Cytoskeleton, Nucleus and Integrin Recycling in Cell Migration

细胞迁移中的细胞骨架、细胞核和整合素回收

基本信息

批准号：
10396505
负责人：
Gregg G Gundersen
金额：
$ 60.48万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2025-04-30
项目状态：
未结题

项目摘要

SUMMARY Metazoan cell migration contributes to developmental and homeostatic processes, including formation of tissues and organs, wound healing, tissue renewal and immune responses. Alterations in cell migration lead to developmental defects, inflammation, persistent wounds and metastasis. Deciphering mechanisms of cell migration thus has broad significance for understanding both physiological and pathological processes. Considerable effort has led to deep understanding of some of the critical processes that contribute to cell migration, including protrusion of the leading edge and formation of adhesions. Other important aspects of cell migration have received scant attention and there is a gap in our understanding of their mechanism and contribution to cell migration. This proposal seeks to fill this knowledge gap for two such processes: nuclear positioning and integrin recycling. Both these processes are essential for cell migration, but mechanistic understanding of how they do so is lacking. This proposal will build on results previously obtained in two previous projects in the Gundersen laboratory supported by NIGMS. For nuclear positioning, we will examine how the linker of nucleoskeleton and cytoskeleton (LINC) complex is mechanically reinforced to resist the large forces necessary to move the nucleus and will use and develop new tension sensors to directly measure forces on the nucleus. We will determine how the LINC complex selects actin filament or microtubules for nuclear movement and the functional consequences of these interactions for different modes of cell migration. We will test new hypotheses that the nucleus functions as a “tension resistor” for actin filaments and as a polarity factor for microtubule trafficking. For integrin recycling, we will test the overall hypothesis that recycled integrins travel in an active conformation and that this seeds new adhesion formation in a polarized manner near the leading edge. We will use new integrin probes and a new integrin recycling system to identify sites of integrin exocytosis and their relationship to newly formed adhesions. In migrating cells, we will determine whether recycled integrin derives from the cell rear and contributes to nascent adhesion formation near the leading edge. We will identify the microtubule machinery that we hypothesize is responsible for the polarized reformation of adhesions from recycled integrins and test the possibility that the recycled integrin plays a role in integrin signaling. Our proposed studies on these two processes will advance understanding of the basic mechanisms of cell migration and potential identify new targets for intervening in cases when cell migration goes awry. The proposed studies will also provide fertile ground for postdoctoral fellows and graduate students to advance their training and develop their own projects.

概括后生动物细胞迁移有助于发育和稳态过程，包括组织和细胞的形成器官、伤口愈合、组织更新和免疫反应的改变会导致发育缺陷，因此，破译细胞迁移的机制具有广泛的意义。为了理解生理和病理过程付出了巨大的努力。一些有助于细胞迁移的关键过程，包括前缘的突出和形成细胞迁移的其他重要方面很少受到关注，我们的理解存在差距。该提案旨在填补两个这样的知识空白。过程：核定位和整合素回收这两个过程对于细胞迁移都是必不可少的，但都是机械性的。目前还缺乏对他们如何做到这一点的了解。该提案将建立在之前两个项目所取得的成果的基础上。在 NIGMS 支持的 Gundersen 实验室中，我们将研究连接器如何进行核定位。核骨架和细胞骨架 (LINC) 复合物经过机械强化，可抵抗移动所需的巨大力量我们将确定原子核上的力，并使用和开发新的张力传感器来直接测量原子核上的力。 LINC 复合体如何选择肌动蛋白丝或微管进行核运动以及其功能后果我们将测试细胞核作为细胞迁移的新假设。对于肌动蛋白丝的“张力电阻器”以及作为微管运输的极性因子，我们将测试整合素回收。总体假设是，回收的整合素以活性构象移动，这会形成新的粘附我们将在前沿附近以极化方式使用新的整合素探针和新的整合素回收系统。确定整合素胞吐作用的位点及其与迁移细胞中新形成的粘附的关系。回收的整合素是否来自细胞后部并有助于前缘附近新生粘附的形成。我们将确定我们所勇敢的微管机制负责粘连的极化改造来自回收的整合素并测试回收的整合素在整合素信号传导中发挥作用的可能性。对这两个过程的研究将促进对细胞迁移和潜力的基本机制的理解确定在细胞迁移出现问题时进行干预的新目标。拟议的研究还将提供生育能力。为博士后研究员和研究生提供进一步培训和开发自己项目的平台。