Mass Spectrometry Methods for Probing Metabolic Dynamics in Normal & Cancer Cells

探测正常代谢动态的质谱方法

基本信息

批准号：
7432649
负责人：
JOSHUA D RABINOWITZ
金额：
$ 21.63万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-18 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7432649
关键词：
Advanced Malignant Neoplasm Amino Acids Anabolism Antineoplastic Agents Arts Bacteria Benchmarking Biochemical Pathway Breast Cancer Cell Cancerous Carbon Cells Characteristics Condition Data Data Set Databases Detection Development Dihydrofolate Reductase Electrospray Ionization Fibroblasts Fluorouracil Glucose Glutamine Glycolysis Goals Human Isotope Labeling Kinetics Knowledge Label Lipids Liquid Chromatography Lung Malignant Neoplasms Maps Mass Chromatography Mass Spectrum Analysis Measurement Measures Metabolic Metabolism Methodology Methods Methotrexate Microbe Monitor Neoplasm Metastasis Normal Cell Nucleotide Biosynthesis Nucleotides Numbers Nutrient Pattern Pentosephosphate Pathway Pharmaceutical Preparations Plague Procedures Reaction Resistance Specificity Spectrometry Stable Isotope Labeling Systems Biology Technology Thymidine Thymidylate Synthase Toxic effect Tracer Treatment Protocols Yeasts anticancer research base bone cancer cell cancer therapy chemotherapeutic agent data integration design fibrosarcoma improved inhibitor/antagonist insight killings lipid biosynthesis mass spectrometer metabolomics neoplastic cell novel novel strategies nucleotide metabolism penis foreskin response solvent extraction synovial sarcoma tissue tropism tool tumor

Advanced Malignant Neoplasm Amino Acids Anabolism Antineoplastic Agents Arts Bacteria Benchmarking Biochemical Pathway Breast Cancer Cell Cancerous Carbon Cells Characteristics Condition Data Data Set Databases Detection Development Dihydrofolate Reductase Electrospray Ionization Fibroblasts Fluorouracil Glucose Glutamine Glycolysis Goals Human Isotope Labeling Kinetics Knowledge Label Lipids Liquid Chromatography Lung Malignant Neoplasms Maps Mass Chromatography Mass Spectrum Analysis Measurement Measures Metabolic Metabolism Methodology Methods Methotrexate Microbe Monitor Neoplasm Metastasis Normal Cell Nucleotide Biosynthesis Nucleotides Numbers Nutrient Pattern Pentosephosphate Pathway Pharmaceutical Preparations Plague Procedures Reaction Resistance Specificity Spectrometry Stable Isotope Labeling Systems Biology Technology Thymidine Thymidylate Synthase Toxic effect Tracer Treatment Protocols Yeasts anticancer research base bone cancer cell cancer therapy chemotherapeutic agent data integration design fibrosarcoma improved inhibitor/antagonist insight killings lipid biosynthesis mass spectrometer metabolomics neoplastic cell novel novel strategies nucleotide metabolism penis foreskin response solvent extraction synovial sarcoma tissue tropism tool tumor

展开

项目摘要

DESCRIPTION (provided by applicant): Many of the most clinically important chemotherapeutic agents inhibit the metabolism of tumor cells. Our overarching goal is to develop a complete and quantitative understanding of the metabolic differences between normal and cancer cells, and to use this knowledge to guide the rational design of novel anticancer regimens. To achieve this goal, we are developing methods that apply state-of-the-art liquid chromatography- electrospray ionization-triple quadrupole mass spectrometry technology to probe cellular metabolism in a dynamic, quantitative, and comprehensive manner. To date, we have succeeded in developing methods for measuring metabolite concentrations and fluxes from several microbes. Here we propose to extend these methods to enable reliable measurement of metabolite concentrations and fluxes in normal and cancer cells. Specifically, we aim to enable quantitation of the concentrations of at least 150 different known, structurally-defined intracellular metabolites. We also aim to measure, using isotopic tracers, the fluxes through central carbon, lipid, amino acid, and nucleotide metabolism. We will apply the analytical technology that we develop to map major metabolic differences between normal and cancer cells and to study the dynamic response of these cells to treatment with anti-metabolite anticancer drugs. The methods developed here will have long-term value for understanding the mechanism of action (and toxicity) of anti-metabolite anticancer drugs, for characterizing the metabolic differences between drug-responsive and resistant cancer cells, and for suggesting new approaches to inhibiting metabolism that will specifically kill cancer cells.

描述（由申请人提供）：许多临床上最重要的化学治疗剂抑制了肿瘤细胞的代谢。我们的总体目标是对正常细胞和癌细胞之间的代谢差异有完整而定量的理解，并利用这些知识来指导新型抗癌方案的合理设计。为了实现这一目标，我们正在开发采用最先进的液相色谱法 - 电喷雾电离 - 四极杆质谱技术来以动态，定量和全面的方式探测细胞代谢。迄今为止，我们已经成功开发了测量几种微生物的代谢产物浓度和通量的方法。在这里，我们建议扩展这些方法，以实现正常和癌细胞中代谢物浓度和通量的可靠测量。具体而言，我们旨在定量至少150个已知的，结构定义的细胞内代谢物的浓度。我们还旨在使用同位素示踪剂，通过中央碳，脂质，氨基酸和核苷酸代谢的通量进行测量。我们将应用我们开发的分析技术来绘制正常和癌细胞之间的主要代谢差异，并研究这些细胞对用抗代谢物抗癌药物治疗的动态反应。此处开发的方法将具有长期价值，以理解抗代谢物抗癌药物的作用机理（和毒性），以表征药物反应性和抗性癌细胞之间的代谢差异，并提出抑制代谢性的新方法，以抑制特定杀死癌症细胞的代谢。