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

• 批准号: 7860582
• 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
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7860582
• 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

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(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的一个非常重要的过程，众所周知，SMC表型的变化在动脉粥样硬化、高血压和再狭窄等几种突出的心血管疾病状态的进展中起着作用。肌钙蛋白和肌钙蛋白相关转录因子是SMC分化的重要调节因子，明确肌钙蛋白因子活性的调控机制对于我们理解SMC特异性转录具有重要意义。我们已经确定组蛋白去甲基酶jmjd1a是一种肌钙蛋白因子结合蛋白，我们的初步数据表明，jmjd1a对组蛋白3赖氨酸9(H3K9)的去甲基化调节SMC特异性转录。我们也有初步的数据表明，肌钙蛋白因子的稳定性受到泛素介导的蛋白酶体降解的调节，而MURF家族的E3连接酶和LIM结构域蛋白FHL2可能调节SMC中的这一途径。当前建议的目标是进一步描述这些调控SMC表型的新机制。我们的具体目的如下：1)评价jmjd1a在SMC表型调控中的作用。我们将检测H3K9在SMC特异性启动子上的甲基化和乙酰化以及下调jmjd1a的表达，以测试其对SMC特异性基因表达和肌钙蛋白因子活性的影响。我们将检查jmjd1a基因敲除小鼠在发育过程中SMC分化的缺陷，以及血管损伤后SMC表型调节的变化。2)研究蛋白酶体介导的肌钙蛋白因子降解对SMC表型的影响。我们将测量肌钙蛋白和MRTF在SMC中的泛素化，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。我们还将在MuRF1和Murf3基因敲除小鼠中检测Myocardin因子的稳定性和SMC分化。完成这些目标将导致更好地了解SMC中肌钙蛋白因子活性的调节，这可能有助于开发旨在治疗多种心血管疾病的疗法。公共卫生相关性：血管平滑肌细胞分化是血管发育过程中的一个非常重要的过程，众所周知，这个过程中的变化在几种主要的心血管疾病状态的进展中发挥了作用，包括动脉粥样硬化、高血压和再狭窄。我们的建议研究了调节平滑肌分化的分子机制，并应有助于确定这些疾病的治疗靶点。