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.

项目摘要/摘要血管屏障功能的精确和动态调节，这是血管排列血管的内皮细胞的能力为了在血液和周围组织之间提供有选择的渗透障碍，是普遍的对于维持组织稳态至关重要。失调障碍的病理后果很明显在各种心血管疾病中，诸如动脉粥样硬化和慢性缺血以及炎症性和神经退行性疾病。在体内稳态条件下，血液动力学剪切应力，摩擦力阻力血液流动在内皮细胞上施加，通过细胞 - 细胞粘附连接（AJS）和固有的肌动蛋白细胞骨架动力学的重塑和增强。这临界细胞 - 细胞粘附分子血管内皮（VE-）钙粘蛋白（AJS的原理）调节连接稳定性通过其营业额和内在化，并在张力下经历重大变化剪切应力。此外，肌动蛋白细胞骨架通过保持在动态推动力以维持稳定细胞内AJ复合物的VE-钙粘蛋白和拉伸力。但是，将血液动力学应力与机械联系起来的特定分子传感器和换能器 AJS和血管屏障功能的调节仍然很少理解。无处不在的激活最近发现Notch1受体可以根据剪切应力调节血管屏障功能与VE-钙粘蛋白复合并稳定AJ。虽然以前的工作已经确定了这种缺口途径调节血管屏障功能，尚不清楚剪切应力如何激活Notch1受体。基于将Notch1与固有细胞粘合剂和细胞骨架机械联系起来的初步数据，该建议测试了Notch1与VE-钙粘蛋白和皮质肌动蛋白细胞骨架的内在耦合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的来接近：（1）确定VE-钙粘蛋白如何进行时空调节 Notch1及其配体DLL4通过剪切应力协调激活，（2）确定机械在剪切应力下Notch1和固有性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，我将获得人类微脉管系统的3D仿生模型的培训，超分辨率的活细胞显微镜和机械分子方法，同时增强通过科学沟通，指导和教学培训的职业发展。我已经组装了非凡的补充指导团队，以帮助我实现自己的研究和职业目标：马修博士库蒂（Kutys）是器官组织建模和细胞力学专家，将是我的主要赞助商和黛安（Diane）博士理发师是细胞细胞骨架动力学的领导者，将是我的共同赞助商。最终，这些发现将确定通过剪切应力激活Notch1并有可能识别新机制调节屏障功能和牵涉NOTCH1的其他血管病的治疗靶标。