Yamada Yoshiji, Kato Kimihiko, Oguri Mitsutoshi, Horibe Hideki, Fujimaki Tetsuo, Yasukochi Yoshiki, 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 Jul;9(1):8-20. doi: 10.3892/br.2018.1104. Epub 2018 May 29.
Given that substantial genetic components have been shown in ischemic stroke, intracerebral hemorrhage (ICH), and subarachnoid hemorrhage (SAH), heritability may be higher in early-onset than late-onset individuals with these conditions. Although genome-wide association studies (GWASs) have identified various genes and loci significantly associated with ischemic stroke, ICH, or intracranial aneurysm mainly in European ancestry populations, genetic variants that contribute to susceptibility to these disorders remain to be identified definitively. We performed exome-wide association studies (EWASs) to identify genetic variants that confer susceptibility to ischemic stroke, ICH, or SAH in early-onset subjects with these conditions. A total of 6,649 individuals aged ≤65 years were examined. For the EWAS of ischemic or hemorrhagic stroke, 6,224 individuals (450 subjects with ischemic stroke, 5,774 controls) or 6,179 individuals (261 subjects with ICH, 176 subjects with SAH, 5,742 controls), respectively, were examined. EWASs were performed with the use of Illumina Human Exome-12 v1.2 DNA Analysis BeadChip or Infinium Exome-24 v1.0 BeadChip. To compensate for multiple comparisons of allele frequencies with ischemic stroke, ICH, or SAH, we applied a false discovery rate (FDR) of <0.05 for statistical significance of association. The association of allele frequencies of 31,245 single nucleotide polymorphisms (SNPs) that passed quality control to ischemic stroke was examined with Fisher's exact test, and 31 SNPs were significantly (FDR <0.05) associated with ischemic stroke. The association of allele frequencies of 31,253 or 30,970 SNPs to ICH or SAH, respectively, was examined with Fisher's exact test, and six or two SNPs were significantly associated with ICH or SAH, respectively. Multivariable logistic regression analysis with adjustment for age, sex, and the prevalence of hypertension and diabetes mellitus revealed that 12 SNPs were significantly [P<0.0004 (0.05/124)] related to ischemic stroke. Similar analysis with adjustment for age, sex, and the prevalence of hypertension revealed that six or two SNPs were significantly [P<0.0016 (0.05/32)] related to ICH or SAH, respectively. After examination of linkage disequilibrium of identified SNPs and results of previous GWASs, we identified , and as susceptibility loci for ischemic stroke, and as susceptibility loci for ICH, and and as such loci for SAH. Therefore, to the best of our knowledge, we have newly identified nine genes that confer susceptibility to early-onset ischemic stroke, ICH, or SAH. Determination of genotypes for the SNPs in these genes may prove informative for assessment of the genetic risk for ischemic stroke, ICH, or SAH in Japanese.
鉴于缺血性中风、脑出血（ICH）和蛛网膜下腔出血（SAH）已显示出大量遗传成分，这些疾病的早发个体的遗传度可能高于晚发个体。尽管全基因组关联研究（GWAS）已在主要为欧洲血统人群中鉴定出与缺血性中风、ICH或颅内动脉瘤显著相关的各种基因和位点，但导致这些疾病易感性的遗传变异仍有待明确鉴定。我们进行了全外显子组关联研究（EWAS），以鉴定在患有这些疾病的早发个体中导致缺血性中风、ICH或SAH易感性的遗传变异。共检查了6649名年龄≤65岁的个体。对于缺血性或出血性中风的EWAS，分别检查了6224名个体（450名缺血性中风患者，5774名对照）或6179名个体（261名ICH患者，176名SAH患者，5742名对照）。使用Illumina Human Exome-12 v1.2 DNA分析芯片或Infinium Exome-24 v1.0芯片进行EWAS。为了补偿与缺血性中风、ICH或SAH的等位基因频率的多重比较，我们将错误发现率（FDR）应用于<0.05以确定关联的统计学显著性。使用Fisher精确检验检查了31245个通过质量控制的单核苷酸多态性（SNP）的等位基因频率与缺血性中风的关联，31个SNP与缺血性中风显著相关（FDR<0.05）。分别使用Fisher精确检验检查了31253个或30970个SNP与ICH或SAH的等位基因频率的关联，分别有6个或2个SNP与ICH或SAH显著相关。对年龄、性别以及高血压和糖尿病患病率进行调整的多变量逻辑回归分析显示，12个SNP与缺血性中风显著相关[P<0.0004（0.05/124）]。对年龄、性别以及高血压患病率进行调整的类似分析显示，分别有6个或2个SNP与ICH或SAH显著相关[P<0.0016（0.05/32）]。在检查已鉴定SNP的连锁不平衡和先前GWAS的结果后，我们鉴定出 、 作为缺血性中风的易感位点， 、 作为ICH的易感位点，以及 、 作为SAH的此类位点。因此，据我们所知，我们新鉴定出九个导致早发缺血性中风、ICH或SAH易感性的基因。确定这些基因中SNP的基因型可能对评估日本人患缺血性中风、ICH或SAH的遗传风险具有参考价值。
