Epigenomes and Epigenetic Mechanisms in BP-relevant Tissues

BP相关组织的表观基因组和表观遗传机制

• 批准号: 10023346
• 负责人: ANDREW S. GREENE
• 金额: $ 61.19万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10023346
• 关键词: ATAC-seq; Accounting; Base Pairing; Bioinformatics; Biopsy; Blood Pressure; Cardiovascular Diseases; Cell physiology; Cells; Chromatin; Chromatin Loop; Code; DNA; DNA Methylation; DNA Sequence; DNA-Protein Interaction; Data; Data Analyses; Development; Engineering; Enhancers; Environment; Epigenetic Process; Event; Gene Expression; Genes; Genetic; Genome; Goals; Haplotypes; Heritability; Human; Human Genome; Hypertension; Individual; Kidney; Knowledge; Laboratories; Link; Maps; Mediating; Myocardial Ischemia; Nephrons; Pathway interactions; Phenotype; Physiological; Physiology; Pluripotent Stem Cells; Program Research Project Grants; Proteins; Reagent; Research; Research Personnel; Resistance; Resolution; Risk Factors; Role; Single Nucleotide Polymorphism; Stroke; Testing; Tissues; Untranslated RNA; Validation; Variant; arteriole; base; blood pressure regulation; cell type; data exchange; epigenetic regulation; epigenome; epigenomics; experimental study; genome editing; genome wide association study; genome-wide; human tissue; validation studies; whole genome

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.

项目 2 项目摘要 高血压仍然是多种心血管疾病最重要的危险因素 约占全球所有中风事件的 54% 和所有缺血性心脏病事件的 47%。 尽管我们知道有一个重要的遗传成分决定了血液中的水平 压力（血压；血压遗传力的估计范围为 30-70%），我们还远远不了解 导致人类血压变化的遗传因素。最重大的挑战之一 了解 BP 的遗传控制在于绝大多数与 BP 相关的单核苷酸多态性 人类全基因组关联研究 (GWAS) 中已鉴定的单核苷酸多态性 (SNP) 位于非编码区 DNA 区域。许多非编码 SNP (ncSNP) 位于位于数千个碱基的单倍型区域中 远离任何蛋白质编码基因的配对使得几乎不可能将 SNP 与蛋白质编码基因联系起来 或基于序列和我们目前对该机制的了解来调节血压的生理途径 SNP 的行动。在这个项目中，我们将重点测试许多 ncSNP 的具体假设 GWAS 发现通过表观遗传机制影响 BP 相关基因的表达。表观遗传学 “DNA功能的稳定变化”是指不直接由于DNA序列的变化而引起的DNA功能的稳定变化。 表观基因组学是指整个基因组发生的这种变化。因此，我们将使用状态 最先进的表观基因组和表观遗传学方法来探索个体的表观遗传机制 ncSNP 影响人类 GWAS 中与 BP 相关的基因的表达。使用肾单位片段 我们将对新鲜的人肾和从臀肌活检中分离出的人抵抗性小动脉进行显微解剖 以及源自人类诱导多能干细胞 (iPSC) 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血压相关细胞和组织的表观基因组图谱。 使用该图谱并结合我们对人类 GWAS SNP 数据和项目 1 数据的生物信息分析 然后，我们将测试此 PPG（BP ncSNP 对基因表达的影响）的特定表观遗传机制 单个 SNP。最后，我们将进行验证研究，其中精确的基因组编辑和靶向 表观遗传分析用于确定假设的机制是否导致了变化 BP相关细胞类型中的基因表达。该项目与其他项目和核心高度协作 通过共享试剂（细胞和组织）、传输数据以及 研究人员及其实验室的智力环境。