In Vivo Analysis of a Noncoding Susceptibility Region for Coronary Artery Disease

冠状动脉疾病非编码易感区的体内分析

基本信息

批准号：
7713519
负责人：
Len Alexander Pennacchio
金额：
$ 42.15万
依托单位：
UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7713519
关键词：
9p21 Affect Alleles Animal Model Animals Aorta Arterial Fatty Streak Atherosclerosis Binding Binding Sites Biological Assay Biological Process Candidate Disease Gene Cardiac Cardiovascular Diseases Cell Proliferation Chromatin Chromosomes, Human, Pair 4 Chromosomes, Human, Pair 9 Clinical Code Coronary Arteriosclerosis Country Coupled Cyclin-Dependent Kinase Inhibitor DNA Sequence Data Development Diet Disease Disease susceptibility Distant EP300 gene Enhancers Epidemiologic Studies Fatty acid glycerol esters Gene Expression Gene Expression Profiling Genes Genetic Genetic Polymorphism Genetically Engineered Mouse Genome Genomics Genotype Goals Haplotypes Heart Heart Diseases Human Human Chromosomes Incidence Individual Knockout Mice Laboratories Lead Link Linkage Disequilibrium Lipids Liver Maps Molecular Profiling Monitor Morbidity - disease rate Mus Pathway interactions Phenotype Plasma Predisposition Proteins Risk Role Severities Smooth Muscle Myocytes System Testing Time Tissues Transcript Transcription Coactivator Transgenic Mice Variant cardiovascular disorder risk cardiovascular risk factor chromatin immunoprecipitation clinical phenotype clinically relevant deviant disorder risk genome wide association study genome-wide human disease in vivo mRNA Expression mortality mouse genome mouse model public health relevance

项目摘要

DESCRIPTION (provided by applicant): Large epidemiological studies have indicated that genetic factors contribute significantly to the development of coronary atherosclerosis, a major cause of morbidity and mortality in Western countries. Through genome-wide association studies (GWASs), we and others have identified a strong link between a 58kb noncoding region on 9p21 and coronary artery disease (CAD). The mechanism by which sequence variation at this locus influences CAD is not known since the interval does not contain any protein-coding genes. To provide an animal model to study this CAD-associated noncoding region, we have deleted the orthologous interval (chr4?70kb) from the mouse genome and show that it significantly impacts on cardiac expression of two genes (Cdkn2a and Cdkn2b) adjacent to the deletion. Accordingly, the central goal of this application is to use this and other genetically engineered mice to uncover the mechanism by which sequence variation in this noncoding interval confers susceptibility to CAD. The specific aims are to (1) identify genes/pathways abnormally expressed in the chr4?70kb mice, (2) to identify and characterize transcriptional enhancers within the deleted interval through a transgenic mouse enhancer assay, (3) to test CAD-associated sequence variants in the enhancers that we have identified in the linkage disequilibrium interval for their impact on gene expression in vivo, and (4) to determine the effect of the chr4?70kb deletion on murine atherosclerosis susceptibility and cellular proliferation. These studies will explore the mechanism of how the human 9p21 linkage disequilibrium region causes CAD susceptibility and serve as a paradigm for relating disease-associated DNA sequence variants occurring in noncoding regions to clinically relevant phenotypes. PUBLIC HEALTH RELEVANCE: Genome-wide association studies (GWASs) have revealed a number of genomic intervals that contribute to human disease but are devoid of protein-coding genes. Remarkably, many individuals have a substantially increased risk for developing coronary artery disease due to sequence variations in a noncoding region of human chromosome 9p21, yet the disease- causing mechanism remains cryptic. Accordingly, we propose to utilize genetically engineered mouse models to study in a controlled laboratory setting how these noncoding sequence changes lead to heart disease in humans.

描述（由申请人提供）：大型流行病学研究表明，遗传因素对冠状动脉粥样硬化的发展有重大影响，冠状动脉粥样硬化是西方国家发病和死亡的主要原因。通过全基因组关联研究 (GWAS)，我们和其他人发现 9p21 上的 58kb 非编码区域与冠状动脉疾病 (CAD) 之间存在密切联系。该位点的序列变异影响 CAD 的机制尚不清楚，因为该区间不包含任何蛋白质编码基因。为了提供一个动物模型来研究这个与 CAD 相关的非编码区域，我们从小鼠基因组中删除了直系同源间隔 (chr4?70kb)，并表明它对与删除相邻的两个基因（Cdkn2a 和 Cdkn2b）的心脏表达有显着影响。因此，本申请的中心目标是使用该小鼠和其他基因工程小鼠来揭示该非编码区间的序列变异导致 CAD 易感性的机制。具体目标是 (1) 识别 chr4?70kb 小鼠中异常表达的基因/通路，(2) 通过转基因小鼠增强子测定来识别和表征删除间隔内的转录增强子，(3) 测试 CAD 相关序列我们在连锁不平衡区间中鉴定了增强子中的变异体，以了解它们对体内基因表达的影响，以及（4）确定 chr4?70kb 缺失对小鼠的影响动脉粥样硬化易感性和细胞增殖。这些研究将探讨人类 9p21 连锁不平衡区域如何导致 CAD 易感性的机制，并作为将非编码区域中发生的疾病相关 DNA 序列变异与临床相关表型联系起来的范例。公共卫生相关性：全基因组关联研究 (GWAS) 揭示了许多导致人类疾病但缺乏蛋白质编码基因的基因组区间。值得注意的是，由于人类染色体 9p21 非编码区的序列变异，许多人患冠状动脉疾病的风险显着增加，但致病机制仍然神秘。因此，我们建议利用基因工程小鼠模型在受控实验室环境中研究这些非编码序列变化如何导致人类心脏病。