MicroRNA-204 Deficiency in Human Aortic Valves Elevates Valvular Osteogenic Activity

Abstract

Aortic valve interstitial cells (AVICs) play a major role in valvular calcification associated with calcific aortic valve disease (CAVD). Although AVICs from diseased valves display a pro-osteogenic phenotype, the underlying mechanism causing this remains unclear. MicroRNA-204 (miR-204) is a negative regulator of osteoblast differentiation. We sought to analyze miR-204 expression in diseased human aortic valves and determine the role of this miR in AVIC osteogenic activity associated with CAVD pathobiology. In situ hybridization and PCR analysis revealed miR-204 deficiency in diseased valves and in AVICs from diseased valves. MiR-204 mimic suppressed alkaline phosphatase (ALP) expression and calcium deposition in AVICs from diseased valves. MiR-204 antagomir enhanced ALP expression in AVICs from normal valves through induction of Runx2 and Osx, and expression of miR-204 antagomir in mouse aortic valves promoted calcium deposition through up-regulation of Runx2 and Osx. Further, miR-204 mimic suppressed the osteogenic responses to TGF-β1 in AVICs of normal valves. In conclusion, miR-204 deficiency contributes to the mechanism underlying elevated osteogenic activity in diseased aortic valves, and miR-204 is capable of reversing the pro-osteogenic phenotype of AVICs of diseased valves and suppressing AVIC osteogenic response to stimulation. Exogenous miR-204 may have therapeutic potential for inhibiting valvular calcification associated with CAVD progression.

MicroRNAs Modulate Signaling Pathways in Osteogenic Differentiation of Mesenchymal Stem Cells

“…It also showed the ability to target CXCL12, coding for SDF-1α, an angiogenic factor and, notably, the silencing of miR-137-3p could favor both osteogenesis and angiogenesis [ 181 ]. MiR-204 deficiency has been found to be involved in the increased osteogenic activity associated with calcific aortic valve disease [ 182 ]. This result agrees with several studies reporting that in different mesenchymal progenitor cell lines, such as C3H10T1/2, C2C12, ST2 and primary BM-MSCs, miR-204, targeting RUNX2, negatively controls osteogenic differentiation [ 94 , 183 ].…”

NcRNAs in Vascular and Valvular Intercellular Communication

“…Furthermore, miR-125b was found to be downregulated in an in vivo model of atherosclerosis and aortic calcification in mice (Goettsch et al, 2011;Rangrez et al, 2012;Coffey and Jones, 2014), while miR-223 was overexpressed in calcified murine aortas (Rangrez et al, 2012). Human calcific aortic valve disease is associated with significantly reduced miR-204 levels and miR-204 mimics suppressed the osteogenic activity of interstitial cells from diseased valves (Song et al, 2020).…”

miRNA Expression Profiling Uncovers a Role of miR-139-5p in Regulating the Calcification of Human Aortic Valve Interstitial Cells

“…Because the expression level of hsa-miR-374c-3p in valves was lower than the other five miRNAs, we removed it from the candidate miRNAs. Among the five candidate miRNAs, it was previously reported that the expression of miR-204-5p was decreased in calcified aortic valves and inhibited the osteogenic differentiation of VICs and aortic valve calcification (Yu et al, 2018;Song et al, 2019). Then, we calculated the correlation of five candidate miRNAs with the osteogenic genes RUNX2.…”

“…Compared with normal human aortic valves, the miRNA expression profiles in calcified aortic valve tissues have changed (Wang et al, 2017). Several miRNAs [miR-204-5p (Yu et al, 2018;Song et al, 2019), miR-143 (Fiedler et al, 2019), miR-125b (Ohukainen et al, 2015), miR-34a (Toshima et al, 2020), etc. ] have been reported to participate in the osteogenic differentiation of VICs and valve calcification.…”