Systems biology of glutamine utilization in melanoma

描述(由申请人提供)：开发针对黑色素瘤的新疗法是公认的医学需求。黑色素瘤是一种几乎无法治愈的疾病，如果允许进展到致命的转移阶段。与所有其他癌症一样，黑色素瘤的肿瘤发生和肿瘤进展伴随着中央碳代谢的实质性重新连接，为诊断和治疗打开了未知的机会。最近对这一应用的研究人员的研究表明，谷氨酰胺作为一种基本的碳源和能量来源以及人类黑色素瘤细胞系的潜在治疗靶点，其利用率大大提高。这些研究的初步结果导致了关于谷氨酰胺起源的碳单位从三羧酸循环中退出的关键重要性的具体假设，这可能提供独特的靶向机会。拟议的研究将使用一种已建立的综合方法来解决这一假设，该方法结合了多方面的代谢分析，包括稳定同位素跟踪，以及在一组具有代表性的人类黑色素瘤株上对选定的靶酶进行基因抑制，然后在小鼠异种移植模型中测试和验证主要发现。除了获得基本的机理洞察外，该项目的预期交付成果将包括经过验证的生物标记物(标志性代谢物和/或焊剂)和潜在的靶标酶。拟议的研究计划包括两个主要的具体目标，包括体外和体内研究：(I)通过靶向代谢组学、生理学和细胞生物学等技术的协同结合，阐明各种黑色素瘤细胞系中谷氨酰胺代谢的关键方面的机制并确定潜在的诊断和治疗靶点；(Ii)验证原发黑色素瘤肿瘤和转移的异种小鼠模型中谷氨酰胺代谢的关键方面、生物标志物和初步确定的靶点。如果成功完成，该项目将为高级翻译研究和治疗开发提供一个强有力的起点

DESCRIPTION (provided by applicant): Development of new therapies against melanoma, a nearly incurable disease if allowed to progress to the deadly metastatic stage, is a well-recognized medical need. As in all other cancers, tumorigenesis and tumor progression in melanoma is accompanied by substantial rewiring of central carbon metabolism opening unexplored opportunities for diagnostics and treatment. Recent studies of the investigators of this application implicated a substantially enhanced utilization of glutamine as an essential anaplerotic source of carbon and energy and a potential therapeutic target in human melanoma cell lines. The preliminary results of these studies led to the specific hypothesis about the critial importance of the cataplerotic exit of glutamine- originating carbon units from the tricarboxylic acid cycle, which may provide unique targeting opportunities. The proposed studies will address this hypothesis using an established integrative approach, which combines multifaceted metabolic analyses, including stable-isotope tracing, with genetic suppression of selected target enzymes over a panel of representative human melanoma lines followed by testing and validation of main finding in mouse xenograft models. In addition to gaining fundamental mechanistic insights, anticipated deliverables of the project would include validated biomarkers (signature metabolites and/or fluxes) and potential target enzymes. The proposed research plan includes two major specific aims featuring in vitro and in vivo studies: (I) to elucidate mechanisms and identify potential diagnostic and therapeutic targets involved in the key aspects of glutamine metabolism in a range of melanoma cell lines by a synergistic combination of techniques including targeted metabolomics, physiology and cell biology; (II) to validate the key aspects of glutamine metabolism, biomarkers and tentatively identified targets in the xenograft mouse models of primary melanoma tumors and metastases. If successfully accomplished, this project would provide a strong starting point for advanced translational studies and therapeutic developments