Endothelial focal adhesions in microvascular barrier dysfunction during ischemia-

缺血期间微血管屏障功能障碍中的内皮粘着斑

基本信息

批准号：
9276101
负责人：
MACK H WU
金额：
$ 37.38万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9276101
关键词：
Actinin Adherens Junction Adhesions Adhesives Antioxidants Basement membrane Binding Biochemical Biological Assay Brain hemorrhage Bypass CD47 gene Cell Adhesion Molecules Cell physiology Cell-Cell Adhesion Cells Cellular biology Clinical Complement Complex Cytoskeleton Development Disease Disseminated Malignant Neoplasm Dissociation Effectiveness Endothelial Cells Endothelium Evaluation Extracellular Matrix Extravasation Fibrinogen Focal Adhesion Kinase 1 Focal Adhesions Functional disorder Future Gene Mutation Gene Silencing Goals Hydrogen Peroxide Hypoxia Image Analysis Immune Impairment Individual Inflammation Inflammation Mediators Inflammatory Inflammatory Response Injury Integrin alpha5beta1 Integrins Intercellular Junctions Intestines Investigation Ischemia Knock-out Knowledge Leukocytes Liquid substance Maintenance Measurement Mediating Mesentery Microcirculation Microscopic Microvascular Dysfunction Microvascular Permeability Modality Molecular Mus Myocardial Infarction Nuclear Translocation Operative Surgical Procedures Organ Organ Transplantation Oxidants Oxidative Stress Pathway interactions Patients Peptides Permeability Pharmaceutical Preparations Pharmacology Physiological Plasma Play Plug-in Prevention Process Property Proteins Reaction Reactive Oxygen Species Regulation Reperfusion Injury Reperfusion Therapy Resuscitation Role Series Signal Transduction Stimulus Structure Superior mesenteric artery structure Talin Techniques Testing Therapeutic Therapeutic Effect Three-Dimensional Imaging Thrombosis Tissues Trauma Vinculin angiogenesis artery occlusion beta catenin clinical application comparative endothelial dysfunction experimental study in vivo injured innovation insight intravital microscopy knock-down knockout gene microvascular pathology mouse model neutrophil novel paxillin prevent public health relevance receptor release factor research and development response stem targeted treatment therapeutic evaluation venule

项目摘要

DESCRIPTION (provided by applicant): Ischemia/reperfusion (I/R) injury triggers a series of inflammatory responses in the microcirculation characterized by plasma leakage and leukocyte diapedesis. The microvascular pathology largely stems from endothelial barrier dysfunction, a complex cellular process that has not been fully understood at the molecular level. Emerging evidence indicates that the barrier property of the microvascular wall is controlled by dynamic interactions of endothelial cell-cell junctions and cell-matrix focal adhesions; the latter are maily composed of transmembrane integrin receptors and associated proteins such as kindlins and focal adhesion kinase (FAK). These molecules not only provide structural support for endothelial barrier integrity but also transmit biochemical signals that regulate barrier function. The goal of this project is to elucidate the molecular mechanisms underlying FAK-mediated microvascular hyperpermeability during I/R injury. We hypothesize that I/R-elicited oxidative stress upregulates FAK signaling in the microvascular endothelium, inducing integrin (α5β1, αvβ3) internalization and kindlin- targeted β-catenin nuclear translocation and junction dissociation, leading to weakened cell-matrix and cell-cell adhesions thereby impairing barrier integrity. This novel pathway will be tested in a series of mechanistic studies centered on intravital microscopic quantification of mesenteric microcirculation, leukocyte dynamics, and fluid/protein permeability in mice subjected to superior mesenteric artery occlusion followed by reperfusion. A newly developed mouse model of endothelial-specific conditional FAK knockout will be used in comparison with pharmacological inhibition of FAK. The in vivo studies will be complemented with imaging analyses and molecular assays in microvascular endothelial cells exposed to hypoxia/reoxygenation or oxidative stress. The studies will provide new mechanistic insights into I/R-induced microvascular injury contributing to the future development of targeted therapies to prevent tissue damage following resuscitation or reperfusion. Knowledge gained from this project may have broad implications in other diseases associated with microvascular barrier injury.

描述（由适用提供）：缺血/再灌注（I/R）损伤触发了以血浆泄漏和白细胞尿症为特征的微循环中的一系列炎症反应。微血管病理学很大程度上始于内皮屏障功能障碍，这是一种复杂的细胞过程，在分子水平尚未完全了解。新兴的证据表明，微血管壁的屏障特性受到内皮细胞 - 细胞连接和细胞矩阵焦点粘合剂的动态相互作用的控制。后者是由跨膜整合素受体和相关蛋白（例如Kindlins和焦点粘合剂激酶（FAK））组成的邮件。这些分子不仅为内皮屏障完整性提供结构支持，而且还传递调节屏障功能的生化信号。目标该项目是为了阐明I/R损伤过程中FAK介导的微血管过敏性的分子机制。我们假设I/r-催化的氧化应激上调微血管内皮中的FAK信号传导，诱导整联蛋白（α5β1，αVβ3）内在化和Kindlin靶向的β-catenin核易位和连接分离，从而导致细胞 - 晶体质量和细胞纤维粘附在此处的脑型较弱的核心性。这一新途径将在一系列机械研究中进行测试，该研究以肠内微循环，白细胞动力学和液/蛋白质渗透性的插入式微观定量为中心，在受到上肠系膜上动脉闭塞的小鼠中，然后再灌注。与FAK的药物抑制相比，将使用新开发的内皮特异性条件FAK敲除敲除的小鼠模型。体内研究将通过暴露于低氧/氧化或氧化应激的微血管内皮细胞中的成像分析和分子测定完成。这些研究将为I/R诱导的微血管损伤提供新的机械见解，这有助于未来的靶向疗法发展，以防止复苏或再灌注后组织损伤。从该项目中获得的知识可能对与微血管屏障损伤相关的其他疾病具有广泛的影响。