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

Fan Zhang,Haibo Zhang,Yingchun Han,Congcong Zhang,Naixuan Cheng

doi:10.3389/fgene.2021.722564

Abstract

Calcific aortic valve disease (CAVD) is the most common structural heart disease, and the morbidity is increased with elderly population. Several microRNAs (miRNAs) have been identified to play crucial roles in CAVD, and numerous miRNAs are still waiting to be explored. In this study, the miRNA expression signature in CAVD was analyzed unbiasedly by miRNA-sequencing, and we found that, compared with the normal control valves, 152 miRNAs were upregulated and 186 miRNAs were downregulated in calcified aortic valves. The functions of these differentially expressed miRNAs were associated with cell differentiation, apoptosis, adhesion and immune response processes. Among downregulated miRNAs, the expression level of miR-139-5p was negatively correlated with the osteogenic gene RUNX2, and miR-139-5p was also downregulated during the osteogenic differentiation of primary human aortic valve interstitial cells (VICs). Subsequent functional studies revealed that miR-139-5p overexpression inhibited the osteogenic differentiation of VICs by negatively modulating the expression of pro-osteogenic gene FZD4 and CTNNB1. In conclusion, these results suggest that miR-139-5p plays an important role in osteogenic differentiation of VICs via the Wnt/β-Catenin pathway, which may further provide a new therapeutic target for CAVD.

Highlights

Calcific aortic valve disease (CAVD), which results in aortic valve stenosis, affects 25% of the population >65 years of age (Coffey et al, 2014; Lindman et al, 2016)
Through the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, we found that the target genes of differentially expressed miRNAs (DEMs) were significantly enriched in a few signaling pathways, especially the phosphatidylinositol 3ʹkinase (PI3K)–protein kinase B (AKT) signaling pathway, miR-139-5p and Aortic Valve Calcification mitogen-activated protein kinase (MAPK) signaling pathway, and cytokine–cytokine receptor interaction pathway (Figure 2B)
We found that the expression levels of FZD4 and CTNNB1 were increased in osteogenically differentiated valve interstitial cells (VICs), while they were repressed by miR-139-5p overexpression (Figures 5F,G)

Summary

Introduction

Calcific aortic valve disease (CAVD), which results in aortic valve stenosis, affects 25% of the population >65 years of age (Coffey et al, 2014; Lindman et al, 2016). CAVD confers a high clinical and economic burden, because no effective pharmacological therapy exists (Hutcheson et al, 2014). Owing to its high morbidity and mortality, a strong incentive exists to identify the key molecular drivers contributing to the development of this disease, which could provide a new target for clinical treatment in its earlier stages before cardiac damage. CAVD is marked by inflammatory infiltration, fibrotic extracellular matrix (ECM) synthesis by activated valve interstitial cells (VICs), increased leaflet thickening and stiffness and calcific mineral deposition (Peeters et al, 2018).

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Results

Conclusion