Regulation of endothelial cell invasion, migration and cell junction plasticity

内皮细胞侵袭、迁移和细胞连接可塑性的调节

• 批准号: 10406685
• 负责人: ANDREI V KARGINOV
• 金额: $ 39.98万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10406685
• 关键词: 3-Dimensional; Adherens Junction; Affect; Blood Vessels; Cell Communication; Cell-Cell Adhesion; Cells; Development; Endothelial Cells; Endothelium; Engineering; Environment; Event; Extracellular Matrix; Extracellular Matrix Degradation; Future; Goals; Growth Factor Receptors; Integral Membrane Protein; Intercellular Junctions; Laboratories; Light; Location; Mediating; Permeability; Phosphorylation; Physiologic Neovascularization; Physiological; Process; Regulation; Research; Role; Signal Transduction; Sphingosine-1-Phosphate Receptor; Stimulus; Structure; Therapeutic; Time; Vascular System; Work; angiogenesis; cadherin 5; cell motility; human disease; migration; novel; optogenetics; spatiotemporal; src-Family Kinases; tool

Project Summary One of the main research directions of my laboratory focuses on regulation of the endothelial barrier and endothelial cell migration. These processes are critical for physiological function of vascular system and they are often dysregulated in human diseases. A lot of progress has been made in understanding signaling that regulates endothelial barrier and cell migration. However, stimulation of endothelial cell migration during angiogenesis is a highly localized and transient event. Defining the role of the local and temporal components of angiogenic signaling has been challenging due to limitations of current tools. Furthermore, spatiotemporal regulation of the endothelial barrier by these stimuli has been poorly understood. Our proposed work will focus on determining how the location and duration of migratory signals direct endothelial cell invasion and migration through extracellular matrix, and how they affect the organization and permeability of the endothelial barrier. The endothelial barrier is controlled at the level of adherens junctions (AJs), cell-cell adhesion structures mediated by the transmembrane protein VE-cadherin. Phosphorylation-mediated signaling regulates the structure and permeability of AJs. In our recent studies, we described a dual role of tyrosine kinase Src and its phosphorylation of VE-cadherin in regulation of endothelial permeability. Our results demonstrated that Src- mediated phosphorylation induces formation of dynamic AJs that still retain their barrier function. This suggests a mechanism for the regulation of AJ plasticity that does not compromise barrier permeability during endothelial cell migration. In parallel studies, we dissected a mechanism of Src-regulated degradation of the extracellular matrix by the endothelial cell and discovered a novel cytoskeletal component that mediates formation of matrix-degrading podosomes. The studies proposed here will continue to build on our previous findings and focus on dissecting how phosphorylation of VE-cadherin and angiogenic signaling by Vascular Growth Factor Receptor 2 (VEGFR2), Sphingosine-1-phosphate Receptor 1 (S1PR1), and Src regulate plasticity of AJs as well as invasion and migration of endothelial cells. We will employ novel optogenetic tools that will allow us to interrogate these processes with precise spatial and temporal control. We will use engineered light-regulated VEGFR2, S1PR1, and Src to determine the effects of locally and temporally controlled angiogenic signals and dissect mechanisms that mediate regulation of AJs and migration of endothelial cells in three dimensional environment. Our long-term goal is to define the processes that control migration of endothelial cells and endothelial barrier function during angiogenesis.

项目概要 我实验室的主要研究方向之一是内皮屏障的调控和 内皮细胞迁移。这些过程对于血管系统的生理功能至关重要，它们 在人类疾病中经常失调。在理解信号方面已经取得了很大进展 调节内皮屏障和细胞迁移。然而，刺激内皮细胞迁移 血管生成是高度局部化且短暂的事件。定义局部和时间组件的作用 由于当前工具的限制，血管生成信号传导的研究一直具有挑战性。此外，时空 人们对这些刺激对内皮屏障的调节知之甚少。我们提议的工作将重点 确定迁移信号的位置和持续时间如何指导内皮细胞侵袭和迁移 通过细胞外基质，以及它们如何影响内皮屏障的组织和通透性。 内皮屏障在粘附连接 (AJ)、细胞间粘附结构水平上受到控制 由跨膜蛋白VE-钙粘蛋白介导。磷酸化介导的信号传导调节 AJ 的结构和渗透性。在我们最近的研究中，我们描述了酪氨酸激酶 Src 的双重作用及其 VE-钙粘蛋白的磷酸化对内皮通透性的调节。我们的结果表明，Src- 介导的磷酸化诱导动态 AJ 的形成，但仍保留其屏障功能。这表明 一种调节 AJ 可塑性的机制，在过程中不会影响屏障渗透性 内皮细胞迁移。在平行研究中，我们剖析了 Src 调节的降解机制 内皮细胞通过细胞外基质，发现了一种介导的新型细胞骨架成分 基质降解足体的形成。这里提出的研究将继续建立在我们之前的基础上 研究结果并重点剖析 VE-钙粘蛋白的磷酸化和血管生成信号传导的机制 生长因子受体 2 (VEGFR2)、1-磷酸鞘氨醇受体 1 (S1PR1) 和 Src 调节 AJ 的可塑性以及内皮细胞的侵袭和迁移。我们将采用新颖的光遗传学工具 这将使我们能够通过精确的空间和时间控制来询问这些过程。我们将使用 设计光调节 VEGFR2、S1PR1 和 Src，以确定局部和暂时的影响 控制血管生成信号并剖析介导 AJ 调节和迁移的机制 三维环境中的内皮细胞。我们的长期目标是定义控制流程 血管生成过程中内皮细胞的迁移和内皮屏障功能。