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

探测正常代谢动态的质谱方法

• 批准号: 7615679
• 负责人: JOSHUA D RABINOWITZ
• 金额: $ 18.11万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 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/7615679
• 关键词: Advanced Malignant Neoplasm; Amino Acids; Anabolism; Antineoplastic Agents; Arts; Bacteria; Benchmarking; Biochemical Pathway; Breast Cancer Cell; Cancerous; Carbon; Cells; Characteristics; Chromatography; 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 Spectrum Analysis; Measurement; Measures; Metabolic; Metabolism; Methodology; Methods; Methotrexate; Microbe; Monitor; Neoplasm Metastasis; Normal Cell; Nucleotide Biosynthesis; Nucleotides; Nutrient; Pattern; Pentosephosphate Pathway; Pharmaceutical Preparations; Plague; Procedures; Reaction; Resistance; Specificity; 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

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疗药物抑制肿瘤细胞的代谢。我们的首要目标是对正常细胞和癌细胞之间的代谢差异进行全面和定量的了解，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，我们正在开发应用最先进的液相色谱-电喷雾电离-三重四极杆质谱技术以动态、定量和全面的方式探测细胞代谢的方法。到目前为止，我们已经成功地开发了用于测量几种微生物的代谢物浓度和通量的方法。在这里，我们建议扩展这些方法，使正常和癌细胞中的代谢物浓度和通量的可靠测量。具体而言，我们的目标是能够定量至少150种不同的已知的，结构确定的细胞内代谢物的浓度。我们的目标也是测量，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。我们将应用我们开发的分析技术来绘制正常细胞和癌细胞之间的主要代谢差异，并研究这些细胞对抗代谢抗癌药物治疗的动态反应。本文开发的方法将具有长期价值，用于了解抗代谢抗癌药物的作用机制（和毒性），用于表征药物反应性和耐药癌细胞之间的代谢差异，并提出抑制代谢的新方法，从而特异性杀死癌细胞。