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SKP-SC-EVs-derived miRNA-30a-5p promote angiogenesis in peripheral nerve regeneration by targeting Lif and ANGPT2

ABSTRACT: Ischemia and hypoxia caused by vascular injury aggravate nerve injury. The extracellular vesicles from skin precursor-derived Schwann cells (SKP-SC-EVs) have proven to be beneficial to the regeneration of peripheral nerve injury, but whether SKP-SC-EVs can repair injured peripheral nerves by promoting angiogenesis remains unclear. Here, we found that SKP-SC-EVs enhanced the angiogenesis, which were verified by the transparency of tissue engineered nerve graft (TENG) and ultrasonic blood flow imaging. In vitro we verified that SKP-SC-EVs promote the proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs), which were traditionally used for the evaluating angiogenesis capacity. Furthermore, the expression pattern of microRNA (miRNA) in SKP-SC-EVs was profiled and functional experiments were screened. Notably, miR-30a-5p was selected because of the remarkably pro-angiogenic ability in vivo and in vitro, which was similar to the effects of SKP-SC-EVs. Luciferase reporter assay and functional experiments revealed miR-30a-5p contained in SKP-SC-EVs promoted angiogenesis by targeting ANGPT2 and LIF in the insufficient of VEGFa. Taken together, miR-30a-5p enriched in SKP-SC-EVs may be a key regulator of angiogenesis for the cell-free treatment for peripheral nerve injury.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE245328 | GEO | 2025/02/12

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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SKP-SC-EVs-derived miRNA-30a-5p promote angiogenesis in peripheral nerve regeneration by targeting Lif and ANGPT2

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