Identifying functional variants in COPD GWAS loci

项目摘要 识别COPD等复杂疾病的新易感基因可能会改变我们的 了解疾病的病理生理学，为治疗提供新的靶点。这个项目的主要目标是 项目是识别五个已建立的COPD Gwas基因座中的功能性遗传变异；识别 受这些功能变异影响的关键基因；并评估这些关键基因和 COPD发病机制中的功能遗传变异。在最近通过国际合作的全球气候变化网络 COPD遗传学联合会，我们发现了22个全基因组意义上与COPD相关的基因组座位。 在这项提案中，我们将把我们的研究重点放在五个COPD基因座上，这些基因座包括多个候选人 基因，其可能的功能变异尚未确定。我们假设一个完整的 利用全基因组测序进行精细定位、基因表达数据、生物信息学的方法 方法和新的实验室评估将使关键基因和功能的识别成为可能 这五个COPD基因座上的变异。此外，我们假设，专注于基于细胞的研究 这些关键基因及其功能变异将为COPD的发病机制提供新的见解。致信地址 在这些假说中，我们将首先使用大规模平行的方法识别这五个基因座内的功能变异 记者分析了多种细胞类型，使用公共和最近生成的Omics数据的生物信息学方法， 和遗传关联分析。然后我们将确定哪些基因或哪些基因受到这些基因的影响 通过进行染色体构象捕获(4C-Seq)，确认调控的 功能变异体对相关基因内源性启动子的影响 利用CRISP-Cas9对调控区域进行基因编辑对相关基因表达的影响 接近了。最后，我们将使用细胞模型来确定关键基因失活的影响及其 与慢性阻塞性肺疾病相关的细胞死亡、凋亡和细胞激活读数的功能变体。要完成 这些目标，是一种独特的高度集成的方法，结合了遗传关联分析、分子 对调节元件的研究，以及基于细胞的功能分析已经开发出来，可能会提供 对COPD发病机制的重要见解。

Project Summary The identification of novel susceptibility genes for complex diseases like COPD could transform our understanding of disease pathophysiology and provide new targets for treatment. The primary goals of this project are to identify the functional genetic variants within five well-established COPD GWAS loci; to identify the key genes influenced by these functional variants; and to assess the impact of these key genes and functional genetic variants on COPD pathogenesis. In recent collaborative GWAS through the International COPD Genetics Consortium, we found 22 genomic loci associated with COPD at genome-wide significance. We will focus our studies in this proposal on five of these COPD GWAS loci that include multiple candidate genes and for which a likely functional variant has not been identified. We hypothesize that an integrated approach that utilizes whole genome sequencing for fine mapping, gene expression data, bioinformatic approaches, and new laboratory assessments will enable the identification of the key genes and functional variants within these five COPD GWAS loci. Moreover, we hypothesize that focused cell-based studies of these key genes and their functional variants will provide novel insights into COPD pathogenesis. To address these hypotheses, we will start by identifying functional variants within these five loci using massively parallel reporter assays in multiple cell types, bioinformatic approaches with public and recently generated Omics data, and genetic association analysis. We will then determine which gene or genes are influenced by these functional variants by performing chromosome conformation capture (4C-Seq), confirming the regulatory effects of the functional variants on the endogenous promoter of the implicated gene, and demonstrating effects on gene expression of the implicated gene by gene editing the regulatory region using CRISP-Cas9 approaches. Finally, we will use cellular models to determine the effects of inactivating the key gene and its functional variants on COPD-related read-outs of cell death, apoptosis, and cell activation. To accomplish these goals, a unique and highly integrated approach combining genetic association analysis, molecular studies of regulatory elements, and functional cell-based assays has been developed that will likely provide important insights into COPD pathogenesis.