Regulation and Function of Intermediate Filaments in Cell Mechanics

细胞力学中中间丝的调节和功能

• 批准号: 10227010
• 负责人: Gaudenz Danuser
• 金额: $ 177.12万
• 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/10227010
• 关键词: 3-Dimensional; Actins; Arteries; Biological; Biological Assay; Biological Process; Cell Polarity; Cell Shape; Cells; Cellular biology; Cues; Cyclic AMP-Dependent Protein Kinases; Cytoskeletal Modeling; Cytoskeleton; Defect; Development; Disease; Environment; Fishes; Foundations; Funding; Gene Expression Profiling; Genes; Grant; Human; Impairment; In Vitro; Innate Immune Response; Intermediate Filament Proteins; Intermediate Filaments; Knockout Mice; Length; Literature; Measures; Mechanical Stress; Mechanics; Mediating; Membrane; Mesenchymal; Methods; Microfilaments; Microfluidics; Microscopic; Microtubules; Morphogenesis; Motor; Movement; Mus; Mutation; Myosin ATPase; Organelles; Pennsylvania; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Play; Polymers; Preparation; Program Research Project Grants; Property; Protein Biosynthesis; Protein Family; Proteins; Reagent; Regulation; Reporting; Research Personnel; Resistance; Resolution; Role; Scientist; Sequence Homology; Signal Transduction; Structural Protein; Structure; Sum; Supporting Cell; System; Techniques; Testing; Time; Tissue Engineering; Tissues; Traction; Transducers; Tubulin; Universities; Vascular Endothelium; Vimentin; Xenopus; animal tissue; base; cell motility; cell type; data sharing; design; experience; improved; insight; knockout animal; leukocyte homing; lymph nodes; macrophage; mechanical properties; mechanotransduction; member; mouse model; polarized cell; polymerization; programs; protein degradation; reconstitution; response; shear stress; tool; wound healing

The overall hypothesis of this P01 Grant is that the unique properties of vimentin intermediate filaments (VIF) play a key role in regulating cytoskeletal organization and modulate the micromechanical properties of cells as well as a diverse set of cellular activities, including cell polarization, cell migration, or tissue morphogenesis. The Project Investigators and Core Leaders are all leaders in the field of cell biology and cell mechanics. Over the past funding period collaborative studies have established unique cell reagents and assays leading to key insights into the properties and functions of VIF. These insights provide a strong foundation for this renewal application. In preparation of this application further preliminary results have been collected supporting the feasibility of each of the projects. The projects are interactive conceptually, technically and programmatically, making the aggregate of the projects much greater than the sum of its parts. In Project #1, Dr. Goldman, Northwestern University, will determine the structural interactions among VIF, microtubules (MT) and actin microfilaments (MF) using high resolution microscopic techniques; determine the role of the assembly and disassembly of VIF in wound healing and motility assays; and determine the role of VIF phosphorylation in cellular signal transduction. In Project #2, Dr. Gelfand, Northwestern University will determine the dynamic mechanisms regulating VIF-MT interactions; determine the mechanisms responsible for the dynamic interactions between VIF and MF; and determine how VIF modulate the transport and distribution of membrane-bound organelles. In Project #3, Dr. Danuser, UTSW Dallas, will examine mechanisms by which VIF control MT organization and cell polarity; investigate mechanisms by which VIF control cell traction; and examine mechanisms by which VIF respond to cell-external guidance cues. In Project #4, Dr. Weitz, Harvard University, will determine the properties of reconstituted networks of VIF as well as composite networks comprised either of VIF, MF and myosin motors, or VIF, MT and their associated motors; study the micromechanical properties of VIF networks in living cells in 3D settings and in reconstituted networks derived from these cells. In Project #5, Dr. Janmey, University of Pennsylvania, will determine the mechanisms that regulate force-dependent VIF assembly in cells; study the mechanics of VIF networks under compression in vitro; and determine how VIF regulate the response of cells and tissues to compression loading. Interactions among members of all projects and data sharing will allow for integration of physical characterizations made by different groups using methods unique to their labs that cover a wide range of time and length scales. These efforts will be supported by the Cell and Tissue Core which, under the guidance of Dr. Ridge, Northwestern University, will support all PIs by maintaining the required WT and vimentin null mouse models; by engineering tissue and cell type-specific vimentin knockout animals; by isolating primary cells from various tissues of these animals; and by analyzing gene expression patterns and providing purified proteins as required.

