Cellular Mechanics and Microvascular Interactions

细胞力学和微血管相互作用

基本信息

批准号：
8006834
负责人：
Richard E Waugh
金额：
$ 57.1万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8006834
关键词：
Address Adhesions Adhesives Affect Affinity Antibodies Area Atherosclerosis Avidity Behavior Biological Models Biophysics Blocking Antibodies Calcium Cell Adhesion Cell Adhesion Molecules Cell Communication Cell Line Cell membrane Cell physiology Cell surface Cells Characteristics Chemicals Chemistry Clinical Complex Computer Simulation Cues Cytoskeleton Data Diffuse Diffusion Disease Distal Effectiveness Endothelium Environment Event Fibrinogen Fluorescence Fluorescence Microscopy Fluorescence Recovery After Photobleaching Fluorescence Resonance Energy Transfer Funding Genetically Engineered Mouse Germ Cells Goals Guanine Nucleotide Exchange Factors HL-60 Cells Health Heart Diseases Human IL8 gene Infection Inflammation Inflammatory Inflammatory Response Integrin Binding Integrins Intercellular adhesion molecule 1 Interphase Cell Investigation Knockout Mice Knowledge L-Selectin Label Lateral Lead Leukocytes Ligands Ligation Liquid substance Literature Macrophage-1 Antigen Malignant Neoplasms Measures Mechanics Mediating Membrane Membrane Proteins Molecular Conformation Movement Mus Neoplasm Metastasis Outcome Study Physiology Play Publications Regulation Relative (related person)Reporter Role Signal Pathway Signal Transduction Signaling Molecule Site Small Interfering RNA Spatial Distribution Speed Stimulus Stress Structure Surface System TNF-alpha converting enzyme Testing Time Translating Travel Up-Regulation Vascular Endothelium Work adhesion receptor cell behavior cell motility chemokine chemokine receptor computer framework density fluorescence imaging imaging modality improved in vivo migration neuronal cell body neutrophil physical property programs protein distribution receptor response shear stress surface coating tool

项目摘要

Our goal in this project is to understand the role that specific physical characteristics of the adhesive interface have on adhesion and subsequent cell behavior. In particular, our investigation focuses on the microtopography of the cell membrane, the distribution and mobility of receptors, and changes in adhesion molecule affinity, as well as how these attributes change as a result of chemokine stimulus and bond formation between neutrophils and the endothelium. Micromechanical manipulation of single cells into contact with artificial substrates with well-defined adhesion molecule presentation provides unparalleled ability to control both the chemistry and the mechanical forces in relation to adhesive interactions. This approach, combined with newly implemented fluorescence imaging methods, enables us to determine the specific role that cellular mechanics, surface chemistry, and membrane topography play in the formation of adhesive contacts. Building on knowledge of the fundamental contributions of these factors obtained In the previous period, we will extend our investigations to determine how the physical and chemical characteristics of the cell surface and the underiying substrate work to effect changes in adhesive behavior and cell migration. Specifically, we will determine how contact with surfaces presenting immobilized IL8 and adhesion receptors (principally ICAM-1) induces changes in surface topography, and leads via key signaling intermediates (e.g., calcium, RAP-1 and the calcium-dependent guanine nucleotide exchange factor CalDAG GEFl) to integrin activation and adhesion. We will also measure the effects of chemokine stimulus on the distribution, mobility and activation state of adhesive ligands and the stability of the membrane cytoskeletal interface. Finally, we will determine how changes in cell surface microtopgraphy at the interface with its substrate enhance haptotactic signals, and how the distribution and concentration of those haptotactic signals lead to cell spreading and directed cell crawling. These studies will result in a clearer understanding of the mechanisms of neutrophil adhesion and migration on endothelium and its regulation, and thus result in a clearer and more detailed understanding of the inflammatory response in health and disease.

我们在这个项目中的目标是了解粘合剂的特定物理特性的作用界面对粘附和随后的细胞行为有影响。特别是，我们的调查重点是细胞膜的微形貌、受体的分布和移动性以及粘附的变化分子亲和力，以及这些属性如何因趋化因子刺激和键而改变中性粒细胞和内皮细胞之间的形成。单细胞的微机械操纵与具有明确粘附分子呈现的人造基质接触提供了无与伦比的控制与粘合剂相互作用相关的化学力和机械力的能力。这方法与新实施的荧光成像方法相结合，使我们能够确定细胞力学、表面化学和膜形貌在形成中发挥的特定作用粘性接触。基于对这些因素的基本贡献的了解前期，我们将扩大调查范围，以确定物理和化学特性如何细胞表面和底层基质的作用会影响粘附行为和细胞的变化迁移。具体来说，我们将确定如何与呈现固定化 IL8 和粘附受体（主要是 ICAM-1）诱导表面形貌的变化，并通过关键信号传导引导中间体（例如钙、RAP-1 和钙依赖性鸟嘌呤核苷酸交换因子 CalDAG GEF1)对整联蛋白激活和粘附。我们还将测量趋化因子刺激对粘附配体的分布、移动性和活化状态以及膜细胞骨架的稳定性界面。最后，我们将确定与其接触的界面处的细胞表面微形貌如何变化。底物增强趋触信号，以及这些趋触信号的分布和浓度如何信号导致细胞扩散和定向细胞爬行。这些研究将带来更清晰的认识中性粒细胞在内皮细胞粘附和迁移的机制及其调控，从而导致对健康和疾病中的炎症反应有更清晰、更详细的了解。