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: 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
• 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

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，并且众所周知，SMC表型的改变在包括动脉粥样硬化、高血压和再狭窄在内的几种主要心血管疾病状态的进展中发挥作用。心肌素和心肌素相关转录因子（MRTF）是 SMC 分化的极其重要的调节因子，调节心肌素因子活性的机制的识别对于我们理解 SMC 特异性转录非常重要。我们已经鉴定出组蛋白去甲基化酶 jmjd1a 是一种心肌素因子结合蛋白，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。我们还有初步数据表明，心肌素因子稳定性受泛素介导的蛋白酶体降解调节，并且 E3 连接酶的 MURF 家族和 LIM 结构域蛋白 FHL2 可能调节 SMC 中的这一途径。当前提案的目标是进一步表征 SMC 表型调节中的这些新机制。我们的具体目标如下： 1)评估jmjd1a对SMC表型调节的贡献。我们将测量 SMC 特异性启动子处的 H3K9 甲基化和乙酰化以及敲低 jmjd1a 表达，以测试其对 SMC 特异性基因表达和心肌因子活性的影响。我们将检查 jmjd1a 敲除小鼠发育过程中 SMC 分化的缺陷以及血管损伤后 SMC 表型调节的变化。 2）研究蛋白体介导的心肌素因子降解对SMC表型的作用。我们将测量 SMC 中的心肌素和 MRTF 泛素化，识别泛素化的赖氨酸残基，并确定各个 MURF 家族 E3 连接酶和 FHL2 对心肌素因子稳定性的作用。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。完成这些目标应该可以更好地了解 SMC 中心肌素因子活性的调节，这可能有助于开发旨在治疗多种心血管疾病的疗法。公众健康相关性：血管平滑肌细胞分化是血管发育过程中非常重要的过程，众所周知，这一过程的改变在几种主要心血管疾病状态（包括动脉粥样硬化、高血压和再狭窄）的进展中发挥着重要作用。我们的建议研究了调节平滑肌分化的分子机制，并应有助于确定治疗这些疾病的治疗靶点。