Long noncoding RNAs expression in mature and immature dendritic cells-derived exosomes
Ontology highlight
ABSTRACT: Dendritic cells release bioactive exosomes that are involved in immune regulation. Long non-coding RNAs (lncRNAs) have been demonstrated to be implicated in many immunoregulatory mechanisms. However, the roles of lncRNAs in dendritic cells-derived exosomes remain unknown. To investigate the roles of lncRNAs in mature and immature dendritic cells-derived exosomes and find specific lncRNAs with immunoregulatory function, this study showed the expression profiles of lncRNAs in bone marrow dendritic cells-derived exosomes from C57 mice.
Project description:Shifts in the gut microbiota composition, called dysbiosis, have been directly associated with acute and chronic diseases. However, the underlying biological systems connecting gut dysbiosis to systemic inflammatory pathologies are not well understood. Phospholipids (PLs) act as precursors of both, bioactive inflammatory and resolving mediators. Their dysregulation is associated with chronic diseases including cancer. Gut microbial-derived lipids are structurally unique and capable of modulating host’s immunity. Lactobacillus johnsonii N6.2 is a Gram-positive gut symbiont with probiotic characteristics. L. johnsonii N6.2 reduces the incidence of autoimmunity in animal models of Type 1 Diabetes and improves general wellness in healthy volunteers by promoting, in part, local and systemic anti-inflammatory responses. By utilizing bioassay-guided fractionation methods with bone marrow-derived dendritic cells (BMDCs), we report here that L. johnsonii N6.2 purified lipids induce a transcriptional signature that resembles that of migratory (mig)DCs. RNAseq-based analysis showed that BMDCs stimulated with L. johnsonii N6.2 total lipids upregulate maturation-mig related genes Cd86, Cd40, Ccr7, Icam1 along with immunoregulatory genes including Itgb8, Nfkbiz, Jag1, Adora2a, IL2ra, Arg1, and Cd274. Quantitative reverse transcription (qRT)-PCR analysis indicated that PLs are the bioactive lipids triggering the BMDCs response. Antibody-blocking of surface Toll-like receptor (TLR)2 resulted in boosted PL-mediated upregulation of pro-inflammatory Il6. Chemical inhibition of the IKKα kinase from the non-canonical NF-κB pathway specifically restricted upregulation of Il6 and Tnf. Phenotypically, PL-stimulated BMDCs display an immature like-phenotype with significantly increased surface ICAM-1. This study provides insight into the immunoregulatory capacity of Gram-positive, gut microbial-derived phospholipids on innate immune responses.
Project description:Dendritic cells (DCs) are the most potent antigen (Ag)-presenting cells. Whereas immature DCs down-regulate T cell responses to induce/maintain immunological tolerance, mature DCs promote immunity. To amplify their functions, DCs communicate with neighboring DCs through soluble mediators, cell-to-cell contact and vesicle exchange. Transfer of nanovesicles (<100nm) derived from the endocytic pathway (termed exosomes) represents a novel mechanism of DC-to-DC communication. The facts that exosomes contain exosome-shuttle microRNAs (miRNAs), and DC functions can be regulated by exogenous miRNAs, suggest that DC-to-DC interactions could be mediated through exosome-shuttle miRNAs, an hypothesis that remains to be tested. Importantly, the mechanism of transfer of exosome-shuttle miRNAs from the exosome lumen to the cytosol of target cells is unknown. Here, we demonstrate that DCs release exosomes with different miRNAs depending on the maturation of the DCs. By visualizing spontaneous transfer of exosomes between DCs, we demonstrate that exosomes fused with the target DCs, the latter followed by release of the exosome content into the DC cytosol. Importantly, exosome-shuttle miRNAs are functional, as they repress target mRNAs of acceptor DCs. Our findings unveil a mechanism of transfer of exosome-shuttle miRNAs between DCs and its role as a means of communication and post-transcriptional regulation between DCs. The study has analyzed the microRNA content of 4 samples of immature exosomes, 4 samples of matures exosomes, 2 samples of immature bone-marrow-derived DCs, and 2 samples of mature bone marrow-derived DCs.
