The construction and analysis of m6A-related ceRNA network and patterns of immune infiltration in calcified aortic valve disease

Abstract

Background Despite being the most prevalent valve heart disease among the elderly, calcified aortic valve disease (CAVD) is not adequately addressed based on its current mechanisms. N6-methyladenosine (m6A) modification is increasingly being studied in cardiovascular disease. Nonetheless, the biological role of m6A in CAVD remains to be determined. Methods We obtained the differentially expressed m6A based on difference analysis, and identified the target genes regulated by key m6A through co-expression analysis and m6A2Target database. The enrichment analysis of targeted genes was performed via Metascape. Immunocyte infiltration analysis was performed by R-package. The tools such as miRDB, Targetscan, miRTarBase, and Cytoscape were applied for the construction of competitive endogenous RNAs (ceRNAs) network. Quantitative real-time PCR (qRT-PCR) was utilized to verify whether the expression of components in the ceRNA network is consistent with the public database. Results The ceRNA network consists of one m6a (KIAA1429), one mRNA (ZC3H12C), three miRNAs (miR-17-5p, miR-20b-5p and miR-137), and two lncRNAs (HCG11 and PRICKLE2-AS3). KIAA1429 is a down-regulated methyltransferase in CAVD. The genes modified by KIAA1429 were primarily enriched in the metabolic process of RNA, viral process, and immune system process et al. There was a rising infiltration of macrophages m0, and a decreasing infiltration of macrophages m2, dendritic cells resting in CAVD. KIAA1429 was negatively correlated with macrophages m0 and positively correlated with macrophages m2. KIAA1429(m6a), ZC3H12C (mRNA), and HCG11(lncRNA) showed lower expression levels in CAVD than those in normal tissue, whereas miR-17-5p, miR-20b-5p (miRNA) showed higher expression levels. Conclusion Potential pathways associated with KIAA1429 in CAVD were identified, in which ZC3H12C and miR-20b-5p might participate in CAVD progression via the nuclear factor kappa-B (NF-κB) pathway.