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: 7194287
• 负责人: Dean Sheppard
• 金额: $ 51.96万
• 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/7194287
• 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

DESCRIPTION (provided by applicant): 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 ¿5 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¿5 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¿5 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¿5 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¿5 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¿5 for assembly and which regions of the ¿5 subunit are required by reconstituting endothelial cells from ¿5 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¿5 and any other component found to be critical in three additional in vivo models - intratracheal bleomycin, intratracheal endotoxin and the combination of low dose endotoxin and moderate volume ventilation. Through the proposed studies we hope to determine whether av¿5 or other components of this pathway are attractive targets for intervention in acute lung injury.

描述（由适用提供）：急性肺损伤是一种常见的疾病，没有已知的有效，药物治疗。急性肺损伤发展的主要因素之一是肺脉管系统的渗透性增加。主要研究者实验室中产生的整合素中无效突变的小鼠在呼吸机诱导的肺损伤的模型中受到保护不受肺水肿的保护再灌注。 AV¿5整联蛋白的封锁或遗传消融可保护培养的内皮细胞免受单层渗透性的增加，并免受Rhoa诱导的肌动蛋白胁迫纤维的形成以及内皮粘附连接处的成分的磷酸化。在当前的建议中，我们将探索将AV¿5整合素连接起来的分子途径，以更详细地调节血管通透性的增加。我们将使用敲除小鼠的主动性和组成型活性构建体，siRNA敲低和内皮细胞来测试该积分，即该整合蛋白通过与小GTPase，GA12和GA13和已知的G蛋白激活剂整合蛋白（IAP）相互作用来调节血管通透性。然后，我们将通过研究呼吸机诱导的每种蛋白质全球或条件敲除的小鼠中呼吸机诱导的肺损伤来检查这些蛋白质中每一种的体内作用。 Because our preliminary data suggest that AV¿ 5 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¿ 5 for assembly and which regions of the ¿ 5 subunits are required by reconstituting endothelial cells from ¿ 5 knockout mice with wild type, truncated and chimeric forms of this整合素。最后，为了确定我们正在研究的途径是否与非心脏病性肺水肿广泛相关，我们将进一步研究Av？5和其他任何其他成分在三种其他体内模型中至关重要的作用 - 运动型波勒霉素，气管内气管毒素内毒素和低剂量内毒素内毒素的组合。通过拟议的研究，我们希望确定该途径的Av考5或其他成分是干预急性肺损伤的有吸引力的靶标。