Stable Isotope amd Metabolomics Core

稳定同位素和代谢组学核心设施(Core Xx)将提供该计划项目的至少两个项目一个全面的代谢物组学设施，帮助研究人员进行全球代谢物和生物合成速率分析，这是研究人员分析辐射损伤过程的代谢“特征”所需的，并评估这些“特征”如何因辐射防护和敏化所涉及的重要信号通路的中断而改变。这些代谢“信号”将通过以下方式与相关的病理生理学相关 个别项目的研究人员将与代谢组学核心密切合作，完善代谢物图谱和生物合成速率图谱，以阐明代谢影响和对潜在病理生理学的贡献。 该核心将就方案设计、样品制备和处理以及不同研究可用的技术和程序提供咨询意见。核心提供的服务可以评估： 1.96孔板形式的细胞/细胞器线粒体呼吸和糖酵解率(海马生物科学通量分析仪)；2.代谢物定量(基于GC/MS和/或LC/MS)；3.稳定同位素(氚水)评估 蛋白质和脂肪的生物合成速率。请注意，通量特征和代谢物特征分析都产生了可用于FISE模型经验分类的细胞代谢‘特征’。当相互结合使用时，这些通量和代谢物的经验性“信号”也可以产生对细胞代谢的强大生理评估，并可以指出代谢途径中的补偿，这对维持肿瘤细胞的新陈代谢至关重要，可能受到治疗的攻击。 核心的Kev人员将是董事和代谢组学核心技术人员。主任将负责监督核心的所有活动，就不同的方法和程序提供建议，解释结果，并生成代谢组学报告和数据解释。技术人员将 为手头的研究准备和处理适当的组织样本，并执行一些专门的技术和程序，例如代谢物发现和定量(使用液相色谱(LC)/串联质谱仪(Waters Xevo三重四极杆)、LTQ-Orbitrap和气相色谱(GC)-FIME of Flight(Waters 用气相色谱-质谱仪(GCMS)，以及用氚水评估脂肪和总蛋白的生物合成速率。 意义：代谢组学“特征”的评估对本计划项目中的体内辐射诱导损伤项目至关重要，因为它允许研究受辐射诱导损伤影响的代谢过程中的干扰在体内的影响，以及为影响辐射诱导损伤结果而开发的技术。

The Stable Isotope and Metabolomics Core Facility (Core xx) will provide at least two projects of this Program Project a comprehensive metabolomics facility that will assist the investigators in global metabolite and biosynthetic rate analyses required for the investigators to analyze the metabolomic "signatures" ofthe radiation injury process, as well as assess how these "signatures" are modified by disruptions of significant signaling pathways involved in radioprotecfion and sensitization. These metabolomic "signatures" will be related to the pertinent pathophysiology by the investigators ofthe individual projects, who will work closely with the Metabolomics Core to refine metabolite profiles, and biosynthetic rate profiles, needed for the elucidafion of the metabolic effects, and contributions to, the underlying pathophysiology. The core will provide advice on protocol design, preparation and processing of samples, as well as on the techniques and procedures available for the different studies. The services offered by the Core can evaluate : 1. Cellular/Organelle mitochondrial respiration and glycolysis rates in a 96 well plate format (Seahorse Biosciences Flux Analyzer); 2. Metabolite quantification (GC/MS and/or LC/MS based); 3. Stable isotope (deuterated water) evealuafion of protein and lipid biosynthefic rates. Note that both flux characterizafion, and metabolite profiling yield cellular metabolic 'signatures' that can be used for empirical classification of fissue models. When used in conjunction with each other, these empirical 'signatures' of flux and metabolites can also yield powerful physiologic assessments of cellular metabolism, and can point to compensations in metabolic pathways that are crucial to the maintenance of tumor cell metabolism that may be amenable to therapeutic attack. The kev personnel of the core will be the director and the Metabolomics Core technical staff. The director will be responsible for supervising all the acfivifies of the core, offering advice on the different methods and procedures, interpretation ofthe results, and generating Metabolomics reports and interpretations ofthe data. The technical staff will prepare and process tissue samples as appropriate for the study at hand, and perform some of the specialized techniques and procedures, such as metabolite discovery and quantificafion (using liquid chromatography (LC)/tandem mass spectrometry (Waters Xevo triple quadrapole), LTQ-Orbitrap, and gas chromatography (GC)-fime of flight (Waters GCT) mass spectrometry(MS)), and lipid and total protein biosynthefic rate assessment using deuterated water, by GCMS. Significance: Evaluation of metabolomics "signatures" is essential to the in vivo radiation-induced injury projects in this Program Project as it allow the study of the in vivo impact of disturbances in metabolic processes affected by radiation-induced injury, as well technologies developed to effect the outcome of radiation-induced injury.