Systematic analysis of the actin cytoskeleton and directed cell migration

肌动蛋白细胞骨架和定向细胞迁移的系统分析

• 批准号: 10090476
• 负责人: JAMES E BEAR
• 金额: $ 58.11万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10090476
• 关键词: Actins; Address; Area; Biochemical; Biological Process; Cardiovascular Diseases; Cause of Death; Cells; Cellular biology; Characteristics; Chemicals; Chemotaxis; Complex; Cues; Cytoskeleton; Disease; Embryonic Development; Environment; Fibroblasts; Fibrosis; Funding; Genes; Goals; Health; Heart; Human; Image Analysis; Immune response; Knowledge; Mechanics; Mesenchymal; Microfluidic Microchips; Microfluidics; Microscope; Molecular; Morphogenesis; Neoplasm Metastasis; Paper; Pathologic Processes; Physiological Processes; Play; Process; Publishing; RNA Interference; Research; Role; Structure; Therapeutic; Work; active control; cell behavior; cell motility; cell type; conditional knockout; interdisciplinary approach; microscopic imaging; migration; mouse model; programs; response; success; tool; tumor; wound healing

Abstract: Cell motility is arguably the oldest problem in cell biology as the movement of cells was one of the first things noted when the microscope was developed in the 17th century. The last sixty years of work revealed that the molecular underpinnings of motility involve the active control of the cytoskeleton. However, many questions remain unanswered. While we have identified many of the components of the cytoskeleton and know a great deal about their biochemical and structural characteristics, we lack a systematic understanding of how the parts interact to produce coordinated cytoskeletal function such as during cell migration. Perhaps the most important problem in cell motility is understanding how cells perceive various cues in their environment and convert this information into a directed migration response. A deeper understanding of these two inter-related problems will inform higher order biological processes such as embryogenesis, immune response and wound healing, as well as diseases such as cancer metastasis. In 2012, our published a watershed paper for our research program that has shaped the course of our work ever since (Wu et al, Cell). In this paper, we discovered that mammalian fibroblasts could be nearly completely depleted of Arp2/3 by RNAi without significantly compromising viability. This allowed us to go the heart of the problem and study the function of the Arp2/3 complex directly. Using these cells and microfluidic devices to produce long-lasting gradients, we made the surprising observation that cells did not need the Arp2/3 complex for chemotaxis, but absolutely required it for haptotaxis. Inspired by our success with RNAi, we shifted to a conditional knockout mouse model where the gene encoding the critical Arpc2 (p34) subunit of Arp2/3 could be deleted on command. Using these tools, we have pursued two goals: 1) Investigate how cells can build an actin cytoskeleton without the Arp2/3 complex (funded by RO1 GM111557) and 2) Interrogate the molecular mechanisms of directed cell migration of mesenchymal cell and other cell types (funded by RO1 GM110155). Building on our progress in the last five years, we propose to extend our work in these two areas by defining the roles of Arp2/3 and non-Arp2/3 actin networks in contributing to cell behaviors such as directed migration towards chemical (chemotaxis), substrate- bound (haptotaxis) and mechanical (durotaxis) cues.

抽象的： 细胞运动可以说是细胞生物学中最古老的问题，因为细胞的运动是第一件事之一 注意到显微镜在17世纪开发时。过去的六十年的工作表明 运动性的分子基础涉及细胞骨架的主动控制。但是，很多问题 保持没有答复。虽然我们已经确定了细胞骨架的许多成分，并且知道了一个很棒的 谈论他们的生化和结构特征，我们对如何有系统地了解 零件相互作用以产生协调的细胞骨架功能，例如在细胞迁移过程中。也许最多 细胞运动中的重要问题是了解细胞如何感知其环境中的各种线索和 将此信息转换为有向的迁移响应。对这两个相互关联的更深入了解 问题将为高阶生物学过程提供信息，例如胚胎发生，免疫反应和伤口 康复以及癌症转移等疾病。 2012年，我们出版了一份流域论文 从那以后就塑造了我们工作进程的研究计划（Wu等人，Cell）。在本文中，我们 发现哺乳动物成纤维细胞几乎可以被RNAi完全耗尽ARP2/3 显着损害了生存能力。这使我们能够成为问题的核心，并研究 ARP2/3复合物。使用这些细胞和微流体设备产生持久的梯度，我们做到了 令人惊讶的观察到细胞不需要ARP2/3复合物进行趋化性，但绝对需要它 用于触觉。受到RNAi成功的启发，我们转移到了条件敲除鼠标模型中 可以在命令上删除编码ARP2/3的关键ARPC2（p34）亚基的基因。使用这些工具，我们 已经实现了两个目标：1）研究细胞如何在没有ARP2/3复合物的情况下构建肌动蛋白细胞骨架 （由RO1 GM111557资助）和2）询问定向细胞迁移的分子机制 间充质细胞和其他细胞类型（由RO1 GM110155资助）。基于我们最近五个进步 几年，我们建议通过定义ARP2/3和非ARP2/3肌动蛋白的角色来扩展这两个领域的工作 促进细胞行为的网络，例如针对化学（趋化性），底物迁移的网络 绑定（haptotaxis）和机械（durotaxis）线索。