CORE D - Proteomics Core

摘要 蛋白质组学核心设施(核心D)将支持拟议的项目，以解决 计划：蛋白糖基化和糖蛋白重塑改变败血症的凝血障碍和炎症。 核心D使用最先进的质谱仪分析脓毒症患者的血浆和细胞蛋白质组。核心D具有 在第一个供资周期为P01的进展作出了重要贡献，提供了重要的调查结果 以支持所有三个研究项目，以及导致两篇论文发表的发现，其中一篇发表在《自然通讯》杂志上。 核心D的具体目标是：1)评估血浆蛋白质组的时间和病原体依赖的变化 由脓毒症引起，2)表征时间和病原体依赖的血管表面重塑 蛋白质组，以及3)确定在革兰氏病过程中表现出的增强性消耗性凝血障碍的分子基础。 阴性败血症。 目标1专注于所有三个项目的血浆蛋白质组学。项目1将使用血浆蛋白质组学来识别 特异性糖苷酶、其糖蛋白底物和胞内凝集素受体，并确定它们对 它们对脓毒症凝血障碍和炎症的动态平衡调节。项目2将使用血浆 蛋白质组学鉴定决定毒力的病原体和寄主的成分和机制 脓毒症的发病机制及其在实验和人类分层和预后中的作用 败血症。项目3将使用血浆蛋白质组学来确定血管表面标志物在血液循环中的脱落 脓毒症的小鼠模型及其在脓毒症的凝血和炎症中的影响 可能使人类疾病分层和预后得以实现的机械联系。 目的2重点描述支持项目3的血管表面蛋白质组。这项工作将表征 血管细胞表面蛋白质组的器官特异性变化，并确定它们在凝血障碍和 败血症的炎症及其在人类疾病分层和预后中的作用。 AIM 3将为项目2提供支持，以确定在以下情况下表现出的增强型消耗性凝血障碍的特征 革兰氏阴性败血症。这项工作将使用基于活性的蛋白质组学和标准蛋白质分级分析 鉴定在SC/ST/EC中观察到的FXI刺激活性的细菌功能(S) 上清液；并确定其在革兰氏阴性脓毒症和 在人类疾病的分层和预后方面的可能效用。

SUMMARY The Proteomics Core facility (Core D) will support the proposed projects to address the central hypothesis of the program; protein glycosylation and glycoprotein remodeling alter the coagulopathy and inflammation of sepsis. Core D uses state-of-the-art mass spectrometry to analyze plasma and cellular proteomes in sepsis. Core D has made essential contributions to the progress of the P01 during the first funding cycle, providing important findings in support of all three research projects, and findings that led to two publications, one in Nature Communications. The Specific Aims of Core D are to: 1) Assess time and pathogen-dependent changes to the Plasma Proteome resulting from sepsis, 2) Characterize the time and pathogen-dependent remodeling of the vascular surface proteome, and 3) Determine the molecular basis of enhanced consumption coagulopathy exhibited during Gram- negative sepsis. Aim 1 focuses on plasma proteomics for all three projects. Project 1 will use plasma proteomics to identify specific glycosidases, their glycoprotein substrates, and endocytic lectin receptors and determine their impact of their homeostatic regulation on the coagulopathy and inflammation of sepsis. Project 2 will use plasma proteomics to identify components and mechanisms of both the pathogen and host determining virulence and sepsis pathogenesis and determine their utility in the stratification and prognosis of experimental and human sepsis. Project 3 will use plasma proteomics to determine vascular surface marker shedding into circulation in murine models of sepsis, and their impact in the coagulopathy and inflammation of sepsis while identifying mechanistic links that may enable human disease stratification and prognosis. Aim 2 focuses on characterizing the vascular surface proteome in support of Project 3. This work will characterize organ-specific changes to vascular cell surface proteomes and determine their impact in the coagulopathy and inflammation of sepsis and their utility in the stratification and prognosis of human disease. Aim 3 will support Project 2 in seeking to characterize the enhanced consumption coagulopathy exhibited during Gram-negative sepsis. This work will use activity-based proteomics and standard protein fractionation analysis to identify bacterial function(s) responsible for the observed FXI-stimulatory activity present in SC/ST/EC supernatants; and determine its role in the elevated consumption coagulopathy in Gram-negative sepsis and possible utility in the stratification and prognosis of human disease.