METTL14 regulates M6A methylation-modified primary miR-19a to promote cardiovascular endothelial cell proliferation...

Area of Science:

• Molecular Biology
• Epigenetics
• Cardiovascular Research

Background:

• N6-methyl-adenosine (M6A) is increasingly recognized for its role in various pathophysiological processes.
• Methylases facilitate mature microRNA (miRNA) processing in an M6A-dependent manner, influencing disease development.
• The specific regulatory mechanisms of M6A in atherosclerosis (AS) remain largely undefined.

Purpose of the Study:

• To elucidate the role of M6A modification and the methyltransferase METTL14 in the pathogenesis of atherosclerosis.
• To investigate the impact of METTL14 on miRNA processing and cellular behavior in atherosclerotic vascular endothelial cells (ASVEC).
• To identify the potential therapeutic targets within the METTL14/M6A/miR-19a signaling axis for AS treatment.

Main Methods:

• Quantification of M6A, METTL14, demethylase, and miR-19a levels in ASVEC using qRT-PCR.
• Assessment of ASVEC proliferation (CCK8), PCNA expression (WB, qRT-PCR), and invasion (Transwell assays).
• Investigation of protein interactions using Co-immunoprecipitation (METTL14-DGCR8) and RNA interactions using RNA Immunoprecipitation (METTL14-miR-19a).

Main Results:

• ASVEC exhibited significantly elevated M6A modification levels and METTL14 expression.
• METTL14 silencing reduced ASVEC proliferation and invasion, suppressed DGCR8 binding, and decreased mature miR-19a while increasing pre-miR-19a.
• METTL14 overexpression enhanced DGCR8 and M6A levels; miR-19a silencing inhibited ASVEC proliferation and invasion.

Conclusions:

• METTL14 promotes ASVEC proliferation and invasion by increasing M6A modification of pri-miR-19a and enhancing mature miR-19a processing.
• The METTL14/M6A/miR-19a signaling pathway represents a potential novel therapeutic target for atherosclerosis.
• Understanding M6A regulation in ASVEC provides insights into epigenetic mechanisms driving vascular disease.