Abstract：
Hypertension is a crucial risk factor for developing cardiovascular disease and reducing life expectancy. We aimed to detect DNA methylation (DNAm) variants potentially related to systolic blood pressure (SBP) and diastolic blood pressure (DBP) by conducting epigenome-wide association studies in 60 and 59 Chinese monozygotic twin pairs, respectively.
Genome-wide DNA methylation profiling in whole blood of twins was performed using Reduced Representation Bisulfite Sequencing, yielding 551,447 raw CpGs. Association between DNAm of single CpG and blood pressure was tested by applying generalized estimation equation. Differentially methylated regions (DMRs) were identified by comb-P approach. Inference about Causation through Examination of Familial Confounding was utilized to perform the causal inference. Ontology enrichment analysis was performed using Genomic Regions Enrichment of Annotations Tool. Candidate CpGs were quantified using Sequenom MassARRAY platform in a community population. Weighted gene co-expression network analysis (WGCNA) was conducted using gene expression data.
The median age of twins was 52 years (95% range 40, 66). For SBP, 31 top CpGs (p < 1 × 10-4) and 8 DMRs were identified, with several DMRs within NFATC1, CADM2, IRX1, COL5A1, and LRAT. For DBP, 43 top CpGs (p < 1 × 10-4) and 12 DMRs were identified, with several DMRs within WNT3A, CNOT10, and DAB2IP. Important pathways, such as Notch signaling pathway, p53 pathway by glucose deprivation, and Wnt signaling pathway, were significantly enriched for SBP and DBP. Causal inference analysis suggested that DNAm at top CpGs within NDE1, MYH11, SRRM1P2, and SMPD4 influenced SBP, while SBP influenced DNAm at CpGs within TNK2. DNAm at top CpGs within WNT3A influenced DBP, while DBP influenced DNAm at CpGs within GNA14. Three CpGs mapped to WNT3A and one CpG mapped to COL5A1 were validated in a community population, with a hypermethylated and hypomethylated direction in hypertension cases, respectively. Gene expression analysis by WGCNA further identified some common genes and enrichment terms.
We detect many DNAm variants that may be associated with blood pressure in whole blood, particularly the loci within WNT3A and COL5A1. Our findings provide new clues to the epigenetic modification underlying hypertension pathogenesis.
Genome-wide DNA methylation profiling in whole blood of twins was performed using Reduced Representation Bisulfite Sequencing, yielding 551,447 raw CpGs. Association between DNAm of single CpG and blood pressure was tested by applying generalized estimation equation. Differentially methylated regions (DMRs) were identified by comb-P approach. Inference about Causation through Examination of Familial Confounding was utilized to perform the causal inference. Ontology enrichment analysis was performed using Genomic Regions Enrichment of Annotations Tool. Candidate CpGs were quantified using Sequenom MassARRAY platform in a community population. Weighted gene co-expression network analysis (WGCNA) was conducted using gene expression data.
The median age of twins was 52 years (95% range 40, 66). For SBP, 31 top CpGs (p < 1 × 10-4) and 8 DMRs were identified, with several DMRs within NFATC1, CADM2, IRX1, COL5A1, and LRAT. For DBP, 43 top CpGs (p < 1 × 10-4) and 12 DMRs were identified, with several DMRs within WNT3A, CNOT10, and DAB2IP. Important pathways, such as Notch signaling pathway, p53 pathway by glucose deprivation, and Wnt signaling pathway, were significantly enriched for SBP and DBP. Causal inference analysis suggested that DNAm at top CpGs within NDE1, MYH11, SRRM1P2, and SMPD4 influenced SBP, while SBP influenced DNAm at CpGs within TNK2. DNAm at top CpGs within WNT3A influenced DBP, while DBP influenced DNAm at CpGs within GNA14. Three CpGs mapped to WNT3A and one CpG mapped to COL5A1 were validated in a community population, with a hypermethylated and hypomethylated direction in hypertension cases, respectively. Gene expression analysis by WGCNA further identified some common genes and enrichment terms.
