Contribution of Endothelial Planar Cell Polarity pathways in Blood Flow Direction Sensing

内皮平面细胞极性通路在血流方向传感中的贡献

• 批准号: 10750690
• 负责人: Shaka X
• 金额: $ 4.77万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750690
• 关键词: Adaptor Signaling Protein; Address; Anti-Inflammatory Agents; Arterial Fatty Streak; Arteries; Atherosclerosis; Basic Science; Binding; Blood Vessels; Blood flow; Cadherins; Cardiovascular Diseases; Cell Line; Cell Polarity; Cell Shape; Cells; Cellular Morphology; Complex; Cytoskeleton; Development; Disease; Endothelial Cells; Endothelium; GPSM2 gene; Health; Human; Immunoprecipitation; Inflammation; Inflammatory; KDR gene; Link; Liquid substance; Mediating; Mediator; Molecular; Mus; Mutation; Output; PECAM1 gene; Pathway interactions; Pattern; Phenotype; Physiology; Predisposition; Research; Resistance; Role; Signal Transduction; Site; Variant; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; atheroprotective; cadherin 5; in vivo; link protein; new therapeutic target; planar cell polarity; polarized cell; prevent; protective pathway; response; shear stress; thrombotic; vascular inflammation

PROJECT SUMMARY Vascular function and development are largely mediated by vascular endothelial cells (VECs) that line the inner wall of blood vessels. Fluid shear stress (FSS) generated by blood flow is a major determinant of their function and phenotype with major roles in development, physiology, and disease. VECs in healthy regions of arteries are under unidirectional laminar flow, where they align in the direction of flow and activate anti-inflammatory pathways which confers resistance to atherosclerosis. By contrast, VECs in curved or branched regions of arteries develop disturbances in flow patterns. These disturbed flow patterns fail to align VECs and activate inflammatory pathways, which correlates with susceptibility to form atherosclerotic plaque. FSS direction with respect to cell alignment also regulates inflammatory signaling outputs, which suggests cell polarity and flow direction sensing is important for the differential atheroprotective and atheroprone responses. Thus, how VECs sense and respond to flow direction is an important basic science question pertinent to human health, but the mechanism is unclear. A junctional flow-dependent complex comprising of VE-Cahderin, PECAM1, and VEGF is critical for integrating endothelial cell flow responses. Our lab recently discovered that the polarity adaptor protein, LGN, which binds directly to VE-Cahderin is important for proper endothelial cell alignment. Since LGN directly interacts with a flow dependent mechanosensitive complex and has an established role regulating cytoskeletal dynamics, I hypothesize LGN is important for flow direction sensing. I plan to address this hypothesis using the following specific aims: Aim 1: Characterize the mechanism by generating mutations in LGN’s functional domains to determine which sites are important for mediating endothelial cell flow dependent signaling. I will similarly examine the effects of known LGN interactors if they are shown to be crucial for flow mediated signaling. Aim 2: Examine the role of cell polarity in flow signaling. I will do this first by tracking the intracellular localization of LGN in response to flow and use patterned substrates to separately constrain cell and cytoskeletal polarity to determine which of these variables is important for LGN polarity and inflammatory vs. anti-inflammatory signaling. Aim 3: Determine the role of LGN in vivo by analyzing mice with endothelial deletion of LGN which will address the role of LGN in VEC alignment, inflammation, and resulting atherosclerosis in vivo. Together, these aims will reveal new mechanisms for endothelial flow sensing, vascular inflammation, and atherosclerotic disease.

项目摘要 血管功能和发育主要由血管内皮细胞（VEC）介导 那条血管内壁。血流产生的流体剪应力（FSS）是 主要确定其功能和表型，在发育，生理学，生理学， 和疾病。健康动脉健康区域的VEC在单向层流下，其中 它们在流动方向并激活抗炎途径的方向 对动脉粥样硬化的抵抗力。相比之下，动脉弯曲或分支区域中的VEC 在流动模式中发展出令人惊讶的形式。这些受影响的流动模式无法对齐VEC，并且 激活炎症途径，这与形成动脉粥样硬化的易感性相关 牌匾。 FSS方向相对于细胞比对也调节炎症信号传导 输出表明细胞极性和流动方向灵敏度对于差分很重要 动脉保护性和动脉op骨反应。那是如何感知和回应流动的 方向是与人类健康有关的重要基本科学问题，但机制 不清楚。 VE-Cahderin，PECAM1和 VEGF对于整合内皮细胞流动反应至关重要。我们的实验室最近发现 直接与Ve-Cahderin结合的极性适配器蛋白LGN对于适当的 内皮细胞对齐。由于LGN直接与流动依赖机制相互作用 复合物，并且具有调查细胞骨架动力学的确定作用，我假设LGN是 对于流动方向灵敏度很重要。我计划使用以下特定的特定来解决这一假设 目的：目标1：通过在LGN功能域中产生突变来表征机制 确定哪些位点对于介导内皮细胞流量依赖信号传导很重要。我 如果证明对已知的LGN相互作用者的效果类似 流动介导的信号传导。 AIM 2：检查细胞极性在流动信号传导中的作用。我会这样做 首先，通过跟踪LGN的细胞内定位，以响应流动和使用图案 底物分别限制细胞和细胞骨架极性以确定其中哪一个 变量对于LGN极性和炎症与抗炎信号传导很重要。目标3： 通过用LGN的内皮缺失分析的小鼠来确定LGN在体内的作用 解决了LGN在体内的VEC比对，炎症和导致的动脉粥样硬化中的作用。 这些目标在一起将揭示内皮流动感，血管的新机制 炎症和动脉粥样硬化疾病。