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

• 批准号: 10613943
• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10613943
• 关键词: Adhesions; Attention; Cell Nucleus; Cells; Complex; Cytoskeleton; Defect; Development; Exocytosis; Immune response; Inflammation; Integrins; Intervention; Knowledge; Laboratories; Lead; Measures; Mechanics; Microfilaments; Microtubules; Molecular Conformation; Movement; National Institute of General Medical Sciences; Neoplasm Metastasis; Nuclear; Organ; Pathologic Processes; Physiological Processes; Positioning Attribute; Postdoctoral Fellow; Process; Property; Recycling; Signal Transduction; Site; System; Testing; Tissues; Training; Travel; adhesion receptor; cell motility; graduate student; non-healing wounds; sensor; tissue regeneration; trafficking; wound healing

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.

总结 后生动物的细胞迁移有助于发育和稳态过程，包括组织的形成， 器官、伤口愈合、组织更新和免疫反应。细胞迁移的改变导致发育缺陷， 炎症、持续伤口和转移。因此，破译细胞迁移的机制具有广泛的意义 来理解生理和病理过程。经过大量的努力， 一些有助于细胞迁移的关键过程，包括前缘的突出和 粘连细胞迁移的其他重要方面很少受到关注，我们的理解也存在差距 它们的机制和对细胞迁移的贡献。该提案旨在填补这两个知识空白， 过程：核定位和整合素再循环。这两个过程对于细胞迁移都是必不可少的，但机制 他们如何做到这一点是缺乏了解。这一建议将以先前两个项目取得的成果为基础 在NIGMS支持下的Gundersen实验室。对于核定位，我们将研究 核骨架和细胞骨架（LINC）复合体被机械地加强，以抵抗移动所需的巨大力量。 并将使用和开发新的张力传感器来直接测量原子核上的力。我们将确定 LINC复合体如何选择肌动蛋白丝或微管进行核运动，以及 这些相互作用导致了不同的细胞迁移模式。我们将测试新的假设，即细胞核的功能是一个 作为肌动蛋白丝的“张力电阻器”和微管运输的极性因子。对于整合素回收，我们将测试 总的假设是，再循环的整合素以活性构象移动，并且这导致新的粘附形成， 以靠近前缘的极化方式。我们将使用新的整合素探针和新的整合素回收系统， 鉴定整合素胞吐作用的位点及其与新形成的粘连的关系。在迁移的细胞中，我们将确定 再循环的整联蛋白是否来源于细胞后部并有助于前缘附近的新生粘附形成。 我们将确定微管机制，我们假设是负责极化重建粘连 并测试再循环的整合素在整合素信号传导中起作用的可能性。我们提出的 对这两个过程的研究将促进对细胞迁移的基本机制的理解， 确定新的目标，以干预细胞迁移出错的情况。拟议的研究还将提供肥沃的土壤， 为博士后研究员和研究生提供基础，以促进他们的培训和开发自己的项目。