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 程序;蛋白质糖基化和糖蛋白重塑改变了脓毒症的凝血病和炎症。 核心D使用最先进的质谱法来分析败血症中的血浆和细胞蛋白质组。核心有 在第一个资金周期中为P01的进度做出了重要贡献，提供了重要的发现 支持所有三个研究项目，以及导致两个出版物的发现，其中一项是自然通信。 核心D的具体目的是：1）评估血浆蛋白质组的时间和病原体依赖性变化 由败血症产生的，2）表征血管表面的时间和病原体依赖性重塑 蛋白质组和3）确定革兰氏阴性升高消耗凝血症的分子基础 负败血症。 AIM 1专注于所有三个项目的血浆蛋白质组学。项目1将使用等离子体蛋白质组学识别 特定的糖苷酶，其糖蛋白底物和内吞凝集素受体，并确定它们的影响 他们对败血症的凝血病和炎症的稳态调节。项目2将使用等离子体 蛋白质组学鉴定病原体和宿主的组成部分和机制，以决定毒力和 败血症的发病机理并确定它们在实验和人类的分层和预后中的效用 败血症。项目3将使用等离子体蛋白质组学来确定在循环中脱落的血管表面标志物 败血症的鼠模型及其对脓毒症的凝血病和炎症的影响 可能使人类疾病分层和预后的机械联系。 AIM 2专注于表征支持项目3的血管表面蛋白质组。这项工作将表征 器官特异性变化对血管细胞表面蛋白质组，并确定其对凝血病的影响 败血症的炎症及其在人类疾病的分层和预后中的效用。 AIM 3将支持项目2，以寻求表征在 革兰氏阴性败血症。这项工作将使用基于活动的蛋白质组学和标准蛋白分馏分析 确定负责SC/ST/EC中观察到的FXI刺激活性的细菌功能 上清液；并确定其在革兰氏阴性败血症和 人类疾病分层和预后的可能实用性。