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Human Fibroblast-Like Synoviocytes Derived Extracellular Vesicles miRNA sequencing

ABSTRACT: Purpose:The goal of this study is to compare the miRNA expressions in LPS stimulated RA FLSs derived EVs between trauma FLSs derived EVs. Methods:Total RNA of EVs was extracted with trizol LS (Invitrogen, USA), and quantified with Nanodrop. Agarose electrophoresis was used for quality inspection, and library was constructed after passing the quality inspection. Then, Agilent 2100 Bioanalyzer was used to determine the quality of the library. RNA was denatured with 0.1 M NaOH and sequenced on Illumina NextSeq500. Results:In total of 24 miRNAs were changed in the 650 scanned miRNAs in LPS stimulated RA FLSs derived EVs, compared to Trauma FLSs derived EVs, with 13 up-regulated miRNAs and 11 down-regulated miRNAs(P<0.05; Fold change>=1.5).Next, we analyzed the possible biological functions of the expression changed miRNAs using GO analysis. Of note, the results showed that expression altered miRNAs may principally correlate with blood vessel morphogenesis, cardiovascular system development, vasculature development and tube development. Conclusions:RNA parts of LPS-RA FLSs derived EVs may play key role in stimulating angiogenesis of ECs.

ORGANISM(S): Homo sapiens

PROVIDER: GSE185672 | GEO | 2022/02/21

REPOSITORIES: GEO

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Project description:Inflammation-mediated oncogenesis has been implicated in a variety of cancer types. Rheumatoid synovial tissues can be viewed as a tumor-like mass, consisting of hyperplastic fibroblast-like synoviocytes (FLSs). FLSs of rheumatoid arthritis (RA) patients have pro-migratory and invasive characteristics, which may be caused by chronic exposure to genotoxic stimuli, including hypoxia and growth factors. We tested whether a transformed phenotype of RA-FLSs is associated with placental growth factor (PlGF), a representative angiogenic growth factor induced by hypoxia. Here, we identified PlGF-1 and PlGF-2 as the major PlGF isoforms in RA-FLSs. Global gene expression profiling revealed that cell proliferation, apoptosis, angiogenesis, and cell migration were mainly represented by differentially expressed genes in RA-FLSs transfected with siRNA for PlGF. Indeed, PlGF-deficient RA-FLSs showed a decrease in cell proliferation, migration, and invasion, but an increase in apoptotic death in vitro. PlGF gene overexpression resulted in the opposite effects. Moreover, exogeneous PlGF-1 and PlGF-2 increased survival, migration, and invasiveness of RA-FLSs by binding their receptors, Flt-1 and NP-1, up-regulating the expression of anti-apoptotic molecules, pErk and Bcl2. Knock-down of PlGF transcripts reduced RA-FLS proliferation in a xeno-transplantation model. Collectively, in addition to their role for neovascularization, PlGF-1 and -2 promote proliferation, survival, migration, and invasion of RA-FLSs in an autocrine and paracrine manner. These results demonstrated how primary cells of mesenchymal origin acquired an aggressive and transformed phenotype. PlGF and its receptors thus offer new targets for anti-FLS therapy. The FLSs were prepared from the synovial tissues of RA patients and incubated in DMEM supplemented with 10% FBS. Cells were cultured in RPMI supplemented with 10% FBS. There are 4 FLS samples treated with PlGF siRNA, and 4 FLS samples treated with control siRNA.

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Human Fibroblast-Like Synoviocytes Derived Extracellular Vesicles miRNA sequencing

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