Vascular Oxidases in Migration

迁移中的血管氧化酶

• 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
• 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; 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

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）迁移在血管成形术和动脉粥样硬化病变形成后对新内膜形成至关重要。 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反过来磷酸化P21-活化的激酶-1（PAK1），并且两个激酶均需要迁移。然而，细胞迁移是一个多步骤过程，涉及层状脂蛋白的形成，在细胞前部创建局灶性粘附，细胞体的收缩以及尾缘在尾边缘的溶解。 ROS和PDK1在这些步骤中的每个步骤中的作用尚不清楚。在这一赠款期间，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刺激的细胞中ROS的来源，重点是NAD（P）H氧化酶NOX1和NOX4。我们将使用RNA沉默技术研究这2个重要氧化酶在形成片状脂蛋白和局灶性粘附中的单独和关节作用。其次，我们将研究导致NOX1激活和PDK1介导的薄片层状形成的信号传导过程。第三，我们将研究NOX4和PDK1在paxillin介导局灶性激酶激活和磷酸化中的作用，这又调节了局灶性接触到粘着斑的粘连的转化。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-或过氧化氢酶超过表达器以及NOX1 - / - 小鼠）来研究NOX1和ROS在VSMC迁移体内模型中的作用。颈动脉连接或股线损伤将诱导迁移，然后使用形态计量学技术。这些目标将共同提供新的见解，即NAD（P）H氧化酶如何介导VSMC迁移，从而介导病变形成。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。