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

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 虽然严重创伤后的初始生存率有所提高，但长期恢复经常因多器官功能障碍综合征（MODS）的发展而受损。 创伤后多器官功能不全综合征（MODS）通常是由感染和相关因素如脂多糖（LPS）引起的，其是脓毒症的特征。 创伤后和脓毒症期间，循环单核细胞和组织固定的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子（如血小板活化因子（PAF）、氧化应激和补体5a（C5 a））以及耐受诱导在体外模拟。 细胞重编程与炎症介质的显著释放无关。 相反，细胞重编程导致响应于随后的感染刺激的炎症介质产生失调，从而增加器官功能障碍的风险。 负责的机制与鞘磷脂酶的初始神经酰胺生成有关，导致称为脂筏的专门膜成分内的流动性改变。 由于这种凝胶相流动性，动力学发生改变，导致筏蛋白的重组。 蛋白质含量的这种改变与Toll样受体（TLR）组分的预组装相关，其导致TLR复合物响应于随后的刺激而加速组装和活化。 为了探索负责的分子机制，我们将使用蛋白质组学技术对筏和非筏蛋白分离物进行细胞重编程和激活过程中发生这些复合物形成的假设。 该项目的具体目标是： 1)确定LPS刺激后单个核细胞的脂筏特征 2)确定严重创伤诱导因子重编程后单个核细胞的脂筏特征