Project description:MiRNA microarray analysis was performed on exosomes secreted by mouse MSC cells under two different conditions of normal oxygen and hypoxia, in order to find out the different miRNAs in exosomes secreted by MSC under two different conditions.
Project description:BACKGROUND The therapeutic potential of endothelial colony-forming cells (ECFCs) may be impaired in an ischemic environment. Direct injection of ECFCs is not an effective method of rescuing the ischemic heart, but exosomes derived from these cells may be a promising therapeutic tool. However, exosomes produced under normoxia and hypoxia may not be identical. Therefore, the purpose of this study was to investigate alterations in the anti-fibrotic effects of hypoxia-treated ECFC-derived exosomes and the underlying mechanism involved. MATERIAL AND METHODS ECFCs were isolated from peripheral blood and exosomes were collected from ECFCs treated with normoxia (nor-exo) or hypoxia (hyp-exo). Effects of exosomes on cardiac fibroblast activation were evaluated in vitro. MicroRNAs (miRNAs) inside the exosomes were extracted and compared using next-generation RNA sequencing. Predicted target mRNAs of miR-10b-5p were validated using a dual-luciferase reporter gene assay method. RESULTS Nor-exo significantly ameliorated cardiac fibroblast activation in vitro. These effects were attenuated in the hyp-exo-treated group. miR-10b-5p was enriched in nor-exo but not in hyp-exo. Dual-luciferase reporter gene assay found that both SMAD-specific E3 ubiquitin protein ligase 1 (Smurf1) and histone deacetylase 4 (HDAC4) mRNAs were inhibited by miR-10b-5p. The expression of neutral sphingomyelinase 2 (N-SMase2) was decreased in hypoxia ECFCs, and this result was consistent with the changes in miR-10b-5p in hyp-exo. CONCLUSIONS Due to a reduction of miR-10b-5p, which targets the fibrotic genes Smurf1 and HDAC4, the anti-fibrotic effects of hyp-exo were abolished.
Project description:We recently showed that exosomes from primary AML cells and cell lines have potent regulatory capacity and we hypothesized that leukemia cell exosome trafficking might account for the suppression of residual hematopoietic stem and progenitor cells (HSPC) in the leukemic niche. Here we studied Molm-14 cells in in vitro experiments using purified exosomes under carefully calibrated low-oxygen conditions. This approach revealed the active regulation of stromal- and hematopoietic progenitor cell function. Systematic analysis of AML exosomes identified a panel of differentially enriched hypoxia-responsive miRNAs, including miR-210, -155, and -146a. We cultured cells under normoxic and hypoxic conditions. Exosomes were released from the parental cells and captured. Total RNA was isolated from both exosomes and cells. Please note that the sample numbers (in the titles) do differ from pair identification (i.e. pair ID) as the pair is only in reference to the paired-t test and parental vs. released pairs.
Project description:microRNA profiles of Exosomes from Pooled NPC Patients serum comparing Control Exosomes from Healthy donors serum Two-condition experiment, Exosomes from Pooled Healthy donors serum vs. Exosomes from Pooled NPC Patients serum. Biological replicates: 1 Exosomes from Pooled Healthy donors serum, 1 Exosomes from Pooled NPC Patients serum,
Project description:Small vesicles, known as exosomes, are secreted from various cell types. Exosomes secreted by mesenchymal stem cells have therapeutic effects against a variety of diseases, and may be able to partially replace stem cell therapies. Previously, we established and characterized human leukocyte antigen (HLA) haplotype homo (HHH) dental pulp cell (DPC) lines from human wisdom teeth. In this report, we purified the exosomes secreted from HHH-DPCs and evaluated their therapeutic potential in a periodontitis model. The exosomes purified from HHH-DPCs showed homogeneous and spherical membrane structures, and showed low but significant expression of HLA class I molecules. The exosomes further promoted proliferation and migration in DPCs. A comparison of miRNAs revealed that the HHH-DPC exosomes contained higher levels of multiple Let-7 family miRNAs compared to HHH-induced pluripotent stem cell (iPSC)-derived exosomes. Finally, the HHH-DPC exosomes showed preventive effects in a mouse model of periodontitis induced by lipopolysaccharides (LPS). In summary, HHH-DPC exosomes expressed HLA molecules which may induce an immune response in HLA-mismatched transplantations. However, they successfully stimulated the proliferation and migration of cells and showed suppressive effects on LPS-induced periodontitis. Therefore, HHH-DPC exosomes show great potential for applications in periodontal treatments.
Project description:We recently showed that exosomes from primary AML cells and cell lines have potent regulatory capacity and we hypothesized that leukemia cell exosome trafficking might account for the suppression of residual hematopoietic stem and progenitor cells (HSPC) in the leukemic niche. Here we studied Molm-14 cells in in vitro experiments using purified exosomes under carefully calibrated low-oxygen conditions. This approach revealed the active regulation of stromal- and hematopoietic progenitor cell function. Systematic analysis of AML exosomes identified a panel of differentially enriched hypoxia-responsive miRNAs, including miR-210, -155, and -146a.
Project description:Hypoxic cancer cells are known to acquire malignance like invasiveness and therapy resistance. In order to identify the candidate genes responsible for the malignant properties of hypoxic cancer cells, gene expression profile of hypoxic cancer cells is investigated herein.
Project description:we studied the role of exosomes isolated from M.tb infected macrophages in modulating the macrophage response to IFN-γ. Nimblegen microarray gene expression studies were used to compare the suppression of IFN-γ inducible genes by exosomes relative to the virulent strain of M.tuberculosis. Overall our study suggest that exosomes, as carriers of M.tb pathogen associated molecular patterns (PAMPs), may provide a mechanistic link by which M.tb may exert its suppression of host immune response beyond the infected cell, and implies a physiological role for exosomes in immune surveillance of TB. Macrophages were treated with exosomes, infected with M.tb H37Rv or left untreated for 18 hours followed by +/- IFN-γ for an additional 18 hours. Cells were harvested and RNA was isolated and converted to double stranded cDNA and subsequently labeled and hybridized onto Mus musculus 4×72 Nimblegen microarray using Nimblegen Hybridization system 4 according to manufacturer’s instructions (Roche)