Development and Implementation of Advanced Technologies for Cancer Metabolism Research

项目总结 这项提议要求支持我作为桑福德癌症代谢中心主任的职位 伯纳姆·普雷比医学发现研究所(SBP)。该中心是NCI指定的癌症中心的一部分，网址为 SBP。Ze‘ev Ronai博士，我和他一起工作了13年，是癌症中心主任和该单位 此应用程序的主管。核心通过提供基于GC-MS的稳定同位素来支持癌症研究 (13C)代谢物追踪和量化服务。我曾在SBP从事稳定同位素示踪技术的工作 自2008年以来，第一次是在研究实验室，过去六年是在癌症代谢核心。我已经发展出 使用不同衍生化方案和MS电离方法对极性代谢物、脂肪酸进行分析 (长链和短链)、胆固醇、糖和糖磷酸盐，包括代谢物的方法 量化和质量控制。在过去的五年里，Core至少贡献了18篇来自实验室的论文 在NCI项目拨款的支持下，我已经成为其中13篇论文的合著者。我在这里描述了这些计划 稳定同位素示踪方法和代谢物定量方法继续扩展的应用 新获得的GC-MS/MS将通过三重四元组技术和 更易于使用的化学电离。我还计划研究癌症研究所需的具体方法， 包括用于胆汁酸、氧甾醇、氨基己糖途径中间体以及用于分离细胞群体的方法 和细胞器代谢组学。来自R50奖金的资金将进一步使我能够致力于 针对不能用GC-MS测定的代谢物进行靶向LC-MS分析。通过加强以下设施 对癌细胞和肿瘤内代谢状态变化的分析，这一奖项将推进卓越的目标 在SBP癌症中心的翻译多学科研究中。

PROJECT SUMMARY This proposal requests support for my position as the Director of the Cancer Metabolism Core at Sanford Burnham Prebys Medical Discovery Institute (SBP). The Core is part of the NCI-designated Cancer Center at SBP. Dr. Ze’ev Ronai, with whom I have worked for thirteen years, is the Cancer Center Director and the Unit Director for this application. The Core supports cancer research by providing a GC-MS-based stable-isotope (13C) metabolite tracing and quantification service. I have worked at SBP on stable-isotope tracer technology since 2008, first in a research lab and for the last six years in the Cancer Metabolism Core. I have developed assays using differing derivatization protocols and MS ionization approaches for polar metabolites, fatty acids (long- and short-chain), cholesterol, sugars and sugar-phosphates, including methods for metabolite quantification and quality control. In the past five years, the Core has contributed to at least 18 papers from labs with NCI project grant support, and I have been a co-author on 13 of these papers. I describe plans in this application for continued expansion of stable isotope tracing methods and metabolite quantification using a newly-acquired GC-MS/MS that will improve detection sensitivity both through the triple quad technology and by being easier to use with chemical ionization. I also plan to work on specific methods needed for cancer research, including for bile acids, oxysterols, hexosamine pathway intermediates, and methods for isolated cell populations and organelle metabolomics. Funding from this R50 award will further allow me to work on the development of targeted LC-MS assays for metabolites that cannot be measured using GC-MS. By strengthening facilities for analysis of altered metabolic states within cancer cells and tumors, this award will advance the goal of excellence in translational multi-disciplinary research in the SBP Cancer Center.