Mechano-dynamics of the Transition to Firm Adhesion and MoIotility in Neutrophils

中性粒细胞向牢固粘附和运动性转变的机械动力学

• 批准号: 8006825
• 负责人: Daniel A Hammer
• 金额: $ 23.69万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 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/8006825
• 关键词: Adhesions; Adhesives; Affect; Antibodies; Atomic Force Microscopy; Behavior; Biological Assay; Blood; Boston; Cell Adhesion Molecules; Cell Polarity; Cell surface; Cells; Cellular Stress; Chemicals; Chemotactic Factors; Collaborations; Complex; Computing Methodologies; Cues; Data; Defect; Disease; Endothelial Cells; Endothelium; Engineering; Environment; Extravasation; GTP-Binding Proteins; Generations; Goals; HL-60 Cells; Health Sciences; Homing; Image; Inflammation; Inflammatory; Inflammatory Response; Integrin Binding; Integrins; Intervention; Kinetics; Knockout Mice; Laboratories; Lead; Learning; Leukocytes; Location; Macrophage-1 Antigen; Malignant Neoplasms; Measurable; Measures; Mechanics; Mediating; Methods; Microfluidics; Modeling; Molecular; Molecular Probes; Motion; Neutrophil Activation; Pharmacologic Substance; Property; Reaction; Research; Role; Shapes; Signal Transduction; Simulate; Small Interfering RNA; Source; Speed; Stress; System; Techniques; Technology; Testing; Time; Tissues; Traction; Transfection; Universities; Work; adhesion process; adhesion receptor; base; cell motility; chemokine; density; inhibitor/antagonist; insight; migration; neutrophil; research study; response; role model; simulation; therapy design; tool

Neutrophils are the first line of defense in the cellular inflammatory response, and studying their behavior can lead to strategies for treating inflammatory disorders. Using quantitative tools and assays, we propose to investigate the fundamental mechano-chemical processes of adhesion and motility that control neutrophil extravasation from blood into tissue. In aim 1, we will simulate integrin-mediated firm adhesion of a neutrophil by merging Adhesive Dynamics - a mechanically accurate method for modeling adhesion - with a stochastic simulation of inside-out signal transduction. These models will predict how the rate and extent of neutrophil firm adhesion are controlled by a neutrophil's internal molecular machinery. Furthermore, we will use stochastic signaling methods to analyze experiments performed in Project 3 in which a neutrophil held in a pipette is stimulated by impingement with a moleculariy-coated bead. In aim 2, we will test predictions from the modeling in aim 1 by performing flow chamber adhesion experiments in which external and internal variables are systematically varied, to confirm that our quantitative understanding of the molecular control of adhesion is correct. Working closely with Projects 2 and 5, we will use pharmacological intervention, antibodies, and sIRNA technology to adjust neutrophil components and examine their effect on the transition to firm adhesion. In aim 3, we will work with Core C and use traction force microscopy to examine the motility of neutrophils in well-defined gradients of chemoattractant. We have built a chamber that combines microfluidics, to impose well-defined chemoattractant gradients across cells, with traction force microscopy. The goal is to understand how speed and direction in neutrophil motility is related to force generation, and to understand how the molecular components in neutrophils control contractility. Previous work has shown that neutrophil directional motion is accompanied by strong loci of contractile traction stress in the uropod. Using pharmacological inhibition, antibodies and sIRNA technology, we will measure how key molecular players affect neutrophil polarity, the generation of traction stresses, and cell motion. In summary, our work will provide fundamental insights as to how molecular components control neutrophil function in inflammation.

中性粒细胞是细胞炎症反应中的第一道防线，研究其行为可以 导致治疗炎症性疾病的策略。使用定量工具和测定，我们建议 研究控制中性粒细胞的粘附和运动的基本机械化学过程 从血液到组织的渗出。在AIM 1中，我们将模拟整联蛋白介导的公司粘附 中性粒细胞通过合并粘附动力学 - 一种机械精确的建模粘附方法 - 与A 内而外信号转导的随机模拟。这些模型将预测 中性粒细胞粘附受嗜中性粒细胞的内部分子机制控制。此外，我们会的 使用随机信号传导方法分析中性粒细胞的项目3中进行的实验 用分子涂层珠子撞击在移液器中。在AIM 2中，我们将测试预测 从AIM 1中的建模通过进行流动室粘附实验，其中外部和内部 变量是系统的，以确认我们对分子控制的定量理解 粘附是正确的。与项目2和5紧密合作，我们将使用药理学干预措施， 抗体和siRNA技术调整中性粒细胞成分并检查其对过渡的影响 坚定的粘附。在AIM 3中，我们将使用Core C并使用牵引力显微镜检查 中性粒细胞在定义明确的趋化剂梯度中的运动性。我们建造了一个结合的室 微流体，以牵引力显微镜施加了跨细胞的明确定义的趋化梯度。 目的是了解中性粒细胞运动中的速度和方向与力产生以及与 了解中性粒细胞中的分子成分如何控制收缩力。以前的工作表明 嗜中性粒细胞方向运动伴随着uropod中收缩牵引力的强大基因座。使用 药理抑制作用，抗体和siRNA技术，我们将如何衡量关键分子参与者如何 影响中性粒细胞极性，牵引力的产生和细胞运动。总而言之，我们的工作将 提供有关分子成分如何控制中性粒细胞在炎症中的功能的基本见解。