Development of an Integrated Intermediary Metabolomics and Metabolic Flux Core

集成中间代谢组学和代谢通量核心的开发

• 批准号: 10419697
• 负责人: Richard G Kibbey
• 金额: $ 36.63万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2023-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10419697
• 关键词: Biochemical; Bioinformatics; Cell Culture Techniques; Cells; Custom; Data; Development; Disease; Drosophila genus; Funding; Genetic Screening; Industry; Intuition; Label; Laboratories; Life; Liquid Chromatography; Malignant Neoplasms; Maps; Measures; Metabolic; Metabolic Pathway; Metabolism; Microbiology; Movement; Nutrient; Organism; Oxygen Consumption; Pathway interactions; Physiology; Plague; Plants; Plasma; Positioning Attribute; Process; Proteomics; Resolution; Spectrum Analysis; Speed; System; Techniques; Technology; Time; Tissues; Universities; cloud based; cost; design; genomic data; human tissue; improved; in vivo; instrument; islet; mass spectrometer; metabolomics; open source; phosphoproteomics; service providers; stable isotope; transcriptomics

Project Summary This proposal seeks funding for a Liquid Chromatography (LC), Differential Mobility Spectroscopy (DMS), Quadrupole Time of Flight (LC-DMS-QToF) SCIEX 6600 mass spectrometer (AB Sciex LLC, Framingham, MA) to build an integrated system describing the metabolic traffic in cells, plasma and tissues. The essential endogenous biochemicals of life are confined to enzymatically delineated metabolic pathways shared across most species. The collective flow of nutrient substrates into synthetic or catabolic pathways that best describes key functions, transformations, differentiations, adaptations, or disease processes of a cell, tissue, or organism. The challenge then is interpreting the flow form concentration data, since the concentration of a metabolite may have little to do with its flow through a metabolic pathway. The lack of robust high-throughput analytical techniques to assess the flow of metabolic traffic through these pathways is a major limitation to improving our understanding of cellular and organismal physiology. Yale lacks a dedicated metabolomics core. The acquisition of the LC-DMS-QToF integrated system will provide a core with: 1) a dedicated intermediary metabolomics platform on the KEGG metabolic map (CoMBI) that through integrated omics can be reinforced by orthogonal transcriptomics, proteomics, phosphoproteomics or genomics data sets, 2) an expansion of Mass Isotopomeric MultiOrdinate Flux Analysis (MIMOSA) developed by my laboratory to quantitatively measure intracellular fluxes through sequential position-specific label transfer of stable isotopes, and 3) an in vivo inter-tissue metabolic flux analysis designed for genetic screens of metabolism. The combination of DMS with the high resolution and speed of ToF and mass fragmentation minimize isobaric interference which otherwise plague stable isotope studies. We have had the opportunity of having the requested instrument consigned to our laboratory at Yale through an academic/industry technology alliance with SCIEX as a way to generate preliminary proof-of-principle data and establish our pipelines. A bioinformatic alliance with Elucidata has developed a customized, open-sourced, cloud-based platform with an intuitive user interface to support our unique computational demands. The instrument operates in our existing Islet, Oxygen consumption, Mass Isotopomer flux Core analytical core (IOMIC) through chargebacks and with institutional support at cost under an established, university-approved Internal Service Provider (ISP) account. This unique and transformative platform will integrate the tens of 1000s of mass spectroscopic features, correct for natural abundance, and track positional movement of mass labels through cells to generate a high resolution “Google maps” of metabolism where the rate and direction of metabolic flow are quantified with demonstrated application to microbiology, plants, fruit flies, cell culture, cancer and human tissue. These capabilities are absolutely critical in order to sustain, and increase, the current pace of understanding metabolism in various tissues.

项目摘要 该提案寻求用于液相色谱法（LC），差异迁移率（DMS）的资金， 飞行时间（LC-DMS-QTOF）SCIEX 6600质谱仪（AB Sciex LLC，弗雷明厄姆，马萨诸塞州） 建立一个描述细胞，血浆和组织中代谢流量的集成系统。必不可少的 生命的内源性生化物仅限于跨酶学的代谢途径 大多数物种。营养基质的集体流动到最能描述的合成或分解代谢途径 细胞，组织或生物体的关键功能，转化，区分，适应或疾病过程。 那时的挑战是解释流动浓度数据，因为代谢物的浓度可能 与它通过代谢途径的流动无关。缺乏强大的高通量分析 评估通过这些途径的代谢流量流的技术是改善我们的主要限制 了解细胞和有机生理学。 耶鲁大学缺乏专用的代谢组学核心。 LC-DMS-QTOF集成系统的获取将 提供一个核心：1）在KEGG代谢地图（Combi）上的专用中间代谢组平台 通过正交转录组学，蛋白质组学，磷光蛋白质组学可以通过综合的OMICS加强。 或基因组数据集，2）开发的质量同位素多层次通量分析（MIMOSA）的扩展 通过我的实验室，通过连续位置特异性标签转移定量测量细胞内通量 稳定的同位素和3）旨在用于遗传筛查的体内组织间代谢通量分析 代谢。 DM与TOF和质量碎片的高分辨率和速度的组合 最小化同质干扰，否则会损害稳定的同位素研究。我们有机会 通过学术/行业技术将要求的工具委托给耶鲁大学的实验室 与SCIEX联盟是生成初步原则数据并建立我们的管道的一种方式。一个 与Eluciodata的生物信息学联盟已经开发了一个定制的，开源的，基于云的平台， 直观的用户界面支持我们独特的计算需求。该仪器在我们现有的 胰岛，氧气消耗，大量同位素通量核心分析核心（IOMIC） 根据已建立的大学批准的内部服务提供商（ISP）帐户，机构支持。 这个独特而变革的平台将整合数十万的质谱特征，正确 对于自然丰度，并跟踪质量标签通过细胞的位置运动以产生高分辨率 代谢的“ Google Maps”，其中代谢流量的速率和方向被证明 应用于微生物学，植物，果蝇，细胞培养，癌症和人体组织。这些功能是 为了维持和增加各种理解新陈代谢的空间，绝对至关重要 组织。