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: 10685981
• 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/10685981
• 关键词: 3-Dimensional; Adherens Junction; Affect; Blood Vessels; Cell Communication; Cell-Cell Adhesion; Cells; Cytoskeleton; Development; Endothelial Cells; Endothelium; Engineering; Environment; Event; Extracellular Matrix; Extracellular Matrix Degradation; Future; Goals; Growth Factor Receptors; Integral Membrane Protein; Intercellular Junctions; Invaded; KDR gene; 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.

项目摘要 我的实验室的主要研究方向之一是内皮屏障的调节和 内皮细胞迁移。这些过程对血管系统的生理功能至关重要，它们 在人类疾病中经常是不受调控的。在理解信号方面已经取得了很大的进展 调节内皮屏障和细胞迁移。然而，刺激内皮细胞迁移在 血管生成是一种高度局部性和暂时性的事件。定义本地和临时组件的角色 由于目前工具的局限性，血管生成信号的研究一直是具有挑战性的。此外，时空 这些刺激对内皮屏障的调节一直知之甚少。我们提议的工作将集中在 确定迁移信号的位置和持续时间如何指导内皮细胞的入侵和迁移 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。 内皮屏障被控制在黏附连接(AJs)的水平，即细胞-细胞黏附结构 由跨膜蛋白VE-钙粘附素介导。磷酸化介导的信号调节 AJs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的双重作用和它的 VE-钙粘蛋白的磷酸化在内皮通透性调节中的作用。我们的结果表明，Src- 介导的磷酸化诱导动态AJ的形成，这些AJ仍然保留其屏障功能。这表明 在不损害屏障通透性的情况下调节AJ可塑性的机制 内皮细胞迁移。在平行研究中，我们剖析了Src调节的一种降解机制 细胞外基质通过内皮细胞并发现了一种新的细胞骨架成分 形成可降解基质的足体。这里提出的研究将继续建立在我们以前的基础上 VE-钙粘蛋白磷酸化与血管生成信号转导机制的研究 生长因子受体2(VEGFR2)、鞘氨醇-1-磷酸受体1(S1PR1)和Src调节 AJs的可塑性以及内皮细胞的侵袭和迁移。我们将使用新的光遗传工具 这将使我们能够通过精确的空间和时间控制来审问这些过程。我们将使用 工程化的光调节VEGFR2、S1PR1和Src，以确定局部和暂时的影响 调控血管生成信号及其调控AJs和迁移的机制剖析 三维环境中的内皮细胞。我们的长期目标是定义控制 血管生成过程中内皮细胞的迁移和内皮屏障功能。