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: 10799051
• 负责人: Gregg G Gundersen
• 金额: $ 24.88万
• 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/10799051
• 关键词: Adhesions; Administrative Supplement; Cell Nucleus; Cells; Complex; Coupled; Cytoskeleton; Development; Exocytosis; Fluorescence Resonance Energy Transfer; Goals; Grant; Image; Immune response; Inflammation; Integrins; Intervention; Laboratories; Link; Measures; Mechanics; Microfilaments; Microscope; Microtubules; Molecular Conformation; Movement; National Institute of General Medical Sciences; Neoplasm Metastasis; Nuclear; Parents; Pathway interactions; Positioning Attribute; Postdoctoral Fellow; Process; Property; Recycling; Signal Transduction; Site; System; Testing; Total Internal Reflection Fluorescent; Training; Travel; adhesion receptor; cell motility; graduate student; migration; sensor; trafficking; wound healing

SUMMARY This application for an Administrative Supplement requests an epifluorescence, FRET and TIRF microscope to pursue the goals of the parent R35 grant to understand two aspects of cell migration for which we have limited understanding: nuclear positioning and integrin recycling. Both these processes are essential for cell migration, yet how they contribute at a mechanistic level to cell migration is lacking. The parent R35 grant seeks to 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 FRET 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 coupled with TIRF microscopy to identify the intracellular pathway of recycling integrins, their sites of integrin exocytosis relative to newly formed adhesions. We will also use TIRF microscopy to determine whether recycled integrin derives from the cell rear during migration 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. We will explore whether these two aspects of cell migration can be linked by determining with FRET imaging of tension sensors whether the nucleus mechanically engages substrate adhesions. 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 此行政补充申请要求使用落射荧光、FRET 和 TIRF 显微镜来追踪 母体 R35 资助的目标是了解细胞迁移的两个方面，对此我们了解有限：核 定位和整合素回收这两个过程对于细胞迁移都是至关重要的，但它们如何在细胞迁移中发挥作用。 缺乏细胞迁移的机械水平，母公司 R35 资助旨在建立在先前获得的两个结果的基础上。 Gundersen 实验室先前由 NIGMS 支持的项目对于核定位，我们将研究如何 核骨架和细胞骨架 (LINC) 复合物的连接体经过机械强化，可抵抗所需的巨大作用力 我们将使用和开发新的 FRET 张力传感器来直接测量原子核上的力。 将决定 LINC 复合体如何选择肌动蛋白丝或微管进行核运动以及功能 我们将测试细胞核的新假设。 充当肌动蛋白丝的“张力电阻器”和整合素微管运输的极性因子。 回收，我们将测试总体假设，即回收的整合素以活性构象移动，并且该种子 在前缘附近以极化方式形成新的粘附我们将使用新的整合素探针和新的整合素。 回收系统与 TIRF 显微镜相结合，以确定回收整合素的细胞内途径及其位点 与新形成的粘连相关的整合素胞吐作用我们还将使用 TIRF 显微镜来确定是否回收。 整合素在迁移过程中源自细胞后部，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 我们将探索回收的整合素在整合素信号传导中发挥作用的可能性。 通过使用张力传感器的 FRET 成像确定是否可以将细胞迁移的这两个方面联系起来 我们提出的对这两个过程的研究将取得进展。 了解细胞迁移的基本机制和潜力，确定新的干预目标 细胞迁移出错。拟议的研究也将为博士后研究员和毕业生提供肥沃的土壤。 学生推进他们的培训并开发自己的项目。

项目成果

Apoptotic stress induces Bax-dependent, caspase-independent redistribution of LINC complex nesprins.

• DOI: 10.1038/s41420-020-00327-6
• 发表时间: 2020
• 期刊: Cell death discovery
• 影响因子: 7
• 作者: Lindenboim L;Grozki D;Amsalem-Zafran AR;Peña-Blanco A;Gundersen GG;Borner C;Hodzic D;Garcia-Sáez AJ;Worman HJ;Stein R
• 通讯作者: Stein R

FHODs: Nuclear tethered formins for nuclear mechanotransduction.

• DOI: 10.3389/fcell.2023.1160219
• 发表时间: 2023
• 期刊: Frontiers in cell and developmental biology
• 影响因子: 5.5

LINC complex protein nesprin-2 has pro-apoptotic activity via Bcl-2 family proteins.

• DOI: 10.1038/s41420-023-01763-w
• 发表时间: 2024-01-15
• 期刊: CELL DEATH DISCOVERY
• 影响因子: 7
• 作者: Lindenboim, Liora;Zohar, Hila;Gundersen, Gregg G.;Worman, Howard J.;Stein, Reuven
• 通讯作者: Stein, Reuven