Fluid shear stress mechanotransduction at endothelial cell-cell junctions

内皮细胞-细胞连接处的流体剪切应力机械转导

基本信息

批准号：
10559534
负责人：
Chenxiang Lin
金额：
$ 53.34万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10559534
关键词：
Address Adhesions Adult Area Arterial Fatty Streak Arteries Atherosclerosis Biochemical Biology Blood Vessels Blood flow Cardiac Cell Line Cells Collaborations Cryoelectron Microscopy Cytoskeleton DNA Data Development Devices Diameter Disease Elements Embryonic Development Endoglin Endothelial Cells Endothelium Event Extracellular Domain Family Foundations G-Protein-Coupled Receptors Genes Heart Valves In Vitro Inflammation Intercellular Junctions KDR gene Knock-out Lead Learning Liquid substance Mammals Mechanics Mechanoreceptors Mediating Mediator Medicine Modeling Molecular Molecular Conformation Mus Myosin ATPase Nanotechnology PECAM1 gene Pathway interactions Pattern Physiological Physiology Play Predisposition Process Protein Conformation Proteins Publishing Role Signal Pathway Signal Transduction Specific qualifier value Testing Trees Vascular Endothelial Cell Visualization Work alpha-latrotoxin receptor bone morphogenetic protein receptors cadherin 5 conformational conversion experimental study in vivo lymphatic valve malformation mechanical force mechanical load mechanotransduction member nanodevice notch protein novel postnatal protein complex response shear stress

项目摘要

Project Summary This project aims to understand in molecular detail how fluid shear stress acting on endothelial cells triggers mechanical activation of signaling pathways at cell-cell junctions. Published data show that shear stress activates a PECAM1-dependent signaling pathway, Notch signaling and Alk1-Endoglin-Smad1/5 signaling, all of which occur at and depend on cell-cell contacts. These pathways play major roles in vascular embryonic development, postnatal physiology and adult disease. However, much remains to be learned about molecular mechanisms. The proposed work is based on two recent advances in our labs. First, we have recently identified latrophilins (LPHNs, also known as ADGRLs), members of the adhesion G protein coupled receptors family, as key upstream mediators of shear activation of all three of these pathways. Second, we have developed a new nanodevice that utilizes DNA origami to apply defined mechanical tension to proteins. Aim 1 will investigate (1) the molecular mechanisms by which LPHNs mediate the effects of shear stress on junctional signaling and (2) determine the role of LPHN2 in vascular development and function in vivo by doing endothelial-specific knockout in mice. Aim 2 will use the DNA origami device to apply defined tension to PECAM1 and visualize protein conformation change via cryoEM. These experiments will allow us to determine the effect of applied force on PECAM’s structural transitions. Together, the project will provide new understanding at unprecedented depth concerning how endothelial cell-cell junctional proteins respond to mechanical force generated by shear stress. .

项目概要该项目旨在从分子角度详细了解流体剪切应力如何作用于内皮细胞细胞触发细胞-细胞连接处信号通路的机械激活。显示剪切应力激活 PECAM1 依赖性信号通路、Notch 信号通路和 Alk1-Endoglin-Smad1/5 信号传导，所有这些都发生并依赖于细胞与细胞的接触。通路在血管胚胎发育、产后生理和成人中发挥重要作用然而，关于分子机制仍有很多知识需要了解。这项工作基于我们实验室的两项最新进展，首先，我们最近发现了懒虫蛋白。（LPHN，也称为 ADGRL），粘附 G 蛋白偶联受体的成员家族，作为所有这三个途径的剪切激活的关键上游介体。我们开发了一种新的纳米设备，利用 DNA 折纸来应用定义的机械目标 1 将研究 (1) LPHN 的分子机制。介导剪切应力对连接信号传导的影响并 (2) 确定 LPHN2 的作用通过在小鼠体内进行内皮特异性敲除来影响体内血管发育和功能。目标 2 将使用 DNA 折纸装置对 PECAM1 施加规定的张力并可视化通过冷冻电镜改变蛋白质构象。这些实验将使我们能够确定该项目将共同提供新的作用力对 PECAM 结构转变的影响。以前所未有的深度了解内皮细胞-细胞连接蛋白如何响应剪切应力产生的机械力。。