VERSICAN PROTEOLYSIS AND REGULATION OF VASCULAR SMOOTH MUSCLE

血管平滑肌的 VERSICAN 蛋白水解和调节

• 批准号: 8494083
• 负责人: SUNEEL S APTE
• 金额: $ 33.42万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8494083
• 关键词: ADAMTS; ADAMTS1 gene; ADAMTS9 gene; Address; Adult; Alleles; Aneurysm; Area; Atherosclerosis; Behavior; Biology; Blood Vessels; Candidate Disease Gene; Carbohydrates; Cardiac; Cardiovascular system; Cell physiology; Cells; Cellular biology; Chemicals; Chondroitin Sulfates; Cleaved cell; Collagen; Complement; Complex; Data; Defect; Development; Elastin; Embryo; Extracellular Matrix; Fibroblasts; Genes; Genetic; Genetic Predisposition to Disease; Glycocalyx; Heart; Homeostasis; Hyaluronan; Hyperplasia; Individual; Instruction; Knockout Mice; Left; Length; Malignant Neoplasms; Mediating; Medicine; Metalloproteases; Modeling; Modification; Molecular; Mus; Muscle function; Mutant Strains Mice; Myocardial; Organ; Pathology; Peptide Hydrolases; Phenotype; Physiological; Process; Property; Proteoglycan; Proteolysis; Publishing; Regulation of Proteolysis; Research; Resources; Role; Signal Transduction; Smooth Muscle Myocytes; Stenosis; Tissues; Transforming Growth Factor beta; Transgenic Mice; Vascular Diseases; Vascular Smooth Muscle; Work; base; cell behavior; combinatorial; gain of function; insight; link protein; novel therapeutic intervention; research study; versican

instmctions): (Project 5, Apte) Versican Proteolysis and Regulation of Vascular Smooth Muscle Function -Versican, an extracellular matrix (ECM) proteoglycan, is a prominent product of vascular smooth muscle cells (VSMCs) and the developing heart. Its proteolytic turnover by ADAMTS metalloproteases is a major mechanism underlying cardiac outflow tract and myocardial development. In contrast, the role of ADAMTS-mediated versican proteolysis in the vascular wall is poorly understood. Our preliminary data demonstrate that ADAMTSI, ADAMTS5 and ADAMTS9 are expressed by embryonic and adult VSMCs, and we propose a cooperative mode of action of these proteases in versican turnover. Preliminary studies identified aortic wall anomalies associated with versican accumulation in Adamts9+/- mice . We found that ADAMTS proteolytic activity influenced the volume of the hyaluronan-based pericellular matrix of cultured fibroblasts, of which versican is a critical component. Associated with this change, we saw enhanced TGF-beta signaling and acquisition of a myofibroblastic phenotype. The hypothesis underlying the proposal is that ADAMTS proteases work cooperatively in VSMCs to regulate the structural and pericellular matrix. Through this physiological function, they are proposed to influence VSMC behavior and the development and integrity of the vasculature. The hypothesis will be addressed by taking a combined genetic and cell biology approach. Using mice deficient In Adamtsi, Adamts5, and Adamts9, we will use combinatorial genetics in aim 1 to define the individual and cooperative roles of these genes in the vasculature. Using VSMCs isolated from these mouse mutants, we will determine the role of ADAMTS proteases in influencing VSMC function in aim 2. We will complement genetic experiments with approaches that use wild-type VSMCs in conjunction with chemical ADAMTS inhibition, as well as gain-of-function strategies (increased expression of versican and ADAMTS) to comprehensively define the underlying mechanisms. Since versican in the pericellular matrix is complexed to hyaluronan (HA), we will undertake quantitative analysis of the effect of ADAMTS proteases on the dynamics of the versican-HA glycocalyx using resources available in Core B. RELEVANCE (See instructions): These experiments address a critical area of vascular biology and medicine, with potential relevance for developmental defects, aneurysms, re-stenosis and atherosclerosis. The research is expected to provide new candidate genes for genetic predisposition to these conditions, and the mechanisms that are uncovered are likely to offer potentially new therapeutic approaches.

Instmctions）： （项目5，APTE）Versican蛋白水解和血管平滑肌功能的调节 - 细胞外基质（ECM）蛋白聚糖是血管平滑肌细胞（VSMC）的重要产物 还有发展的心。 ADAMTS金属蛋白酶的蛋白水解周转率是主要机制 基础心脏流出道和心肌发育。相反，Adamts介导的作用 血管壁中的versican蛋白水解知之甚少。我们的初步数据表明 ADAMTSI，ADAMTS5和ADAMTS9由胚胎和成人VSMC表示，我们建议 这些蛋白酶在versican周转率中的合作作用方式。初步研究确定了主动脉壁 与ADAMTS9 +/-小鼠中versican积累相关的异常。我们发现Adamts蛋白水解 活性影响培养的成纤维细胞的基于透明质酸的周围基质基质的体积，其中 Versican是关键组成部分。与这种变化相关，我们看到了增强的TGF-beta信号传导和 获得肌纤维细胞表型。该提议的基础假设是Adamts 蛋白酶在VSMC中协同工作，以调节结构和周围的基质。通过这个 生理功能，他们被提议影响VSMC的行为以及 脉管系统。该假设将通过采用遗传和细胞生物学方法来解决。 使用缺乏ADAMTSI，ADAMTS5和ADAMTS9的小鼠，我们将在AIM 1中使用组合遗传学 定义这些基因在脉管系统中的个体和合作作用。使用与从中隔离的VSMC 这些小鼠突变体，我们将确定ADAMTS蛋白酶在影响VSMC功能中的作用 2。我们将通过使用野生型VSMC结合结合的方法来补充遗传实验 化学ADAMTS抑制以及功能获得策略（versican和 ADAMTS）全面定义了基本机制。由于细胞基质中的versican是 复合到透明质酸（HA），我们将对Adamts蛋白酶对蛋白酶的影响进行定量分析 使用CoreB中可用的资源，Versican-Ha Glycocalyx的动力学。 相关性（请参阅说明）： 这些实验涉及血管生物学和医学的关键领域，并具有潜在的相关性 发育缺陷，动脉瘤，再生史病和动脉粥样硬化。该研究有望提供 新的候选基因，用于对这些条件的遗传倾向以及发现的机制 可能会提供潜在的新治疗方法。