Physical Approaches for Probing the Mechanical Properties of Intermediate Filaments

探测中间丝机械性能的物理方法

• 批准号: 10227017
• 负责人: DAVID A WEITZ
• 金额: $ 26.13万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10227017
• 关键词: 3-Dimensional; Actins; Behavior; Biological; Biological Models; Blood flow; Bundling; Cell Extracts; Cell Nucleus; Cells; Collaborations; Complement; Cytoskeleton; Disease; Environment; Exhibits; F-Actin; Foundations; Gel; Goals; Image; Individual; Intermediate Filaments; Ions; Knowledge; Liquid substance; Measurement; Measures; Mechanics; Microfluidic Microchips; Microscopy; Microtubules; Modulus; Monitor; Motor; Motor Activity; Mutation; Myosin ATPase; Nature; Null Lymphocytes; Organelles; Pharmacotherapy; Phosphorylation; Play; Post-Translational Protein Processing; Property; Proteins; Regulation; Role; Stimulus; Stress; Structure; Surface; System; Techniques; Time; Tissues; Vesicle; Vimentin; Work; behavior measurement; cell behavior; cell motility; in vivo; laser tweezer; mechanical behavior; mechanical properties; mutant; particle; pressure; programs; protein expression; reconstitution; response; single-cell RNA sequencing; therapy development; wound healing

Abstract Cell must support forces, must exert forces, and must respond to forces. All these behaviors are fundamentally built on mechanical properties of the cells, which are largely determined by three filamentous networks within the cytoskeleton, actin, microtubules, and intermediate filaments (IF). While actin and microtubules are widely cited as the crucial components determining the mechanical properties of the cells and are well studied, the importance of intermediate filaments is increasingly being recognized. For example, cells without vimentin intermediate filaments are much weaker and more susceptible to break up upon application of small shear forces comparable in magnitude to those encountered in blood flow; moreover, they are much less stable, with the nucleus and organelles lacking positional stability and being easier to move. The presence of vimentin IF networks mechanically stabilize cells; however, they must work in concert with the other filamentous networks. The overarching goal of this Program Project is to elucidate the roles of the vimentin IF network and how it operates in concert with other filamentous networks to determine the mechanical properties of cells. We will accomplish this through studies of reconstituted mixed networks with and without motor proteins to isolate the mechanics of the IF networks in the presence of the other networks. The results of these studies will be compared to the mechanical behavior of cells grown on flat substrates, both isolated and in confluent layers, and for cells from vimentin-related diseases. Ultimately, we will build new systems to enable studies of the properties of cells grown in three-dimensional networks that mimic environments in vivo. The knowledge gained in this study will establish foundational scientific understanding of roles of vimentin IFs in cell mechanics, which will ultimately provide guidance to the development of treatments for vimentin related diseases.

抽象的 细胞必须支撑力，必须施加力，并且必须对部队做出反应。所有这些行为从根本上都是 基于单元的机械性能，这些特性在很大程度上由三个丝状网络确定 细胞骨架，肌动蛋白，微管和中间丝（如果）。而肌动蛋白和微管广泛 被认为是确定细胞机械性能的关键组成部分，并经过了很好的研究， 中间细丝的重要性越来越被认识到。例如，没有波形蛋白的细胞 中间丝弱得多，并且更容易在使用小剪切力时破裂 与血流中遇到的人相当；而且，它们不那么稳定， 核和细胞器缺乏位置稳定性，更容易移动。波形蛋白的存在 网络机械稳定细胞；但是，他们必须与其他丝状网络协同合作。 该计划项目的总体目标是阐明Vimentin If网络的角色及其如何 与其他丝状网络协同工作，以确定细胞的机械性能。我们将 通过有或没有运动蛋白的重构混合网络的研究来实现此目的 IF网络的力学在其他网络存在的情况下。这些研究的结果将被比较 对于在平坦基材上生长的细胞的机械行为，无论是孤立的还是在汇合层中，以及细胞的机械行为 来自与波形蛋白有关的疾病。最终，我们将建立新的系统来研究细胞的性质 在模仿体内环境的三维网络中生长。这项研究中获得的知识将 建立对波形蛋白IFS在细胞力学中的作用的基本科学理解，最终将 为开发波形蛋白相关疾病的治疗提供指导。