权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Stable isotopomer analysis of anabolic metabolic pathways in breast cancer

乳腺癌合成代谢途径的稳定同位素分析

基本信息

批准号：
7876986
负责人：
ANDREW N. LANE
金额：
$ 16.28万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7876986
关键词：
Accounting Aerobic Amino Acids Anabolism Apoptosis Biochemical Biochemical Pathway Biochemistry Biomass Breast Breast Cancer Cell Breast Cancer Detection Cancerous Cell Culture Techniques Cell Cycle Cell Proliferation Cells Characteristics Citric Acid Cycle Clinical Data Development Diagnostic Neoplasm Staging Disease Early Diagnosis Environment Epithelial Cells Estrogens Fatty acid glycerol esters Female Future Genomics Genotype Glucose Glutamine Glycolysis Goals Human Hypoxia Intake Knowledge Label Laboratories Lead MCF7 cell Malignant Neoplasms Mammary Neoplasms Mammary gland Mass Spectrum Analysis Measures Messenger RNA Metabolic Metabolic Pathway Metabolism Modeling Monitor Mus Nude Mice Nutrient Oncogenes Oxygen Pathway interactions Pentosephosphate Pathway Pharmaceutical Preparations Phenotype Phospholipid Metabolism Phospholipids Plasma Production Proteins Reaction Regulation Relative (related person)Role Sampling Solid Neoplasm Source Staging System TP53 gene Techniques Technology Testing Time Tissues Tracer Tumor Suppressor Proteins Tumor stage Tumor-Derived Warburg Effect Xenograft Model Xenograft procedure aminoacid biosynthesis cancer cell cell transformation cell type design human subject improved in vivo interest lifetime risk malignant breast neoplasm metabolomics mouse model neglect nucleobase outcome forecast public health relevance response stable isotope tumor tumor progression tumorigenesis uptake

项目摘要

DESCRIPTION (provided by applicant): Despite the numerous advances in technology and screening breast cancer remains the major form of human cancers (lifetime risk ca. 1/8). A neglected aspect of tumorigenesis is the metabolic profile of cells, which expresses the biochemical phenotype. Transformed cells differ from their normal counterparts by loss of critical regulatory functions that lead to immortalization, avoidance of apoptosis and deregulated cell cycling. A dividing cell must double its contents during each cycle, requiring intake of materials both for biosynthesis, and to provide the energy to drive endergonic anabolic reactions. Thus, cancer cells must increase the activity of metabolic pathways for energy production and anabolism. Cancer cells of different origin should display metabolism characteristic of the cell type, and reflect specific aspects of proliferation, and how these depend on the external conditions such as the supply of nutrients and oxygen. We propose to determine the relative importance of specific cancer-relevant metabolic pathways in the development and progression of breast cancer cells (four lines) both in cell culture and in an orthotopic mouse tumor model (xenografts), using stable isotope assisted metabolomics by NMR and MS, as follows. Aim 1. To determine the relative importance of energy producing pathways and anabolic metabolism by quantitative isotopomer analysis of normal and cancerous human breast epithelial cells in culture in response to hypoxia and specific nutrients. The precursors ([U-13C]-glucose, [13C-1] and [13C-2] glucose), and 13C/15N glutamine can differentiate relative fluxes through glycolysis, pentose phosphate pathways, citric acid cycle and major anaplerotic reactions, amino acid metabolism, nucleobase and phospholipid biosynthesis. Relative fluxes through these major pathways in non-estrogen-sensitive MDAMB231 and estrogen sensitive ZR-75-1 and MCF-7 cells will be compared with normal human mammary epithelial cells (HMEC). Aim 2. To determine the extent that the specific pathway activities identified in cell culture are present in actual tumors in a xenograft mouse model. Female nude mice bearing mammary fat pad tumors deriving from non MDAMB231 and ZR-75-1 cells are given 13C glucose or 13C/15N glutamine i.v. and the tumors excised after sufficient time has elapsed for metabolism to occur. Plasma is analyzed to monitor uptake and redistribution of labeled precursors. Metabolites are extracted and analyzed by the same techniques as in Aim 1. The comparison of the tumor model with cell culture shows how the tumor environment and tissue interactions determine the relative importance of different pathways to breast tumor progression. The biochemical information will be used to interpret the metabolic profiles of human breast cancers. The development of these approaches will provide basic biochemical information about breast cancer, and is essential for the design of future studies with human subjects, with the goal of improved diagnosis of early stage tumors. PUBLIC HEALTH RELEVANCE: Despite the numerous advances in technology and screening breast cancer remains the major form of human cancers (lifetime risk ca. 1/8). However, although oncogenes such as Myc and Ras or tumor suppressors such as p53 are frequently upregulated in cancers, the precise roles of these factors in disease initiation and progression is not completely clear. A deeper understanding of the biochemical phenotype of cancerous cells and their normal counterparts under defined conditions is an essential counterpart to genomics analyses of cancer cells. We are developing the technology to measure the activity of biochemical pathways in cancer cells that ultimately can be applied to human tumors in a clinical setting. The comparison of the in vivo tumor model with cell culture will show how the tumor environment and tissue interactions may determine the relative importance of different metabolic pathways to breast tumor progression. This fundamental knowledge of tumor biochemistry then provides a rich source of hypotheses about tumor progression that can be tested directly in actual breast cancer tissue. Application of this approach to a clinical setting will set the stage for identifying sets of markers for early diagnosis, prognosis and ultimately drug responses.

