CORE D - Proteomics Core

CORE D - 蛋白质组学核心

• 批准号: 10641845
• 负责人: Jeffrey W Smith
• 金额: $ 58.95万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641845
• 关键词: Address; Blood Coagulation Disorders; Blood Vessels; Cell surface; Circulation; Communication; Core Facility; Disease stratification; Disseminated Intravascular Coagulation; Exhibits; Fractionation; Funding; Glycoproteins; Glycoside Hydrolases; Human; Inflammation; Lectin Receptors; Link; Mass Spectrum Analysis; Molecular; Nature; Organ; Pathogenesis; Plasma; Prognosis; Protein Glycosylation; Proteins; Proteome; Proteomics; Publications; Regulation; Research Project Grants; Role; Sepsis; Stratification; Surface; Time; Vascular remodeling; Virulence; Work; disease prognosis; gram-negative sepsis; human disease; mouse model; pathogen; programs

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.

概括 蛋白质组学核心设施（核心 D）将支持拟议的项目，以解决以下问题的中心假设： 程序;蛋白质糖基化和糖蛋白重塑改变脓毒症的凝血障碍和炎症。 Core D 使用最先进的质谱分析脓毒症中的血浆和细胞蛋白质组。核心D有 在第一个资助周期内为 P01 的进展做出了重要贡献，提供了重要发现 支持所有三个研究项目以及导致发表两篇论文的研究结果，其中一篇发表在《自然通讯》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性变化 脓毒症引起的，2) 表征血管表面的时间和病原体依赖性重塑 蛋白质组，以及 3) 确定革兰氏染色期间表现出的消耗性凝血病增强的分子基础 阴性败血症。 目标 1 重点关注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 特定糖苷酶、其糖蛋白底物和内吞凝集素受体，并确定它们的影响 它们对脓毒症的凝血病和炎症的稳态调节。项目2将使用等离子 蛋白质组学可识别病原体和宿主决定毒力的成分和机制 脓毒症发病机制并确定其在实验和人类分层和预后中的效用 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脱落到循环中 脓毒症小鼠模型及其在鉴定时对脓毒症凝血障碍和炎症的影响 可能实现人类疾病分层和预后的机制联系。 目标 2 侧重于表征血管表面蛋白质组以支持项目 3。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对凝血病和 脓毒症的炎症及其在人类疾病分层和预后中的作用。 目标 3 将支持项目 2，以寻求描述期间表现出的增强消耗性凝血病的特征。 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 鉴定对 SC/ST/EC 中观察到的 FXI 刺激活性负责的细菌功能 上清液；并确定其在革兰氏阴性败血症和消耗性凝血病升高中的作用 在人类疾病的分层和预后中可能有用。