Towards a Molecular Signature of Neutrophil Priming

中性粒细胞启动的分子特征

• 批准号: 7708311
• 负责人: ARTHUR Robert SALOMON
• 金额: $ 20.66万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-17 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7708311
• 关键词: Address; Affect; Applications Grants; Arts; Biochemical; Bone Marrow; Cell Line; Cells; Cessation of life; Collaborations; Complex; Data; Detection; Development; Disease; Dose; Environmental Risk Factor; Eukaryotic Cell; Event; Experimental Designs; Exposure to; Functional disorder; Future; Global Change; Goals; Host Defense; Human; Imagery; Infection; Inflammation; Inflammatory; Inflammatory Response; Injury; Laboratories; Lead; Life; Lipopolysaccharides; Mass Spectrum Analysis; Mediator of activation protein; Methods; Modeling; Molecular; Molecular Profiling; Noise; Organ failure; Pathway interactions; Patients; Peptides; Phosphorylation; Phosphorylation Site; Platelet Activating Factor; Protein Array; Proteins; Proteome; Publishing; Reactive Oxygen Species; Reagent; Receptor Activation; Relative (related person); Reperfusion Injury; Reporting; Rest; Role; Sampling; Sepsis; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Molecule; Site; System; Systems Analysis; T-Lymphocyte; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Trauma; Tyrosine Phosphorylation; Tyrosine Phosphorylation Site; Work; cytokine; design; in vivo; injury and repair; mast cell; methionyl-leucyl-phenylalanine; microbial; network models; neutrophil; novel diagnostics; public health relevance; response

DESCRIPTION (provided by applicant): Neutrophil priming is an integral aspect of a well-regulated inflammatory response and is the first step in controlling microbial infection and repair of injury. In this model, an initial injury "primes" neutrophils such that a secondary insult, which would otherwise be tempered in an unprimed cell, now results in an exaggerated inflammatory response. The excessive and unregulated release of cytokines and toxic oxygen radicals causes damage to adjacent tissues leading to tissue dysfunction, organ failure and death. The precise molecular pathways initiated upon priming of human neutrophils are poorly understood. The present proposal focuses on the application of quantitative phosphoproteomic technology to understand the unique proteome-wide tyrosine phosphorylation events associated with human neutrophil priming by exposure to low dose N-formyl-met-leu-phe, lipopolysaccharide or platelet activating factor. Using currently available quantitative phosphoproteomic technology and expertise with primary human neutrophils in our laboratories, we will sample primed neutrophils at various time points and quantify relative changes in wide-scale tyrosine phosphorylation from total cell lysates of cellular peptides. Information to be derived from this project include: global identification of tyrosine phosphorylation events that occur over time after neutrophil priming; visualization of the network of signal transduction pathways that are activated in the primed neutrophils; comparison of activation events that take place over time from neutrophils primed with different proinflammatory mediators. This type of quantitative analysis of global tyrosine phosphorylation sites during a time course of neutrophil priming along with traditional biochemical analysis will permit the construction of a signaling pathway network model for neutrophil priming. Together these findings will test the hypothesis that the primed state of the neutrophil includes signaling events that are common among proinflammatory mediators and can be presented as a molecular signature of this fundamental cellular response. Given that responding neutrophils must interpret a number of signals at a site of inflammation, the results from these studies may assist in the development of novel diagnostic or therapeutic reagents aimed at limiting the auto-inflammatory tissue damage patients suffer as a result of sepsis and trauma. PUBLIC HEALTH RELEVANCE: Neutrophil priming is an integral aspect of a well-regulated inflammatory response and is the first step in controlling microbial infection and repair of injury. The precise molecular pathways initiated upon priming of human neutrophils are poorly understood. In this proposal we introduce modern methods in quantitative mass spectrometry to facilitate the characterization of the cellular signaling pathways initiated upon neutrophil priming by a variety of agents by providing a global view of the phosphorylation state of normal and primed cells.

描述（由申请人提供）：中性粒细胞启动是炎症反应良好的组成部分，是控制微生物感染和修复损伤的第一步。在此模型中，最初的损伤“素”中性粒细胞，使得次要的损伤（否则将在未刺激的细胞中都会降低），现在导致夸张的炎症反应。细胞因子和有毒氧自由基的过度和不调节的释放会损害相邻组织，导致组织功能障碍，器官衰竭和死亡。在启动人类嗜中性粒细胞时引发的精确分子途径知之甚少。本提案的重点是应用定量磷酸蛋白质组学技术，以了解与人类嗜中性粒细胞启动相关的独特蛋白质酪氨酸磷酸化事件，通过暴露于低剂量N-甲基甲基甲基 - leu-phe，脂多糖或血小板激活因子。使用当前可用的定量磷酸化蛋白质学技术和实验室中原代人性中性粒细胞的专业知识，我们将在各个时间点采样启动的嗜中性粒细胞，并量化来自细胞肽的总细胞裂解物的广泛酪氨酸磷酸化的相对变化。从该项目中得出的信息包括：中性粒细胞启动后随着时间的时间发生的酪氨酸磷酸化事件的全球鉴定；在引发性中性粒细胞中激活的信号转导途径网络的可视化；从中性粒细胞随着不同促炎性介体引发的激活事件的比较。这种类型的定量分析在中性粒细胞启动期间全球酪氨酸磷酸化位点以及传统的生化分析将允许构建中性粒细胞启动的信号通路网络模型。这些发现共同检验了以下假设：中性粒细胞的启动状态包括在促炎介质中常见的信号传导事件，并且可以作为这种基本细胞反应的分子特征表示。鉴于中性粒细胞的反应必须在炎症部位解释许多信号，因此这些研究的结果可能有助于开发新型的诊断或治疗试剂，旨在限制自身炎症组织损害患者因脓毒症和创伤而遭受的损失。公共卫生相关性：中性粒细胞启动是炎症反应良好的组成部分，是控制微生物感染和损伤修复的第一步。在启动人类嗜中性粒细胞时引发的精确分子途径知之甚少。在此提案中，我们引入了定量质谱法中的现代方法，以促进通过各种药物在中性粒细胞启动时通过各种药物启动的细胞信号传导途径的表征，通过提供正常细胞和引发细胞的磷酸化状态的全局视图。