Biophysical Control of Cell Form and Function by Single Actomyosin Stress Fibers

单个肌动球蛋白应力纤维对细胞形态和功能的生物物理控制

基本信息

批准号：
10445792
负责人：
Sanjay Kumar
金额：
$ 33.06万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-20 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10445792
关键词：
3-Dimensional Actins Actomyosin Address Adopted Architecture Award Back Biological Biophysics Cell Nucleus Cell Shape Cells Cellular Structures Clinical Collaborations Complex Computer Models Confined Spaces Cues Dependence Disease Dorsal Engineering Environment Extracellular Matrix Fiber Foundations Genetic Genetic Transcription Geometry Glioblastoma Glioma Image Individual Knowledge Lasers Maintenance Malignant neoplasm of brain Measures Mechanics Mediating Mesenchymal Methodology Microfluidic Microchips Microfluidics Molecular Morphogenesis Myosin ATPase Neurosurgeon Nuclear Pathway interactions Pattern Phosphatidylinositol 4,5-Diphosphate Phosphorylation Physiology Play Process Publications Regulation Resistance Role Shapes Signal Transduction Stress Fibers Surface Technology Testing Tissues Transcription Coactivator Translating Width Work base bevacizumab cell motility cofilin design human disease in vivo innovation insight migration mimetics monolayer mouse model multi-scale modeling nanosurgery network architecture novel optogenetics reconstitution recruit stem cells tool two-dimensional

3-Dimensional Actins Actomyosin Address Adopted Architecture Award Back Biological Biophysics Cell Nucleus Cell Shape Cells Cellular Structures Clinical Collaborations Complex Computer Models Confined Spaces Cues Dependence Disease Dorsal Engineering Environment Extracellular Matrix Fiber Foundations Genetic Genetic Transcription Geometry Glioblastoma Glioma Image Individual Knowledge Lasers Maintenance Malignant neoplasm of brain Measures Mechanics Mediating Mesenchymal Methodology Microfluidic Microchips Microfluidics Molecular Morphogenesis Myosin ATPase Neurosurgeon Nuclear Pathway interactions Pattern Phosphatidylinositol 4,5-Diphosphate Phosphorylation Physiology Play Process Publications Regulation Resistance Role Shapes Signal Transduction Stress Fibers Surface Technology Testing Tissues Transcription Coactivator Translating Width Work base bevacizumab cell motility cofilin design human disease in vivo innovation insight migration mimetics monolayer mouse model multi-scale modeling nanosurgery network architecture novel optogenetics reconstitution recruit stem cells tool two-dimensional

