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（由申请方提供）：血管发育和伤口愈合均依赖于内皮细胞粘附的动态控制。然而，不适当的粘连损失会导致炎症状态下的病理学。细胞-细胞连接的调节控制内皮细胞粘附，了解连接调节的机制将有助于了解正常的血管发育和疾病。在这个应用程序中，我们试图调查的作用，内吞作用的调节血管内皮钙粘蛋白（VE-钙粘蛋白），在内皮粘附连接的主要粘附分子。尽管VE-钙粘蛋白对血管发育、血管生成和炎症的重要性，但对其调控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。我们实验室以前的工作表明，p120-catenin（p120），一种与VE-钙粘蛋白胞质尾结合的Armadillo家族蛋白，保护VE-钙粘蛋白免于快速内化和降解。我们假设，破坏p120与VE-钙粘蛋白的结合揭示了一个内吞衔接子结合位点，导致VE-钙粘蛋白的内化和内皮细胞粘附力的丧失。为了检验这一假设，我们将追求以下具体目标。首先，我们将鉴定VE-钙粘蛋白胞质尾区中介导接头结合和内吞作用的氨基酸序列。我们还将测试VE-钙粘蛋白突变的功能后果，防止内吞作用。其次，我们将使用泛素连接酶K5，这导致VE-钙粘蛋白的快速下调，作为一个模型，以确定是否破坏p120结合被用作一种机制，以控制VE-钙粘蛋白水平。我们将确定目前未知的机制，其中K5导致VE-钙粘蛋白的下调和它的关系，内源性途径VE-钙粘蛋白的控制。我们的长期目标是揭示内皮细胞如何调节细胞-细胞粘附的强度，从而更好地理解血管生成和炎症性疾病的新治疗方法。 公共卫生相关性：控制血管中的细胞-细胞粘附对于血管发育和伤口愈合至关重要。不适当的粘连损失会导致炎症和心脏病、中风和严重感染等疾病引起的组织损伤。研究血管中细胞粘附的调节将为这些疾病确定潜在的新治疗方法。