Yamada Yoshiji, Yasukochi Yoshiki, Kato Kimihiko, Oguri Mitsutoshi, Horibe Hideki, Fujimaki Tetsuo, Takeuchi Ichiro, Sakuma Jun
Department of Human Functional Genomics, Advanced Science Research Promotion Center, Mie University, Tsu, Mie 514-8507, Japan.
CREST, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan.
Biomed Rep. 2018 Nov;9(5):383-404. doi: 10.3892/br.2018.1152. Epub 2018 Sep 17.
Early-onset coronary artery disease (CAD) has a strong genetic component. Although genome-wide association studies have identified various genes and loci significantly associated with CAD mainly in European populations, genetic variants that contribute toward susceptibility to this condition in Japanese patients remain to be definitively identified. In the present study, exome-wide association studies (EWASs) were performed to identify genetic variants that confer susceptibility to early-onset CAD in Japanese. A total of 7,256 individuals aged ≤65 years were enrolled in the present study. EWAS were conducted on 1,482 patients with CAD and 5,774 healthy controls. Genotyping of single nucleotide polymorphisms (SNPs) was performed using Illumina Human Exome-12 DNA Analysis BeadChip or Infinium Exome-24 BeadChip arrays. The association between allele frequencies for 31,465 SNPs that passed quality control and CAD was examined using Fisher's exact test. To compensate for multiple comparisons of allele frequencies with CAD, a false discovery rate (FDR) of <0.05 was applied for statistically significant associations. The association between allele frequencies for 31,465 SNPs and CAD, as determined by Fisher's exact test, demonstrated that 170 SNPs were significantly (FDR <0.05) associated with CAD. Multivariable logistic regression analysis with adjustment for age, sex, and the prevalence of hypertension, diabetes mellitus and dyslipidemia revealed that 162 SNPs were significantly (P<0.05) associated with CAD. A stepwise forward selection procedure was performed to examine the effects of genotypes for the 162 SNPs on CAD. The 54 SNPs were significant (P<0.05) and independent [coefficient of determination (R), 0.0008 to 0.0297] determinants of CAD. These SNPs together accounted for 15.5% of the cause of CAD. Following examination of results from previous genome-wide association studies and linkage disequilibrium of the identified SNPs, 21 genes ( and ) and five chromosomal regions (2p13, 4q31.2, 5q12, 13q34 and 20q13.2) that were significantly associated with CAD were newly identified in the present study. Gene ontology analysis demonstrated that various biological functions were predicted in the 18 genes identified in the present study. The network analysis revealed that the 18 genes had potential direct or indirect interactions with the 30 genes previously revealed to be associated with CAD or with the 228 genes identified in previous genome-wide association studies. The present study newly identified 26 loci that confer susceptibility to CAD. Determination of genotypes for the SNPs at these loci may prove informative for assessment of the genetic risk for CAD in Japanese patients.
早发性冠状动脉疾病（CAD）具有很强的遗传因素。尽管全基因组关联研究已经在欧洲人群中确定了与CAD显著相关的各种基因和基因座，但在日本患者中导致这种疾病易感性的基因变异仍有待明确鉴定。在本研究中，进行了全外显子组关联研究（EWAS）以确定在日本人中赋予早发性CAD易感性的基因变异。本研究共纳入了7256名年龄≤65岁的个体。对1482例CAD患者和5774名健康对照进行了EWAS。使用Illumina Human Exome - 12 DNA分析微珠芯片或Infinium Exome - 24微珠芯片阵列对单核苷酸多态性（SNP）进行基因分型。使用Fisher精确检验检查了31465个通过质量控制的SNP的等位基因频率与CAD之间的关联。为了补偿等位基因频率与CAD的多重比较，对具有统计学显著关联的应用了<0.05的错误发现率（FDR）。由Fisher精确检验确定的31465个SNP的等位基因频率与CAD之间的关联表明，170个SNP与CAD显著相关（FDR <0.05）。对年龄、性别以及高血压、糖尿病和血脂异常患病率进行调整的多变量逻辑回归分析显示，162个SNP与CAD显著相关（P<0.05）。进行了逐步向前选择程序以检查这162个SNP的基因型对CAD的影响。54个SNP是CAD的显著（P<0.05）且独立的[决定系数（R），0.0008至0.0297]决定因素。这些SNP共同占CAD病因的15.5%。在检查先前全基因组关联研究的结果以及所鉴定SNP的连锁不平衡后，本研究新鉴定出21个基因（和）以及五个染色体区域（2p13、4q31.2、5q12、13q34和20q13.2）与CAD显著相关。基因本体分析表明，在本研究中鉴定出的18个基因预测了各种生物学功能。网络分析显示，这18个基因与先前揭示的与CAD相关的30个基因或先前全基因组关联研究中鉴定的22个基因具有潜在的直接或间接相互作用。本研究新鉴定出26个赋予CAD易感性的基因座。确定这些基因座处SNP的基因型可能对评估日本患者CAD的遗传风险具有参考价值。
