Coordination of PRKCA/PRKCA-AS1 interplay facilitates DNA methyltransferase 1 recruitment on DNA methylation to affect protein kinase C alpha transcription in mitral valve of rheumatic heart disease

In the present study, mitral valve tissues from three mitral stenosis patients with RHD by valve replacement and two healthy donors were harvested and conducted DNA methylation signature on PRKCA by MeDIP-qPCR. The presence of hypomethylated CpG islands at promoter and 5MODIFIER LETTER PRIME terminal of PRKCA was observed in RHD accompanied with highly expressed PRKCA and down-regulated antisense long non-coding RNA (lncRNA) PRKCA-AS1 compared to health control. Furthermore, the enrichments of DNMT1/3A/3B on PRKCA were detected by ChIP-qPCR assay in vivo and in human cardiomyocyte AC16 and RL-14 cells exposed to TNF-alpha in vitro, and both demonstrated that DNMT1 substantially contributed to DNA methylation. Additionally, PRKCA-AS1 was further determined to bind with promoter of PRKCA via 5MODIFIER LETTER PRIME terminal and interact with DNMT1 via 3MODIFIER LETTER PRIME terminal. Taken together, our results illuminated a novel regulatory mechanism of DNA methylation on regulating PRKCA transcription through lncRNA PRKCA-AS1, and shed light on the molecular pathogenesis of RHD occurrence.

Automated summary

In this study, mitral valve tissues from rheumatic heart disease patients showed hypomethylated CpG islands at the PRKCA promoter, along with increased PRKCA expression and decreased PRKCA-AS1 expression. DNMT1 was found to play a significant role in DNA methylation on PRKCA, both in vivo and in vitro. PRKCA-AS1 was shown to interact with both the PRKCA promoter and DNMT1, revealing a novel regulatory mechanism in DNA methylation and the molecular pathogenesis of rheumatic heart disease.