Association between the Extents of Methylation in Subjects with Coronary Artery Disease By Pyrosequencing

OBJECTIVES: DNA methylation, one of the most stable forms of epigenetic modification is associated with the development and progression of CAD. Top 5 significant CpGs from Discovery phase by microarray were analysed by the gold standard, Pyrosequencing(PSQ). We aimed to standardise and associate the...

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Published inIndian journal of clinical biochemistry Vol. 36; no. S1; p. S47
Main Authors Banerjee, Shyamashree, Paradkar, Minal U, Ponde, Chandrashekhar K, Rajani, Rajesh M, Pillai, Sudhir, Ashavaid, Tester F
Format Journal Article
LanguageEnglish
Published Springer 24.05.2022
Subjects
Analysis
Coronary heart disease
DNA sequencing
Epigenetic inheritance
Histocompatibility antigens
HLA histocompatibility antigens
Methylation
Nucleotide sequencing
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Summary:OBJECTIVES: DNA methylation, one of the most stable forms of epigenetic modification is associated with the development and progression of CAD. Top 5 significant CpGs from Discovery phase by microarray were analysed by the gold standard, Pyrosequencing(PSQ). We aimed to standardise and associate the methylation% in an independent CAD cohort. METHODOLOGY: 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 MannWhitney U; Kruskal-Wallis' rank test and two way ANOVA by MedCalc(vl9.6). RESULTS: The methylation levels in HLA-DQA1 for cgl0217052-78.5(37-85) and 76.5(2484) and cg09411910-81(72.0 to 93.0) and 81.5(50.0 to 89.0) in cases and controls respectively. In HLA-DQB1-cg03344051, levels in cases were 28.88+9.41 and 30.36+9.37 in controls. For HLA-DRBl-cg07889003, in cases and controls-15.5(5.00-39.00) and 10.5(5.00 29.0); while in cg08269402-52(16-65) and 42.5(17-61) was seen respectively. No association was observed between methylation levels and their respective lipid profile. Significant difference was obtained in methylation% in double or triple vessel disease(DVD or TVD) as compared to single vessel disease(SVD) in 3 sites-cg03344051,cg07889003 and cg08269402. This suggests an increase in the extent of methylation with the increase in CAD severity. CONCLUSION: PSQ was standardised for analysis of methylation levels for 5 CpG sites. There was a marked increase in the extent of methylation in 3 CpG sites in DVD/TVD cases as compared to SVD cases. A novel site, cg07889003 identified in our Discovery phase has shown association with the severity of CAD. Keywords: Epigenetics, CAD, Methylation, Pyrosequencing, HLA
ISSN:0970-1915