TR&D 2: Integrated Metabolomics

项目总结/摘要 尽管对代谢途径有广泛的了解，但了解基本的代谢机制 突出的疾病是有限的能力，掌握复杂的代谢系统的调节。遗传 功能获得/丧失的方法提供了大量关于代谢如何可以 这些方法受到监管，但在确定复杂疾病的原因方面并不十分成功。理想情况下，自上而下 方法将被用来检查代谢的功能，使用定量的方法，然后指导 功能增益/损失研究。科学界正在迅速转向稳定同位素，因为它的高 信息产量、便利性和在人类受试者、啮齿动物模型之间翻译方法的能力 和细胞制剂。为了实现这一目标，有必要使用计算扩展早期的概念， 方法，并将质谱（MS）与核磁共振（NMR）结合起来。但在 分子生理学家/遗传学家或感兴趣的临床科学家采用的方法的顺序 在疾病生理学中，它们必须是灵活、可靠和易于使用的方法， 条件在这个TR & D项目中，我们将进行细胞，啮齿动物和计算研究，以开发翻译 方法来满足这些需求。首先，我们将整合示踪通量方法和代谢组学， 在疾病期间调节代谢或其失调的靶酶。其次，我们将利用 NMR同位素异构体分析的特异性和质量同位素异构体分析测量通量的灵敏度， 毫克级样品的高置信度。最后，我们将开发和分发一个免费的，开源的 基于通量/示踪剂输入模拟NMR和MS数据并从 实验数据在单一的，易于使用的图形界面完成。实现这些目标将提供 为科学界提供新的工具，指导药理学、遗传学和临床 科学

Project Summary/Abstract Despite extensive knowledge of metabolic pathways, understanding the basic metabolic mechanisms of prominent diseases is limited by the ability to grasp the regulation of complex metabolic systems. Genetic gain/loss of function approaches have provided a wealth of information about how metabolism can be regulated, but have not been very successful at identifying the cause of complex disease. Ideally, top-down approaches would be used to examine the function of metabolism using quantitative approaches, then to guide gain/loss of function studies. The scientific community is moving quickly to stable isotopes because of high information yield, convenience, and the capacity to translate methods between human subjects, rodent models and cell preparations. To accomplish this goal, it is essential to extend earlier concepts using computational methods, and to integrate mass spectrometry (MS) with nuclear magnetic resonance (NMR). However, in order for the approaches to be embraced by the molecular physiologist/geneticist or clinical scientist interested in disease physiology, they must be flexible, reliable and easy-to-use methods relevant in a wide range of conditions. In this TR&D project we will perform cell, rodent and computational studies to develop translation methods to address these needs. First, we will integrate tracer flux approaches and metabolomics to identify target enzymes in the regulation of metabolism or its dysregulation during disease. Secondly, we will leverage the specificity of NMR isotopomer analysis and the sensitivity of mass isotopomer analysis to measure flux with high confidence on milligram scale samples. Finally, we will develop and distribute a free, open source software platform that simulates NMR and MS data based on flux/tracer input and calculates flux from experimental data in a single, easy to use graphical interface complete. Accomplishing these aims will provide the scientific community with new tools that will guide tactical studies in pharmacology, genetics and clinical science.