miR-214 Attenuates Aortic Valve Calcification by Regulating Osteogenic Differentiation of Valvular Interstitial Cells

Abstract

Calcific aortic valve disease (CAVD) is a common heart valve disease in aging populations, and aberrant osteogenic differentiation of valvular interstitial cells (VICs) plays a critical role in the pathogenesis of ectopic ossification of the aortic valve. miR-214 has been validated to be involved in the osteogenesis process. Here, we aim to investigate the role and mechanism of miR-214 in CAVD progression. miR-214 expression was significantly downregulated in CAVD aortic valve leaflets, accompanied by upregulation of osteogenic markers. Overexpression of miR-214 suppressed osteogenic differentiation of VICs, while silencing the expression of miR-214 promoted this function. miR-214 directly targeted ATF4 and Sp7 to modulate osteoblastic differentiation of VICs, which was proved by dual luciferase reporter assay and rescue experiment. miR-214 knockout rats exhibited higher mean transvalvular velocity and gradient. The expression of osteogenic markers in aortic valve leaflets of miR-214 knockout rats was upregulated compared to that of the wild-type group. Taken together, our study showed that miR-214 inhibited aortic valve calcification via regulating osteogenic differentiation of VICs by directly targeting ATF4 and Sp7, indicating that miR-214 may act as a profound candidate of targeting therapy for CAVD.

Basic Fibroblast Growth Factor Inhibits Aortic Valvular Interstitial Cells Calcification via Notch1 Pathway

“…The harvested VICs were collected by centrifugation and then seeded on culture dish in DMEM containing 10% fetal bovine serum, and the medium was changed every 2–3 days. The VICs isolation and immunophenotype identification were reported previously 12. The VICs at passages 3–5 were used for experiment.…”

Rho A/ROCK1 signaling-mediated metabolic reprogramming of valvular interstitial cells toward Warburg effect accelerates aortic valve calcification via AMPK/RUNX2 axis

“…To stimulate calcification, human VICs were cultured with inorganic phosphate-osteogenic medium (IP-OM) (complete media supplemented with 2 mM sodium dihydrogen phosphate (Beyotime Biotech, Jiangsu, China), 75 mM ascorbic acid (Beyotime Biotech), 100 nM dexamethasone (Beyotime Biotech) and 10 −7 mM insulin (Beyotime Biotech)) for 7–9 days, modified from previously described approaches [ 4 , 37 , 38 ]. Calcific nodules could be observed by day 3.…”

The Complex Interplay of Inflammation, Metabolism, Epigenetics, and Sex in Calcific Disease of the Aortic Valve

“…In particular, performing enzymatic assays, they detected a higher expression level of NF-κB, TLR-4, and miR-214 in the blood and aortic valve tissue samples of patients with CAVD compared to control ones. Furthermore, knockdown of miR-214 was able to inhibit the secretion of IL-6, IL-8, ICAM-1 ( 100 , 101 ). Up-regulation of human myeloid differentiation factor 88 (MyD88) promoted the secretion of pro-inflammatory factors and increased calcium deposition over the leaflets and increased the expression of RUNX-2 and BMP-2 ( 100 ).…”