as the target genes in the chondrocytes-derived exosomes. Seven clusters were labelled by cell type according to the expression of previously described markers. 315 common genes were found among serum/chondrocytes-derived exosomes DE-miRNA target genes and DEGs identified by scRNA-seq analysis. We firstly integratly analyzed the serum and chondrocytes exosomes miRNA with single-cell RNA sequencing (scRNA-seq) data of KBD chondrocyte, the results showed that DE-miRNAs in exosomes might play a potential role in regulating genes expression in different KBD chondrocytes clusters by exosomes mediating cell-cell communications functions, which could improve the new diagnosis and treatment methods for KBD.
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