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Metabolic Prolifing & Biochemical Analysis for Anti-Cancer Target Identification

代谢增殖

基本信息

批准号：
6871273
负责人：
HARVEY W BLANCH
金额：
$ 11.78万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-03-15 至 2006-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Because cellular enzymes, transcription factors, and their respective genes interact in a complex and interdependent manner, cellular physiology is not dictated by the genome alone. Targeting the genetic aberrations of transformed cells may therefore not be as effective as directly targeting the altered physiology of tumors. Metabolic profiling, by determining the substrate fluxes in major metabolic pathways, can reveal phenotypic transformations in cancer cells, and provide more detailed information than signal transduction and genetic studies alone. Transforming agents typically induce high pentose pathway activity. Rapidly proliferating breast cancer cells have high rates of aerobic glycolysis and glutaminolysis, and enhanced fatty acid synthesis activity. These altered metabolic fluxes suggest that enzymes and pathways identified as critical for cancer cell proliferation may be excellent targets for drug development. In the proposed project, the concentrations and fluxes of intracellular metabolites of human breast cancer cells will be determined from 13C NMR spectroscopy of cells grown on 13C glucose and glutamine. Biochemical systems analysis, by providing a quantitative description of metabolite flows, will be employed to reveal the link between growth signaling and metabolic processes. Metabolic profiling of MCF7 breast cancer cells will be accomplished by determination of intracellular fluxes using 13C NMR, under conditions mimicking those found in tumors. The hypothesis is that the crucial enzymes responsible for the altered metabolic fluxes in transformed human cells provide targets for new anti-cancer drugs. Specific aims are to: (1) Identify the primary metabolic pathways that are operative in estrogen receptor positive (ER+) and negative cells (ER-) breast cancer cells under conditions that mimic those of a tumor. (2) To quantify the primary and secondary metabolic fluxes and enzymes control coefficients in breast cancer cells in the presence and absence of tamoxifen (TAM) and estrogen, including those fluxes which may be reversed by increased concentrations of estrogen ("estrogen rescue"), and those effects of TAM that are not related to ER binding. (3) To examine modes of estrogen and tamoxifen action by investigating the relationship between fatty acid synthesis, pentose phosphate pathway activity, and other pathways requiring NADP+/NADPH, and to determine the effects of impaired fatty acid synthesis (resulting from the addition of the fatty acid synthase inhibitor, cerulenin) on other metabolic pathways in the presence of estrogen and tamoxifen.

描述（由申请人提供）：由于细胞酶、转录因子及其各自的基因以复杂和相互依赖的方式相互作用，因此细胞生理学不仅仅由基因组决定。因此，靶向转化细胞的遗传畸变可能不如直接靶向肿瘤的改变的生理学那样有效。通过测定主要代谢途径中的底物通量，代谢谱可以揭示癌细胞的表型转化，并提供比单独的信号转导和遗传研究更详细的信息。转化剂通常诱导高戊糖途径活性。快速增殖的乳腺癌细胞具有高速率的有氧糖酵解和β-内酰胺分解，以及增强的脂肪酸合成活性。这些改变的代谢通量表明，被确定为对癌细胞增殖至关重要的酶和途径可能是药物开发的极好靶点。在拟议的项目中，人乳腺癌细胞的细胞内代谢物的浓度和通量将从13 C NMR光谱测定13 C葡萄糖和谷氨酰胺上生长的细胞。生化系统分析，通过提供代谢物流的定量描述，将揭示生长信号和代谢过程之间的联系。MCF 7乳腺癌细胞的代谢谱将通过在模拟肿瘤中发现的条件下使用13 C NMR测定细胞内通量来完成。这一假说是，在转化的人类细胞中负责改变代谢通量的关键酶为新的抗癌药物提供了靶点。具体目标是：（1）鉴定在模拟肿瘤的条件下在雌激素受体阳性（ER+）和阴性细胞（ER-）乳腺癌细胞中起作用的主要代谢途径。(2)在存在和不存在他莫昔芬（TAM）和雌激素的情况下，量化乳腺癌细胞中的初级和次级代谢通量和酶控制系数，包括那些可能被雌激素浓度增加逆转的通量（“雌激素拯救”），以及TAM与ER结合无关的效应。(3)通过研究脂肪酸合成、戊糖磷酸途径活性和其他需要NADP+/NADPH的途径之间的关系，检查雌激素和他莫昔芬的作用模式，并确定在存在雌激素和他莫昔芬的情况下，脂肪酸合成受损（由添加脂肪酸合成酶抑制剂浅蓝菌素引起）对其他代谢途径的影响。