Epigenomes and Epigenetic Mechanisms in BP-relevant Tissues

项目2项目总结 高血压仍然是多种心血管疾病最重要的危险因素 全球约54%的中风和47%的缺血性心脏病事件。 尽管我们知道有一种重要的遗传成分决定了血液的水平 压力(BP；对BP遗传率的估计在30%-70%之间)我们远不能理解无数 导致人类血压变化的遗传因素。最重大的挑战之一是 了解BP的遗传控制是指绝大多数与BP相关的单核苷酸多态性 在人类基因组广泛关联研究中已确定的(SNPs)位于非编码 DNA的区域。许多非编码SNP(NcSNPs)位于位于数千个碱基的单倍型区域 远离任何蛋白质编码基因的配对，使得几乎不可能将SNPs与蛋白质编码基因联系起来 或根据序列和我们目前对该机制的了解来调节BP的生理途径 为SNPS采取的行动。在这个项目中，我们将重点检验许多ncSNPs的具体假设 通过表观遗传机制影响BP相关基因的表达。表观遗传学 指的是DNA功能的稳定变化，而不是直接由于DNA序列的变化。 表观基因组学是指当它们发生在整个基因组时的这种变化。因此，我们将使用国家- 最先进的表观基因组学和表观遗传学方法，探索个体 NcSNPs影响人类GWABP相关基因的表达。使用肾单位节段 我们将从新鲜的人肾和从臀部活检中分离的人的阻力小动脉中进行显微解剖。 以及来自人类可诱导多能干细胞(IPSCs)的相关细胞类型，我们将 以前所未有的组织分辨率创建人类BP相关细胞和组织的表观基因组图谱。 将该地图与我们对人类GWASSNP数据和来自项目1的数据的生物信息学分析相结合 这个PPG(BP ncSNPs对基因表达的影响)，我们将测试特定的表观遗传机制 单个SNPs。最后，我们将进行验证研究，其中精确的基因组编辑和目标 表观遗传学分析被用来确定假设的机制是否对 BP相关细胞类型中的基因表达。该项目与其他项目和核心具有高度的协作性 在该计划中，通过共享试剂(细胞和组织)、数据传输和 调查人员及其实验室的智力环境。

PROJECT 2 PROJECT SUMMARY Hypertension remains the most important risk factor for a wide variety of cardiovascular diseases accounting for approximately 54 percent of all strokes and 47 percent of all ischemic heart disease events globally. Despite the fact that we know there is a significant genetic component that determines the level of blood pressure (BP; estimates of heritability of BP range from 30-70%) we are far from understanding the myriad of genetic factors that account for BP variation in humans. One of the most significant challenges for understanding genetic control of BP is that the vast majority of BP-associated single nucleotide polymorphisms (SNPs) that have been identified in human genome wide association studies (GWAS) are located in noncoding regions of DNA. Many of the noncoding SNPs (ncSNPs) are in haplotype regions located thousands of base pairs away from any protein-coding gene making it nearly impossible to link the SNPs to a protein-coding gene or a physiological pathway that regulates BP based on sequence and our current knowledge of the mechanism of action for the SNPs. In this project we will focus on testing the specific hypothesis that many ncSNPs identified in GWAS impact the expression of BP relevant genes through epigenetic mechanisms. Epigenetics refers to stable changes in DNA function that are not directly due to changes in the sequence of DNA. Epigenomics refers to such changes when they occur across the entire genome. Therefore, we will use state- of-the-art epigenomic and epigenetic approaches to probe the epigenetic mechanisms by which individual ncSNPs impact the expression of genes associated with BP in human GWAS. Using nephron segments that we will micro-dissect from fresh human kidney and human resistance arterioles isolated from gluteal biopsies as well as related cell types that will be derived from human inducible pluripotent stem cells (iPSCs), we will create an epigenomic map of human BP related cells and tissues at an unprecedented tissue resolution. Using that map combined with our bioinformatic analysis of human GWAS SNP data and data from Project 1 in this PPG (effects of BP ncSNPs on gene expression) we will then test specific epigenetic mechanisms for individual SNPs. Finally, we will perform validation studies in which precision genome editing and targeted epigenetic analysis are utilized to determine if the hypothesized mechanisms are responsible for changes in gene expression in BP-relevant cell types. This project is highly collaborative with the other projects and cores in this Program Project Grant through the sharing of reagents (cells and tissues), transfer of data, and the intellectual environment of the investigators and their laboratories.