Shear stress-mediated Notch1 activation by intrinsic cell adhesive and cytoskeletal activity

通过内在细胞粘附和细胞骨架活性剪切应力介导的 Notch1 激活

基本信息

批准号：
10683710
负责人：
Tania Singh
金额：
$ 3.94万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-15 至 2025-02-14
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10683710
关键词：
3-Dimensional Ablation Actins Adherens Junction Adhesives Architecture Atherosclerosis Barbering Binding Biomimetics Biosensor Blocking Antibodies Blood Blood Circulation Blood Plasma Volume Blood Vessels Blood flow C-terminal Cardiovascular Diseases Cell Adhesion Molecules Cell Line Cell membrane Cell-Cell Adhesion Cells Chronic Clathrin Clinical Clustered Regularly Interspaced Short Palindromic Repeats Communication Complex Coupling Cytoskeleton Data Disease Edema Educational process of instructing Endothelial Cells Endothelium Equilibrium Family Friction Genetic Transcription Goals Homeostasis Human Human Development Inflammatory Intervention Ischemia Knowledge Lead Ligands Link Liquid substance Measures Mechanics Mediating Mentors Modeling Molecular Morphogenesis Myocardial Infarction Natural regeneration Neurodegenerative Disorders Nutrient Pathogenesis Pathologic Pathway interactions Permeability Phosphorylation Polymers Process Proteins Proteomics Receptor Activation Regulation Research Resistance Role Signal Transduction Stroke Testing Tissue Model Tissues Training Transducers Vascular Cell Adhesion Molecule-1 Vascular Diseases Vascular Endothelium Vascular Permeabilities Work beta catenin cadherin 5 career career development endothelial dysfunction experience hemodynamics human model improved inhibitor insight live cell microscopy mechanical force mutant new therapeutic target notch protein novel polymerization prevent receptor repaired response sensor shear stress spatiotemporal superresolution microscopy ultra high resolution

项目摘要

PROJECT SUMMARY/ABSTRACT Precise and dynamic regulation of vascular barrier function, the ability of endothelial cells that line blood vessels to provide a selectively permeable barrier between the bloodstream and surrounding tissue, is universally important for maintaining tissue homeostasis. The pathological consequences of dysregulated barrier are evident in various cardiovascular diseases like atherosclerosis and chronic ischemia as well as inflammatory and neurodegenerative diseases. In homeostatic conditions, hemodynamic shear stress, the frictional drag force exerted by the flow of blood on endothelial cells, promotes vascular homeostasis and barrier function through remodeling and enhancement of cell-cell adherens junctions (AJs) and intrinsic actin cytoskeletal dynamics. The critical cell-cell adhesion molecule vascular endothelial (VE-) cadherin, the principle component of AJs, regulates junctional stability through its turnover and internalization and experiences significant changes in tension under shear stress. Additionally, the actin cytoskeleton regulates vascular barrier by maintaining a balance between dynamic pushing forces to maintain VE-cadherin and tensile forces which stabilize intracellular AJ complexes. However, the specific molecular sensors and transducers that link hemodynamic shear stress to the mechanical regulation of AJs and vascular barrier function remain poorly understood. Activation of the ubiquitously important Notch1 receptor was recently been found to modulate vascular barrier function in response to shear stress by complexing with VE-cadherin and stabilizing AJs. While previous work has determined how this Notch1 cortical pathway modulates vascular barrier function, it remains unclear how shear stress activates the Notch1 receptor. Building on preliminary data linking Notch1 to intrinsic cellular adhesive and cytoskeletal machinery, this proposal tests the hypothesis that intrinsic coupling of Notch1 to VE-cadherin and the cortical actin cytoskeleton regulates shear stress-mediated Notch1 activation. Interrogation of Notch1 activation in response to shear stress will be approached by completing two specific aims: (1) determine how VE-cadherin spatiotemporally regulates Notch1 and its ligand Dll4 to coordinate activation by shear stress and (2) identify the mechanical interplay between Notch1 and intrinsic actin cytoskeletal dynamics under shear stress. Throughout the course of the proposed research, I will gain training in 3D biomimetic models of the human microvasculature, super-resolution live cell microscopy, and mechanistic molecular approaches, while simultaneously enhancing career development through training in scientific communication, mentoring, and teaching. I have assembled an exceptional, complementary mentoring team to help me achieve my research and career goals: Dr. Matthew Kutys, an expert in organotypic tissue modeling and cell mechanics will be my primary sponsor and Dr. Diane Barber, a leader in cellular cytoskeletal dynamics, will be my co-sponsor. Ultimately, these findings will identify new mechanisms by which Notch1 is activated in response to shear stress and potentially identify new therapeutic targets for modulating barrier function and other vasculopathies where Notch1 is implicated.

项目概要/摘要精确动态调节血管屏障功能、血管内皮细胞的能力在血流和周围组织之间提供选择性渗透屏障，是普遍的做法对于维持组织稳态很重要。屏障失调的病理后果是显而易见的各种心血管疾病，如动脉粥样硬化、慢性缺血以及炎症和神经退行性疾病。在稳态条件下，血流动力学剪切应力、摩擦阻力血流对内皮细胞产生作用，通过促进血管稳态和屏障功能细胞-细胞粘附连接（AJ）的重塑和增强以及内在肌动蛋白细胞骨架动力学。这关键的细胞间粘附分子血管内皮 (VE-) 钙粘蛋白是 AJ 的主要成分，可调节通过其周转和内化实现连接稳定性，并在压力下经历显着变化剪切应力。此外，肌动蛋白细胞骨架通过维持之间的平衡来调节血管屏障。维持 VE-钙粘蛋白的动态推力和稳定细胞内 AJ 复合物的张力。然而，将血流动力学剪切应力与机械应力联系起来的特定分子传感器和换能器 AJ 和血管屏障功能的调节仍知之甚少。激活无处不在的重要最近发现 Notch1 受体可以通过以下方式调节血管屏障功能以响应剪切应力与 VE-钙粘蛋白复合并稳定 AJ。虽然之前的工作已经确定了 Notch1 皮质如何虽然剪切力如何调节血管屏障功能，但剪切应力如何激活 Notch1 受体仍不清楚。该提案基于将 Notch1 与内在细胞粘附和细胞骨架机制联系起来的初步数据检验 Notch1 与 VE-钙粘蛋白和皮质肌动蛋白细胞骨架的内在耦合调节的假设剪切应力介导的 Notch1 激活。响应剪切应力的 Notch1 激活的询问将是通过完成两个具体目标来实现：（1）确定 VE-钙粘蛋白如何时空调节 Notch1 及其配体 Dll4 通过剪切应力协调激活，并 (2) 识别机械剪切应力下 Notch1 和内在肌动蛋白细胞骨架动力学之间的相互作用。整个在拟议的研究过程中，我将获得人体微脉管系统 3D 仿生模型的培训，超分辨率活细胞显微镜和机械分子方法，同时增强通过科学传播、指导和教学培训进行职业发展。我已经组装了一个卓越、互补的指导团队帮助我实现我的研究和职业目标：Matthew 博士 Kutys 是器官组织建模和细胞力学方面的专家，他将是我的主要赞助商，Diane 博士将是我的主要赞助商巴伯是细胞骨架动力学领域的领军人物，他将成为我的共同发起人。最终，这些发现将确定 Notch1 响应剪切应力而被激活的新机制，并可能识别新的调节屏障功能和其他涉及 Notch1 的血管疾病的治疗靶点。