Hydrodynamic Interactions and Cell Deformation in Neutrophil Adhesion

中性粒细胞粘附中的流体动力学相互作用和细胞变形

• 批准号: 8006838
• 负责人: Michael R. King
• 金额: $ 32.57万
• 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/8006838
• 关键词: ABL1 gene; Address; Adhesions; Adhesives; Affinity; Appearance; Arts; Behavior; Binding; Binding Proteins; Biological Assay; Biophysics; Blood flow; Cell Adhesion; Cell Communication; Cell Line; Cell Shape; Cells; Cellular Morphology; Chemotactic Factors; Collaborations; Communities; Complex; Computer Simulation; Cytoplasmic Tail; Cytoskeleton; Data; Dose; Endothelial Cells; Endothelium; Engineering; Environment; Exposure to; Extravasation; Flow Cytometry; G Protein-Coupled Receptor Genes; G-Protein-Coupled Receptors; Goals; Guanine Nucleotide Dissociation Inhibitors; HL-60 Cells; Hematopoietic; Human; Immune System Diseases; Immune response; In Vitro; Inflammation; Inflammatory; L-Selectin; Lateral; Length; Leukocyte Rolling; Leukocytes; Life; Ligands; Liquid substance; Measurable; Measurement; Measures; Mechanics; Mediating; Modeling; Molecular; Mus; P-selectin ligand protein; Paper; Peptides; Phase; Physics; Physiological; Platelet Activating Factor; Platelet Activating Factor Activation; Probability; Process; Protein Kinase; Proteins; Regulation; Reperfusion Injury; Research; Resistance; Retinal Cone; Role; Selectins; Shapes; Signal Transduction; Signaling Molecule; Site; Small Interfering RNA; Stream; Surface; Suspension substance; Suspensions; TNF-alpha converting enzyme; Testing; Tissues; Work; c-abl Proto-Oncogenes; cell motility; cell type; crosslink; immune function; in vivo; inhibitor/antagonist; mitogen-activated protein kinase p38; neutrophil; novel; platelet activating factor receptor; programs; protein aminoacid sequence; receptor; receptor expression; research study; response; shear stress; sialyl Lewis x; simulation; single molecule; small molecule; surface coating; venule

The central goal of Project 5 is to understand the interplay between fluid shear stress, cell morphology, and L-selectin expression on the dynamics of neutrophil tethering and rolling on the endothelium. We will use a combination of state-of-the-art computational simulations of receptor-mediated cell adhesion under flow, in vitro experiments with isolated human neutrophils and neutrophil-like cell lines in well-deflned fluid shear environments, and collaborative invesfigafion with other projects. The mulfiparticle adhesive dynamics simulafion developed by the PI, enables the invesfigafion of previously unaddressed problems such as the influence of non-spherical shape on the physics of leukocyte rolling, and computational and experimental study of L-selecfin shedding and mechanosensing. The multitude of physical determinants combining to control neutrophil inflammatory recruitment, including receptor expression, activation state, cell shape, local flow environment, and cell-cell collisions are highly complex and nonlinear and so we have taken a systematic integrated engineering approach to elucidate these behaviors. The proposed work is organized around three specific aims. Aim 1: Selecfin-Mediated Tethering and Rolling of Activated Leukocytes: In this aim we will use mulfiparticle adhesive dynamics simulafions of acfivated cell shapes, and detailed analysis of in vivo observations of activated cell rolling, to study the dynamics of non-spherical cell adhesion. Aim 2: Mechanisms of L-Selecfin Mechanotransducfion and Shedding During Rolling. This aim will explore the molecular mechanisms of mechanical shedding in flow chamber experiments with primary neutrophils and an altered neutrophil-like cell line. Aim 3: Shear-Induced Resistance to Activation via Chemoattractant GPCRs. In this aim, we will study the quantitafive dynamics of the shear stress-dependent GPCR-mediated response of neutrophils to fMLP and platelet activating factor (PAF). Together, the proposed research will determine for the first fime how the physics of nonspherical leukocyte shape, and the mechanical response of neutrophil receptors at the single molecule level, influence the dynamics of cell tethering and rolling to selecfin-presenfing endothelium under physiological flow.

项目 5 的中心目标是了解流体剪切应力、细胞形态和 L-选择素表达对中性粒细胞在内皮上束缚和滚动动力学的影响。我们将使用一个 流下受体介导的细胞粘附的最先进的计算模拟相结合， 在明确的流体剪切中使用分离的人中性粒细胞和中性粒细胞样细胞系进行体外实验 环境以及与其他项目的协作调查。多颗粒粘合剂动力学 PI 开发的仿真能够研究以前未解决的问题，例如 非球形形状对白细胞滚动物理的影响以及计算和实验 L-选择蛋白脱落和机械传感的研究。众多的物理决定因素结合在一起 控制中性粒细胞炎症募集，包括受体表达、激活状态、细胞形状、局部 流动环境和细胞间碰撞是高度复杂和非线性的，因此我们采取了 系统集成工程方法来阐明这些行为。建议的工作已组织好 围绕三个具体目标。目标 1：Selecfin 介导的活化白细胞的束缚和滚动：在此 目标我们将使用活化细胞形状的多粒子粘附动力学模拟，并详细分析 体内观察激活的细胞滚动，以研究非球形细胞粘附的动力学。目标 2： 轧制过程中 L-Selecfin 力传导和脱落的机制。这一目标将探索 原代中性粒细胞流室实验中机械脱落的分子机制 改变的中性粒细胞样细胞系。目标 3：剪切诱导的化学引诱剂抗激活性 GPCR。为此，我们将研究剪切应力依赖性 GPCR 介导的定量动力学 中性粒细胞对 fMLP 和血小板激活因子 (PAF) 的反应。总之，拟议的研究将 首次确定非球形白细胞形状的物理原理以及机械响应 中性粒细胞受体在单分子水平上的作用，影响细胞束缚和滚动的动力学 生理流下的选择性内皮细胞。