Systems biology of glutamine utilization in melanoma

描述（由申请人提供）：黑色素瘤是一种几乎无法治愈的疾病，如果允许其发展到致命的转移阶段，开发新的治疗方法是一种公认的医疗需求。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枢碳代谢的大量重新布线，为诊断和治疗开辟了尚未开发的机会。该应用的研究人员最近的研究表明，谷氨酰胺作为碳和能量的重要来源和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源碳单位从三羧酸循环中突变退出至关重要的具体假设，这可能提供独特的靶向机会。拟议的研究将采用一种既定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，包括稳定同位素示踪，以及在一组具有代表性的人类黑色素瘤系上选择的靶酶的遗传抑制，然后在小鼠异种移植模型中测试和验证主要发现。除了获得基本的机制见解外，该项目的预期交付成果将包括验证的生物标志物（标志性代谢物和/或通量）和潜在的目标酶。拟议的研究计划包括两个主要的具体目标，即体外和体内研究：(I)通过靶向代谢组学、生理学和细胞生物学等技术的协同组合，阐明在一系列黑色素瘤细胞系中涉及谷氨酰胺代谢关键方面的机制并确定潜在的诊断和治疗靶点；（II）在原发性黑色素瘤和转移瘤的异种移植小鼠模型中验证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，该项目将为先进的转化研究和治疗发展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