VERSICAN PROTEOLYSIS AND REGULATION OF VASCULAR SMOOTH MUSCLE

血管平滑肌的 VERSICAN 蛋白水解和调节

基本信息

批准号：
8378609
负责人：
SUNEEL S APTE
金额：
$ 36.89万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
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/8378609
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

项目摘要

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.

指令）：（项目 5，Apte）Versacan 蛋白水解和血管平滑肌功能调节 -Versacan，一种细胞外基质 (ECM) 蛋白聚糖是血管平滑肌细胞 (VSMC) 的重要产物和正在发育的心脏。 ADAMTS 金属蛋白酶的蛋白水解转换是其主要机制潜在的心脏流出道和心肌发育。相反，ADAMTS 介导的作用对血管壁中多功能蛋白水解作用的了解还很少。我们的初步数据表明 ADAMTSI、ADAMTS5 和 ADAMTS9 由胚胎和成人 VSMC 表达，我们提出这些蛋白酶在多功能蛋白转换中的合作作用模式。初步研究确定主动脉壁 Adamts9+/- 小鼠中与多功能蛋白聚糖积累相关的异常。我们发现 ADAMTS 蛋白水解活性影响培养成纤维细胞的基于透明质酸的细胞周基质的体积，其中多功能蛋白聚糖是一个关键成分。与这一变化相关的是，我们看到 TGF-β 信号传导增强，获得肌成纤维细胞表型。该提案的假设是 ADAMTS 蛋白酶在 VSMC 中协同作用，调节结构和细胞周基质。通过这个生理功能，它们被认为影响 VSMC 行为以及 VSMC 的发育和完整性脉管系统。该假设将通过采用遗传和细胞生物学相结合的方法来解决。使用具有 Adamtsi、Adamts5 和 Adamts9 缺陷的小鼠，我们将在目标 1 中使用组合遗传学来定义这些基因在脉管系统中的个体和协作作用。使用隔离的 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 与化学 ADAMTS 抑制，以及功能获得策略（增加多功能蛋白聚糖和 ADAMTS）来全面定义底层机制。由于细胞周基质中的多功能蛋白聚糖是与透明质酸 (HA) 复合后，我们将定量分析 ADAMTS 蛋白酶对使用 Core B 中可用的资源进行 versican-HA 糖萼的动态变化。相关性（参见说明）：这些实验涉及血管生物学和医学的一个关键领域，具有潜在的相关性发育缺陷、动脉瘤、再狭窄和动脉粥样硬化。该研究预计将提供这些疾病遗传易感性的新候选基因以及已发现的机制可能会提供潜在的新治疗方法。