Association between NPPA promoter methylation and hypertension: results from Gusu cohort and replication in an independent sample

Abstract

Background: Atrial natriuretic peptide (ANP), one of the main members of the natriuretic peptides system, has been associated with hypertension and related complications, but the underlying molecular mechanisms are not very clear. Here, we aimed to examine whether DNA methylation, a molecular modification to the genome, of the natriuretic peptide A gene (NPPA), the coding gene of ANP, was associated with hypertension. Methods: Peripheral blood DNA methylation of NPPA promoter was quantified by target bisulfite sequencing in 2498 community members (mean aged 53 years, 38% men) as a discovery sample and 1771 independent participants (mean aged 62 years, 54% men) as a replication sample. In both samples, we conducted a single CpG association analysis, followed by a gene-based association analysis, to examine the association between NPPA promoter methylation and hypertension, adjusting for age, sex, education level, cigarette smoking, alcohol consumption, obesity, fasting glucose, and lipids. Multiple testing was controlled by the false discovery rate approach. Results: Of the 9 CpG loci assayed, hypermethylation at 5 CpGs (CpG1, CpG3, CpG6, CpG8, and CpG9) was significantly associated with a lower odds of prevalent hypertension in the discovery sample, and one CpG methylation (CpG1 located at Chr1:11908353) was successfully replicated in the replication sample (OR = 0.82, 95%CI 0.74-0.91, q = 0.002) after adjusting for covariates and multiple testing. The gene-based analysis found that DNA methylation of the 9 CpGs at NPPA promoter as a whole was significantly associated with blood pressure and prevalent hypertension in both samples (all P < 0.05).

Serum Atrial Natriuretic Peptide, NPPA Promoter Methylation, and Cardiovascular Disease: A 10-year Follow-Up Study in Chinese Adults

“…DNA methylation levels in the promoter region of the NPPA gene were quantified by target bisulfite sequencing as previously described [ 31 ], using genomic DNA isolated from peripheral blood mononuclear cells. In brief, based on the genomic coordinates of the NPPA promoter in Genome Reference Consortium Human Build 37 (GRCh37), we carefully designed the primers to detect the maximum CpG loci within the CpG islands.…”

“…In brief, based on the genomic coordinates of the NPPA promoter in Genome Reference Consortium Human Build 37 (GRCh37), we carefully designed the primers to detect the maximum CpG loci within the CpG islands. The targeted sequence (Chr1: 11908117 – 11908380, reverse strand, relative to TSS: –540 bp to –277 bp) was illustrated in Supplementary Figure S1 and presented previously [ 31 ]. Following primer validation, genomic DNA was bisulfite-treated using the EZ DNA Methylation-Gold Kit (Zymo Research, Inc., CA, United States) according to the manufacturer’s protocol, which converts unmethylated cytosine into uracil and leaves methylated cytosine unchanged.…”

<b><i>NPPA</i></b> Promoter Hypomethylation Predicts Central Obesity Development: A Prospective Longitudinal Study in Chinese Adults

“…In brief, based on the genomic coordinates of the NPPA promoter in Genome Reference Consortium Human Build 37 (GRCh37), we carefully designed the primers to detect the maximum CpG loci within the CpG islands. The targeted sequence (Chr1:11908117–11908380, reverse strand, relative to TSS: −540 bp to −277 bp) was illustrated previously [31]. Following primer validation, genomic DNA was bisulfite-treated using the EZ DNA Methylation-Gold Kit (Zymo Research, Inc., Irvine, CA, USA) according to the manufacturer’s protocol, which converts unmethylated cytosine into uracil and leaves methylated cytosine unchanged.…”

Roles and Mechanisms of DNA Methylation in Vascular Aging and Related Diseases

“…In addition, a study surveyed the association between blood pressure and DNA methylation among Europeans, Hispanics, and African Americans and identified 14 relevant CpG sites ( Richard et al, 2017 ). Moreover, DNA methylation of the natriuretic peptide-A gene was decreased in patients diagnosed with hypertension among the Chinese community ( Li J. et al, 2020 ). As we all know, the renin–angiotensin–aldosterone system (RAAS) is vital to the occurrence and progression of hypertension ( Drummond et al, 2019 ).…”