Stable Isotope amd Metabolomics Core

稳定同位素和代谢组学核心设施（Core xx）将提供本计划项目的至少两个项目，一个综合代谢组学设施，将协助研究人员进行研究人员分析辐射损伤过程代谢组学“特征”所需的全球代谢物和生物合成率分析，以及评估这些“特征”是如何通过破坏参与辐射防护和致敏的重要信号通路而被改变的。这些代谢组学“特征”将通过以下方式与相关病理生理学相关： 个别项目的研究人员，他们将与代谢组学核心密切合作，以完善代谢产物谱和生物合成速率谱，这是阐明代谢效应和对基础病理生理学的贡献所需的。 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孔板形式中的细胞/细胞器线粒体呼吸和糖酵解速率（Seahorse Biosciences Flux Analyzer）; 2.代谢物定量（基于GC/MS和/或LC/MS）; 3.稳定同位素（氘代水）评价 蛋白质和脂质的生物合成率。请注意，通量表征和代谢物谱分析产生细胞代谢“签名”，可用于裂隙模型的经验分类。当相互结合使用时，这些流量和代谢物的经验“签名”也可以产生对细胞代谢的强大生理评估，并且可以指出代谢途径中的补偿，这对维持可能适合治疗攻击的肿瘤细胞代谢至关重要。 核心的kev人员将是负责人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 准备和处理适合当前研究的组织样本，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（沃茨Xevo三重四极杆）、LTQ-轨道阱和气相色谱（GC）-飞行时间（沃茨 GCT）质谱（MS）），以及使用氘代水通过GCMS评估脂质和总蛋白质生物合成速率。 重要性：代谢组学“特征”的评价对于本方案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的体内影响，以及为影响辐射诱导损伤的结果而开发的技术。

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.