Revealing cancer metabolism via mass spectrometry and isotope tracers

项目总结/摘要 代谢在癌症生长、诊断（如FDG-PET）和治疗（如放射治疗）中起着重要作用。 抗叶酸剂、天冬酰胺酶）。在过去的十年里，对癌症代谢的研究蓬勃发展， 由于认识到癌症中的代谢变化是由癌基因信号传导引发的， 新的测量工具的出现加速了这一进程。这个NCI研究专家奖将研究 癌症代谢使用最重要的现代工具：质谱和同位素示踪剂。工作 与单位主任约书亚拉宾诺维茨和他的实验室，以及来自罗格斯大学的各种合作者一起， 新泽西癌症研究所和其他地区癌症中心（宾夕法尼亚大学，MSKCC，纽约大学），我的目标是在未来 5年内实现以下目标：（1）开发基于质谱的分析方法，使更多 完整和准确的代谢组定量。我将特别侧重于开发分析解决方案， 很难测量与癌症高度相关的代谢物。这些包括不稳定的物种，如氧化还原 辅因子（例如，NADPH）和活化的叶酸（例如5，10-亚甲基-THF）。(2)应用这些方法 癌症代谢组的测量，从培养细胞到人类肿瘤。这将包括研究 快速冷冻的去识别患者肿瘤标本，来自我们已经处于领先地位的癌症 （胰腺导管腺癌，嗜酸细胞瘤）和那些仍然研究不足的代谢 透视（甲状腺，神经母细胞瘤）。基于我们之前成功发现的癌代谢物2- 羟基戊二酸，一个特别的重点将是确定意想不到的或新的代谢物，改变， 癌(3)联合收割机将这些方法与同位素示踪剂相结合，以确定癌细胞和体内肿瘤中的通量。 我们在开发同位素示踪方法以定量代谢方面具有长期的领导地位。 通量，包括最近使用2 H示踪剂测量NADPH生产途径的显著成功。我们有 最近开发的用于将各种各样的13 C和2 H-示踪剂输注到小鼠中的方案，目的是 从而能够定量肿瘤代谢流。总的来说，这些示踪剂覆盖了中央碳代谢，一个- 碳代谢、蛋氨酸和谷胱甘肽代谢。我们将应用它们来研究代谢， 基因工程小鼠模型，包括有和没有Myc活化的胰腺癌和肺 有和没有自噬缺失的癌症。由此产生的数据将提供关键的见解， 在天然肿瘤微环境中的癌症的代谢病理生理学，从而将告知治疗 选择策略和新疗法的开发。

Project Summary/Abstract Metabolism plays a fundamental role in cancer growth, diagnosis (e.g. FDG-PET), and treatment (e.g. antifolates, asparaginase). Over the past decade, research into cancer metabolism has flourished, contextualized by the realization that metabolic changes in cancer are triggered by oncogene signaling and accelerated by the emergence of new measurement tools. This NCI Research Specialist Award will study cancer metabolism using the most important modern tools: mass spectrometry and isotope tracers. Working together with Unit Director Joshua Rabinowitz and his lab, and a diverse set of collaborators from Rutgers Cancer Institute of New Jersey and other regional cancer centers (U Penn, MSKCC, NYU), I aim over the next 5 years to achieve the following: (1) Develop mass spectrometry-based analytical methods that enable more complete and accurate metabolome quantitation. I will focus in particular on developing analytical solutions for hard to measure metabolites of high relevance to cancer. These include unstable species such as redox cofactors (e.g., NADPH) and activated folates (e.g. 5,10-methylene-THF). (2) Apply these methods to measurement of the cancer metabolome, from cultured cells to human tumors. This will include studies on flash-frozen de-identified patient tumor specimens, from cancers where we already have a leadership position (pancreatic ductal adenocarcinoma, oncocytoma) and those that remain understudied for a metabolic perspective (thyroid, neuroblastoma). Building on our prior success in discovering the oncometabolite 2- hydroxyglutarate, a particular focus will be identifying unexpected or novel metabolites that are altered in cancer. (3) Combine these methods with isotope tracers to determine fluxes in cancer cells and in vivo tumors. We have a long-standing leadership position in developing isotope tracer methods for quantitating metabolic fluxes, including notable recent success in measuring NADPH production pathways using 2H-tracers. We have recently developed protocols for infusing a wide variety of 13C and 2H-tracers into mouse, with the goal of enabling quantitation of tumor metabolic flux. Collectively, these tracers cover central carbon metabolism, one- carbon metabolism, and methionine and glutathione metabolism. We will apply them to study metabolism in genetically engineered mouse models including of pancreatic cancer with and without Myc activation and lung cancer with and without autophagy deletion. Resulting data will provide critical insights into the actual metabolic pathophysiology of cancer in the native tumor microenvironment and will thereby inform treatment selection strategies and the development of novel therapeutics.