Machine Learning and Multi-Omics Integration Reveal TRPV2 as a Central Regulator in Bicuspid Aortic Valve Calcification

Bicuspid aortic valve (BAV), the most common congenital heart defect, is strongly predisposed to early calcification, yet the molecular drivers remain poorly defined. This study aims to identify the functional role of transient receptor potential vanilloid 2 (TRPV2) in valve interstitial cell (VIC) calcification and to explore its upstream and downstream regulatory network.

Transcriptomic data from calcified BAV tissues (GSE83453, n = 18) were analyzed using gene set variation analysis, weighted gene co-expression network analysis, and multiple machine learning algorithms. A ceRNA regulatory network was constructed by integrating miRNA–mRNA–lncRNA interactions. Findings were validated in murine VICs using osteogenic induction. TRPV2 function was evaluated by small interfering RNA-mediated knockdown and overexpression assays. Alizarin Red staining, quantitative real-time PCR, Western blot, and dual-luciferase reporter assays were used to assess calcification phenotypes and molecular mechanisms.

Multi-omics integration and machine learning identified TRPV2 as a hub gene associated with calcified BAV tissue. TRPV2 expression is significantly upregulated during osteogenic induction of VICs, while its knockdown reduces calcium nodule formation and suppresses osteogenic markers Runx2 and ALP. Bioinformatics and luciferase assays confirm that miR-3619–5p directly targets the 3′ UTR of TRPV2. TRPV2 overexpression rescued the miR-3619–5p inhibitory effects, restoring calcification phenotypes. A competing endogenous RNA (ceRNA) axis involving TRPV2 is proposed as a novel driver of BAV calcification.

TRPV2 emerges as a potential pro-osteogenic regulator of VIC calcification operating within a ceRNA-linked regulatory axis. These findings nominate TRPV2 and its upstream microRNA interactions as candidate therapeutic targets for mitigating calcific progression in BAV.