P01 Grant的总体假设是波形蛋白中间丝（VIF）的独特性质 在调节细胞骨架组织和调节细胞的微观力学性质方面起关键作用， 以及多种细胞活动，包括细胞极化、细胞迁移或组织形态发生。 项目研究者和核心领导者都是细胞生物学和细胞力学领域的领导者。超过 在过去的资助期间，合作研究已经建立了独特的细胞试剂和测定， 深入了解VIF的性质和功能。这些见解为这一更新提供了坚实的基础 应用程序.在本申请的准备过程中，已经收集了进一步的初步结果， 每个项目的可行性。这些项目在概念上、技术上和程序上都是互动的， 使项目的总和远远大于其各部分的总和。在第一个项目中，戈德曼博士， 美国西北大学，将确定VIF，微管（MT）和肌动蛋白之间的结构相互作用 使用高分辨率显微镜技术的微丝（MF）;确定组件的作用， 在创伤愈合和运动性测定中VIF的分解;并确定VIF磷酸化在创伤愈合中的作用。 细胞信号转导在项目#2中，西北大学的Gelfand博士将确定 调节VIF-MT相互作用的机制;确定负责动态 VIF和MF之间的相互作用;并确定VIF如何调节 膜结合的细胞器。在项目#3中，UTSW达拉斯的Danuser博士将研究 VIF控制MT组织和细胞极性;研究VIF控制细胞牵引的机制; 研究VIF响应细胞外部指导线索的机制。在项目4中，哈佛的韦茨博士 大学，将确定VIF以及复合网络的重构网络的属性 包括VIF，MF和肌球蛋白马达，或VIF，MT及其相关马达;研究 活细胞中VIF网络在3D环境中的微观力学特性和在重构网络中的微观力学特性 从这些细胞。在项目#5中，宾夕法尼亚大学的Janmey博士将确定 调节细胞中依赖于力的VIF组装;研究VIF网络在压缩下的力学， 体外;并确定VIF如何调节细胞和组织对压缩负荷的反应。相互作用 在所有项目的成员之间和数据共享将允许整合的物理特性， 不同的小组使用其实验室特有的方法，涵盖广泛的时间和长度尺度。这些 这些努力将得到细胞和组织核心的支持，在西北大学里奇博士的指导下， 大学，将通过维持所需的WT和波形蛋白缺失小鼠模型来支持所有PI;通过工程 组织和细胞类型特异性波形蛋白敲除动物;通过从这些动物的各种组织中分离原代细胞 动物;以及分析基因表达模式并根据需要提供纯化的蛋白质。

项目成果

Withaferin a alters intermediate filament organization, cell shape and behavior.

• DOI: 10.1371/journal.pone.0039065
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Grin B;Mahammad S;Wedig T;Cleland MM;Tsai L;Herrmann H;Goldman RD
• 通讯作者: Goldman RD

Probing the stochastic, motor-driven properties of the cytoplasm using force spectrum microscopy.

• DOI: 10.1016/j.cell.2014.06.051
• 发表时间: 2014-08-14
• 期刊: Cell
• 影响因子: 64.5
• 作者: Guo M;Ehrlicher AJ;Jensen MH;Renz M;Moore JR;Goldman RD;Lippincott-Schwartz J;Mackintosh FC;Weitz DA
• 通讯作者: Weitz DA

Vimentin intermediate filaments and filamentous actin form unexpected interpenetrating networks that redefine the cell cortex.

• DOI: 10.1073/pnas.2115217119
• 发表时间: 2022-03-08
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Wu H;Shen Y;Sivagurunathan S;Weber MS;Adam SA;Shin JH;Fredberg JJ;Medalia O;Goldman R;Weitz DA
• 通讯作者: Weitz DA

Intermediate filament mechanics in vitro and in the cell: from coiled coils to filaments, fibers and networks.

体外和细胞中的中间细丝力学：从盘绕线圈到丝，纤维和网络。

• DOI: 10.1016/j.ceb.2015.01.001
• 发表时间: 2015-02
• 期刊: Current opinion in cell biology
• 影响因子: 7.5
• 作者: Köster S;Weitz DA;Goldman RD;Aebi U;Herrmann H
• 通讯作者: Herrmann H

Microfluidic Generation of Monodisperse, Structurally Homogeneous Alginate Microgels for Cell Encapsulation and 3D Cell Culture.

• DOI: 10.1002/adhm.201500021
• 发表时间: 2015-08-05
• 期刊: Advanced healthcare materials
• 影响因子: 10
• 作者: Utech S;Prodanovic R;Mao AS;Ostafe R;Mooney DJ;Weitz DA
• 通讯作者: Weitz DA