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MicroRNA profile of extracellular vesicles derived from control vehicle- and LIPUS-induced stem cells from the apical papilla

ABSTRACT: Low intensity pulsed ultrasound (LIPUS) is shown to promote osteogenic differentiation and antiinflammation of periodontal mesenchymal stromal cells. We found extracellular vesicles (EVs) derived from LIPUS-induced stem cells from apical papilla (SCAPs) also have stronger anti-inflammatory and osteogenic effects in vitro and in vivo. To obtain clues on whether microRNA (miR) of EVs contribute to stronger anti-inflammatory and osteogenic effects, we analyzed miR expression in EVs from control- and LIPUS-induced SCAPs by deep sequencing. RNA-sequencing suggests that the LIPUS-EVs induce stronger osteogenic differentiation and antiinflammation by using their cargos, including upregulated osteogenic miRNAs to activate the PI3K/Akt and MAPK signaling pathways, and upregulated anti-inflammatory miRNAs to inhibit the TNF-α signaling pathways.

ORGANISM(S): Homo sapiens

PROVIDER: GSE188347 | GEO | 2022/07/02

REPOSITORIES: GEO

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Project description:Malignant germ-cell-tumours (GCTs) are characterised by microRNA (miRNA/miR-) dysregulation, with universal over-expression of miR-371~373 and miR-302/367 clusters regardless of patient age, tumour site, or subtype (seminoma/yolk-sac-tumour/embryonal carcinoma). These miRNAs are released into the bloodstream, presumed within extracellular-vesicles (EVs) and represent promising biomarkers. Here, we comprehensively examined the role of EVs, and their miRNA cargo, on (fibroblast/endothelial/macrophage) cells representative of the testicular GCT (TGCT) tumour microenvironment (TME). Small RNA next-generation-sequencing was performed on 34 samples, comprising representative malignant GCT cell lines/EVs and controls (testis fibroblast [Hs1.Tes] cell-line/EVs and testis/ovary samples). TME cells received TGCT co-culture, TGCT-derived EVs, and a miRNA overexpression system (miR-371a-OE) to assess functional relevance. TGCT cells secreted EVs into culture media. MiR-371~373 and miR-302/367 cluster miRNAs were overexpressed in all TGCT cells/subtypes compared with control cells and were highly abundant in TGCT-derived EVs, with miR-371a-3p/miR-371a-5p the most abundant. TGCT co-culture resulted in increased levels of miRNAs from the miR-371~373 and miR-302/367 clusters in TME (fibroblast) cells. Next, fluorescent labelling demonstrated TGCT-derived EVs were internalised by all TME (fibroblast/endothelial/macrophage) cells. TME (fibroblast/endothelial) cell treatment with EVs derived from different TGCT subtypes resulted in increased miR-371~373 and miR-302/367 miRNA levels, and other generic (eg, miR-205-5p/miR-148-3p) and subtype-specific (seminoma, eg, miR-203a-3p; yolk-sac-tumour, eg, miR-375-3p) miRNAs. MiR-371a-OE in TME cells resulted in increased collagen contraction (fibroblasts) and angiogenesis (endothelial cells), via direct mRNA downregulation and alteration of relevant pathways. TGCT cells communicate with nontumour stromal TME cells through release of EVs enriched in oncogenic miRNAs, potentially contributing to tumour progression.

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MicroRNA profile of extracellular vesicles derived from control vehicle- and LIPUS-induced stem cells from the apical papilla

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