Targeted Sequencing Study of GWAS-implicated Blood Pressure Loci

项目摘要(研究项目1) 拟议研究的总体目标是识别新的基因和功能性遗传变异。 与血压(BP)相关的9个基因座在最近的亚洲人群中具有全基因组意义 遗传流行病学网络(AGEN)-BP全基因组关联研究(GWAS)使用NEXT的荟萃分析 世代排序技术。这项拟议的研究将在5000名汉族人中进行 亚洲心血管疾病国际合作研究(Interasia)的参与者。亚细亚 提供了关于BP相关表型的非凡资源以及已有的足够数量的DNA 储存在我们的实验室用于基因研究。在拟议的研究中，我们将发现新的功能变体 通过对300名Interasia参与者中9个Gwas相关基因座的连续基因组区域进行深度测序 平均动脉压(MAP)最高，300名参与者没有服用降压药 最低的地图。这种不可知的测序方法将使我们能够用 潜在的重要但还不是很清楚的调控功能以及这些基因座上的所有15个已知基因， 它们的平均跨度分别为155千碱基。生物信息学工具将被用于过滤大量的 发现了变异，优先考虑那些与预测的功能相关的变异。为了最大限度地发挥统计力量，罕见的 变体将使用：1)不可知的滑动窗口方法进行分组，该方法将罕见的变体聚集在 整个基因座上相邻和重叠的片段；以及2)基于生物学的方法，聚合 已知功能单位(如保守区域或基因)的罕见变异。我们将使用最先进的统计方法 方法在聚合分析中检验稀有变异的集体效应。新颖的低频和 将使用传统的单标记分析分别检查常见的变种。我们将对这些基因进行分型 在剩下的4,400名Interasia参与者中选择200个最有希望的新变种，并测试这种关联 在5,000名Interasia参与者(4,400个基因分型600个测序)中，每个变异与BP之间存在差异。 我们将在10,000名汉族人的独立随机样本中复制25个最有希望的变体 中国参赛者。此外，我们还将利用现有的GWAS、全外显子组和表型数据 在基因型和表型数据库中评估25名最有希望的人的跨种族相关性 在基于人群的大样本中，多达15,076名非洲裔美国人和30,821名欧洲人- 美国参赛者。然后将进行特定于祖先的和总体的荟萃分析。这些发现 可能具有重要的临床和公共卫生影响。通过帮助阐明生物途径 在BP调控的基础上，这些发现可能被用来开发新的基于基因的策略，用于 预防和治疗高血压。此外，拟议的科布雷少年所获得的经验 学院调查员应该是她作为独立研究员取得成功的重要踏脚石。

Project Summary (Research Project 1) The overall objective of the proposed study is to identify novel genes and functional genetic variants associated with blood pressure (BP) at 9 loci which attained genome-wide significance in the recent Asian Genetic Epidemiology Network (AGEN)-BP genome-wide association study (GWAS) meta-analysis using next generation sequencing technology. The proposed study will be carried out among 5,000 Han Chinese participants of the International Collaborative Study of Cardiovascular Disease in Asia (InterASIA). InterASIA provides an extraordinary resource on BP-related phenotypes as well as sufficient quantities of DNA already stored at our laboratories for genetic research. In the proposed study, we will discover novel functional variants by deep sequencing contiguous genomic regions of 9 GWAS-implicated loci among 300 InterASIA participants with the highest mean arterial pressure (MAP) and 300 participants not taking antihypertension medication with the lowest MAP. This agnostic sequencing approach will allow us to examine intergenic variants with potentially important but not well-understood regulatory functions as well as all 15 known genes at these loci, which span an average of 155 kilobases each. Bioinformatic tools will be used to filter the large number of discovered variants, prioritizing those with predicted functional relevance. To maximize statistical power, rare variants will be grouped using: 1) an agnostic sliding window approach which aggregates rare variants in adjacent and overlapping segments across the entire loci; and 2) a biology-based approach which aggregates rare variants by known functional units (e.g. conserved region or gene). We will use state-of-the-art statistical methods to examine the collective effects of rare variants in aggregate analyses. Novel low-frequency and common variants will be examined separately using traditional single-marker analyses. We will genotype the 200 most promising novel variants among the remaining 4,400 InterASIA participants and test the association between each variant and BP among the 5,000 (4,400 genotyped + 600 sequenced) InterASIA participants. We will replicate each of the 25 most promising variants in an independent random sample of 10,000 Han Chinese participants. In addition, we will leverage existing GWAS, whole-exome, and phenotype data available in the database of Genotypes and Phenotypes to assess the trans-ethnic relevance of the 25 most promising BP variants within large, population-based samples of up to 15,076 African-American and 30,821 European- American participants. Ancestry-specific and overall meta-analyses will then be carried out. These findings may have important clinical and public health implications. By helping to elucidate the biological pathways underlying BP regulation, these findings may be used to develop novel gene-based strategies for the prevention and treatment of hypertension. Furthermore, the experience gained by the proposed COBRE junior faculty investigator should serve as an important stepping stone to her success as an independent researcher.