描述（由申请人提供）：尽管技术和筛查取得了巨大进步，但乳腺癌仍然是人类癌症的主要形式（终生风险约为 1/8）。肿瘤发生的一个被忽视的方面是细胞的代谢特征，它表达生化表型。转化细胞与正常细胞的不同之处在于，它们丧失了导致永生化、避免细胞凋亡和细胞周期失调的关键调节功能。分裂细胞在每个周期中其含量必须加倍，需要摄入物质进行生物合成，并提供驱动吸能合成代谢反应的能量。因此，癌细胞必须增加能量产生和合成代谢的代谢途径的活性。不同来源的癌细胞应表现出细胞类型的代谢特征，并反映增殖的特定方面，以及这些如何依赖于外部条件（例如营养物和氧气的供应）。我们建议通过 NMR 和 MS 使用稳定同位素辅助代谢组学来确定细胞培养物和原位小鼠肿瘤模型（异种移植物）中乳腺癌细胞（四系）发育和进展中特定癌症相关代谢途径的相对重要性，如下。目标 1. 通过对培养物中正常和癌性人类乳腺上皮细胞对缺氧和特定营养物的反应进行定量同位素异构体分析，确定能量产生途径和合成代谢的相对重要性。前体（[U-13C]-葡萄糖、[13C-1] 和 [13C-2] 葡萄糖）和 13C/15N 谷氨酰胺可以通过糖酵解、戊糖磷酸途径、柠檬酸循环和主要回补反应、氨基酸代谢、核碱基和磷脂生物合成来区分相对通量。将非雌激素敏感的 MDAMB231 和雌激素敏感的 ZR-75-1 和 MCF-7 细胞中通过这些主要途径的相对通量与正常人乳腺上皮细胞 (HMEC) 进行比较。目标 2. 确定细胞培养物中鉴定的特定途径活性在异种移植小鼠模型的实际肿瘤中存在的程度。携带源自非 MDAMB231 和 ZR-75-1 细胞的乳腺脂肪垫肿瘤的雌性裸鼠静脉注射 13C 葡萄糖或 13C/15N 谷氨酰胺。在经过足够的时间进行新陈代谢后切除肿瘤。分析血浆以监测标记前体的吸收和重新分布。采用与目标 1 相同的技术提取和分析代谢物。肿瘤模型与细胞培养物的比较显示肿瘤环境和组织相互作用如何决定不同途径对乳腺肿瘤进展的相对重要性。生化信息将用于解释人类乳腺癌的代谢特征。这些方法的开发将提供有关乳腺癌的基本生化信息，对于设计未来以人类为对象的研究至关重要，其目标是改善早期肿瘤的诊断。公共卫生相关性：尽管技术和筛查取得了巨大进步，但乳腺癌仍然是人类癌症的主要形式（终生风险约为 1/8）。然而，尽管 Myc 和 Ras 等癌基因或 p53 等肿瘤抑制基因在癌症中经常上调，但这些因素在疾病发生和进展中的确切作用尚不完全清楚。更深入地了解癌细胞及其正常细胞在特定条件下的生化表型是癌细胞基因组学分析的重要对应部分。我们正在开发测量癌细胞生化途径活性的技术，最终可应用于临床环境中的人类肿瘤。体内肿瘤模型与细胞培养的比较将显示肿瘤环境和组织相互作用如何决定不同代谢途径对乳腺肿瘤进展的相对重要性。这种肿瘤生物化学的基础知识提供了关于肿瘤进展的丰富假设来源，可以直接在实际乳腺癌组织中进行测试。这种方法在临床环境中的应用将为识别早期诊断、预后和最终药物反应的标志物组奠定基础。