Functional evaluation of GWAS loci for cardiovascular intermediate phenotypes

心血管中间表型的 GWAS 位点的功能评估

• 批准号: 7713461
• 负责人: Calum A. MacRae
• 金额: $ 54.44万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7713461
• 关键词: Alleles; Antihypertensive Agents; Biological; Biological Assay; Biological Models; Biology; Blood Vessels; Cardiac; Cardiovascular Diseases; Cardiovascular system; Complement; Complex; Data; Data Set; Disease; Disease model; Drug Interactions; Environment; Evaluation; Exposure to; Foundations; Framingham Heart Study; Genes; Genetic; Gonadal Steroid Hormones; Heritability; Heterogeneity; Human; Human Genetics; Human Genome; Human Genome Project; Hypertrophy; Investigation; Left; Left Ventricular Hypertrophy; Left Ventricular Mass; Lipids; Low-Density Lipoproteins; Methods; Modeling; Molecular Profiling; Myocardial; Organ; Pathology; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phenotype; Physiology; Production; Reporter; Resolution; Series; Signal Transduction; Structure; System; Testing; Tissues; Vascular calcification; Ventricular; Whole Organism; Work; Zebrafish; base; calcification; clinical phenotype; disease phenotype; gene environment interaction; gene function; gene interaction; genetic association; genetic manipulation; genome wide association study; genome-wide; human data; human disease; in vivo; in vivo Model; indexing; insight; interest; loss of function; metabolomics; multidisciplinary; novel; novel strategies; public health relevance; research study; trait

DESCRIPTION (provided by applicant): The simultaneous identification of multiple novel disease loci in genome wide association studies (GWAS) has highlighted a need for efficient model systems both to define the causal gene(s) at each locus, and to explore disease mechanisms 2, 3. Many GWAS have explained only a small proportion of the heritability for the traits of interest, and for some key disease phenotypes no loci have been identified2-4. There is a clear need for rapid in vivo approaches to vertebrate gene function, but also an imperative to begin to identify intermediate phenotypes more proximate in the disease causal pathways11-13. We have taken an integrative approach combining quantitative intermediate phenotypes for major cardiovascular traits, specifically; left ventricular LV hypertrophy, arterial calcification scores and key metabolites, from the Framingham Heart Study (FHS) with cognate validated functional assays in the intact zebrafish14-20. We will explore the use of high throughput zebrafish genetics for gene identification at GWAS loci, for prioritization of loci of borderline significance, and for investigation of gene-gene and gene-drug as well phenotypic interactions in the following Specific Aims: Aim 1 Identify the causal genes at GWAS loci for cardiovascular traits in FHS using in vivo modeling in the zebrafish: All genes at each GWAS locus (within 500kb of the original genetic signal) for three core phenotypes in the FHS will be tested using a range of multiple cognate assays in the zebrafish and loss of function and over-expression alleles for each gene. The human phenotypes and their zebrafish counterparts are; LV mass: in vivo myocardial mass, myocardial hypertrophy reporters and LVH expression profiles;Arterial calcification indices: in vivo vascular structure and physiology, aortic LDL transport and NO production, Wnt and Id reporters of vascular calcification pathways; and Metabolomics: direct comparisons of human and zebrafish metabolites. These experiments will define the major effect gene(s) at many of the loci for each phenotype, the direction of biologic effect and potential relations across shared intermediate phenotypes 11,12, 22. Aim 2 Defining gene-gene and gene-environment interactions: Exploiting the large effect sizes and the scalability of the zebrafish, we will test gene-gene and gene-environment (lipid lowering or antihypertensive drugs, and sex hormones) interactions that are beyond the power of the available human datasets. The results of quantitative zebrafish analyses will then be directly tested in the original human data. This application will rigorously test the utility of combining GWAS data with in vivo zebrafish biology for the prioritization and functional evaluation of genes within loci. The work will allow the exploration of gene-gene and gene-drug interactions. We also anticipate that the multi-system exploration of the effects of genetic manipulations in the zebrafish will also inform our understanding of clinical phenotypes. Finally, these studies will lay the foundation for exploration of the gene networks underlying common cardiac and vascular phenotypes, and establish high-throughput biology in the zebrafish as a platform to complement GWAS. PUBLIC HEALTH RELEVANCE: With the completion of the Human Genome Project there has been a wave of genetic studies, generating large numbers of new potential disease genes. Understanding how these genes work, alone, together or in concert with the environment, to cause each disease requires new approaches which allow the study of complex effects on different tissues or organs, but offer sufficient throughput to deal with literally hundreds of genes. The zebrafish can model many human diseases, and in this application a multidisciplinary team will integrate insights from zebrafish biology and the Framingham Heart Study to explore the genetics of major cardiovascular diseases.

