LncRNA OIP5-AS1 inhibits osteoblast differentiation of valve interstitial cells via miR-137/TWIST11 axis.

OIP5-AS1, a highly abundant imprinted long non-coding RNA (lncRNA), has been implicated in calcific aortic valve disease (CAVD). However, the function and underlying mechanism of OIP5-AS1 in CAVD progression remains unknown. In this study, osteoblastic differentiation of valve interstitial cells (VICs) isolated from human calcific aortic valves was induced by osteogenic medium. The protein levels of osteogenic markers were determined by immunofluorescence and western blotting. OIP5-AS1, miR-137 and TWIST-related protein 1 (TWIST1) expressions were detected by quantitative real-time PCR (qRT-PCR). ALP activity was evaluated by spectrophotometry. Mineralized bone matrix formation was assessed by Alizarin Red S staining. The interaction between OIP5-AS1 and miR-137 was studied using luciferase reporter assay, RNA pull-down assay and RNA-binding protein immunoprecipitation (RIP) assay. Luciferase reporter assay was also used to identify the possible interaction between miR-137 and TWIST11. The results showed that downregulated expression of OIP5-AS1 was observed in human aortic VICs after osteogenic induction. In vitro experiments revealed that OIP5-AS1 acted as a negative regulator of osteogenic differentiation. Mechanistically, we further showed that OIP5-AS1 could relieve osteogenic differentiation of VICs via upregulating miR-137 target gene TWIST1. Our study provides novel mechanistic insights into the cross-talk between OIP5-AS1, miR-137, and TWIST11, shedding light on the therapy for CAVD.