We detect many DNAm variants that may be associated with blood pressure in whole blood, particularly the loci within WNT3A and COL5A1. Our findings provide new clues to the epigenetic modification underlying hypertension pathogenesis.
摘要：
背景：高血压是发生心血管疾病和降低预期寿命的重要危险因素。我们旨在通过在60和59对中国单卵双胞胎中进行全基因组关联研究来检测与收缩压（SBP）和舒张压（DBP）潜在相关的DNA甲基化（DNAm）变异。分别。
方法：使用还原代表亚硫酸氢盐测序进行双胞胎全血全基因组DNA甲基化分析，产生551,447个原始CpG。通过应用广义估计方程测试了单个CpG的DNAm与血压之间的关联。通过comb-P方法鉴定差异甲基化区域(DMRs)。通过家族混淆检查推断因果关系被用来进行因果推断。使用基因组区域富集注释工具进行本体富集分析。在社区人群中使用SequenomMassARRAY平台对候选CpG进行定量。使用基因表达数据进行加权基因共表达网络分析(WGCNA)。
结果：双胞胎的中位年龄为52岁（95％范围为40、66）。对于SBP，确定了31个顶部CpG（p<1×10-4）和8个DMRs，NFATC1、CADM2、IRX1、COL5A1和LRAT内有多个DMR。对于DBP，确定了43个顶部CpG（p<1×10-4）和12个DMRs，WNT3A中有几个DMRs，CNOT10和DAB2IP。重要途径，如Notch信号通路，p53途径通过葡萄糖剥夺，和Wnt信号通路，显著富集了SBP和DBP。因果推断分析表明，NDE1，MYH11，SRRM1P2和SMPD4中最高CpG处的DNAm影响了SBP，而SBP影响TNK2内CpG的DNAm。WNT3A内最高CpG处的DNAm影响了DBP，而DBP影响GNA14内CpG处的DNAm。在社区人口中验证了映射到WNT3A的三个CpG和映射到COL5A1的一个CpG，在高血压病例中具有高甲基化和低甲基化的方向，分别。通过WGCNA的基因表达分析进一步鉴定了一些常见的基因和富集术语。
结论：我们在全血中检测到许多可能与血压有关的DNAm变异，特别是WNT3A和COL5A1内的基因座。我们的发现为高血压发病机制的表观遗传修饰提供了新的线索。
方法：使用还原代表亚硫酸氢盐测序进行双胞胎全血全基因组DNA甲基化分析，产生551,447个原始CpG。通过应用广义估计方程测试了单个CpG的DNAm与血压之间的关联。通过comb-P方法鉴定差异甲基化区域(DMRs)。通过家族混淆检查推断因果关系被用来进行因果推断。使用基因组区域富集注释工具进行本体富集分析。在社区人群中使用SequenomMassARRAY平台对候选CpG进行定量。使用基因表达数据进行加权基因共表达网络分析(WGCNA)。
结果：双胞胎的中位年龄为52岁（95％范围为40、66）。对于SBP，确定了31个顶部CpG（p<1×10-4）和8个DMRs，NFATC1、CADM2、IRX1、COL5A1和LRAT内有多个DMR。对于DBP，确定了43个顶部CpG（p<1×10-4）和12个DMRs，WNT3A中有几个DMRs，CNOT10和DAB2IP。重要途径，如Notch信号通路，p53途径通过葡萄糖剥夺，和Wnt信号通路，显著富集了SBP和DBP。因果推断分析表明，NDE1，MYH11，SRRM1P2和SMPD4中最高CpG处的DNAm影响了SBP，而SBP影响TNK2内CpG的DNAm。WNT3A内最高CpG处的DNAm影响了DBP，而DBP影响GNA14内CpG处的DNAm。在社区人口中验证了映射到WNT3A的三个CpG和映射到COL5A1的一个CpG，在高血压病例中具有高甲基化和低甲基化的方向，分别。通过WGCNA的基因表达分析进一步鉴定了一些常见的基因和富集术语。
结论：我们在全血中检测到许多可能与血压有关的DNAm变异，特别是WNT3A和COL5A1内的基因座。我们的发现为高血压发病机制的表观遗传修饰提供了新的线索。