描述（由申请人提供）：在基因组广泛关联研究（GWAS）中同时鉴定多个新型疾病基因座的表明，需要在每个基因座定义有效的模型系统来定义因果基因（s），探索疾病机制2，3。许多GWS仅解释了对遗传性的一小部分，而不是某些特征的遗传性，并且是某些特征的比例，并且是某些特征的特征。显然需要快速的体内方法来实现脊椎动物基因功能，但也必须开始在疾病因果途径中识别中间表型更接近的中间表型11-13。我们采取了一种综合方法，将主要心血管特征的定量中间表型结合在一起，特别是。左心室LV肥大，动脉钙化评分和关键代谢产物，来自Framingham心脏研究（FHS），在完整的斑马鱼114-20中具有认知验证的功能测定。 We will explore the use of high throughput zebrafish genetics for gene identification at GWAS loci, for prioritization of loci of borderline significance, and for investigation of gene-gene and gene-drug as well phenotypic interactions in the following Specific Aims: Aim 1 Identify the causal genes at GWAS loci for cardiovascular traits in FHS using in vivo modeling in the zebrafish: All FHS中三种核心表型的每个GWAS基因座的基因（在原始遗传信号的500kb之内）将使用斑马鱼中的一系列多个同源测定以及每个基因的功能和过表达等位基因的损失进行测试。人类的表型及其斑马鱼是； LV质量：体内心肌肿块，心肌肥大报告和LVH表达谱；动脉钙化指标：体内血管结构和生理学，主动脉LDL转运，无产生，Wnt和ID的血管钙化途径的记者；和代谢组学：人类和斑马鱼代谢产物的直接比较。这些实验将定义每个表型的许多基因型的主要效应基因，生物学效应的方向以及共享中间表型的潜在关系11,12，22。目标2定义基因基因和基因 - 环境相互作用：利用Zeebrafish和Gene-Gene和Gene-Gene的较大效应和延伸性（我们）降压药物和性激素）相互作用超出了可用人类数据集的功能。然后，定量斑马鱼分析的结果将直接在原始人类数据中进行测试。该应用程序将严格测试将GWAS数据与体内斑马鱼生物学相结合的实用性，以便对基因座内基因的优先级和功能评估。这项工作将允许探索基因基因和基因 - 毒品相互作用。我们还预计，对斑马鱼中遗传操纵影响的多系统探索也将为我们对临床表型的理解提供信息。最后，这些研究将为探索共同心脏和血管表型的基因网络奠定基础，并在斑马鱼中建立高通量生物学，作为补充GWAS的平台。公共卫生相关性：随着人类基因组项目的完成，遗传研究已经产生了大量新的潜在疾病基因。了解这些基因如何单独，与环境共同起作用，以引起每种疾病，需要新的方法来研究对不同组织或器官的复杂作用，但提供了足够的吞吐量来处理几百个基因。斑马鱼可以对许多人类疾病进行建模，在此应用中，多学科团队将整合斑马鱼生物学和弗雷明汉心脏研究的见解，以探索主要心血管疾病的遗传学。