Vascular Oxidases in Migration

迁移中的血管氧化酶

• 批准号: 8108578
• 负责人: Kathy K Griendling
• 金额: $ 38.75万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8108578
• 关键词: Abbreviations; Accounting; Acetylcysteine; Actin-Binding Protein; Actins; Acute; Affect; Agonist; Angioplasty; Antioxidants; Arterial Fatty Streak; Arterial Injury; Arteries; Atherosclerosis; Balloon Angioplasty; Binding; Blood Pressure; Blood Vessels; Cardiovascular Pathology; Caveolae; Cell Nucleus; Cells; Cessation of life; Cytoskeletal Modeling; Cytoskeleton; Data; Development; Devices; Disease; Electron Spin Resonance Spectroscopy; Endothelial Cells; Enzymes; Event; F-Actin; Focal Adhesion Kinase 1; Focal Adhesions; G Actin; GTP-Binding Proteins; Genetically Modified Animals; Grant; Green Fluorescent Proteins; Growth; Growth Factor; Guanine Nucleotide Exchange Factors; Healed; Hydrogen Peroxide; Hyperplasia; Hypertension; Immigration; Immunoprecipitation; In Vitro; Injury; Integrins; Investigation; Knockout Mice; Lead; Lesion; Location; Mechanics; Mediating; Medical; Metalloproteases; Microfilaments; Modeling; Molecular; Molecular Weight; Mus; NADPH Oxidase; Nitric Oxide; Normal tissue morphology; Oxidases; Oxidative Stress; PTK2 gene; Phosphoric Monoester Hydrolases; Platelet-Derived Growth Factor; Play; Polymerase; Production; Protein Kinase C; Protein Tyrosine Phosphatase; Proteins; ROS1 gene; Reactive Oxygen Species; Reduced Glutathione; Regulation; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle; Smooth Muscle Myocytes; Stem cells; Stress Fibers; Superoxides; Testing; Transforming Growth Factors; Vascular remodeling; Work; atherogenesis; cell growth; cofilin; coronin protein; depolymerization; design; diphenyleneiodonium; genetic regulatory protein; healing; human EMS1 protein; in vivo; in vivo Model; insight; interest; migration; novel therapeutics; overexpression; oxidation; polymerization; repaired; response; restenosis; rho; rho GTP-Binding Proteins; vascular factor; vascular smooth muscle cell migration; web site

DESCRIPTION (provided by applicant): Vascular smooth muscle cell (VSMC) migration is critically important in neointimal formation following vascular injury and atherosclerotic lesion formation. During the last grant period, we showed that VSMC migration in response to platelet-derived growth factor (PDGF) is mediated by reactive oxygen species (ROS) derived from the Nox1 NADPH oxidase via regulation of cofilin, an actin binding and severing protein. We also found that the Nox4 NADPH oxidase and its regulator Poldip2 are required for migration via regulation of the small molecular weight G-protein Rho and focal adhesion turnover. In this grant period, we propose to extend our investigation of the signaling pathways by which these two NADPH oxidases separately and specifically regulate the reorganization of the cytoskeleton that is required for VSMC migration. We propose that the subcellular location of ROS production dictates the cellular response. Thus, production of ROS by PDGF-induced activation of Nox1 in lamellipodia regulates acute actin polymerization/depolymerization during the cytoskeletal reorganization that accompanies lamellipodial protrusion, while the production of H2O2 by integrin-mediated Nox4 activation in focal adhesions regulates focal adhesion turnover and stabilizes actin filaments via oxidation. To test this hypothesis, three specific aims will be accomplished. First, we will define the signaling pathways by which Nox1 regulates PDGF- induced lamellipodial formation in VSMCs. We will investigate factors controlling actin polymerization in the lamellipodia, including coronin 1b, Arp2/3 and slingshot 1L phosphatase (SSH1L), and their relationship to Nox1. Second, we plan to determine the role of Nox4 in integrin-mediated actin oxidation and focal adhesion turnover during migration. We will test the hypothesis that Nox4 critically regulates focal adhesion turnover and migration by virtue of its ability to oxidize actin, thus leading to changes in the binding of actin regulatory proteins, and to facilitate RhoA-mediated focal adhesion formation. Finally, we will test this model in vivo by determining the role of Nox4/Poldip2-derived H2O2 production in neointimal formation. We plan to use genetrap mice in which Poldip2 expression is reduced, as well as Nox4 knockout mice, to investigate the role of Nox4/Poldip2 and their targets using an in vivo model of wire injury-induced neointimal formation that is dependent upon VSMC migration. Together, these aims will provide new insight into how NADPH oxidases mediate VSMC migration and therefore lesion formation. Such information may lead to the development of new therapeutic strategies that can be carefully and specifically targeted to the critically important events in disease initiation. PUBLIC HEALTH RELEVANCE: This proposal will investigate the roles of the vascular smooth muscle cell (VSMC) NADPH oxidases, Nox1 and Nox4, in PDGF-induced migration, and the molecular mechanisms by which reactive oxygen species derived from these enzymes mediate the early cellular events that lead to VSMC migration. VSMC migration is clinically important during neointimal hyperplasia after vascular injury and atherosclerotic lesion formation, but despite advances in the design of the mechanical devices, and despite new adjunctive medical therapies, complications from atherosclerosis and restenosis account for >36% of all deaths (per the AHA website). Understanding the mechanisms underlying migration may lead to the development of new therapeutic strategies that can be carefully and specifically targeted to the critically important events in disease initiation.

