ENDOTOXIN EXPOSURE IN THE MACROPHAGE: ANALYSIS OF LIPID RAFT PROTEOMICS

巨噬细胞中的内毒素暴露：脂筏蛋白质组学分析

• 批准号: 7721400
• 负责人: Joseph Cuschieri
• 金额: $ 2.4万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-08 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721400
• 关键词: Cells; Ceramides; Characteristics; Complement 5a; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; Endotoxins; Functional disorder; Funding; Gel; Generations; Grant; In Vitro; Infection; Inflammatory; Injury; Institution; Kinetics; Lipopolysaccharides; Mediator of activation protein; Membrane; Membrane Microdomains; Molecular; Mononuclear; Multiple Organ Failure; Multiple Trauma; Organ; Oxidants; Phase; Platelet Activating Factor; Production; Proteins; Proteomics; Recovery; Research; Research Personnel; Resources; Risk; Sepsis; Source; Sphingomyelinase; Stimulus; Stress; Techniques; Testing; Toll-like receptors; Trauma; United States National Institutes of Health; fluidity; improved; macrophage; monocyte; response; tissue fixing

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. While initial survival following severe trauma has improved, long term recovery is frequently impaired by the development of multiple organ dysfunction syndrome (MODS). Following trauma, MODS is usually in response to infection and associated factors, such as lipopolysaccharide (LPS), which is characteristic of sepsis. Following trauma and during sepsis, the circulating monocyte and tissue-fixed macrophage undergo phenotypic differentiation and reprogramming that results in organ injury. In our background studies, we found that this state can be mimicked in vitro by factors induced by severe trauma, such as platelet activating factor (PAF), oxidant stress and complement 5a (C5a) and by tolerance induction. Cellular reprogramming is not associated with significant liberation of inflammatory mediators. Rather, cellular reprogramming leads to dysregulated inflammatory mediator production in response to subsequent infectious stimuli, thus increasing the risk for organ dysfunction. The mechanism responsible is associated with initial ceramide generation by sphingomyelinase, resulting in altered fluidity within specialized membrane components termed lipid rafts. Due to this gel-phase fluidity, altered kinetics occur leading to re-organization of raft proteins. This alteration in protein content is associated with the pre-assembly of Toll-like receptor (TLR) components that results in accelerated TLR complex assembly and activation in response to subsequent stimuli. To explore the molecular mechanisms responsible, we will test the hypothesis that formation of these complexes occurs during cellular reprogramming and activation using proteomic techniques on raft and non-raft protein isolates. The specific aims of this project are to: 1) Determine the lipid raft characteristics of mononuclear cells following LPS stimulation 2) Determine the lipid raft characteristics of mononuclear cells following reprogramming by factors induced by severe injury

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 虽然严重创伤后的初始生存率有所改善，但长期恢复常常因多器官功能障碍综合征 (MODS) 的发展而受到损害。 创伤后，MODS 通常是对感染和相关因素（如脂多糖 (LPS)）的反应，脂多糖 (LPS) 是脓毒症的特征。 创伤后和脓毒症期间，循环单核细胞和组织固定巨噬细胞经历表型分化和重编程，导致器官损伤。 在我们的背景研究中，我们发现这种状态可以通过严重创伤诱导的因素在体外模拟，例如血小板活化因子（PAF）、氧化应激和补体5a（C5a）以及耐受诱导。 细胞重编程与炎症介质的显着释放无关。 相反，细胞重编程会导致炎症介质的产生失调，以应对随后的感染刺激，从而增加器官功能障碍的风险。 其机制与鞘磷脂酶最初产生神经酰胺有关，导致称为脂筏的特殊膜成分内的流动性改变。 由于这种凝胶相流动性，发生动力学改变，导致筏蛋白重组。 这种蛋白质含量的改变与 Toll 样受体 (TLR) 组件的预组装有关，从而导致 TLR 复合物组装加速并激活以响应后续刺激。 为了探索相关的分子机制，我们将使用蛋白质组技术对筏和非筏蛋白分离物检验这些复合物的形成发生在细胞重编程和激活过程中的假设。 该项目的具体目标是： 1) 确定LPS刺激后单核细胞的脂筏特征 2) 确定严重损伤诱导因素重编程后单核细胞的脂筏特征