Mechanotransduction Pathways of Endothelial Barrier Regulation

内皮屏障调节的力传导途径

基本信息

批准号：
8214991
负责人：
Konstantin Birukov
金额：
$ 30.06万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-02-01 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8214991
关键词：
Actin-Binding Protein Actinin Actins Adherens Junction Adhesives Affect Agonist Alveolar Animal Model Atomic Force Microscopy Beryllium Biochemical Blood Platelets Blood Vessels Cell Shape Cell membrane Cells Cellular Structures Cellular biology Cholesterol Complex Critical Illness Cytoskeletal Proteins Cytoskeleton DNA Sequence Rearrangement Development Elements Endothelial Cells Endothelium Event Floods Fluorescence Resonance Energy Transfer Functional disorder Generations Genomics Goals Imaging Techniques In Situ In Vitro Inflammatory Laboratories Lead Link Lung MYLK gene Maintenance Mechanics Mediating Membrane Membrane Microdomains Microfilaments Microscopic Modeling Molecular Molecular Target Mus Pathway interactions Patients Peripheral Permeability Phospholipids Play Principal Investigator Process Protein Array Proteins Recombinant Proteins Regulation Role Signal Transduction Site Stimulus Stretching Structure Supportive care Syndrome Techniques Therapeutic Intervention Tight Junctions Vascular Endothelial Growth Factors Vascular Permeabilities Work base clinically relevant clinically significant extracellular human EMS1 protein in vivo lung injury monolayer novel polymerization programs pulmonary vascular permeability radixin protein receptor repaired response restoration sphingosine 1-phosphate therapeutic target tool

Actin-Binding Protein Actinin Actins Adherens Junction Adhesives Affect Agonist Alveolar Animal Model Atomic Force Microscopy Beryllium Biochemical Blood Platelets Blood Vessels Cell Shape Cell membrane Cells Cellular Structures Cellular biology Cholesterol Complex Critical Illness Cytoskeletal Proteins Cytoskeleton DNA Sequence Rearrangement Development Elements Endothelial Cells Endothelium Event Floods Fluorescence Resonance Energy Transfer Functional disorder Generations Genomics Goals Imaging Techniques In Situ In Vitro Inflammatory Laboratories Lead Link Lung MYLK gene Maintenance Mechanics Mediating Membrane Membrane Microdomains Microfilaments Microscopic Modeling Molecular Molecular Target Mus Pathway interactions Patients Peripheral Permeability Phospholipids Play Principal Investigator Process Protein Array Proteins Recombinant Proteins Regulation Role Signal Transduction Site Stimulus Stretching Structure Supportive care Syndrome Techniques Therapeutic Intervention Tight Junctions Vascular Endothelial Growth Factors Vascular Permeabilities Work base clinically relevant clinically significant extracellular human EMS1 protein in vivo lung injury monolayer novel polymerization programs pulmonary vascular permeability radixin protein receptor repaired response restoration sphingosine 1-phosphate therapeutic target tool

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项目摘要

Disruption of the endothelial cell (EC) barrier that lines the pulmonary vasculature is a central pathophysiologic event in inflammatory lung injury syndromes and results in clinically significant oxygenation derangements in critically ill patients. The goal of Project #2 is to mechanistically characterize linkage between key peripheral EC structures (dynamic cortical actin, lipid rafts, tight junctions, adherens junctions) and the cytoskeletal elements critical to the regulation of lung EC barrier function. The EC cytoskeleton is a complex array of proteins intimately involved in the cell shape changes necessary for regulation of EC barrier function. We have identified essential roles for multiple cytoskeletal linker and effector proteins in the regulation of vascular permeability through modulation of these EC cytoskeletal rearrangements: MLCK, cortactin, actinin, radixin, ZO-1. To assess the importance of these cytoskeletal elements at the periphery of the cell, Specific Aim #1 will define the structure and the regulation of dynamic cortical actin filaments rapidly polymerized at the EC periphery in response to multiple barrier enhancing stimuli. Specific Aim #2 will rigorously characterize the cytoskeletal linkage to membrane-associated lipid rafts, cholesterol-enriched microdomains within the plasma membrane that are important sites for concentrating transmembrane receptors and transducing their signals into the cell. Peripheral cell-cell contacts along the EC monolayer are essential components of barrier maintenance that serve as linkages to the actin cytoskeleton to provide both mechanical stability as well as transduction of extracellular signals into the cell. In the EC, these intercellular contacts consist primarily of two types of junctional complexes¿tight junctions (the focus of Specific Aim #3) and adherens junctions (the focus of Specific Aim #4),whose cytoskeletal linkages will be precisely defined. This project will utilize biochemical, molecular, and cell biology approaches, murine models, translational genomic techniques, and highly novel atomic force microscopy approaches to focus intensely on characterizing the regulation of key peripheral structures by cytoskeletal linker/effector proteins. By mechanistically characterizing key membrane cytoskeletal activators, key cytoskeletal effectors, and the junctional targets affected in these processes, we expect to provide a basis for development of effective therapeutic interventions for pulmonary vascular permeability dysfunction and All.

肺脉管系统的内皮细胞（EC）屏障的破坏是一种中心病理生理炎症性肺损伤综合征的事件，并导致临床上显着的氧合重症患者的进化。项目＃2的目标是机械地表征联系在关键的外围EC结构（动态皮质肌动蛋白，脂质筏，紧密连接，粘附连接）之间以及对调节肺EC屏障功能至关重要的细胞骨架元素。 EC细胞骨架是一个密切涉及细胞形状的复杂蛋白质阵列调节EC屏障所需的变化功能。我们已经确定了多个细胞骨架接头和效应蛋白的重要作用通过调节这些EC细胞骨架重排来调节血管通透性：MLCK， cortactin，阳阳激素，radixin，ZO-1。评估这些细胞骨架元素在周围的重要性细胞，特定的目标＃1将迅速定义动态皮质肌动蛋白丝的结构和调节响应多个屏障增强刺激的响应在EC外围聚合。特定的目标＃2将严格地表征了与膜相关的脂质筏的细胞骨架链接，富含胆固醇质膜内的微区域是浓缩跨膜的重要位点接收器并将其信号转换为单元格。沿EC单层的外围细胞细胞触点是屏障维护的基本组成部分，可作为与肌动蛋白细胞骨架的联系，以提供两者机械稳定性以及细胞外信号转换为细胞。在EC中，这些细胞间触点是两种类型的连接络合物的一致的一致的主要连接连接（特定目的＃3的重点）和粘附连接（特定目标＃4的重点），其细胞骨架链接将得到精确定义。该项目将利用生化，分子和细胞生物学方法，鼠模型，翻译基因组技术和高度新颖的原子力显微镜方法，以积极关注通过细胞骨架接头/效应蛋白来表征关键的周围结构的调节。经过机械地表征关键膜细胞骨架活化剂，关键细胞骨架效应和在这些过程中受影响的连接目标，我们希望为开发有效的发展提供基础肺血管通透性功能障碍的治疗干预措施以及全部。