Plasticity of the Vascular Smooth Muscle Cytoskeleton

血管平滑肌细胞骨架的可塑性

• 批准号: 7329374
• 负责人: KATHLEEN G MORGAN
• 金额: $ 37.79万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7329374
• 关键词: Actins; Actomyosin; Address; Agonist; Antisense Technology; Biochemical; Biology; Blood Vessels; Cardiovascular Diseases; Cell Shape; Cells; Centrifugation; Collection; Color; Complement; Cytoskeleton; Data; Development; Disease; Dominant-Negative Mutation; Fibroblasts; Filament; Fluorescence Resonance Energy Transfer; Future; G Actin; Goals; Hand; Immunoblotting; In Vitro; Label; Lead; Literature; Maintenance; Mass Spectrum Analysis; Mechanics; Methodology; Microfilaments; Molecular; Monitor; Muscle; Muscle Cells; Muscle function; PTK2 gene; Peptides; Pharmaceutical Preparations; Plastics; Play; Population; Protein Fragment; Proteins; Receptor Protein-Tyrosine Kinases; Review Literature; Role; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Standards of Weights and Measures; Stretching; Striated Muscles; Structure; System; Techniques; Testing; Time; Tissues; Vascular Smooth Muscle; base; cell type; crosslink; drug development; in vivo; novel; novel therapeutics; profilin; protein protein interaction; tool; transmission process; vasodilator-stimulated phosphoprotein

Proliferative cells, such as fibroblasts, are well known to have a very "plastic" cytoskeleton. In contrast, striated muscle cells are known to have a rigid cytoskeleton. Differentiated vascular smooth muscle cells (dVSMCs), based on limited existing literature and our preliminary data, appear to function somewhere in between these two cell types. The long-term goal of this project is to determine to what degree and by what mechanisms the "non-muscle" cytoskeleton modulates smooth muscle contractility during agonist-, hydrodynamic and disease-induced remodeling of its structure. The Specific Aims over the next 5 years are: 1. To test the hypothesis, based on preliminary data, that actin filaments remodel during agonist stimulation and stretch in dVSMCs and to determine whether this alters contractility. 2. To test the hypothesis that specific nucleation/elongation and stabilizing proteins are involved in cytoskeletal remodeling of dSMC, focusing first on profilin, N-WASP, and VASP (nucleating/elongation), and CaD, Tm, and CaP (stabilizing). We will also test the hypothesis that the distribution of smooth muscle putative filament stabilizer proteins, CaD, Tm, and CaP, play a role in defining the localizationof the less labile actin filaments. 3. To test the hypothesis that remodeling of the dense plaques and/or dense bodies and/or their attachements to actin filaments occurs in dVSMCs, and, that the non-receptor tyrosine kinases FAK and Src regulate cytoskeleton/plaque remodeling in dVSMC. Techniques will include differential centrifugation, and in vivo labeling to monitor F and G actin populations, 2-color immunoblot, mass spectrometry, in-cell photoaffinity cross-linking and FRET, cell permeant decoy peptides, dominant negative protein fragments, antisense technology and standard cellular, biochemical and molecular methodologies. The results from this project will further our understanding of the, yet unclear mechanisms offeree maintenance in the dVSMC and will increase our fundamental understanding of the role of the nonmuscle cytoskeleton in the control of vascular function. LAY SUMMARY: The limitations of current drugs to treat cardiovascular disease is due, in part, to our limited understanding of how the muscle cells of blood vessels function. This project will expand our understanding of the VSMC and has the potential of identifying entirely novel targets for drug development.

众所周知，增殖细胞，例如成纤维细胞，具有非常“可塑”的细胞骨架。相比之下， 已知横纹肌细胞具有刚性的细胞骨架。分化的血管平滑肌细胞 （dVSMC），基于有限的现有文献和我们的初步数据，似乎在 这两种细胞类型之间。该项目的长期目标是确定在何种程度上以及以什么方式 “非肌肉”细胞骨架在激动剂期间调节平滑肌收缩力的机制， 流体动力学和疾病引起的结构重塑。未来5年的具体目标是： 1. 根据初步数据检验肌动蛋白丝在激动剂刺激期间重塑的假设 并拉伸 dVSMC，以确定这是否会改变收缩性。 2. 检验假设 特定的成核/延伸和稳定蛋白参与 dSMC 的细胞骨架重塑， 首先关注 Profilin、N-WASP 和 VASP（成核/伸长）以及 CaD、Tm 和 CaP（稳定）。 我们还将测试平滑肌假定的细丝稳定蛋白的分布的假设， CaD、Tm 和 CaP 在定义不太不稳定的肌动蛋白丝的定位方面发挥着作用。 3. 测试 假设致密斑块和/或致密体和/或其与肌动蛋白的附着物的重塑 细丝发生在 dVSMCs 中，非受体酪氨酸激酶 FAK 和 Src 调节 dVSMC 中的细胞骨架/斑块重塑。技术包括差速离心和体内 用于监测 F 和 G 肌动蛋白群体的标记、2 色免疫印迹、质谱、细胞内光亲和 交联和 FRET、细胞渗透诱饵肽、显性失活蛋白片段、反义 技术和标准细胞、生化和分子方法。该项目的结果将 进一步我们对 dVSMC 中尚不明确的受要约人维护机制的理解，并将 增加我们对非肌肉细胞骨架在血管控制中的作用的基本理解 功能。 简单总结：目前治疗心血管疾病的药物的局限性部分是由于我们的有限 了解血管的肌肉细胞如何发挥作用。这个项目将扩大我们的理解 VSMC 的研究具有确定药物开发全新靶点的潜力。