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.

项目摘要 我的实验室的主要研究方向之一是调节内皮障碍和 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 通常在人类疾病中失调。在理解信号传导方面已经取得了很多进展 调节内皮屏障和细胞迁移。但是，刺激内皮细胞迁移 血管生成是一个高度局部和瞬态事件。定义本地和时间组件的作用 由于当前工具的局限性，血管生成信号传导一直具有挑战性。此外，时空 这些刺激对内皮屏障的调节知之甚少。我们提出的工作将集中 确定迁移信号的位置和持续时间如何直接内皮细胞入侵和迁移 通过细胞外基质，以及它们如何影响内皮屏障的组织和渗透性。 内皮屏障在粘附连接（AJS），细胞细胞粘附结构的水平上受到控制 由跨膜蛋白VE-钙粘着蛋白介导。磷酸化介导的信号传导调节 AJS的结构和渗透性。在最近的研究中，我们描述了酪氨酸激酶SRC及其ITS的双重作用 VE-钙粘着蛋白在调节内皮渗透性方面的磷酸化。我们的结果表明SRC- 介导的磷酸化诱导仍保留其屏障功能的动态AJ的形成。这暗示着 一种调节AJ可塑性的机制，该机制不会损害屏障的渗透率 内皮细胞迁移。在平行研究中，我们阐述了SRC调节的机制 内皮细胞的细胞外基质，发现了一种新型的细胞骨架成分，可介导 基质降解足体的形成。这里提出的研究将继续基于我们以前的 发现并着重于剖析VE-钙粘蛋白和血管生成信号的磷酸化如何通过血管磷酸化 生长因子受体2（VEGFR2），鞘氨酸1-磷酸受体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 这将使我们能够以精确的空间和时间控制来审问这些过程。我们将使用 设计的光调节VEGFR2，S1PR1和SRC确定本地和时间上的影响 受控的血管生成信号和剖析机制，介导AJ的调节和迁移的机制 内皮细胞在三维环境中。我们的长期目标是定义控制的过程 血管生成过程中内皮细胞和内皮屏障功能的迁移。