Project description:Microarray analysis of exosomal miRNAs vs the miRNAs of their respective donor cells. To determine the miRNA repertoires of exosomes secreted by immune cells, we isolated exosomes from cell supernatants of the Raji B cell line, the Jurkat-derived J77 T cell line, and primary dendritic cells (DCs) derived from human monocytes. Exosomes were isolated by a series of microfiltration and ultracentrifugation steps
Project description:Dendritic cells (DCs) are the most potent antigen (Ag)-presenting cells. Whereas immature DCs down-regulate T cell responses to induce/maintain immunological tolerance, mature DCs promote immunity. To amplify their functions, DCs communicate with neighboring DCs through soluble mediators, cell-to-cell contact and vesicle exchange. Transfer of nanovesicles (<100nm) derived from the endocytic pathway (termed exosomes) represents a novel mechanism of DC-to-DC communication. The facts that exosomes contain exosome-shuttle microRNAs (miRNAs), and DC functions can be regulated by exogenous miRNAs, suggest that DC-to-DC interactions could be mediated through exosome-shuttle miRNAs, an hypothesis that remains to be tested. Importantly, the mechanism of transfer of exosome-shuttle miRNAs from the exosome lumen to the cytosol of target cells is unknown. Here, we demonstrate that DCs release exosomes with different miRNAs depending on the maturation of the DCs. By visualizing spontaneous transfer of exosomes between DCs, we demonstrate that exosomes fused with the target DCs, the latter followed by release of the exosome content into the DC cytosol. Importantly, exosome-shuttle miRNAs are functional, as they repress target mRNAs of acceptor DCs. Our findings unveil a mechanism of transfer of exosome-shuttle miRNAs between DCs and its role as a means of communication and post-transcriptional regulation between DCs.
Project description:Communication between the maternal uterus and the embryo is vital for a successful pregnancy. Exosomes, subtypes of extracellular vesicles comprising many bioactive factors regulate the early stages of pregnancy, specifically during embryo implantation. Nevertheless, the mechanism by which exosomal microRNAs (miRNAs) derived from placental trophoblasts regulate embryo implantation remains elusive. Herer, we isolated and identified exosomes derived from placental trophoblasts cells (HTR8/SVneo). Subsequently, we evaluated the loading miRNA in exosomes by small RNA sequencing. This study provides novel insights into the mechanism of trophoblasts cells-derived exosomes during embryo implantation.
Project description:Exosomes are cell-released bioactive nanovesicles now considered as new partners in the inter-organ cross-talks. The possibility that skeletal muscle (SkM)-derived exosomes act as a mode of systemic communication has hitherto never been described. Thus, in this study we have tested the hypothesis that through the exosomal route, muscle cells might transmit specific signals during insulin-resistance.
Project description:Long non-coding RNAs (lncRNAs) form the largest transcript class in the human transcriptome. These lncRNA are expressed not only in the cells, but they are also present in the cell-derived extracellular vesicles such as exosomes. Our study examines the expression of lncRNAs in several prostate cancer cell lines but also measures the levels of these lncRNAs in the released exosomes. We show that specific lncRNAs are enriched in cancer exosomes and furthermore these sequences harbour miRNA seed regions and appear to be enriched for specific RNA binding motifs.
Project description:small RNAseq was preformed on Wt bone marrow-derived dendritic cells (BMDC) and miR-155 and miR-146a double knockout (DKO) BMDCs that received Wt exosomes to investigate the differences in transferred miRNA Small RNA profiles were generated from Wt donor BMDCs and DKO BMDCs given Wt exosomes 3 replicates in each group
Project description:The sensitivity and specificity of traditional non-invasive diagnostic markers for gastric cancer are insufficient. Long-chain noncoding RNAs (lncRNAs) in circulating exosomes are a new class of promising cancer biomarkers. However, the expression profile of long-chain noncoding RNAs (lncRNAs) in exosomes derived from gastric high-grade intraepithelial neoplasia (GHGIN) has not been reported.The aim of this study was to investigate the expression profile of long non-coding RNAs (lncRNAs) in the plasma exosomes of patients with gastric high-grade intraepithelial neoplasia. Peripheral blood samples were collected from five patients with GHGIN and five healthy donors, and the exosomes were isolated. We used high-throughput sequencing to detect differently expressed lncRNAs (DE lncRNAs) in these individuals. Real-time quantitative polymerase chain reaction (RT-qPCR) assay was performed to verify the sequencing results. The potential roles of the DE lncRNAs in GHGIN were speculated by performing Gene ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) analysis. We constructed a lncRNAs–miRNA–mRNA network diagram and performed association analyses to explore the underlying molecular mechanisms. A total of 25145 lncRNAs were identified in all samples; 83 lncRNAs that showed significant differential expression were further screened, including 76 up‐regulated and 7 down‐regulated lncRNAs. RT-qPCR further confirmed these results. GO and KEGG analyses predicted that these lncRNAs played important roles in protein and macromolecule glycosylation, regulation of protein ubiquitination, and renin-angiotensin system and MAPK signaling pathways. We further constructed a lncRNA–miRNA–mRNA interaction network. This study may help in discovering new molecular changes, enrich the expression profile of lncRNAs in human GHGIN, and help us better understand the molecular mechanisms of GHGIN and gastric cancer.