Regulation of Pulmonary Vascular Permeability by Integrin AlphaVBeta5

整合素 AlphaVbeta5 对肺血管通透性的调节

• 批准号: 7384374
• 负责人: Dean Sheppard
• 金额: $ 52.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7384374
• 关键词: Ablation; Actins; Acute Lung Injury; Adenoviruses; Adherens Junction; Agonist; Antibodies; Bleomycin; Blocking Antibodies; CD47 Antigen; Cell-Free System; Cells; Coagulation Process; Complex; Cytoplasmic Tail; Data; Development; Disease; Dominant-Negative Mutation; Dose; Endopeptidases; Endothelial Cells; Endotoxins; Environmental air flow; GTP-Binding Proteins; Genes; Genetic; Growth Factor; In Vitro; Integrins; Intervention; Ischemia; Knock-out; Knockout Mice; Laboratories; Link; Lung; Lung diseases; Maps; Mechanical ventilation; Modeling; Molecular; Monoclonal Antibodies; Monomeric GTP-Binding Proteins; Mus; Pathway interactions; Patients; Peptide Hydrolases; Permeability; Phosphorylation; Physiological reperfusion; Prevention; Principal Investigator; Protein Binding; Proteins; Pulmonary Edema; Rattus; Regulation; Reperfusion Therapy; Research Personnel; Rho-associated kinase; Role; Signal Pathway; Signaling Protein; Small Interfering RNA; Stimulus; Stress Fibers; Testing; Thrombin; Vascular Endothelial Growth Factors; Vascular Permeabilities; Ventilator; Ventilator-induced lung injury; Wild Type Mouse; Work; base; cadherin 5; extracellular; in vivo; in vivo Model; integrin alphaVbeta5; lung ischemia; monolayer; null mutation; prevent; programs; protective effect; pulmonary vascular permeability; reconstitution; response

Acute lung injury is a common disorder with no known, effective, pharmacologic treatment. One of the central contributing factors to the development of acute lung injury is an increase in the permeability of the pulmonary vasculature. Mice homozygous for a null mutation in the integrin (35 subunit gene, generated in the principal investigator's laboratory, are protected from pulmonary edema in a model of ventilator induced lung injury, and a blocking monoclonal antibody against the av(35 integrin protects wild type mice in this same model and also protects rats from pulmonary edema induced by unilateral lung ischemia and reperfusion. Blockade or genetic ablation of the av(35 integrin protects cultured endothelial cells from increases in monolayer permeability and from both the formation of rhoA-induced actin stress fibers and the phosphorylation of components of the endothelial adherens junction. In the current proposal, we will explore the molecular pathways linking the av(35 integrin to regulated increases in vascular permeability in more detail. We will use dominant negative and constitutively active constructs, siRNA knockdown and endothelial cells from knockout mice to test the hypotheses that this integrin regulates vascular permeability through interaction with the small GTPases, Ga12 and Ga13 and the known G protein activator integrin associated protein (IAP). We will then examine the in vivo roles for each of these proteins by studying ventilator-induced lung injury in mice expressing global or conditional knockouts of each of these proteins. Because our preliminary data suggest that av(35 is a central component of a multi-protein complex that contains Ga13, and many of the components of the rhoA dependent signaling pathway, we will examine which components of this complex depend on av(35 for assembly and which regions of the (35 subunit are required by reconstituting endothelial cells from (35 knockout mice with wild type, truncated and chimeric forms of this integrin. Finally, to determine whether the pathways we are studying are broadly relevant to non-cardiogenic pulmonary edema, we will further examine the role of av(35 and any other component found to be critical in three additional in vivo models - intracheal bleomycin, intratracheal endotoxin and the combination of low dose endotoxin and moderate volume ventilation. Through the proposed studies we hope to determine whether avp5 or other components of this pathway are attractive targets for intervention in acute lung injury.

急性肺损伤是一种常见的疾病，没有已知的有效的药理治疗。中的一个 急性肺损伤发展的主要因素是渗透率的提高 肺脉管系统。在整联蛋白（35个亚基基因）中纯合子的小鼠纯合子 主要研究者的实验室在呼吸机诱导的模型中受到保护免受肺水肿的保护 肺损伤和针对AV的封闭单克隆抗体（35个整合素保护野生型小鼠在此中 相同的模型，还保护大鼠免受单侧肺部缺血诱导的肺水肿和 再灌注。 AV的封锁或遗传消融（35个整合素保护培养的内皮细胞免受 单层渗透率的增加，以及从RhoA诱导的肌动蛋白应激纤维的形成和 内皮粘附连接的成分的磷酸化。在当前的建议中，我们将探索 将AV连接的分子途径（35个整合素与调节的血管通透性增加 细节。我们将使用主要的负面和组成型积极的构造，siRNA敲低和内皮 敲除小鼠的细胞以测试该整合蛋白调节血管通透性的假设 与小的GTPases GA12和GA13以及已知的G蛋白激活蛋白相关的小GTPase相互作用 蛋白质（IAP）。然后，我们将通过研究呼吸机诱导的每种蛋白质的体内角色 表达每种蛋白质的全球或条件敲除小鼠的肺损伤。因为我们 初步数据表明AV（35是包含GA13， 以及RhoA依赖性信号通路的许多组件，我们将检查哪些组件 该复合物的取决于AV（组装35和哪个区域（35个亚基 从野生型，截短和嵌合的形式重建内皮细胞（35只敲除小鼠 整合素。最后，确定我们正在研究的途径是否与非心脏病有关 肺水肿，我们将进一步检查AV的作用（35个和任何其他成分在 其他三个体内模型 - 气内博来霉素，气管内内毒素和低的组合 剂量内毒素和中等体积通气。通过拟议的研究，我们希望确定 AVP5还是该途径的其他组件是干预急性肺损伤的有吸引力的目标。