Authors:

Ashlin Monish

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About Paper:

MicroRNAs are the predominant exosomal cargo that can regulate a variety of cellular processes. Previously, we have shown that Dentin Matrix Protein 1 (DMP1) stimulated periodontal ligament stem cells (PDLSCs) release exosomes containing miRNA cargo that promoted osteogenic differentiation. In this study, we demonstrate that DMP1-stimulated PDLSC- exosomes carry miRNA-rich cargo that facilitates cytokine signaling.Exosomes from PDLSCs cultured with and without DMP1 were isolated using ultracentrifugation. A Qiagen miRNome PCR array was used to identify miRNA differentially expressed in exosomes of DMP1 treated PDLSCs. Predicted gene targets of differentially expressed miRNAs were evaluated using Gene Ontology to identify pathways related to cell differentiation, immune and inflammatory signaling. RT-PCR was performed on the total RNA of PDLSCs to identify the expression of miRNA targets.Nanosight analysis of the isolated exosomes showed sizes in the 50-150nm range. Western blot analysis confirmed the presence of the exosomal marker CD63. The miRNA array showed 45 differentially expressed miRNAs. 5 miRNA were further analyzed including miR-106a-5p (fc = 0.33), miR-145-5p (fc = 0.22), miR-27a-3p (fe = 0.4), miR-92a-3p (fe = 0.32), and let-7a-5p (fe = 0.21). Gene Ontology analysis of these miRNA targets, with a predication score >90, demonstrated enrichment of MAPK and TNFa signaling pathways, cell differentiation and cytoskeletal alterations. qRT-PCR results of gene targets indicate a positive correlation between miRNA and gene target expression. Exosomal miRNAs enriched in DMP1 stimulated PDLSCs may function in cellular differentiation and paracrine immunomodulation via cytokine signaling. Future study on miR-106a-5p and its role in regulating targets, such as CTSK and DUSP10, could provide insight into the downstream effects of DMP1 on miRNA regulated immunomodulatory signaling in PDLSCs. POSS GOSH OHM OOH SHS SOHO OSO OO HKO COSC OOCOHCOH OSC OC OOCOOCO PBDSDSDDDISDIDIDDIPSSDDIDIDFTIDADIDIIIAIDDDSIDSDIOOSD

Source:

University of Illinois Chicago

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Co-authors:

Ashlin Monish