Stable Isotope Resolved Metabolomics Analytical Shared Core

项目摘要(见说明)：稳定同位素解析(SIRM)分析共享核心将为小组提供高信息吞吐量(HIT)生物分析支持，特别强调基于稳定同位素的代谢途径追踪。这一核心的基础已经以NSF发起的UofL监管和环境分析代谢组学中心(CREAM)的形式存在，该中心专门为SIRM代谢生物分析服务以及自2002年以来在样品准备和处理、高场核磁共振、质谱学和生物化学信息学方面的合作而建立。奶油屋拥有最先进的仪器，对几个研究小组的实验设计产生了深远的影响。因此，许多核心互动本质上都是长期的、协作的努力。SIRM共享核心的主要目标是为所有三个研究项目提供一套共享资源，并无缝集成跨越组织培养、小鼠模型和人类受试者的实验设计、数据收集和数据分析。Core将提供一系列实验方法，使用我们最先进的核磁共振和质谱学分析平台来确定癌症和非癌症肺细胞和组织的代谢表型。在与个别项目负责人协商后，稳定同位素分析的具体实验设计将根据个别项目的需要量身定做，但各项目之间要保持一致，以促进综合分析。我们有三个具体目标来实现我们的目标：具体目标1.提供基础设施和专家样品处理，以确保所有三个研究项目的样品处理、处理和存档的一致性；具体目标2.为三个研究项目提供各种核磁共振和质谱学技术方面的访问和专业知识：A)为基于SIRM的稳健和一致的分析性实验设计提供仪器和专业知识；以及B)提供各种核磁共振和质谱学技术的专家数据收集和分析。具体目标3.为研究项目提供数据分析支持，包括数据、代谢途径和生化机制分析，其中包括：A)实施一个基于网络的平台，用于数据减少、质量控制和数据分析支持；B)提供识别和量化代谢物的方法；C)在GORE A的项目、数据采集和生化分析之间提供桥梁。

PROJECT SUMMARY (See instructions): The Stable Isotope Resolved (SIRM) Analytical Shared Core will provide high information throughput (HIT) bioanalytical support for the groups, with special emphasis on stable isotope-based metabolic pathway tracing. The basis for this Core already exists in the form of the NSF-initiated, UofL Center for Regulatory and Environmental Analytical Metabolomics (CREAM), established expressly for SIRM metabolomic bioanalytical services and collaborations on sample preparation and handling, high field NMR, mass spectrometry, and biochemoinformatics since 2002. CREAM houses state of the art instrumentation that has profoundly influenced experimental design for several research groups. Therefore, many Core interactions are by their nature long-standing, collaborative efforts. The main goals of the SIRM Shared Core are to provide a set of shared resources for all three Research Projects and seamlessly integrate experimental design, data collection, and data analysis spanning tissue culture, mouse models and human subjects. The Core will provide a combination of experimental methods to determine metabolic phenotypes of cancerous and non-cancerous lung cells and tissue using our state of the art analytical platforms in both NMR and mass spectrometry. In consultation with individual project leaders, specific experimental design for stable isotope analysis will be tailored to individual project needs, but with consistency across projects to facilitate integrated analyses. We have 3 specific aims to achieve our goals: Specific Aim 1. Provide infrastructure and expert sample handling that ensures consistent handling, processing, and archiving of samples across all three Research Projects; Specific Aim 2. Provide access and expertise across a wide range of NMR and MS technologies to the three Research Projects: A) Provide access to instrumentation and expertise for robust and consistent SIRM-based analytical experimental design and B) Provide expert data collection and analysis across a wide range of NMR and MS technologies. Specific Aim 3. Provide data analytical support to the Research Projects, including data, metabolic pathway, and biochemical mechanism analyses, that include: A) Implement a web-based platform for data reduction, quality control and data analytical support; B) Provide methodologies for identifying and quantifying metabolites; C) Provide a bridge between the Projects, data acquisition and biochemical analysis in Gore A. Core B provides extensive interoperability, handling and advising all aspects of sample preparation, data collection, data analysis & quality control, ensuring consistency of data across all 3 research projects.