Multiplexed UPLC-MS/MS System for Advanced Target Metabolomics

用于高级目标代谢组学的多重 UPLC-MS/MS 系统

• 批准号: 9075163
• 负责人: DANIEL RAFTERY
• 金额: $ 59.61万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-15 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9075163
• 关键词: Basic Science; Biological; Biological Assay; Chromatography; Clinical Research; Coupled; Development; Disease; Growth; Injection of therapeutic agent; Laboratories; Lead; Lipids; Liquid Chromatography; Mass Spectrum Analysis; Measurement; Measures; Metabolism; Methodology; Pacific Northwest; Peer Review; Pump; Reporting; Reproducibility; Research; Research Project Grants; Running; Sampling; System; Systems Biology; Time; Universities; Washington; advanced system; aqueous; base; biomarker discovery; experience; improved; instrument; interest; ionization; mass spectrometer; metabolic profile; metabolome; metabolomics; method development; novel strategies

 DESCRIPTION (provided by applicant): The Northwest Metabolomics Research Center (NW-MRC) at the University of Washington in Seattle is the largest comprehensive metabolic profiling laboratory in the Pacific Northwest Region. We have developed a robust and reproducible targeted mass spectrometry platform that measures more than 200 aqueous metabolites and is in very high demand. This platform is based on an AB-Sciex API5500 Qtrap mass spectrometer coupled to Agilent 1260 dual liquid chromatography pumps that separate aqueous metabolites using HILIC columns under two identical conditions for both positive and negative ionization. Due to its excellent measurement reproducibility and robustness, this assay has attracted a very high interest from basic and clinical researches. For example, since the beginning of 2014, we have analyzed over 3,500 biological samples from more than 60 collaborators on this system, and the results have been reported in 10 peer-reviewed articles. At this point, demand for the system is exceeding our capacity, and with the strong growth we have experienced, we anticipate longer and longer delays in running samples. In addition, we would like to add targeted lipidomics to our assay, but do not have instrument time to devote for method development or for running the additional lipids on this platform. Finally, we have developed a new approach for metabolomics, globally optimized targeted (GOT)-MS, which combines the benefits of global and targeted analysis and promises very broad metabolome coverage with high reproducibility. However, our AB-Sciex system is the only one we have that can perform targeted measurements of over 200 molecules with good long term stability. Given the high demand for this system, we greatly need the addition of a more versatile and sensitive LC-MS/MS system for advanced targeted metabolomics. To further enhance targeted metabolomics capabilities we propose to install a state-of-the-art multiplexed UPLC- MS/MS system capable of performing simultaneous separations on four analytical columns followed by high sensitive targeted MS acquisition. Each column will be independently connected to a 4-port injection auto- sampler, which in turn will allow separations under four different chromatography conditions. The major benefits of this platform will be: (i) much broader coverage of measured metabolites since we will add hundreds of lipids to our targeted assay; (ii) significant enhancement in the throughput capabilities, which in turn will eliminate delays in analysis turnaround time; (iii) additional time available for method development. As a result, we will be able to develop and utilize our new GOT-MS methodology to improve metabolite measurements. The proposed system will be available for collaborative research projects at least 75% of the run time, and its extensive utilization will lead to a better understanding of metabolic processes associated with disease development, biomarker discovery, and systems biology research.

 描述（由申请人提供）：位于西雅图的华盛顿大学的西北代谢组学研究中心（NW-MRC）是太平洋西北地区最大的综合代谢分析实验室。我们开发了一个强大且可重复的靶向质谱平台，可测量200多种水性代谢物，需求量非常大。该平台基于AB-Sciex API 5500 Qtrap质谱仪，该质谱仪与Agilent 1260双液相色谱泵耦合，该双液相色谱泵使用HILIC柱在两种相同的正离子化和负离子化条件下分离水性代谢物。由于其良好的测量重现性和鲁棒性，该方法已引起了基础和临床研究的高度兴趣。例如，自2014年初以来，我们已经在该系统上分析了来自60多个合作者的3,500多个生物样本，并在10篇同行评议文章中报告了结果。在这一点上，对系统的需求超过了我们的能力，随着我们经历的强劲增长，我们预计运行样品的延迟时间会越来越长。此外，我们希望将靶向脂质组学添加到我们的测定中，但没有仪器时间用于方法开发或在该平台上运行额外的脂质。最后，我们开发了一种新的代谢组学方法，即全局优化靶向（GOT）-MS，它结合了全局和靶向分析的优点，并承诺具有高重现性的非常广泛的代谢组覆盖范围。然而，我们的AB-Sciex系统是唯一一个可以对200多个分子进行有针对性的测量，并具有良好的长期稳定性。鉴于对该系统的高需求，我们非常需要增加一个更通用和灵敏的LC-MS/MS系统，用于先进的靶向代谢组学。为了进一步增强靶向代谢组学能力，我们建议安装最先进的多重UPLC-MS/MS系统，该系统能够在四个分析柱上同时进行分离，然后进行高灵敏度靶向MS采集。每个色谱柱将独立连接到4端口进样自动进样器，从而允许在四种不同色谱条件下进行分离。该平台的主要好处将是：（i）由于我们将向我们的靶向测定中添加数百种脂质，因此测量代谢物的覆盖范围更广;（ii）显著提高了通量能力，这反过来又将消除分析周转时间的延迟;（iii）可用于方法开发的额外时间。因此，我们将能够开发和利用我们新的GOT-MS方法来改善代谢物测量。拟议的系统将可用于合作研究项目至少75%的运行时间，其广泛的利用将导致更好地了解与疾病发展，生物标志物发现和系统生物学研究相关的代谢过程。

项目成果

Metabolomics in chronic pain research.

• DOI: 10.1002/ejp.1677
• 发表时间: 2021-03
• 期刊: European journal of pain (London, England)
• 作者: Teckchandani S;Nagana Gowda GA;Raftery D;Curatolo M
• 通讯作者: Curatolo M

Boosting NAD preferentially blunts Th17 inflammation via arginine biosynthesis and redox control in healthy and psoriasis subjects.

• DOI: 10.1016/j.xcrm.2023.101157
• 发表时间: 2023-09-19
• 期刊: CELL REPORTS MEDICINE
• 影响因子: 14.3
• 作者: Han, Kim;Singh, Komudi;Meadows, Allison M.;Sharma, Rahul;Hassanzadeh, Shahin;Wu, Jing;Goss-Holmes, Haley;Huffstutler, Rebecca D.;Teague, Heather L.;Mehta, Nehal N.;Griffin, Julian L.;Tian, Rong;Traba, Javier;Sack, Michael N.
• 通讯作者: Sack, Michael N.

Lipid exposure activates gene expression changes associated with estrogen receptor negative breast cancer.

脂质暴露激活了与雌激素受体阴性乳腺癌相关的基因表达变化。

• DOI: 10.1038/s41523-022-00422-0
• 发表时间: 2022-05-04
• 期刊: NPJ breast cancer
• 影响因子: 5.9
• 作者: Yadav S;Virk R;Chung CH;Eduardo MB;VanDerway D;Chen D;Burdett K;Gao H;Zeng Z;Ranjan M;Cottone G;Xuei X;Chandrasekaran S;Backman V;Chatterton R;Khan SA;Clare SE
• 通讯作者: Clare SE