Nanostructure Devices for Measuring Cell Mechanics

用于测量细胞力学的纳米结构装置

基本信息

批准号：
7876735
负责人：
MICHAEL Patrick SHEETZ
金额：
$ 6.27万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-01 至 2010-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7876735
关键词：
3-Dimensional Address Affect Architecture Area Behavior Binding Sites Cell Adhesion Cell Adhesion Molecules Cell Line Cell physiology Cell surface Cells Cellular Morphology Cellular biology Characteristics Cicatrix Collaborations Collagen Collagen Fiber Complex Contracts Custom Cytoplasmic Protein Development Devices Dimensions Disease Environment Extracellular Matrix Fiber Fibronectins Focal Adhesions Goals Growth Hand Healed Integrin Binding Integrins Length Ligands Link Malignant Neoplasms Measures Mechanics Membrane Methods Molecular Morphology Movement Muscle Rigidity Mutation Myosin ATPase Myosin Type II Nanostructures Neoplasm Metastasis Nonmuscle Myosin Type IIB Pattern Phenotype Polymers Process Protein Binding Proteins Radial Reaction Relative (related person)Resolution Signal Transduction Signaling Protein Structure Surface System Talin Testing Tissue Engineering Tissues Work Wound Healing Writing cell behavior cell motility cell type design healing inhibitor/antagonist interest nanofabrication nanofiber nanometer nanoscale novel numb protein practical application protein complex research study response submicron surface coating tool

项目摘要

Cellular environment is a major factor in determining cell behavior and ultimately cell phenotype. Many recent studies have shown that the mechanical aspects of the environment are important factors in determining the cell response. The mechanical factors that appear critical are the nanometer level spacing, curvature, and rigidity of the extracellular matrix components. We have formed a collaboration between two nanofabrication labs and a cell biology lab to explore how these factors are sensed by cells at the molecular level. Using an e-beam writing system, we will be able to create cellular sized arrays with a range of nanometer level features that will enable us to screen for the critical distances and patterns that trigger cell responses. Once we define the critical distance for a cell response such as cell spreading, we will screen a number of cell lines that are missing critical motility and signaling proteins to determine if the cell spreading response is altered. Of particular interest is talin that has multiple integrin binding sites spaced over about 55 nm. In regard to membrane curvature, we have observed that cells respond to collagen and other fibers by extending lamellipodia with myosin II-B in them and myosin \\-B-l- cells contract fibers at less than 30% efficiency. We will fabricate fibers and 2-D surface features with different radii of curvature and use the assembly of GFP-myosin II-B in lamellipodia as a criterion for defining the range of curvatures that elicit the response. Cells not only follow fibers but also follow surface features through contact guidance and we will explore the important parameters in determining cell polarization and tracking. Recent results indicate that specific mutations will dramatically affect the ability of cells to polarize and move, giving rise to grossly abnormal morphologies that will enable us to understand important cellular parameters involved in contact guidance. Rigidity sensing is critical for cell spreading and growth and we have defined a number of proteins that are needed to sense the difference between rigid and soft surfaces. The development a device for probing the effect of changes in surface rigidity on adjacent regions of the cell surface will enable us to probe the detailed mechanism of rigidity sensing. Knowing both the size and geometry of surface features that are sensed by the cells will enable us to elicit given cell behaviors in a defined way. Knowing the proteins involved will enable us to use targeted pharmacological inhibitors to alter morphology in defined ways. These studies have many practical applications in tissue engineering and in designing potential therapies for wound healing, cancer and a variety of disorders.

细胞环境是确定细胞行为和最终细胞表型的主要因素。许多最近的研究表明，环境的机械方面是重要因素确定细胞响应。似乎关键的机械因素是纳米水平间距，曲率和细胞外基质成分的刚度。我们已经建立了两个之间的合作纳米制作实验室和细胞生物学实验室，以探索分子处细胞如何感知这些因素等级。使用电子束写作系统，我们将能够创建具有一定范围的蜂窝大小数组纳米水平的功能将使我们能够筛选触发单元格的临界距离和图案回答。一旦定义了细胞响应（例如细胞扩散）的临界距离，我们将筛选一个缺少关键运动性和信号蛋白的细胞系数量，以确定细胞是否扩散响应发生了变化。特别感兴趣的是塔林（Talin 55 nm。关于膜曲率，我们观察到细胞对胶原蛋白和其他纤维的反应通过在其中延伸lamellipodia和肌球蛋白II-B，肌球蛋白\\ -b-l-细胞以小于30％的收缩纤维收缩纤维效率。我们将制造具有不同曲率半径的纤维和2-D表面特征，并使用 GFP肌球蛋白II-B组装在薄片中，作为定义引起的曲率范围的标准回复。细胞不仅遵循纤维，而且还通过接触指导遵循表面特征，我们将探索确定细胞极化和跟踪的重要参数。最近的结果表明特定突变将极大地影响细胞极化和移动的能力，从而严重产生异常的形态学将使我们能够理解接触涉及的重要细胞参数指导。刚性传感对于细胞扩散和生长至关重要，我们定义了许多需要感知刚性和软表面之间差异所需的蛋白质。开发a 探测表面刚性变化对细胞表面相邻区域的影响的设备将实现我们要探测刚性感应的详细机制。知道表面的大小和几何形状细胞感知的特征将使我们能够以定义的方式引起给定的细胞行为。会心所涉及的蛋白质将使我们能够使用靶向的药理学抑制剂来改变定义的形态方式。这些研究在组织工程和设计潜力方面具有许多实际应用伤口愈合，癌症和多种疾病的疗法。