Variation in Genotype and DNA Methylation Patterns based on Alcohol Use and CVD in the Korean Genome and Epidemiology Study (KoGES)

Abstract

Background: Many epidemiological researches have shown that alcohol consumption can increase the risk of cardiovascular diseases (CVDs), such as myocardial infarction (MI), coronary artery, dyslipidemia, and hypertension. This study aimed to confirm the association between DNA methylation patterns of HECT domain E3 ubiquitin protein ligase 4 (HECTD4) genes and alcohol consumption and CVDs in the Korean population. Methods: 8,840 genotype and 446 DNA methylation data were collected from a survey during 2001–2002 years in the Ansan-Ansung cohort of the Korean Genome and Epidemiology Study (KoGES). After the base survey, it was followed every two years and total eight follow-up surveys were conducted. As a case–control study, I compared Drinkers and Never Drinkers in the two groups, CVDs (MI, coronary artery, dyslipidemia, and hypertension) and Non-CVD groups. I then identified the frequently detected rs2074356, rs11066280 and rs12229654 in HECTD4 genotypes and epigenotypes. Furthermore, I retrieved Differentially methylated regions (DMRs) from four groups based on sex and CVDs and compared them by alcohol status. Finally, I analyzed the enriched Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and visualized the network, heatmap, and volcano plot of DMR. Results: The pattern of DNA methylation of HECTD4 genes associated with CVD is strongly influenced by alcohol consumption. DMRs of two pairs were screened from each group based on the methylation patterns and were represented as a heatmap, Enriched KEGG pathways etc. After dividing into groups with and without CVD, they were divided into four groups, male and female, and each group was reclassified as alcohol intake. rs2074356 single nucleotide polymorphism (SNP) in HECTD4 gene according to alcohol consumption was significant in GWAS on the both presence (p-value = 1.49 * 10-26; odds ratio = 0.2402; 95% CI = 0.1680 - 0.3434) and absence (p-value = 1.41 * 10-107; odds ratio = 0.3013; 95% CI =0.2598 - 0.3504) of CVDs. rs11066280 SNP in HECTD4 gene according to alcohol consumption was significant in GWAS on the both presence (p-value = 1.15 * 10-24; odds ratio = 0.2919; 95% CI = 0.2109 - 0.3996) and absence (p-value = 1.95 * 10-106; odds ratio = 0.3326; 95% CI =0.2899 - 0.3814) of CVDs. Conclusions: Genome-wide association study (GWAS) revealed single nucleotide polymorphisms (SNPs) rs12229654, rs2074356 and rs11066280 in HECTD4 to be significantly associated with CVDs and without CVD between alcohol status. In the same alcohol status, the risk of CVD onset changed according to the presence or absence of HECTD4 in the gene. This study show that genetic variation in HECTD4 is associated with CVD risk in the presence and absence of alcohol and that alcohol significantly influences the pattern of DMR methylation associated with CVD in DNA prepared from whole blood. In conclusion, there were associations between rs12229654, rs2074356 and rs11066280 of HECTD4 and alcohol intake. In addition, HECTD4 plays a role in lowering the incidence of CVD in Drinkers and Never Drinkers. Keyword: alcohol consumption, cardiovascular disease (CVD), HECTD4, DNA methylation