Caveolin-1 and NO Regulate PMN-mediated Increases in Vascular Permeability

Caveolin-1 和 NO 调节 PMN 介导的血管通透性增加

• 批准号: 7370182
• 负责人: RICHARD D MINSHALL
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-10 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370182
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acute Lung Injury; Address; Adhesions; Albumins; Biochemical; Blood Vessels; Caveolae; Complement; Cultured Cells; Cysteine; Data; Disease; Dynamin 2; Edema; Endothelial Cells; Extravasation; Gene Deletion; Generations; ITGB2 gene; Image Analysis; Inflammation; Inflammatory; Injury; Intercellular adhesion molecule 1; Knockout Mice; Life; Lung; Mediating; Microvascular Permeability; Molecular Target; Mus; Neutrophil Activation; Oxidants; PH Domain; Pathway interactions; Permeability; Peroxonitrite; Phosphoinositide-3-Kinase, Catalytic, Gamma Polypeptide; Phosphorylation; Plasma Proteins; Production; Proteins; Pulmonary Edema; Reaction; Regulation; Reperfusion Injury; Role; Sepsis; Signal Pathway; Signal Transduction; Small Interfering RNA; Superoxides; Testing; Vascular Permeabilities; base; caveolin 1; design; lung injury; monolayer; neutrophil; nitration; novel; protein expression; src-Family Kinases; trafficking; transcytosis; uptake

DESCRIPTION (provided by applicant): Increased lung vascular endothelial permeability induced by activation of polymorphonuclear leukocytes (PMNs) adherent to endothelial cells via ICAM-1 leads to protein-rich pulmonary edema formation and acute lung injury. However, the mechanisms responsible for PMN-induced increased endothelial permeability are incompletely understood. Our supporting data demonstrate that increased caveolae- mediated transcellular transport of albumin may contribute to increased endothelial permeability, and thus may be a factor leading to leaky lung microvessels. In this renewal application, we will address the role of PMNs in inducing the activation of caveolae-dependent transcytosis of albumin and promoting edema formation. The studies will test the hypotheses that (i) fMLP activation of PMNs stimulates caveolae- mediated albumin transcytosis in endothelial cells and mediates the increase in endothelial permeability and lung injury, and (ii) PMN activation of Src kinase in endothelial cells regulates eNOS-mediated NO production via PI3-kinase and Akt signaling and that the NO derived from PMN-endothelial interaction regulates transcellular and junctional permeability pathways to increase lung microvessel permeability. We will delineate the signaling pathways responsible for the activation of endothelial permeability and its consequences in the mechanism of lung edema formation. The approaches to be used include knockout mice, siRNA-induced suppression of protein expression, endothelial permeability assessment of transcellular and paracellular pathways, imaging analysis of caveolae mediated trafficking in live endothelial cells, and biochemical assessments of relevant signaling pathways. Studies in intact mouse lungs will be complemented by studies utilizing mouse lung endothelial cells to provide a more detailed and mechanistic understanding of the signaling basis of PMN-activated increase in endothelial permeability. These studies will provide a novel perspective into the mechanism of increased transendothelial permeability in lungs induced by PMN activation, with the hope of identifying novel molecular targets and better designing strategies directed at treating protein-rich pulmonary edemagenesis and acute lung injury. PROJECT NARRATIVE: Vascular injury and protein-rich pulmonary edema formation are life-threatening complications associated with inflammatory diseases such as ischemia/reperfusion injury and sepsis. We hypothesize that increased transendothelial albumin transport via a caveolar mechanism contributes to neutrophil-induced endothelial hyperpermeability during inflammation. This project therefore addresses the role of caveolae-mediated albumin uptake and transcytosis as compared to paracellular leakage of plasma protein in neutrophil-mediated increase in endothelial permeability and lung injury. We will assess the role of neutrophil activation of Src kinase in endothelial cells in regulating caveolin-1, dynamin-2, and eNOS function, and determine the consequences of increased albumin transport and NO production in regulating the transcellular and junctional permeability pathways.

描述（由申请人提供）：通过ICAM-1激活多形核白细胞（PMN）诱导的肺血管内皮内皮渗透性增加（PMNS）通过ICAM-1依附于内皮细胞，导致富含蛋白质的肺部水肿的形成和急性肺损伤。但是，尚不完全了解导致PMN引起的内皮渗透率增加的机制。我们的支持数据表明，增加的小窝介导的白蛋白的跨细胞转运可能导致内皮渗透性的增加，因此可能是导致肺漏肺微量无丝的因素。在此续签应用中，我们将解决PMN在诱导白蛋白的依赖性跨胞菌病和促进水肿形成的激活中的作用。 The studies will test the hypotheses that (i) fMLP activation of PMNs stimulates caveolae- mediated albumin transcytosis in endothelial cells and mediates the increase in endothelial permeability and lung injury, and (ii) PMN activation of Src kinase in endothelial cells regulates eNOS-mediated NO production via PI3-kinase and Akt signaling and that the NO derived from PMN - 内皮相互作用调节跨细胞和连接性通透性途径，以增加肺微血管通透性。我们将描述负责内皮渗透性激活及其在肺水肿形成机理中的后果的信号通路。要使用的方法包括敲除小鼠，siRNA诱导的蛋白质表达的抑制作用，跨细胞和细胞细胞途径的内皮通透性评估，对生物内皮细胞中的小窝介导的运输的成像分析以及相关信号通路的生物化学评估。完整小鼠肺的研究将通过使用小鼠肺内皮细胞的研究来补充，以对内皮渗透性的PMN激活增加的信号传导基础提供更详细的机械理解。这些研究将为PMN激活诱导的肺中增加的跨内皮渗透性的机制提供新的观点，希望能鉴定出新的分子靶标，并更好地设计旨在治疗富含蛋白质富含蛋白质的肺em耐和急性肺损伤的设计策略。项目叙述：血管损伤和富含蛋白质的肺水肿形成是威胁生命的并发症，与炎症性疾病（如缺血/再灌注损伤和败血症）有关。我们假设通过洞穴机制增加了跨内皮白蛋白的转运，从而有助于炎症期间中性粒细胞诱导的内皮过度透露率。因此，与血浆蛋白在嗜中性粒细胞介导的内皮渗透性和肺损伤中的副细胞泄漏相比，该项目探讨了小窝介导的白蛋白摄取和转胞细胞增多的作用。我们将评估SRC激酶在调节小窝蛋白-1，Dynamin-2和eNOS功能中中性粒细胞激活在内皮细胞中的作用，并确定白蛋白转运增加和NO产生在调节跨细胞和联合通透性途径中的后果。