AUTHOR=Li Zhaoshui , Song Yanjie , Wang Meng , Shen Ruxin , Qin Kun , Zhang Yu , Jiang Ting , Chi Yifan 

TITLE=m6A regulator-mediated RNA methylation modification patterns are involved in immune microenvironment regulation of coronary heart disease

JOURNAL=Frontiers in Cardiovascular Medicine

VOLUME=9

YEAR=2022

URL=https://www.frontiersin.org/journals/cardiovascular-medicine/articles/10.3389/fcvm.2022.905737

DOI=10.3389/fcvm.2022.905737

ISSN=2297-055X

ABSTRACT=<sec><title>Background</title><p>Although the roles of m6A modification in the immune responses to human diseases have been increasingly revealed, their roles in immune microenvironment regulation in coronary heart disease (CHD) are poorly understood.</p></sec><sec><title>Methods</title><p>The <ext-link ext-link-type="NCBI:geo" xlink:href="GSE20680" xmlns:xlink="http://www.w3.org/1999/xlink">GSE20680</ext-link> and <ext-link ext-link-type="NCBI:geo" xlink:href="GSE20681" xmlns:xlink="http://www.w3.org/1999/xlink">GSE20681</ext-link> datasets related to CHD were acquired from the Gene Expression Omnibus (GEO) database. A total of 30 m6A regulators were used to perform LASSO regression to identify the significant genes involved in CHD. Unsupervised clustering analysis was conducted using the m6A regulators to distinguish the m6A RNA methylation patterns in patients with CHD. The differentially expressed genes (DEGs) and biological characteristics, including GO and KEGG enrichment results, were assessed for the different m6A patterns to analyse the impacts of m6A regulators on CHD. Hub genes were identified, and subsequent microRNAs-mRNAs (miRNAs–mRNAs) and mRNAs-transcriptional factors (mRNA-TFs) interaction networks were constructed by the protein and protein interaction (PPI) network method using Cytoscape software. The infiltrating proportion of immune cells was assessed by ssGSEA and the CIBERSORT algorithm. Quantitative real-time PCR (qRT-PCR) was performed to detect the expression of the significant m6A regulators and hub genes.</p></sec><sec><title>Results</title><p>Four of 30 m6A regulators (<italic>HNRNPC, YTHDC2, YTHDF3</italic>, and <italic>ZC3H13</italic>) were identified to be significant in the development of CHD. Two m6A RNA methylation clusters were distinguished by unsupervised clustering analysis based on the expression of the 30 m6A regulators. A total of 491 genes were identified as DEGs between the two clusters. A PPI network including 308 mRNAs corresponding to proteins was constructed, and 30 genes were identified as hub genes that were enriched in the bioprocesses of peptide cross-linking, keratinocyte differentiation. Twenty-seven hub genes were found to be related to miRNAs, and seven hub genes were found to be related to TFs. Moreover, among the 30 hub genes, eight genes were found to be upregulated in CHD, and three were found to be downregulated in CHD compared to the normal people. The high m6A modification pattern was associated with a higher infiltrated abundance of immune cells.</p></sec><sec><title>Conclusion</title><p>Our findings demonstrated that m6A modification plays crucial roles in the diversity and complexity of the immune microenvironment in CHD.</p></sec>