Nanostructure Devices for Measuring Cell Mechanics

用于测量细胞力学的纳米结构装置

• 批准号: 7424241
• 负责人: MICHAEL Patrick SHEETZ
• 金额: $ 3.01万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 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/7424241
• 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; Numbers; Pattern; Phenotype; Polymers; Process; Protein Binding; Proteins; Range; 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; radius bone structure; research study; response; size; submicron; surface coating; tool

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.

细胞环境是决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 确定细胞反应。看来关键的机械因素是纳米级间距， 曲率和细胞外基质成分的刚性。我们已经建立了一个合作， 纳米纤维实验室和细胞生物学实验室，以探索这些因子是如何被细胞在分子水平上感知的。 水平使用电子束写入系统，我们将能够创建细胞大小的阵列， 纳米级的功能，使我们能够筛选的关键距离和模式，触发细胞 应答一旦我们定义了细胞反应（如细胞扩散）的临界距离， 缺失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 反应被改变了。特别令人感兴趣的是具有多个整合素结合位点的talin，所述多个整合素结合位点间隔约 55 nm。关于膜弯曲，我们观察到细胞对胶原和其他纤维有反应， 通过延长含有肌球蛋白II-B的板状伪足，肌球蛋白II-B-1细胞收缩纤维少于30 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在板状伪足中的组装作为定义引起肌球蛋白II的曲率范围的标准。 反应细胞不仅遵循纤维，而且通过接触引导遵循表面特征，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， 特定的突变将极大地影响细胞的生长和移动能力， 异常的形态，这将使我们能够了解参与接触的重要细胞参数， 指导刚性感测对于细胞扩散和生长是至关重要的，并且我们已经定义了许多刚性感测。 这些蛋白质是用来区分硬表面和软表面的。开发出一种 用于探测细胞表面的相邻区域上的表面刚性变化的影响的装置 探讨了刚性感知的具体机制。了解曲面的大小和几何形状 细胞感知到的特征将使我们能够以定义的方式引出给定的细胞行为。知道 所涉及的蛋白质将使我们能够使用靶向药理学抑制剂来改变定义的 的方式这些研究在组织工程和设计潜力方面有许多实际应用 用于伤口愈合、癌症和各种疾病的疗法。