Novel Mechanisms Controlling Endothelial Junctions and Vascular Permeability

控制内皮连接和血管通透性的新机制

• 批准号: 10630183
• 负责人: BARRY M. GUMBINER
• 金额: $ 61万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10630183
• 关键词: Actins; Acute; Acute Respiratory Distress Syndrome; Adherens Junction; Adhesions; Adhesives; Affect; Allosteric Regulation; Angiopoietins; Biochemical; Biophysics; Blood; Blood Vessels; CDH5 gene; Cadherins; Cell surface; Cytoskeleton; Development; Disease; Edema; Endocytosis; Endothelial Cells; Endothelium; Environment; Epithelium; Event; Hemorrhage; Histamine; In Vitro; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Intercellular Junctions; Ischemic Stroke; Leucocytic infiltrate; Leukocytes; Mediating; Microcirculatory Bed; Modeling; Monoclonal Antibodies; Morphogenesis; Mus; Neoplasm Metastasis; Organ; Permeability; Phosphorylation; Physiological Processes; Process; Proteins; Publications; Regulation; Role; Route; Sepsis; Signal Transduction; Stream; Structure; Surface; TIE-2 Receptor; Testing; Therapeutic; Thrombin; Tight Junctions; Tissues; Vascular Endothelial Growth Factors; Vascular Endothelium; Vascular Permeabilities; Work; angiogenesis; cadherin 5; cell assembly; experimental study; in vivo; inhibiting antibody; interest; interstitial; mouse model; novel; novel strategies; prevent; response; tool; tumor; vascular factor

Project Summary The vascular endothelium forms a highly regulated permeability barrier between the blood stream and interstitial tissues. Excessive vascular leakiness contributes to many inflammation related disease processes, including edema, sepsis, acute respiratory distress syndrome (ARDS), ischemic stroke, and hemorrhage. To prevent tissue damage resulting from vascular leakiness during inflammation, there is a need to be able to enhance endothelial barrier function. Intercellular junctions control changes in endothelial paracellular permeability, and Vascular-endothelial cadherin (VE-cadherin, CDH5) is a major regulator of endothelial junctions and paracellular permeability. We will explore a novel hypothesis and approach to understanding its role in permeability regulation, based on our findings of allosteric regulation of other cadherins at the cell surface in response to signaling events. The main hypothesis to be examined is that allosteric regulation of VE- cadherin, in coordination with activities of the actin cytoskeleton, is a key mechanism by which it regulates endothelial permeability in response to a range of vascular factors. We will use two novel activating monoclonal antibodies (mAbs) to VE-cadherin that prevent the increase in endothelial permeability induced by thrombin, VEGF, and TNFa, as major tools to test this hypothesis both in vitro and in vivo and to study the mechanism of regulation. The specific aims are: A. Determine whether VE-cadherin cell surface regulation is a common mechanism for a range of endothelial physiological processes. In vitro experiments will be used to determine whether it is similarly regulated by other key factors, including histamine, angiopoietins, Tie2 receptors, and S1P. Endothelial cells from different microvascular beds will be examined. Collaborative experiments will be done to determine flow and shear forces affect mAb induced barrier function. We will also investigate whether activating mAbs inhibit leukocyte diapedesis or affect endothelial morphogenesis and angiogenesis. B. Elucidate the cellular and biochemical mechanisms underlying VE-cadherin cell surface regulation. We will investigate the structural and biophysical basis of its activation by mAbs and test the roles of the phosphorylation of VE-cadherin and associated catenins in activation. We’ll also explore the relationship between cell surface regulation and endocytosis as well as barrier altering cytoskeletal functions. C. Investigate whether activation of VE-cadherin by mAbs enhances barrier function in vivo in mice in leaky vascular conditions or inflammation. We’ll test the effects of activating mAbs on induced acute induced vascular leak, and leukocyte infiltration. We will also examine their effects on mouse models of inflammatory disease processes involving the vasculature, including sepsis and Inflammatory Bowel Disease. These studies will help us understand regulation of endothelial permeability and develop novel approaches to enhance endothelial barrier function to reduce inflammatory processes and diseases.

项目概要 血管内皮在血流和血管之间形成高度调节的渗透性屏障。 间质组织过度渗漏会导致许多炎症相关的疾病过程， 包括水肿、败血症、急性呼吸窘迫综合征 (ARDS)、缺血性中风和出血。 为了防止炎症期间因血管渗漏而造成的组织损伤，需要能够 增强内皮屏障功能。细胞间连接控制内皮细胞旁细胞的变化。 通透性和血管内皮钙粘蛋白（VE-cadherin，CDH5）是内皮细胞的主要调节因子 我们将探索一种新的假设和方法来理解它。 基于我们对细胞中其他钙粘蛋白的变构调节的发现，在通透性调节中的作用 表面对信号事件的反应 要检验的主要假设是 VE- 的变构调节。 钙粘蛋白与肌动蛋白细胞骨架的活动相协调，是其调节的关键机制 我们将使用两种新颖的激活剂来调节内皮通透性对一系列血管因素的反应。 VE-钙粘蛋白单克隆抗体 (mAb) 可防止 VE-钙粘蛋白引起的内皮通透性增加 凝血酶、VEGF 和 TNFa 作为主要工具在体外和体内检验这一假设并研究 具体目标是： A. 确定 VE-钙粘蛋白细胞表面调节是否是一种调节机制。 一系列内皮生理过程的共同机制将用于体外实验。 确定它是否受到其他关键因素的类似调节，包括组胺、血管生成素、Tie2 受体和 S1P 来自不同微血管床的内皮细胞将被协作检查。 我们还将进行实验以确定流动力和剪切力对 mAb 屏障诱导功能的影响。 研究激活的单克隆抗体是否会抑制白细胞渗出或影响内皮形态发生 B. 阐明 VE-钙粘蛋白细胞表面的细胞和生化机制。 我们将研究单克隆抗体激活其的结构和生物物理基础并测试其作用。 我们还将探讨 VE-钙粘蛋白和相关连环蛋白的磷酸化在激活中的关系。 细胞表面调节和内吞作用以及改变细胞骨架功能的屏障之间的关系。 C. 研究 mAb 激活 VE-钙粘蛋白是否会增强渗漏小鼠体内的屏障功能 我们将测试激活单克隆抗体对诱发急性诱发的影响。 我们还将检查它们对小鼠炎症模型的影响。 涉及脉管系统的疾病过程，包括败血症和炎症性肠病这些研究。 将帮助我们了解内皮通透性的调节并开发新的方法来增强 内皮屏障功能可减少炎症过程和疾病。