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.

总结 本行政补充申请要求使用落射荧光、FRET和TIRF显微镜， 父母R35赠款的目标是了解我们有限了解的细胞迁移的两个方面：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，但是它们如何在一个特定的时间内起作用， 缺乏对细胞迁移的机械水平。父R35赠款旨在建立在以前在两个 NIGMS支持的Gundersen实验室以前的项目。对于核定位，我们将研究 核骨架和细胞骨架（LINC）复合物的连接物被机械地加强以抵抗所需的大的力 移动细胞核，并将使用和开发新的FRET张力传感器，直接测量细胞核上的力。我们 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LINC复合体的功能。 这些相互作用对不同细胞迁移模式的影响。我们将测试新的假设， 作为肌动蛋白丝的“张力电阻器”和作为微管运输的极性因子。对整联蛋白 回收，我们将测试整体假设，回收整合素旅行的积极构象，这种子 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一种新的整合素 再循环系统与TIRF显微镜相结合，以鉴定整合素再循环的细胞内途径， 整合素胞吐作用相对于新形成的粘连。我们还将使用TIRF显微镜，以确定是否回收 整联蛋白在迁移过程中来源于细胞后部，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。我们将探讨 细胞迁移的这两个方面是否可以通过用张力传感器的FRET成像来确定， 核机械地接合基底粘连。我们对这两个过程的拟议研究将取得进展 了解细胞迁移的基本机制，并有可能确定新的干预目标， 细胞迁移就会出错拟议的研究还将为博士后研究员和研究生提供肥沃的土壤。 学生推进他们的培训和开发自己的项目。

项目成果

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

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