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.

项目概要 血管功能和发育主要由血管内皮细胞（VEC）介导 血管内壁的流体剪切应力 (FSS) 是由血流产生的。 其功能和表型的主要决定因素，在发育、生理学、 健康动脉中的 VEC 处于单向层流状态，其中 它们沿着流动方向排列并激活抗炎途径，从而赋予 相比之下，动脉弯曲或分支区域的 VEC 具有抗动脉粥样硬化的作用。 这些扰动的流动模式无法对齐 VEC 和 激活炎症途径，这与形成动脉粥样硬化的易感性相关 FSS 相对于细胞排列的方向也调节炎症信号传导。 输出，这表明细胞极性和流动方向感测对于微分很重要 因此，VEC 如何感知和响应血流。 方向是关系人类健康的重要基础科学问题，但其作用机制 尚不清楚由 VE-Cahderin、PECAM1 和 组成的连接流依赖性复合物。 我们的实验室最近发现，VEGF 对于整合内皮细胞流反应至关重要。 极性接头蛋白 LGN 直接与 VE-Cahderin 结合，对于正确 由于 LGN 直接与流量依赖性机械敏感相互作用。 复杂并且具有调节细胞骨架动力学的既定作用，我导致 LGN 是 对于流向传感很重要，我计划使用以下具体内容来解决这个假设。 目标：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依赖性信号传导很重要。 如果已知的 LGN 相互作用因子被证明对 目标 2：检查细胞极性在血流信号传导中的作用。 首先通过跟踪 LGN 响应流量和使用模式的细胞内定位 分别限制细胞和细胞骨架极性的底物，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非常重要 目标 3： 通过分析 LGN 内皮缺失的小鼠来确定 LGN 在体内的作用，这将 解决 LGN 在 VEC 排列、炎症和由此产生的体内动脉粥样硬化中的作用。 总之，这些目标将揭示内皮血流传感、血管 炎症和动脉粥样硬化疾病。