Involvement of miR-30a-5p and miR-30d in Endothelial to Mesenchymal Transition and Early Osteogenic Commitment under Inflammatory Stress in HUVEC

Abstract

The endothelial to mesenchymal transition (End–MT) can be associated with vascular calcification, by providing mesengenic progenitors. In this study, we investigated a link between End–MT and the osteogenic process and explored the involvement of miR-30a-5p and miR-30d as potential regulators of these processes. End–MT was induced in Human Umbilical Vein Endothelial Cells (HUVEC) through transforming growth factor-β1 (TGF-β1), TGFβ-3 and tumor necrosis factor-α (TNF-α), for 24 h and 6 days. End–MT mediators, mesenchymal and osteo/chondrogenic markers were analyzed through Real-Time PCR, immunofluorescence, flow cytometry and Western Blot. miR-30a-5p and miR-30d over-expression was carried out in HUVEC to explore their effects on End–MT and osteogenic differentiation. HUVEC at 24 h and 6 days gained mesenchymal morphology markers, including matrix metalloproteinase 9 (MMP-9), SLUG, VIMENTIN and α-smooth muscle actin (α-SMA), and a significant migratory potential, notably with TNF-α. After 6 days, the osteo/chondrogenic markers runt-related transcription factor 2 (RUNX-2) and SRY box transcription factor 9 (SOX-9) were upregulated. At this time point, miR-30a-5p and miR-30d decreased. Over-expression of miR-30a-5p and miR-30d affected End–MT mediators and the osteogenic potency in HUVEC, by reducing SLUG, VIMENTIN and RUNX-2. Our data suggest that End–MT represents a key link between inflammation and vascular calcification. Further, miR-30a-5p and miR-30d can regulate both the End–MT and the osteogenic processes, prompting future studies for exploring their potential use as therapeutic targets or biomarkers in vascular diseases.

RNA-seq analysis of extracellular vesicles from hyperphosphatemia-stimulated endothelial cells provides insight into the mechanism underlying vascular calcification

“…The published research that BMP-2 down-regulation miR-30c to increase Runx2 expression in human coronary artery SMCs and promoting mineralization may partly explain our result [ 23 ]. Additionally, the expression of miR-30a was significantly higher in VSMCs during vascular calcification [ 24 – 26 ]. However, miR-30a-3p and miR-30a-5p was significantly reduced in the HP-EC-EVs compared to controls in this study.…”

Profiling Secreted miRNA Biomarkers of Chemical-Induced Neurodegeneration in Human iPSC-Derived Neurons

“…TGF-β signaling pathway was shown to regulate the expression of miR-30s (TGF-β treatment downregulated miR-30s, and this is potentially mediated via Smad2)—shown in human umbilical vein endothelial cells ( Ciavarella et al , 2021 ) and human podocytes ( Shi et al , 2013 )…”

Epigallocatechin-3-Gallate (EGCG) Mitigates Endothelial and Circulating Cells Alterations Following PLLA Electrospun Mat Placement

“…Indeed, the vascular disease context is commonly characterized by inflammation, oxidative stress and hypoxia, pathological conditions that have been shown to stimulate End-MT [ 14 ]. In this regard, we recently demonstrated that End-MT induced by soluble factors, including TNF-α, TGF-β1 and TGF-β3, is associated with increased in vitro calcification [ 15 ]. In the present study, we detected an up-regulation of SLUG, VIMENTIN, MMP-9 and RUNX-2 mRNA levels in HUVEC after being cultured with PLLA for 48 h. The study of the interactions between cells and biomaterials was assessed by an indirect co-culture where a piece of PLLA electrospun mat was placed floating in the growth medium.…”

“…ECs can undergo phenotype transition into mesenchymal cells able to differentiate into the osteogenic lineage by a mechanism known as endothelial-mesenchymal transition (End-MT) [ 13 , 14 ]. This process can be induced by various microenvironment stimuli such as inflammation [ 15 ] and oxidative stress [ 16 , 17 ] conditions that commonly occur in vascular graft procedures, as suggested by the study of Jiang et al, performed on rat aortas. Here, an antioxidant approach through the incorporation of 1,8- octamethylenecitrate-co-cysteine (POCC) into vascular graft was able to reduce the calcification extent and consequent degenerative alterations [ 14 ].…”