Outside-in Mechanotransduced Inflammatory Targets

由外而内的机械传导炎症目标

• 批准号: 9975676
• 负责人: Scott Irwin Simon
• 金额: $ 37.04万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975676
• 关键词: Acute; Adhesions; Adhesives; Affinity; Anatomy; Anti-Inflammatory Agents; Atherosclerosis; Autoimmune Diseases; Avidity; Bacterial Infections; Binding; Biochemical; Blood; Blood Vessels; Blood flow; Calcium; Cell Adhesion Molecules; Cells; Chronic; Code; Cues; Disease; Dissociation; Docking; Drug Screening; Drug Targeting; E-Selectin; Emigrations; Endothelial Cells; Endothelium; Epitopes; Event; Family; G-Protein-Coupled Receptors; Genes; Genetic Diseases; Goals; Homeostasis; Human; ITGB2 gene; Image; Immune; Immunity; Immunofluorescence Immunologic; Immunologic Surveillance; Infection; Inflammation; Inflammatory; Innate Immune Response; Integrins; Intercellular adhesion molecule 1; Interruption; Ischemia; L-Selectin; Lead; Leukocyte Adhesion Deficiency; Leukocytes; Ligand Binding; Ligands; Ligation; Link; Lung; Malignant Neoplasms; Mechanics; Mediating; Membrane; Microfluidics; Molecular; Molecular Target; Motor Activity; Mucous Membrane; Mutation; Necrosis; Neutrophil Activation; Neutrophil Infiltration; Pathway interactions; Patients; Phosphorylation; Play; Process; Property; Psoriasis; Regulation; Regulatory Pathway; Role; Selectins; Shapes; Signal Transduction; Signaling Molecule; Site; Skin; Structure; Surveys; Talin; Technology; Testing; Therapeutic; Time; Tissues; Transgenic Mice; Translations; Up-Regulation; Work; base; chronic infection; chronic inflammatory disease; design; granulocyte; innovation; insight; intercellular cell adhesion molecule; leukocyte activation; mechanotransduction; migration; mimetics; molecular scale; mortality; mouse model; neutrophil; novel; novel therapeutics; outcome forecast; pathogen; receptor; recruit; release of sequestered calcium ion into cytoplasm; screening; shear stress; small molecule; synergism; tool

Abstract Neutrophils mount an early and critical defense in the innate immune response to pathogens, but are also associated with chronic inflammatory diseases such as atherosclerosis, autoimmune disease, and cancer. Recruitment of leukocytes to sites of acute inflammation is a finely orchestrated process initiated by membrane expression and functional activation of leukocyte and endothelial cell adhesion molecules (CAMs) including selectins, integrins, and Ig-super family ligands. A set of unifying themes have emerged over the tenure of this R01 which provide molecular scale insight into how PMNs integrate force as a spatio-mechanical cue in the transition from rolling to firm arrest and transendothelial migration: 1) Selectins are endowed with mechanical and biochemical properties that allow their dissociation lifetime to increase with the rate of tensile loading; 2) Shear stress and transmembrane calcium release-activated Ca2+ (CRAC) channels regulate calcium flux which functions to synchronize integrin mediated arrest and shape orientation under shear; 3) allosteric upregulation in affinity of both LFA-1 and L-selectin in binding ICAM-1 and E-selectin, respectively, facilitates tensile force buildup to trigger key outside-in signals that are cooperative during PMN recruitment. In this competitive renewal we apply innovative vascular mimetic microfluidic channels combined with real-time immunofluorescence imaging to pursue the following specific aims: 1) Establish the ligand binding and mechanotransduction events by which human neutrophils are activated to arrest on E-selectin/ICAM-1 via signaling through L-selectin. 2) Define the mechanotransduction mechanism by which LFA-1 bonds to ICAM-1 elicits outside-in signaling via Kindlin-3 and other as yet unidentified adaptors. 3) Develop and test small molecule antagonists to allosteric domains that interrupt neutrophil activation and recruitment using vascular mimetic microfluidic screening technology. Our strategy entails the use of freshly isolated human neutrophils with the overarching goal of identifying regulatory pathways and molecular targets for prognosis and treatment of granulocytic inflammatory diseases. Keywords Neutrophils, Tensile bond force, Integrins, Selectins, Inflammation, Adhesion, Signaling

抽象的 中性粒细胞在先天免疫反应中发挥早期和关键的防御作用 病原体，但也与慢性炎症性疾病有关，例如 动脉粥样硬化、自身免疫性疾病和癌症。将白细胞募集至以下部位 急性炎症是由膜表达引发的精心策划的过程 以及白细胞和内皮细胞粘附分子 (CAM) 的功能激活 包括选择素、整合素和 Ig-super 家族配体。一组统一的主题 在 R01 的任期内出现，它提供了分子尺度的洞察力 PMN 将力作为从滚动到坚定过渡的空间机械线索 阻滞和跨内皮迁移：1）选择素被赋予机械和 生化特性使其解离寿命随着解离速率的增加而增加 拉伸载荷； 2）剪切应力和跨膜钙释放激活的Ca2+ (CRAC) 通道调节钙通量，其功能是同步整合素介导的 剪切力下的滞留和形状定向； 3) 两者亲和力的变构上调 LFA-1 和 L-选择素分别结合 ICAM-1 和 E-选择素，促进拉伸 力量积聚触发 PMN 期间合作的关键由外向内信号 招聘。在这次竞争更新中，我们应用创新的血管模拟 微流控通道结合实时免疫荧光成像来追踪 具体目标如下： 1）建立配体结合和力转导 人类中性粒细胞被激活并在 E-选择素/ICAM-1 上停滞的事件 通过 L-选择素信号传导。 2）定义机械传导机制 LFA-1 与 ICAM-1 结合，通过 Kindlin-3 和其他途径引发由外向内的信号传导 未识别的适配器。 3) 开发并测试变构小分子拮抗剂 使用血管模拟物中断中性粒细胞激活和募集的结构域 微流控筛选技术。我们的策略需要使用新鲜分离的人类 中性粒细胞的总体目标是确定调控途径和分子 粒细胞炎症性疾病的预后和治疗目标。 关键词 中性粒细胞、拉伸键力、整合素、选择素、炎症、粘附、信号传导