Discovery of Novel Epigenetic Regulators of Heart Failure in a Panel of Mice

项目总结 心力衰竭(HF)是研究人员日益关注的问题，预计到2019年，心力衰竭的发病率将增加25%。 2030年。心力衰竭是一种极其复杂的疾病，具有许多病理特征，包括心肌细胞 肥大、收缩功能障碍和纤维化重塑。心衰病因复杂，有共同风险 高血压和糖尿病等因素本身就是多因素的。因此，有一个 人类群体在疾病发生和发展过程中的巨大异质性掩盖了 常见形式的心衰有很强的遗传成分。由于这种异质性，人类 全基因组关联研究只能恢复少数几个重要的基因座。最近，私家侦探 小组描述了一个小鼠群体，在其中发现了30多个与心力衰竭相关的表型基因座，并且 在人类中，显著重叠(50%)与显著的或暗示的心力衰竭相关基因座。 这项建议概述了这组小鼠的扩展，以探索心脏的表观基因组 儿茶酚胺刺激前后。利用新的实验技术和计算工具， 该提案旨在确定控制DNA甲基化的重要基因和途径。它还寻求与 DNA甲基化与心力衰竭相关表型的变化以及勾勒出广泛的职业发展 计划让劳博士在王博士的指导下完成培训，并过渡到独立的 通过建立心血管遗传学和心血管遗传学的多学科研究计划获得学术地位 基因组学。 在该奖项的K99阶段，劳博士将分析88个品系的小鼠和 儿茶酚胺刺激前后用还原代表性亚硫酸氢盐测序。研究将会 重点使用甲基化和表型数据来确定表观基因组范围内的CpG-表型关联 关联研究基因座。将检查基因-甲基化关联，以确定DNA所在的基因座 突变导致整个基因组中DNA甲基化的巨大差异。通过将这些基因座与 之前在这个小组中收集的大量数据，劳博士将预测因果基因和途径 与心力衰竭有关。根据初步结果，在K99奖项的部分期间，PI还将在 利用CRISPR/Cas9基因敲除系统对两个高置信度候选基因进行体内功能研究： MOSPD3，它调节心脏重量和Serpina3n，它参与了1800多个CPGS的调节。 在R00部分，PI将把在K99部分确定的基因与他在体内的训练结合起来 确认导致心力衰竭的新基因和途径。 拟议研究的总体目标是将系统生物学、表观遗传学和分子生物学结合起来。 分析有助于加深对调控DNA甲基化和HF的遗传途径的理解 相关表型。

PROJECT SUMMARY Heart failure (HF) is a growing concern among researchers, with rates expected to increase by 25% by 2030. HF is an incredibly complex disease with many pathological features including cardiomyocyte hypertrophy, contractile dysfunction and fibrotic remodeling. HF has complex etiologies, with common risk factors such as hypertension and diabetes being in and of themselves multi-factorial. Consequently, there is a tremendous amount of heterogeneity in human populations in both disease onset and progression which mask the demonstrated strong genetic component of common forms of HF. As a result of this heterogeneity, human genome-wide association studies have only been able to recover a handful of significant loci. Recently, the PI's group described a population of mice in which over 30 loci for HF-related phenotypes were identified and which demonstrated significant overlap (50%) with significant or suggestive HF-associated loci in humans. This proposal outlines an extension of this panel of mice to explore the epigenome of the heart both before and after catecholamine stimulation. Using novel experimental techniques and computational tools, the proposal seeks to identify important genes and pathways which control DNA methylation. It also seeks to connect DNA methylation to changes in HF-related phenotypes as well as outlining an extensive career development plan for Dr. Rau to complete his training under the mentorship of Dr. Wang and transition to an independent academic position by establishing a multi-disciplinary research program in cardiovascular genetics and genomics. During the K99 phase of this award, Dr. Rau will analyze the methylomes of 88 strains of mice both before and after catecholamine stimulation using reduced representation bisulfite sequencing. Research will focus on the use of methylation and phenotypic data to identify CpG-phenotype associations at epigenome-wide association study loci. Genotype-methylation associations will be examined to identify loci within which DNA mutations drive large differences in DNA methylation across the genome. By combining these loci with the significant amount of data previously gathered in this panel, Dr. Rau will predict causal genes and pathways implicated in HF. Based on preliminary results, during the K99 portion of the award the PI will also perform in vivo functional studies using the CRISPR/Cas9 gene knockout system of two high-confidence candidate genes: Mospd3, which regulates heart weight and Serpina3n, which is implicated in the regulation of over 1800 CpGs. During the R00 portion, the PI will combine the genes identified during the K99 portion with his training in in vivo validation to identify novel genes and pathways which contribute to heart failure. The overall goal of the proposed studies is to integrate systems biology, epigenetics and molecular analyses to lead to deeper understandings of the genetic pathways which regulate DNA methylation and HF- related phenotypes.