CELLULAR MECHANICS AND MICROVASCULAR INTERACTION

细胞力学和微血管相互作用

基本信息

批准号：
7457993
负责人：
Richard E Waugh
金额：
$ 47.16万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7457993
关键词：
Adhesions Adhesives Affect Affinity Anions Area Atomic Force Microscopy Behavior Blocking Antibodies CXC Chemokines Calcium Cell Adhesion Molecules Cell membrane Cell surface Cells Characteristics Chemicals Chemistry Chimera organism Chloride Ion Chlorides Clinical Endothelial Cells Endothelium Family Flow Cytometry Fluorescence Fluorescence Microscopy Fluorescence Recovery After Photobleaching G-substrate Goals Inflammatory Response Integrins Intercellular adhesion molecule 1 Ions Kinetics Knowledge L-Selectin Lateral Leukocytes Ligands Ligation Lymphocyte Macrophage-1 Antigen Measurement Measures Mechanics Mediating Membrane Methods Micromanipulation Microscopy Molecular Molecular Conformation NCAM1 gene Neutrophil Infiltration Object Attachment Occupations P-Selectin P-selectin ligand protein Play Probability Protein Tyrosine Kinase Rate Reagent Regulation Role Selectins Signal Pathway Signal Transduction Solid Surface Swelling Techniques Testing Time Vascular Cell Adhesion Molecule-1 Work adhesion receptor base cellular microvillus endoglycan fluorescence imaging gastrointestinal microvillus improved migration neutrophil novel particle physical property prevent reaction rate receptor research study simulation

项目摘要

In this project we focus on the physical and chemical mechanisms that determine the kinetics of neutrophil-endothelial adhesion. Our goal is to understand the role that specific physical characteristics of the adhesive interface have on adhesion. In particular, the deformability of the cell, the microtopography of the cell membrane, the distribution and mobility of receptors, and changes in adhesion molecule affinity will be assessed in relation to the formation of adhesive contacts between neutrophils and 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 topology play in the formation of adhesive contacts. Once an understanding of the importance of these different mechanisms is reached, this knowledge will be used as a basis for understanding the mechanisms by which different signaling mechanisms work to effect changes in adhesive behavior. Specifically we will examine the effect of selectin ligation, anion transport inhibition, and exposure of cells to chemokines of the CXC family on cellular deformability, surface topography and compliance, and the distribution and mobility of adhesive ligands in relation to the kinetics of neutrophil adhesion. Finally, we will replace artificial substrates presenting endothelial cell adhesion molecules with cultured endothelial cells to identify additional modulating effects of the endothelium. Lateral mobility and distribution of adhesion receptors, cytoskeletal microrheology, and cellular deformability in the vicinity of neutrophil contact will be measured to assess the influence of physical properties of the endothelium on leukocyte adhesion and migration. These studies will result in a clear understanding of the mechanisms of neutrophil adhesion to endothelium and its regulation, and should serve as a basis for developing novel and improved strategies for clinical therapy.

在这个项目中，我们重点研究决定中性粒细胞-内皮粘附动力学的物理和化学机制。我们的目标是了解粘合界面的特定物理特性对粘合力的作用。特别是，细胞的变形能力、细胞膜的微形貌、受体的分布和移动性以及粘附分子亲和力的变化将根据嗜中性粒细胞和内皮细胞之间粘附接触的形成进行评估。通过微机械操作使单细胞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附相互作用相关的化学力和机械力的能力。这种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、表面化学和膜拓扑结构在粘合接触的形成中发挥作用。一旦了解了这些不同机制的重要性，这些知识将被用作理解不同信号机制影响粘附行为变化的机制的基础。具体来说，我们将检查选择素连接、阴离子转运抑制和细胞暴露于 CXC 家族趋化因子对细胞变形性、表面形貌和顺应性的影响，以及与中性粒细胞粘附动力学相关的粘附配体的分布和迁移性的影响。最后，我们将用培养的内皮细胞替换呈现内皮细胞粘附分子的人造基质，以鉴定内皮的额外调节作用。粘附受体的横向移动性和分布、细胞骨架微流变学以及中性粒细胞接触附近的细胞变形性将被测量以评估内皮物理特性对白细胞粘附和迁移的影响。这些研究将使人们清楚地了解中性粒细胞粘附内皮的机制及其调节，并应作为开发新的和改进的临床治疗策略的基础。