Vascular Oxidases in Migration

迁移中的血管氧化酶

基本信息

批准号：
7822197
负责人：
Kathy K Griendling
金额：
$ 0.76万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-01 至 2009-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7822197
关键词：
1-Phosphatidylinositol 3-Kinase Abbreviations Acetylcysteine Actins Affect Angioplasty Angiotensin II Animal Model Antioxidants Arterial Fatty Streak Arterial Injury Atherosclerosis Balloon Angioplasty Blood Pressure Blood Vessels Cardiovascular Pathology Catalytic Domain Cells Cyclin-Dependent Kinases Cytoskeletal Modeling Data Development Disease Electron Spin Resonance Spectroscopy Event Focal Adhesion Kinase 1 Focal Adhesions GTPase-Activating Proteins Generations Grant Growth Growth Factor Guanine Nucleotide Exchange Factors Healed Hydrogen Peroxide Hypertension Immigration In Vitro Injury JUN gene Joints LIM Domain Kinase 1 Lead Lesion Ligation MAP Kinase Gene Mediating Metalloproteases Mitogen-Activated Protein Kinases Molecular Molecular Target Mus Myosin Heavy Chains Nitric Oxide Normal tissue morphology Oxidases Oxidative Stress PTK2 gene Pathway interactions Phosphorylation Phosphotransferases Platelet-Derived Growth Factor Play Principal Investigator Process Production Protein Binding Domain Protein-Serine-Threonine Kinases Proteins RNA Interference ROS1 gene Reactive Oxygen Species Role Signal Pathway Signaling Molecule Smooth Muscle Smooth Muscle Myocytes Smooth Muscle Myosins Source Superoxide Dismutase Superoxides Techniques Testing Transgenic Organisms Tyrosine Phosphorylation Vascular remodeling atherogenesis catalase cell growth cell motility computerized data processing diphenyleneiodonium extracellular healing in vivo in vivo Model insight knockout animal migration neuronal cell body novel therapeutics overexpression p21-activated kinase 1 paxillin phosphoinositide-dependent kinase 1 programs repaired response restenosis src-Family Kinases vascular smooth muscle cell migration

项目摘要

DESCRIPTION (provided by applicant): Vascular smooth muscle cell (VSMC) migration is critically important in neointimal formation following angioplasty and atherosclerotic lesion formation. During the last grant period, we showed that VSMC migration in response to platelet-derived growth factor (PDGF) requires the production of reactive oxygen species (ROS), which mediate the tyrosine phosphorylation of phosphoinositide-dependent kinase-1 (PDK1), a serine/threonine kinase that mediates actin cytoskeletal reorganization. PDK1, in turn, phosphorylates p21- activated kinase-1 (PAK1), and both kinases are required for migration. However, cell migration is a multistep process that involves formation of lamellipodia, creation of focal adhesions at the front of the cell, contraction of the cell body, and dissolution of focal adhesions at the trailing edge. The role of ROS and PDK1 in each of these steps remains unclear. In this grant period, we propose to investigate the mechanisms leading to ROS formation and the downstream molecular targets that regulate the early steps in migration. 4 specific aims will be accomplished. First, we will investigate the sources of ROS in PDGF-stimulated cells, focusing on the NAD(P)H oxidases Nox1 and Nox4. We will use RNA silencing techniques to investigate the separate and joint roles of these 2 important oxidases in formation of lamellipodia and focal adhesions. Second, we will investigate the signaling processes leading to Nox1 activation and PDK1- mediated lamellipodia formation. Third, we will investigate the role of Nox4 and PDK1 in mediating focal adhesion kinase activation and phosphorylation 'of paxillin, which in turn regulate the conversion of focal contacts to focal adhesions. Finally, we will use unique transgenic and knockout animals (smooth muscle specific nox1- or catalase-overexpressors and nox1-/- mice) to investigate the role of Nox1 and ROS in an in vivo model of VSMC migration. Migration will be induced by carotid ligation or femoral wire injury and followed using morphometric techniques. Together, these Aims will provide new insight into how NAD(P)H 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）迁移对于血管成形术和动脉粥样硬化病变形成后的新内膜形成至关重要。在上一次资助期间，我们表明，响应血小板衍生生长因子 (PDGF) 的 VSMC 迁移需要产生活性氧 (ROS)，其介导磷酸肌醇依赖性激酶 1 (PDK1) 的酪氨酸磷酸化，PDK1 是一种介导肌动蛋白细胞骨架重组的丝氨酸/苏氨酸激酶。 PDK1 反过来磷酸化 p21 激活激酶 1 (PAK1)，并且这两种激酶都是迁移所必需的。然而，细胞迁移是一个多步骤的过程，涉及板状伪足的形成、细胞前部粘着斑的产生、细胞体的收缩以及后缘粘着斑的溶解。 ROS 和 PDK1 在每个步骤中的作用仍不清楚。在此资助期内，我们建议研究导致 ROS 形成的机制以及调节迁移早期步骤的下游分子靶标。将实现4个具体目标。首先，我们将研究 PDGF 刺激细胞中 ROS 的来源，重点关注 NAD(P)H 氧化酶 Nox1 和 Nox4。我们将使用 RNA 沉默技术来研究这两种重要的氧化酶在板状伪足和粘着斑形成中的单独和联合作用。其次，我们将研究导致 Nox1 激活和 PDK1 介导的板状伪足形成的信号传导过程。第三，我们将研究Nox4和PDK1在介导粘着斑激酶激活和桩蛋白磷酸化中的作用，进而调节粘着斑向粘着斑的转化。最后，我们将使用独特的转基因和基因敲除动物（平滑肌特异性 nox1 或过氧化氢酶过表达小鼠和 nox1-/- 小鼠）来研究 Nox1 和 ROS 在 VSMC 迁移体内模型中的作用。迁移将通过颈动脉结扎或股骨线损伤诱导，然后使用形态测量技术。总之，这些目标将为 NAD(P)H 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。