Research Laboratories, P.D Hinduja Hospital & Medical Research Centre, Mumbai, India.
Department of Biochemistry, P.D Hinduja Hospital & Medical Research Centre, Mumbai, India.
Mol Biol Rep. 2024 Jan 22;51(1):173. doi: 10.1007/s11033-023-08974-5.
DNA methylation, one of the most stable forms of epigenetic modification is associated with the development and progression of coronary artery disease (CAD). Our previously reported study on epigenome-wide microarray analysis showed significantly methylated CpG sites. Top 5 significant CpGs from HLA gene were selected and analysed by Pyrosequencing (PSQ) to determine their association with severity of CAD.
Blood samples of 50-age matched angiographically CAD positive male cases with 50 angiographically CAD negative male controls were subjected to lipid profile estimation and PSQ for methylation level analysis. Findings and subgroup analysis were evaluated by Mann-Whitney U; Kruskal-Wallis' rank test and two-way ANOVA by MedCalc (v19.6).
Methylation levels in HLA-DQA1 for cg10217052 was 78.5 (37-85) and 76.5 (24-84); cg09411910 was 81 (72.0 to 93.0) and 81.5 (50.0 to 89.0) in cases and controls respectively. Levels in HLA-DQB1-cg03344051, were 28.88 + 9.41 for cases and 30.36 + 9.37 in controls. For HLA-DRB1-cg07889003, levels in cases and controls were 15.5 (5.00-39.00) and 10.5 (5.00-29.0); while in cg08269402 were 52 (16-65) and 42.5 (17-61) respectively. No association was observed between methylation levels and lipid profile. cg03344051, cg07889003 and cg08269402 were significantly differentiated in double or triple vessel disease (DVD or TVD) as compared to single vessel disease (SVD) suggesting an increase in the extent of methylation with the increase in CAD severity.
The present study shows significant increase in the extent of methylation in 3 CpG sites in DVD/TVD cases as compared to SVD cases. Additionally, a novel site, cg07889003 identified in our discovery phase has shown association with the severity of CAD.
DNA 甲基化是最稳定的表观遗传修饰形式之一，与冠状动脉疾病（CAD）的发展和进展有关。我们之前的全基因组甲基化微阵列分析研究显示出明显甲基化的 CpG 位点。从 HLA 基因中选择前 5 个显著 CpG 并通过焦磷酸测序（PSQ）进行分析，以确定它们与 CAD 严重程度的关系。
对 50 名年龄匹配的经血管造影 CAD 阳性男性病例和 50 名经血管造影 CAD 阴性男性对照的血液样本进行血脂谱估计和 PSQ 甲基化水平分析。采用 Mann-Whitney U 检验、Kruskal-Wallis 秩检验和 MedCalc（v19.6）的双向方差分析评估结果和亚组分析。
HLA-DQA1 中 cg10217052 的甲基化水平为 78.5（37-85）和 76.5（24-84）；cg09411910 为 81（72.0 至 93.0）和 81.5（50.0 至 89.0），病例组和对照组分别为 81（72.0 至 93.0）和 81.5（50.0 至 89.0）。HLA-DQB1-cg03344051 的水平为 28.88+9.41 为病例组，30.36+9.37 为对照组。对于 HLA-DRB1-cg07889003，病例组和对照组的水平分别为 15.5（5.00-39.00）和 10.5（5.00-29.0）；而 cg08269402 则分别为 52（16-65）和 42.5（17-61）。未观察到甲基化水平与血脂谱之间存在相关性。与单支血管疾病（SVD）相比，cg03344051、cg07889003 和 cg08269402 在双支或三支血管疾病（DVD 或 TVD）中差异明显，表明随着 CAD 严重程度的增加，甲基化程度也增加。
本研究显示，在 DVD/TVD 病例中，3 个 CpG 位点的甲基化程度明显增加，而在 SVD 病例中则没有。此外，在我们的发现阶段确定的一个新位点 cg07889003 与 CAD 的严重程度有关。
