Analyses of 120nm Local Contractions Linked to Rigidity Sensing

与刚性传感相关的 120nm 局部收缩分析

• 批准号: 8539049
• 负责人: James C Hone
• 金额: $ 28.31万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8539049
• 关键词: Adhesions; Affect; Area; Behavior; Biochemical Process; Biological; Cell Differentiation process; Cell Line; Cell physiology; Cells; Characteristics; Complex; Devices; Dimensions; Disease; Elastomers; Event; Focal Adhesions; Frequencies; Generations; Goals; Grant; Knock-out; Link; Longitudinal Studies; Measurement; Measures; Mechanics; Mesenchymal Stem Cells; Molecular; Molecular Models; Movement; Mutation; Natural regeneration; Pattern; Pharmaceutical Preparations; Play; Process; Property; Research Personnel; Role; Site; Surface; Techniques; Testing; Time; Ursidae Family; Work; base; chemical property; inhibitor/antagonist; molecular modeling; molecular scale; mutant; nanometer; research study; response; stem cell differentiation; submicron; tool

DESCRIPTION (provided by applicant): For a cell to differentiate properly it must sense both the mechanical and chemical properties of its microenvironment. Sensing of the matrix mechanical properties plays a critical role and influences many aspects of cellular differentiation, regeneration and disease. Matrix mechanosensing is not understood in part because many of the steps involved occur at a molecular scale of nanometers. We have recently shown that submicrometer pillars are displaced at early times by a local contraction of 120 nm. This is not observed with larger (>1 mm) pillars. Because these early nanometer displacements are altered in cells that are unable to respond to matrix rigidity changes, we suggest that the cells use the force needed to produce a 120 nm displacement of matrix as a measure of matrix rigidity as others also believe. To understand how this process occurs will require a combination of submicrometer fabrication and quantitative cell biological techniques that the two investigators can bring to bear upon this problem. We will define the steps in the local contractions at a second and nanometer level using new nanofabricated devices that will test the effects of changes in matrix area, spacing and rigidity even on single cells. From a description of the steps in the process, we will analyze which molecular complexes are involved in each step. These same tools will enable us to analyze the mechanical factors that affect adhesion maturation and to know if local contractions continue in fully spread cells. Using mesenchymal stem cells, we will determine the time course of the contractions that precede differentiation in response to rigid and soft small pillars. This grant will enable us to determine the bases for the mechanochemical steps involved in stem cell differentiation that will enable targeting of the specific steps for manipulations of differentiation or disease processes.

描述（由申请人提供）：细胞要正确分化，必须感知其微环境的机械和化学特性。基质机械特性的传感起着至关重要的作用，并影响细胞分化、再生和疾病的许多方面。矩阵机械传感尚不被理解，部分原因是所涉及的许多步骤发生在纳米分子尺度上。我们最近发现，亚微米柱在早期因 120 nm 的局部收缩而发生位移。对于较大（>1 毫米）的柱子，不会观察到这种情况。由于这些早期纳米位移在无法响应基质刚性变化的细胞中发生了改变，因此我们建议细胞使用产生 120 nm 基质位移所需的力作为基质刚性的度量，正如其他人也认为的那样。要了解这个过程是如何发生的，需要结合亚微米制造和定量细胞生物学技术，两位研究人员可以利用这些技术来解决这个问题。我们将使用新的纳米制造设备来定义秒级和纳米级的局部收缩步骤，这些设备将测试基质面积、间距和刚性变化的影响，甚至对单个细胞也是如此。从过程步骤的描述中，我们将分析每个步骤涉及哪些分子复合物。这些相同的工具将使我们能够分析影响粘附成熟的机械因素，并了解完全扩散的细胞中局部收缩是否持续。使用间充质干细胞，我们将确定分化之前响应刚性和柔软小柱的收缩的时间过程。这笔资助将使我们能够确定干细胞分化中涉及的机械化学步骤的基础，从而能够针对分化或疾病过程操作的特定步骤进行靶向。