Advancing Cancer Research through Comprehensive Proteomics and Metabolomics Analyses

项目摘要 蛋白质组学和代谢组学共享资源（PMSR）提供了一套全面的蛋白质组学和 代谢组学分析的Wistar癌症中心，福克斯大通癌症中心在坦普尔大学，和西德尼 托马斯杰斐逊大学的基梅尔癌症中心作为主要目标。PMSR的资源也 可供其他癌症中心和学术机构的研究人员作为次要目标。唐医生的 作为PMSR常务董事的角色是通过提供专家支持NCI资助的癌症研究项目 咨询和最先进的技术，以可承受的成本实现最高性能， 癌症研究人员和其他生物医学研究人员。总经理将协助实验 设计，执行MS数据分析，并协助结果的生物学解释。董事总 主任还将投入大量精力优化和实施新方法，更新分析和 数据分析方法，并更新仪器，以确保每个项目都使用最先进的技术执行 方法论。这一点至关重要，因为仪器、软件和分析策略都在不断发展 目前和未来的大多数项目都涉及非常具有挑战性的蛋白质组学和代谢组学 问题蛋白质组学项目将包括：1）外泌体的深入，全球定量比较， 分泌体、细胞裂解物、组织和生物液体; 2）翻译后的定量比较， 修饰;和3）具有和不具有化学交联的分离的蛋白质复合物的LC-MS/MS分析。 定量数据将使用积分MS离子电流的无标记定量或使用 稳定同位素如SILAC或TMT同量异位素标记。代谢组学项目将包括量化 极性代谢物、脂质和脂肪酸的稳态水平，以及13 C同位素示踪剂分析。代表 今后发展新的分析方法的具体计划包括：1）改进 蛋白质棕榈酰化，鉴定特异性修饰位点; 2）从头肽测序， HLA肽的鉴定; 3）另一种泛素富集方法; 4）绝对定量 肠道代谢物的含量此外，唐博士将实施其他新的或改进的方法，这些方法很可能是 在未来的癌症项目中。这些蛋白质组学和代谢组学分析将有助于获得关键数据 需要识别生物分子目标，以及产生对成功的假设， 本申请中描述的癌症相关项目。

Project Summary The Proteomics and Metabolomics Shared Resource (PMSR) provides a comprehensive set of proteomics and metabolomics assays to the Wistar Cancer Center, Fox Chase Cancer Center at Temple University, and Sidney Kimmel Cancer Center at Thomas Jefferson University as a primary goal. Resources of the PMSR are also available to investigators in other Cancer Centers and academic institutions as a secondary goal. Dr. Tang’s role as the PMSR Managing Director is to support NCI-funded cancer research projects by providing expert consultation and state-of-the-art technologies that operate at maximum performance at affordable costs to cancer investigators and other biomedical researchers. The Managing Director will assist in experimental design, perform MS data analyses as needed, and assist in the biological interpretation of results. The Managing Director will also devote substantial effort to optimizing and implementing new methods, update analytical and data analyses methods, and update instrumentation to ensure each project is performed using state-of-the-art methodologies. This is critical because instrumentation, software, and analytical strategies continue to evolve rapidly, and most current and anticipated future projects involve very challenging proteomics and metabolomics problems. Proteomics projects will include: 1) in-depth, global quantitative comparisons of exosomes, secretomes, cell lysates, tissues and biological fluids; 2) quantitative comparisons of post-translational modifications; and 3) LC-MS/MS analysis of isolated protein complexes with and without chemical crosslinking. Quantitative data will be obtained either using label-free quantitation of integrated MS ion currents, or using stable isotopes such as SILAC or TMT isobaric tag labeling. Metabolomics projects will include quantifying the steady state levels of polar metabolites, lipids and fatty acids, and 13C isotope tracer analysis. Representative specific plans for future development of new analytical approaches include: 1) improving characterization of protein palmitoylation with identification of specific modification sites; 2) de novo peptide sequencing for identification of HLA peptides; 3) an alternate approach to ubiquitome enrichment; and 4) absolute quantitation of gut metabolites. In addition, Dr. Tang will implement additional new or improved methods that are likely to be needed in future cancer projects. These proteomics and metabolomics analyses will contribute to critical data required to identify biomolecular targets, as well as generate hypotheses that are vital for the success of the cancer-related projects described in this application.