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项目摘要

PROJECT SUMMARY/ABSTRACT Actomyosin stress fibers (SFs) enable cells to tense the extracellular matrix (ECM), a process key to cell shape determination, motility, and morphogenesis. Over the past 15+ years, including the past period of R01 support, we have made significant contributions to the field’s understanding of SF mechanics and contributions to cell structure. Our work is particularly notable for the use of femtosecond laser nanosurgery (FLN), which has enabled us to show that the three canonical SF subtypes – dorsal fibers, transverse arcs, and ventral fibers – collectively enforce a front-back tension gradient that underlies two-dimensional (2D) mesenchymal migration. We also showed that the SF network architecture can mechanically reinforce individual SFs, which has significant implications for symmetry breakage during directed migration and force propagation through cell monolayers. With this intellectual foundation in place, our renewal application turns to two important questions: How is polarization of tension in the SF network encoded by molecular signals classically understood to establish front-back polarity? And how does our knowledge of 2D SF networks translate to confined migration geometries like those found in tissue? We will address these questions through two specific aims, both of which build upon publications from this award. In Specific Aim 1, we will investigate mechanistic contributions of cofilin-1 to establishment and maintenance of SF front-back tension polarization during migration. We hypothesize that cofilin-1 establishes front-back polarization of SF tension by promoting the assembly and contractile maturation of transverse arcs. By combining biophysical, engineering, and cell biological tools, we will identify key molecular and force-based signals that modulate recruitment of cofilin-1 to developing transverse arcs. In an innovative new collaboration with Dr. Bruce Goode (Brandeis) we will reconstitute actin bundles in microfluidic devices and quantify the relationship between tensile force and cofilin- 1 engagement. In Specific Aim 2, we will dissect contributions of SF networks to migration in confined geometries where the ECM imposes axial cues and sterically precludes elaboration of 2D SF networks. We hypothesize that increasing confinement redirects SF assembly from the 2D dorsal fiber-transverse arc-ventral fiber assembly pathway towards de novo parallelized SF assembly. We will combine microengineered culture platforms, single-cell mechanical tools, and superresolution imaging to probe confinement-induced changes in SF assembly, architecture, and mechanics. Aim 2 will leverage two established, productive collaborations: With Dr. Ulrich Schwarz (U. Heidelberg), we will develop multiscale computational models that relate SF network architecture and mechanics to cell migration in confined spaces. With neurosurgeon Dr. Manish Aghi (UCSF), we will test the clinical value of our observations by asking if confined migration of glioblastoma stem cells is retrospectively predictive of in vivo invasion patterns. Our studies will create unprecedented new insight into how SFs contribute to migration, with innovative methodology and close connection to human disease.

项目摘要/摘要肌动蛋白应激纤维（SFS）使细胞能够紧张细胞外基质（ECM），这是细胞形状的过程键确定，运动和形态发生。在过去的15年以上，包括过去的R01 支持，我们为该领域对SF力学和贡献的理解做出了重大贡献我们的工作特别值得注意，用于使用飞秒激光纳米外科手术（FLN），这是使我们能够证明三种规范的SF亚型 - 背纤维，横臂和腹侧纤维 - 集体强制执行二维（2D）间充质的前后张力梯度迁移。我们还表明，SF网络体系结构可以机械加强单个SF，这在定向迁移和通过细胞的力传播期间对称对称破裂具有重要意义单层。有了这个智力基础，我们的续签应用将变成两个重要的问题：SF网络中张力的极化是如何通过分子信号编码的了解建立前后极性吗？以及我们对2D SF网络的了解如何转化为像在组织中发现的那样，受到狭窄的迁移几何形状？我们将通过两个解决这些问题具体目标，这两者都基于该奖项的出版物。在特定目标1中，我们将调查 Cofilin-1对SF前后张力极化的建立和维护的机械贡献在迁移期间。我们假设Cofilin-1通过促进SF张力建立了前后后背极化横向弧的组装和收缩成熟。通过结合生物物理，工程和细胞生物工具，我们将确定关键分子和基于力的信号，以调节cofilin-1募集到发展横向弧。在与Bruce Goode博士（Brandeis）的创新新合作中，我们将在微流体设备中重新构成肌动蛋白束，并量化拉伸力与cofilin-之间的关系 1参与。在特定目标2中，我们将剖析SF网络对迁移的贡献 ECM不可能轴向提示和在空间上排除2D SF网络的阐述的几何形状。我们假设增加限制会从2D背纤维 - 横杆 - 腹侧重定向SF组件纤维组件通向从头并行化的SF组件。我们将结合微工程文化平台，单细胞机械工具和超分辨率成像，以探测完成诱导的变化 SF组装，建筑和力学。 AIM 2将利用两种已建立的产品合作：与Ulrich Schwarz博士（U. Heidelberg），我们将开发与SF相关的多尺度计算模型网络体系结构和机制，以限制空间的细胞迁移。与神经外科医生Manish Aghi博士（UCSF），我们将通过询问胶质母细胞瘤茎的约束迁移来测试观察结果的临床价值细胞回顾性地预测体内侵袭模式。我们的研究将创造前所未有的新见解 SFS如何通过创新的方法和与人类疾病紧密联系的方式促进迁移。