描述（由申请人提供）：血管平滑肌细胞（VSMC）迁移对于血管损伤和动脉粥样硬化病变形成后的新内膜形成至关重要。在最后一个赠款期间，我们表明，响应血小板衍生的生长因子（PDGF）的VSMC迁移是由通过调节cofilin（一种肌动蛋白（一种肌动蛋白）结合和断裂蛋白的NOX1 NADPH氧化酶衍生而来的活性氧（ROS）介导的。我们还发现，通过调节小分子量G蛋白RHO和局灶性粘附周转，NOX4 NADPH氧化酶及其调节剂POLDIP2是必需的。在此赠款期间，我们建议扩展对这两个NADPH氧化酶分别调节的信号传导途径的研究，并特别调节VSMC迁移所需的细胞骨架的重组。我们建议ROS产生的亚细胞位置决定了细胞反应。因此，通过PDGF诱导的NOX1在层状脂蛋白中的激活产生ROS，调节急性肌动蛋白聚合/聚合在细胞骨架重组过程中，而层状膜片突出伴随着层状型的延伸，而整合蛋白介导的NOX4激活在探测器中均通过探针粘连和稳定性调节H2O2的产生。为了检验这一假设，将实现三个具体目标。首先，我们将定义NOX1调节VSMC中PDGF诱导的lamellipodial形成的信号通路。我们将研究控制层状脂蛋白肌动蛋白聚合的因素，包括冠状酸1b，ARP2/3和弹弓1L磷酸酶（SSH1L），以及它们与NOX1的关系。其次，我们计划确定NOX4在整合素介导的肌动蛋白氧化和局灶性粘附过程中的作用。我们将检验以下假设：NOX4因其氧化肌动蛋白的能力而严重调节局灶性粘附周转和迁移，从而导致肌动蛋白调节蛋白的结合变化，并促进RhoA介导的局灶性局灶性粘附形成。最后，我们将通过确定NOX4/POLDIP2衍生的H2O2产生在新形成形成中的作用来测试该模型。我们计划使用降低POLDIP2表达以及NOX4基因敲除小鼠的Genetrap小鼠，以研究NOX4/POLDIP2及其靶标的作用，使用电线损伤诱导的新内膜形成的体内模型，这取决于VSMC迁移。这些目标一起将提供有关NADPH氧化酶如何介导VSMC迁移并因此形成病变的新见解。这些信息可能会导致新的治疗策略的发展，这些策略可以仔细，专门针对疾病开始中至关重要的事件。 公共卫生相关性：该提案将调查PDGF诱导的迁移中的血管平滑肌细胞（VSMC）NADPH氧化酶，NOX1和NOX4的作用，以及从这些酶介导VSMC迁移的早期细胞事件的反应性氧物种的分子机制。 VSMC迁移在血管损伤和动脉粥样硬化病变后的新内膜增生期间在临床上很重要，但是尽管机械设备的设计方面进步了，尽管动脉粥样硬化的并发症，但动脉粥样硬化和再狭窄的并发症占所有死亡的36％（根据AHA网站）> 36％（根据AHA网站）。了解迁移迁移的机制可能会导致新的治疗策略的发展，这些策略可以仔细，专门针对疾病开始中至关重要的事件。