Regulation of Vascular Smooth Muscle Cell Migration

血管平滑肌细胞迁移的调节

• 批准号: 8063024
• 负责人: Satoshi Komatsu
• 金额: $ 24.68万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8063024
• 关键词: Actomyosin; Address; Antibodies; Atherosclerosis; Behavior; Biological Models; Blood Vessels; Cardiovascular Diseases; Cell Shape; Cells; Collagen; Cytoskeleton; Development; Environment; Extracellular Matrix; Fibroblasts; Filament; Foundations; Glass; Goals; Heart Diseases; Lead; Light; Medial; Mediating; Migration Assay; Modeling; Motor; Motor Activity; Movement; Muscle Contraction; Myocardial Infarction; Myosin Type II; Nonmuscle Myosin Type IIA; Phosphorylation; Physiological; Plastics; Platelet-Derived Growth Factor; Play; Process; Protein Isoforms; Proteins; RNA Interference; Regulation; Reporting; Research; Resistance; Role; Smooth Muscle; Smooth Muscle Myocytes; Staging; Stroke; Surface; System; Techniques; Testing; Thick; Tissues; Vascular Diseases; Vascular System; Wound Healing; ZIP kinase; angiogenesis; atherogenesis; base; cell motility; improved; in vitro Model; insight; migration; motor control; non-muscle myosin; public health relevance; restenosis; two-dimensional; vascular smooth muscle cell migration

DESCRIPTION (provided by applicant): Migration of vascular smooth muscle cells (VSMC) plays a key role in both the physiological and the pathophysiological vascular function such as angiogenesis, restenosis and atherosclerosis. Migration of cells in three-dimensional (3D) matrix is a more closely related model system to tissue than conventional two-dimensional (2D) cell migration. However, there is very little research on the mechanisms controlling VSMC migration in 3D matrix. We will use a 3D migration system and VSMC as an in vitro model system for identifying the mechanisms that regulate VSMC migration. It is well established that myosin II is the crucial motor component of muscle contraction and plays a fundamental role in various types of cellular movement. Whereas there are abundant studies of the myosin II motor in VSMC contraction, little is known of the details of how the myosin II motor controls and is regulated in VSMC migration in 3D matrix. In vertebrate, the function of myosin II molecules is dictated by phosphorylation of its regulatory light chain (MLC). Therefore, it has been thought that the phosphorylation of MLC is a key factor in the regulation of numerous cellular movements. In addition to the involvement of MLC phosphorylation in cell motility, non-muscle myosin IIA (NMIIA) and IIB (NMIIB) isoforms have been shown to serve different roles in the regulation of the cell migration in mammalian fibroblasts. Quite recently, we have proposed a new model where myosin II controls cell migration in a negative and positive fashion, which are mediated by cPKC1 and ZIP kinase. Thus, the following Specific Aims will be addressed: Specific Aim 1) To determine the mechanism by which MLC phosphorylation controls the reorganization of the actomyosin cytoskeleton and regulates VSMC migration in the 3D collagen matrix. Analyses will be performed to determine when and where MLC phosphorylation occurs during platelet-derived growth factor (PDGF)-mediated VSMC migration, and to investigate the roles of cPKC1 and ZIP kinase in this process. Specific Aim 2) To determine the roles and mechanisms of non-muscle myosin IIA (NMIIA) and IIB (NMIIB) isoforms in regulating directionality and contractility in VSMC migration in the 3D matrix. RNA interference (RNAi) approaches will be used to examine the roles of different NMII isoforms in VSMC migration in the 3D collagen matrix. Our studies will provide an important foundation of high relevance to motile behavior of VSMC during atherogenesis, which is considered the most critical underlying cause of vascular disease such as heart attack and stroke. Moreover, the studies proposed here will be important for understanding the basic mechanisms by which VSMC migration is achieved in vascular development such as angiogenesis and wound healing. PUBLIC HEALTH RELEVANCE: The proposed project focuses on the regulatory mechanisms of the cellular protein known as myosin II during cell migration. Migration of the vascular smooth muscle cells (VSMC) associated with blood vessels is a fundamental process in the development of atherosclerosis, a major cause of heart attacks and other cardiovascular diseases. Understanding the precise role of myosin II during VSMC migration will help to improve the current understanding of stable blood vessel formation-an essential step toward the treatment of deadly heart disease.

描述（由申请人提供）：血管平滑肌细胞（VSMC）的迁移在生理和病理生理血管功能（如血管生成、再狭窄和动脉粥样硬化）中起关键作用。细胞在三维（3D）基质中的迁移是比常规的二维（2D）细胞迁移与组织更密切相关的模型系统。然而，目前对三维基质中VSMC迁移控制机制的研究还很少。我们将使用三维迁移系统和VSMC作为体外模型系统，用于识别调节VSMC迁移的机制。众所周知，肌球蛋白II是肌肉收缩的关键运动成分，并在各种类型的细胞运动中发挥着重要作用。虽然有大量的研究肌球蛋白II马达在VSMC收缩，很少有人知道的细节，肌球蛋白II马达如何控制和调节VSMC迁移的三维矩阵。在脊椎动物中，肌球蛋白II分子的功能由其调节轻链（MLC）的磷酸化决定。因此，人们认为MLC的磷酸化是调节许多细胞运动的关键因素。除了MLC磷酸化参与细胞运动之外，非肌肉肌球蛋白IIA（NMIIA）和IIB（NMIIB）同种型已被证明在哺乳动物成纤维细胞中的细胞迁移调节中起不同的作用。最近，我们提出了一个新的模型，其中肌球蛋白II控制细胞迁移的负和积极的方式，这是由cPKC 1和ZIP激酶介导的。具体目的1）确定MLC磷酸化控制肌动球蛋白细胞骨架重组和调节VSMC在3D胶原基质中迁移的机制。将进行分析，以确定在血小板衍生生长因子（PDGF）介导的VSMC迁移过程中何时何地发生MLC磷酸化，并研究cPKC 1和ZIP激酶在此过程中的作用。具体目的2）确定非肌肉肌球蛋白IIA（NMIIA）和IIB（NMIIB）亚型在调节VSMC在3D基质中迁移的方向性和收缩性中的作用和机制。RNA干扰（RNAi）方法将被用来检查不同的NMII亚型在VSMC迁移的3D胶原蛋白基质中的作用。我们的研究将为动脉粥样硬化形成过程中VSMC运动行为的高度相关性提供重要的基础，动脉粥样硬化被认为是心脏病发作和中风等血管疾病的最关键的潜在原因。此外，这里提出的研究将是重要的理解的基本机制，通过它VSMC迁移是实现血管发育，如血管生成和伤口愈合。 公共卫生相关性：拟议的项目侧重于细胞迁移过程中称为肌球蛋白II的细胞蛋白质的调节机制。与血管相关的血管平滑肌细胞（VSMC）的迁移是动脉粥样硬化发展的基本过程，动脉粥样硬化是心脏病发作和其他心血管疾病的主要原因。了解肌球蛋白II在VSMC迁移过程中的确切作用将有助于提高目前对稳定血管形成的了解--这是治疗致命心脏病的重要一步。