Regulation of endothelial adhesion by VE-cadherin endocytosis

VE-钙粘蛋白内吞作用对内皮粘附的调节

• 批准号: 8315132
• 负责人: Benjamin Andrew Nanes
• 金额: $ 3.09万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8315132
• 关键词: Adaptor Protein Complex 2; Adaptor Signaling Protein; Adherens Junction; Adhesions; Amino Acid Sequence; Anti-Inflammatory Agents; Anti-inflammatory; Binding; Binding Sites; Blood Vessels; Cadherins; Cell Adhesion; Cell Adhesion Molecules; Cell membrane; Cell-Cell Adhesion; Cells; Clathrin; Cytoplasmic Tail; Development; Disease; Dissociation; Down-Regulation; Dynamin; Edema; Endocytosis; Endothelial Cells; Family; Family Dasypodidae; Functional disorder; Goals; Hemorrhage; Human Herpesvirus 8; Infection; Inflammation; Inflammatory; Intercellular Junctions; Knowledge; Measures; Mediating; Membrane Protein Traffic; Membrane Proteins; Modeling; Mutation; Myocardial Infarction; Pathology; Pathway interactions; Permeability; Protein Binding; Regulation; Role; Site; Stroke; Surface; Testing; Tissues; Tube; Ubiquitination; Vascular Permeabilities; Work; Wound Healing; angiogenesis; cadherin 5; cell motility; drug development; human disease; insight; monolayer; mouse model; prevent; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): Vascular development and wound healing both depend on dynamic control of endothelial cell adhesion. However, inappropriate loss of adhesion contributes to pathology in inflammatory conditions. Modulation of cell-cell junctions controls endothelial adhesion, and understanding the mechanisms of junction regulation will lend insight into normal vascular development and disease. In this application, we seek to investigate the role of endocytosis in the regulation of vascular endothelial cadherin (VE-cadherin), the main adhesion molecule in endothelial adherens junctions. Despite the importance of VE-cadherin to vessel development, angiogenesis, and inflammation, relatively little is known about the mechanisms of its regulation. One way cells regulate the abundance of plasma membrane proteins such as VE-cadherin is through membrane trafficking. Previous work by our lab demonstrates that p120-catenin (p120), an Armadillo family protein that binds to the VE- cadherin cytoplasmic tail, protects VE-cadherin from rapid internalization and degradation. We hypothesize that disrupting p120 binding to VE-cadherin unmasks an endocytic adaptor binding site, leading to internalization of VE-cadherin and loss of adhesion in endothelial cells. To test this hypothesis, we will pursue the following specific aims. First, we wll identify the amino acid sequences in the VE-cadherin cytoplasmic tail which mediate adaptor binding and endocytosis. We will also test the functional consequences of mutations in VE-cadherin which prevent endocytosis. Second, we will use the ubiquitin ligase K5, which causes rapid down-regulation of VE-cadherin, as a model to determine if disrupting p120 binding is used as a mechanism to control VE-cadherin levels. We will identify the currently unknown mechanism by which K5 causes VE- cadherin down-regulation and its relationship to endogenous pathways for VE-cadherin control. Our long-term goal is to reveal how endothelial cells modulate the strength of cell-cell adhesion, leading to a better understanding of angiogenesis and new treatments for inflammatory disease. PUBLIC HEALTH RELEVANCE: Control of cell-cell adhesion in blood vessels is critical for vessel development and wound healing. Inappropriate loss of adhesion contributes to inflammation and tissue damage from diseases like heart attack, stroke, and severe infection. Studying the regulation of cell adhesion in blood vessels will identify potential new treatments for these diseases.

描述（由申请人提供）：血管发育和伤口愈合都依赖于内皮细胞粘附的动态控制。然而，不适当的失去粘连有助于病理炎症条件。细胞-细胞连接的调节控制内皮粘附，了解连接调节的机制将有助于了解正常血管发育和疾病。在这个应用中，我们试图研究内吞作用在血管内皮钙粘蛋白（VE-cadherin）的调节中的作用，钙粘蛋白是内皮粘附连接中的主要粘附分子。尽管ve -钙粘蛋白对血管发育、血管生成和炎症的重要性，但对其调节机制的了解相对较少。细胞调节质膜蛋白如ve -钙粘蛋白丰度的一种方式是通过膜运输。我们实验室之前的工作表明，p120-catenin （p120）是一种犰狳家族蛋白，与VE-钙粘蛋白细胞质尾部结合，保护VE-钙粘蛋白免受快速内化和降解。我们假设，破坏p120与VE-cadherin的结合可以揭示一个内吞接头结合位点，从而导致VE-cadherin内化和内皮细胞的粘附丧失。为了验证这一假设，我们将追求以下具体目标。首先，我们将确定ve -钙粘蛋白细胞质尾部介导适配器结合和内吞作用的氨基酸序列。我们还将测试阻止内吞作用的ve -钙粘蛋白突变的功能后果。其次，我们将使用导致VE-cadherin快速下调的泛素连接酶K5作为模型，以确定破坏p120结合是否作为控制VE-cadherin水平的机制。我们将确定目前未知的K5导致VE-cadherin下调的机制及其与内源性VE-cadherin控制途径的关系。我们的长期目标是揭示内皮细胞如何调节细胞间粘附的强度，从而更好地理解血管生成和炎症疾病的新治疗方法。