CELLULAR MECHANICS AND MICROVASCULAR INTERACTION

细胞力学和微血管相互作用

• 批准号: 6932951
• 负责人: Richard E Waugh
• 金额: $ 24.65万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6932951
• 关键词: atomic force microscopy; cell adhesion; cell differentiation; cell morphology; cell senescence; clinical research; flow cytometry; fluorescence recovery after photobleaching; fluorescent dye /probe; hemodynamics; human subject; integrins; laboratory mouse; leukocyte adhesion molecules; membrane activity; membrane structure; neutrophil; single cell analysis; tissue /cell culture; vascular cell adhesion molecule; vascular endothelium

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，我们着重于确定嗜中性粒细胞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在粘附方面具有特定的物理特征的作用。特别是，将评估细胞的可变形性，细胞膜的微图，受体的分布和迁移率以及粘附分子亲和力的变化，以与中性粒细胞和内皮之间的粘附接触形成有关。将单个细胞的微电机械操作与具有明确定义的粘附分子呈现的人工底物接触，提供了无与伦比的能力，可以控制化学和机械力与粘合性相互作用相关的能力。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 膜拓扑在形成粘合剂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，该知识将被用作理解不同信号传导机制影响粘附行为变化的机制的基础。具体而言，我们将研究选择结扎，阴离子转运抑制以及细胞向CXC家族的趋化因子暴露对细胞变形性，表面形象和依从性的影响，以及粘合剂配体与中性粒细胞粘附动力学有关的粘合剂配体的分布和迁移率。最后，我们将取代用培养的内皮细胞呈现内皮细胞粘附分子的人工底物，以鉴定内皮的其他调节作用。中性粒细胞接触附近的横向迁移率和粘附受体，细胞骨架微流体学和细胞可变形性将是 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对内皮及其调节的机制，并应作为制定新颖和改进的临床治疗策略的基础。