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

• 批准号: 10389629
• 负责人: Tania Singh
• 金额: $ 3.85万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 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/10389629
• 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; Cardiovascular system; 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; 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; Microscopy; Modeling; Molecular; Morphogenesis; Myocardial Infarction; Natural regeneration; Neurodegenerative Disorders; Nutrient; Pathogenesis; Pathologic; Pathway interactions; Permeability; Phosphorylation; Play; Process; Proteins; Proteomics; Receptor Activation; Regulation; Research; Resistance; Resolution; Role; Signal Transduction; Stroke; Testing; Tissue Model; Tissues; Training; Transducers; Ursidae Family; 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

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.

项目总结/摘要 精确和动态调节血管屏障功能，即内衬血管的内皮细胞的能力 为了在血流和周围组织之间提供选择性渗透屏障， 对维持组织内环境稳定很重要。屏障失调的病理后果是显而易见的 在各种心血管疾病如动脉粥样硬化和慢性缺血以及炎症和 神经退行性疾病在稳态条件下，血流动力学剪切力、摩擦阻力 通过血液流动对内皮细胞的作用，促进血管稳态和屏障功能， 细胞-细胞粘附连接（AJs）和内在肌动蛋白细胞骨架动力学的重塑和增强。的 关键的细胞-细胞粘附分子血管内皮（VE-）钙粘蛋白，AJs的主要成分，调节 通过其周转和内在化的连接稳定性，并在张力下经历重大变化， 剪应力此外，肌动蛋白细胞骨架通过维持细胞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， AJs的调节和血管屏障功能仍然知之甚少。激活无处不在的重要 最近发现Notch 1受体通过调节血管屏障功能来响应剪切应力， 与VE-钙粘蛋白复合并稳定AJs。虽然之前的研究已经确定了Notch 1皮层 尽管Notch 1信号通路调节血管屏障功能，但剪切应力如何激活Notch 1受体仍不清楚。 基于Notch 1与细胞内在粘附和细胞骨架机制相关的初步数据， 测试Notch 1与VE-钙粘蛋白和皮质肌动蛋白细胞骨架的内在偶联调节 剪切应力介导的Notch 1激活。响应于剪切应力的Notch 1激活的询问将是 通过完成两个具体目标来接近：（1）确定VE-钙粘蛋白如何时空调节 Notch 1及其配体Dll 4协调剪切应力激活和（2）鉴定机械活化， 剪切应力下Notch 1和内在肌动蛋白细胞骨架动力学之间的相互作用。在整个 在拟议的研究过程中，我将获得人体微血管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 通过科学交流、指导和教学培训实现职业发展。我召集了一个 一个特殊的，互补的指导团队，以帮助我实现我的研究和职业目标：马修博士 Kutys是器官型组织建模和细胞力学方面的专家，他将是我的主要赞助商， 巴伯，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 Notch 1响应剪切应力而被激活的新机制，并可能识别新的 用于调节屏障功能和涉及Notch 1的其它血管病的治疗靶点。