Molecular regulation of the myocardin factors in vascular smooth muscle

血管平滑肌心肌素因子的分子调控

基本信息

批准号：
7583535
负责人：
Christopher P. Mack
金额：
$ 37万
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：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7583535
关键词：
Acetylation Affect Atherosclerosis Binding Proteins Blood Vessels Cardiac Cardiovascular Diseases Cell Differentiation process Cell Lineage Chromatin Structure Complement Factor B Cues Data Defect Degradation Pathway Development Differentiation Antigens Disease Down-Regulation ERG gene Embryo Enhancers FHL2 gene Family Gene Expression Genes Genetic Transcription Goals Hereditary Disease Histones Hypertension In Vitro Individual Injury Knockout Mice LIM Domain Protein Lead Lysine Measures Mediating Methylation Methyltransferase Modeling Modification Molecular Mus Pathway interactions Pattern Phenotype Play Process Proteins Regulation Reporting Research Role Serum Response Factor Smooth Muscle Smooth Muscle Myocytes System Testing Transferase Ubiquitin Ubiquitination Vascular Smooth Muscle angiogenesis cell type demethylation design factor A femoral artery heterochromatin-specific nonhistone chromosomal protein HP-1 in vivo knock-down multicatalytic endopeptidase complex myocardin novel promoter public health relevance restenosis therapeutic development therapeutic target transcription factor ubiquitin-protein ligase vasculogenesis yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): 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. The myocardin and the myocardin-related transcription factors (MRTFs) are extremely important regulators of SMC differentiation, and the identification of the mechanisms that regulate myocardin factor activity will be important for our understanding of SMC-specific transcription. We have identified the histone demethylase, jmjd1a, as a myocardin factor binding protein and our preliminary data indicate that the demethylation of histone 3 lysine 9 (H3K9) by jmjd1a regulates SMC-specific transcription. We also have preliminary data to suggest that myocardin factor stability is regulated by ubiquitin-mediated proteosomal degradation, and that the MURF family of E3 ligases and the LIM domain protein, FHL2, may regulate this pathway in SMC. The goal of the current proposal is to further characterize these novel mechanisms in the regulation of SMC phenotype. Our specific aims are as follows; 1) to evaluate the contributions of jmjd1a to the regulation of SMC phenotype. We will measure H3K9 methylation and acetylation at the SMC-specific promoters and knock-down jmjd1a expression to test its effects on SMC-specific gene expression and myocardin factor activity. We will examine jmjd1a knockout mice for defects in SMC differentiation during development and for alterations in SMC phenotypic modulation following vessel injury. 2) to study the role of proteosomal-mediated degradation of the myocardin factors on SMC phenotype. We will measure myocardin and MRTF ubiquitination in SMC, identify lysine residues that are ubiquitinylated, and determine the roles of individual MURF family E3 ligases and FHL2 on myocardin factor stability. We will also examine myocardin factor stability and SMC differentiation in Murf1 and Murf3 knockout mice. Completion of these aims should lead to a better understanding of the regulation of myocardin factor activity in SMC which could aid in the development of therapeutics designed to treat a number of cardiovascular diseases. 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（SMC）分化是血管生成和血管生成过程中非常重要的过程，人们众所周知，SMC表型的改变在包括几种著名心血管疾病状态的进展中起作用，包括动脉粥样硬化，包括动脉粥样硬化，高血压和再狭窄。心肌蛋白和与心肌相关的转录因子（MRTF）是SMC分化的极为重要的调节剂，并且鉴定调节肌动蛋白因子活性的机制对于我们对SMC特异性转录的理解非常重要。我们已经将组蛋白脱甲基酶JMJD1A确定为肌动蛋白因子结合蛋白，我们的初步数据表明，JMJD1A通过JMJD1A调节SMC特异性转录的组蛋白3赖氨酸9（H3K9）的脱甲基化。我们还拥有初步数据，以表明肌动蛋白因子稳定性受泛素介导的蛋白体降解调节，并且E3连接酶的MURF家族和LIM域蛋白FHL2可能会调节SMC中的该途径。当前建议的目的是进一步在SMC表型调节中进一步表征这些新型机制。我们的具体目标如下； 1）评估JMJD1A对SMC表型调节的贡献。我们将在SMC特异性启动子和敲低JMJD1A表达上测量H3K9甲基化和乙酰化，以测试其对SMC特异性基因表达和肌心蛋白因子活性的影响。我们将检查JMJD1A敲除小鼠的发育过程中SMC分化的缺陷以及血管损伤后SMC表型调节的改变。 2）研究蛋白酶体介导的肌动蛋白因子在SMC表型中的降解的作用。我们将测量SMC中的心肌和MRTF泛素化，鉴定泛素化的赖氨酸残基，并确定单个MURF家族E3连接酶和FHL2对肌钙蛋白因子稳定性的作用。我们还将检查MURF1和MURF3敲除小鼠中的心肌因子稳定性和SMC分化。这些目标的完成应使SMC中肌动蛋白因子活性的调节更好地了解，这可能有助于开发旨在治疗多种心血管疾病的治疗剂。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。