Epigenetic regulation of vascular smooth muscle cell phenotype

血管平滑肌细胞表型的表观遗传调控

基本信息

批准号：
8297230
负责人：
Christopher P. Mack
金额：
$ 36.62万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-02 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8297230
关键词：
Acetylation Adenoviruses Atherosclerosis Binding Bioinformatics Biological Assay Blood Vessels Cardiovascular Diseases Cell Culture Techniques Cell Differentiation process Cell Proliferation Chromatin Chromatin Remodeling Factor Chromatin Structure Collaborations CpG Islands Cultured Cells DNA Methylation Data Deoxyribonucleases Development Differentiation Antigens Disease Electrophoretic Mobility Shift Assay Elements Embryo Enhancers Epigenetic Process Exhibits Family Formaldehyde Gene Activation Gene Expression Gene Silencing Genetic Transcription Genome Goals Heart Histones Human Hypersensitivity Hypertension In Vitro Injury Knockout Mice Location Lysine Maps Measures Mediating Methylation Modeling Molecular Monitor Morphology Mus Pattern Phenotype Play Process Promoter Regions Proteins Protocols documentation Recruitment Activity Regulation Regulatory Element Role Serum Serum Response Factor Signal Transduction Smooth Muscle Smooth Muscle Myocytes Tamoxifen Techniques Testing Tissues Trans-Activators Transcription Repressor/Corepressor Transferase Transgenic Organisms angiogenesis base bisulfite calponin cell type chromatin immunoprecipitation gene repression genome-wide histone modification in vivo in vivo Model injured myocardin notch protein novel promoter restenosis therapeutic target transcription factor vasculogenesis

项目摘要

DESCRIPTION (provided by applicant): Epigenetic regulation of vascular smooth muscle cell phenotype Vascular smooth muscle cell (SMC) differentiation is a very important process during vasculogenesis and angiogenesis, and it is well recognized that alterations in SMC phenotype play a role in the progression of several prominent cardiovascular disease states including atherosclerosis, hypertension, and restenosis. Although serum response factor (SRF) and the myocardin factors are critical for SMC differentiation, recent studies from our lab and others indicate that epigenetic mechanisms are also critical for the overall pattern of SMC-specific gene expression. Indeed, we identified the histone demethylase, jmjd1a, as a myocardin factor interacting protein and have shown that jmjd1a stimulates SMC differentiation marker gene expression in aortic SMC cultures by demethylating H3K9 near the SMC-specific. A major goal of the current proposal will be to closely examine jmjd1a knock-out mice for effects on SMC phenotype, and we will characterize H3K9 methylation in several in vitro and in vivo models of SMC phenotypic modulation. H3K9 methylation is strongly associated with DNA methylation, and interestingly, the SRF binding regions within the SMC-specific promoters are embedded within CpG islands. Based on our preliminary data indicating that the SMC-specific promoters are regulated by methylation, the goal of Aim 2 is to identify the molecular mechanisms involved. Finally, in an attempt to better understand the global chromatin changes that control SMC phenotype, we have established collaborations with Jason Lieb and Terry Furrey to "map" open chromatin regions in SMC using DNase hypersensitivity and Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE). We have generated a genome-wide map from human aortic SMC cultures and have identified a number of previously unexamined promoter regions as potentially important for SMC-specific gene expression. The goals of Aim 3 are to test whether these regions drive expression in vivo and to identify the chromatin modifying and transcription factors that are involved. PUBLIC HEALTH RELEVANCE: Vascular smooth muscle cell differentiation is a very important process during the development of blood vessels and it is well recognized that alterations in this process play a role in the progression of several prominent cardiovascular disease states including atherosclerosis, hypertension, and restenosis. Our proposal examines the molecular mechanisms that regulate smooth muscle differentiation and should help to identify therapeutic targets for the treatment of these diseases.

描述（由申请人提供）：血管平滑肌细胞表型血管平滑肌细胞（SMC）分化是血管生成和血管生成过程中非常重要的过程，人们非常认识到，SMC表型的改变在几种突出的心血管疾病状态的进展中起着作用，包括心血管疾病态的进展，包括高螺旋蛋白局症，高级序列，高级效应，并延伸。尽管血清反应因子（SRF）和心肌素因子对于SMC分化至关重要，但我们实验室的最新研究表明，表观遗传机制对于SMC特异性基因表达的总体模式也至关重要。的确，我们将组蛋白脱甲基酶JMJD1A确定为一种相互作用的肌动蛋白因子，并表明JMJD1A通过在SMC特异性附近脱甲基H3K9在主动脉SMC培养物中刺激SMC分化标记基因表达。当前建议的主要目标是密切检查JMJD1A敲除小鼠对SMC表型的影响，我们将在几种体外和体内SMC表型调节模型中表征H3K9甲基化。 H3K9甲基化与DNA甲基化密切相关，有趣的是，SMC特异性启动子内的SRF结合区域嵌入CpG岛中。基于我们的初步数据，表明SMC特异性启动子受甲基化调节，目标2的目的是确定所涉及的分子机制。最后，为了更好地了解控制SMC表型的全球染色质变化，我们与Jason Lieb和Terry Furrey建立了合作，使用DNase超敏性和甲醛辅助隔离调节元件（公平），以“映射” SMC中的开放染色质区域。我们已经从人主动脉SMC培养物中生成了全基因组图，并确定许多先前未经证实的启动子区域对于SMC特异性基因表达可能很重要。目标3的目标是测试这些区域是否在体内驱动表达并确定所涉及的染色质修饰和转录因子。公共卫生相关性：血管平滑肌细胞分化是血管发展过程中非常重要的过程，人们认识到，这一过程的改变在几种著名的心血管疾病状态的进展中起作用，包括动脉粥样硬化，高血压和再衰变。我们的建议检查了调节平滑肌分化的分子机制，并应有助于确定治疗这些疾病的治疗靶标。