Vascular Oxidases in Migration

迁移中的血管氧化酶

基本信息

批准号：
8452205
负责人：
Kathy K Griendling
金额：
$ 36.89万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
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/8452205
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 public health relevance repaired response restenosis rho rho GTP-Binding Proteins vascular factor vascular smooth muscle cell migration web site

项目摘要

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.

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。在最后的资助期间，我们表明，VSMC 响应血小板源性生长因子 (PDGF) 的迁移是由 Nox1 NADPH 氧化酶衍生的活性氧 (ROS) 通过调节丝切蛋白 (一种肌动蛋白结合和切断蛋白) 介导的。我们还发现，Nox4 NADPH 氧化酶及其调节剂 Poldip2 是通过调节小分子量 G 蛋白 Rho 和粘着斑周转来实现迁移所必需的。在此资助期内，我们建议扩展对这两种 NADPH 氧化酶单独并特异性调节 VSMC 迁移所需的细胞骨架重组的信号通路的研究。我们认为 ROS 产生的亚细胞位置决定了细胞反应。因此，片状伪足中PDGF诱导的Nox1激活产生的ROS调节伴随片状伪足突出的细胞骨架重组过程中的急性肌动蛋白聚合/解聚，而粘着斑中整合素介导的Nox4激活产生的H2O2调节粘着斑周转并稳定肌动蛋白通过氧化形成细丝。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 PDGF 诱导的 VSMC 中板状足形成的信号通路。我们将研究控制板状伪足中肌动蛋白聚合的因素，包括 Coronin 1b、Arp2/3 和 slingshot 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迁移过程中粘着斑周转中的作用。我们将检验这样的假设：Nox4 凭借其氧化肌动蛋白的能力，关键性地调节粘着斑周转和迁移，从而导致肌动蛋白调节蛋白的结合发生变化，并促进 RhoA 介导的粘着斑形成。最后，我们将通过确定 Nox4/Poldip2 衍生的 H2O2 产生在新内膜形成中的作用来测试该模型。我们计划使用 Poldip2 表达降低的基因陷阱小鼠以及 Nox4 敲除小鼠，利用依赖于 VSMC 迁移的线损伤诱导的新内膜形成体内模型来研究 Nox4/Poldip2 及其靶标的作用。总之，这些目标将为 NADPH 氧化酶如何介导 VSMC 迁移进而介导病变形成提供新的见解。这些信息可能会导致新的治疗策略的开发，这些策略可以仔细而具体地针对疾病发生中的至关重要的事件。