Discovery and interrogation of genetic regulatory variation impacting Atrial Fibrillation risk

摘要 这一多主体调查员提案的总体目标是促进从隐含的 在广泛的房颤全基因组关联研究中发现的遗传变异 房颤风险的分子机制。我们假设一条新的基因组和分析管道 询问遗传变异的调控功能将确定候选致病变异体及其目标 基因，使得从简单的关联到导致心律失常的机制转变。在……里面 初步研究，我们已经应用了新的单细胞方法来产生细胞类型分辨的高分辨率 染色体可及性图谱，并利用协调的基因组信号将房颤风险变异与 候选致病基因房颤。这些结果描述了一个高度互联的心脏基因调控网络。 心房基因的表达。在我们的第一个目标中，我们建议生成多模式单细胞基因组数据以提供 不同效果的更高分辨率注释。我们将改进我们的计算程序，以更好地利用 这些数据集用于房颤变异和基因发现。在我们的第二个目标中，我们将审问相互关联的 分子增强子分析中的基因调控网络与基因组染色质构象捕获 实验，以直接检查被提名的遗传变异的影响及其与 候选目标基因。在我们的第三个目标中，我们将研究高置信度变异SNPs在 深度，包括它们对顺式基因调控的影响，它们对人类心肌细胞电生理的影响， 以及它们对反式心肌细胞基因表达和染色质状态的影响。我们已经建立了一个 易于处理的策略，将有助于实现从房颤风险变量向分子机制的转变。我们 预计我们的方法将有助于将房颤遗传学的前景转化为有意义的生物学见解 并为任何系统中的遗传关联研究建立一个分子理解的范例。

Abstract The overall goal of this multi-principal investigator proposal is to facilitate the transition from implication of genetic variants identified in extensive genome wide association studies (GWAS) of Atrial Fibrillation (AF) to the molecular mechanisms underlying AF risk. We hypothesize that a novel genomic and analytic pipeline interrogating regulatory function of genetic variation will identify candidate causative variants and their target genes, enabling the transition from simple associations to causative mechanisms for the arrhythmia. In preliminary studies, we have applied novel single cell approaches to generate cell-type-resolved high-resolution chromosome accessibility maps and taken advantage of coordinated genomic signals to link AF risk variants to candidate causative AF genes. The results describe a highly interconnected gene regulatory network for cardiac atrial gene expression. In our first aim we propose to generate multi-modal single-cell genomics data to provide higher-resolution annotation of variant effects. We will improve our computational procedure to better leverage these datasets for AF variant and gene discovery. In our second aim, we will interrogate the interconnected gene regulatory network in molecular enhancer assays and genomic chromatin conformation capture experiments, to directly examine the impact of nominated genetic variants and their physical association with candidate target genes. In our third aim, we will examine the functionality of high confidence variant SNPs in depth, including their impact on gene regulation in cis, their impact on human cardiomyocyte electrophysiology, and their impact on cardiomyocyte gene expression and chromatin status in trans. We have established a tractable strategy that will help enable the transition from AF risk variants to molecular mechanisms. We anticipate that our approach will help translate the promise of AF genetics into meaningful biological insights for AF and establish a paradigm for the molecular understanding of genetic association studies in any system.