Role of End Binding 3 in Mechanism of vascular permeability

末端结合3在血管通透性机制中的作用

• 批准号: 8050461
• 负责人: Yulia A Komarova
• 金额: $ 39.25万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050461
• 关键词: Actins; Address; Adhesions; Adult Respiratory Distress Syndrome; Affinity; Agonist; Binding; Binding Proteins; Biochemical; Blood Vessels; Calcineurin; Calmodulin; Cell Shape; Cells; Complex; Cytoskeleton; Data; Development; Down-Regulation; Edema; Endoplasmic Reticulum; Endothelium; Event; Exhibits; Extravasation; Functional disorder; Gases; Gene Transfer; Genetic; Growth; Homeostasis; Hypoxemia; Image; Impairment; Inflammation; Inflammation Mediators; Inflammatory; Inositol; Life; Liquid substance; Lung; Lung Inflammation; Maintenance; Mediating; Microtubules; Modeling; Molecular; Mus; Myosin Light Chain Kinase; Pathway interactions; Peptides; Permeability; Phosphorylation; Phosphorylation Inhibition; Physiological; Plasma; Plus End of the Microtubule; Protein Kinase C; Protein Serine/Threonine Phosphatase; Proteins; Pulmonary Edema; Regulation; Role; SRC gene; Sepsis; Signal Transduction; Stimulus; Technology; Testing; Therapeutic; Time; Tissues; Vascular Permeabilities; Work; base; cadherin 5; cellular imaging; design; expectation; insight; mutant; novel; novel therapeutics; prevent; receptor; response

DESCRIPTION (provided by applicant): Increases in lung vascular permeability result in protein rich tissue edema, an important feature of adult respiratory distress syndrome. The role of the microtubule (MT) cytoskeleton in the mechanism of increased endothelial permeability is not well understood. MTs are known to undergo re-organization in response to pro- inflammatory mediators, and may thus contribute to the mechanism of increased endothelial permeability. The proposed studies will address the central role of End Binding protein-3 (EB3), a MT plus-end binding factor, in regulating increased lung vascular permeability. We will test the hypotheses that (i) EB3 is a major component of cross-talk between Vascular Endothelial (VE)-cadherin adhesion complexes and the MT cytoskeleton and (ii) EB3-mediated control of MT dynamics is critical for maintenance of basal permeability of lung microvessels and for permeability increase caused by inflammatory mediators. These studies will address the following Specific Aims: (1) role of VE-cadherin-mediated signaling in the mechanism of EB3 phosphorylation and inhibition of MT growth and, thereby the role of "outside-in" signaling in establishing basal permeability of lung endothelia; and (2) critical role of EB3 in regulating Ca2+ signaling, thus in mediating increased endothelial permeability and development of lung edema. It is our expectation that by understanding how VE-cadherin adhesion signals EB3 phosphorylation and how EB3 thereby elicits the barrier permeability increase will provide novel insights into the mechanisms of dysregulation of lung fluid homeostasis. We will exploit state of the art technologies including live cell imaging, expression of mutant constructs, gene transfer, and murine models of lung inflammation to accomplish the specific aims. PUBLIC HEALTH RELEVANCE: The focus of the work is the lung endothelium, in which the planned studies will establish the relevance of EB3, a microtubule-binding protein, to the pathophysiology of ALI/ ARDS. The work leading up to this proposal has allowed us to design a potential therapeutic peptide that by inhibiting EB3 function prevents lung edema and lethality in sepsis.

描述（由申请人提供）：肺血管通透性增加导致富含蛋白质的组织水肿，这是成人呼吸窘迫综合征的重要特征。微管（MT）细胞骨架在增加内皮通透性的机理中的作用尚不清楚。已知MT会对促炎性介质进行重新组织，因此可能有助于内皮渗透性增加的机制。拟议的研究将解决终端结合蛋白3（EB3）（MT加末端结合因子）在调节肺血管渗透性增加的核心作用。我们将测试（I）EB3是血管内皮（VE） - 钙粘着蛋白粘附复合物与MT细胞骨架和（II）EB3介导的MT动态控制对维持肺部微量弥生和渗透剂导致基础渗透性增加的eB3介导的控制至关重要的假设。这些研究将解决以下特定目的：（1）VE-钙粘蛋白介导的信号传导在EB3磷酸化机制和抑制MT生长的机理中的作用，从而在建立肺内皮层的基础渗透性中的“外部”信号传导的作用； （2）EB3在调节Ca2+信号传导中的关键作用，因此在介导增加的内皮渗透性和肺水肿的发育中。我们期望通过了解VE-钙粘着蛋白的粘附如何信号EB3磷酸化以及EB3如何引起屏障渗透性的增加将提供对肺部稳态失调机制的新见解。我们将利用艺术技术的状态，包括活细胞成像，突变构建体的表达，基因转移和肺部炎症的鼠模型，以实现特定目的。 公共卫生相关性：工作的重点是肺部内皮，在该细胞中，计划的研究将确定微管结合蛋白EB3与ALI/ ARDS的病理生理学的相关性。导致该提案的工作使我们能够设计一种潜在的治疗肽，该肽通过抑制EB3功能可防止败血症中的肺水肿和致死性。