Investigating the glycolytic interactome to understand cancer metabolism

描述（由申请人提供）：与正常组织相比，癌细胞通过有氧糖酵解表现出增加的葡萄糖摄取和代谢，即使在充足的氧气存在下。这个被称为“华宝效应”的观察结果是自相矛盾的，因为它意味着癌细胞使用一种看似效率较低的代谢途径将葡萄糖转化为ATP形式的能量。最近的研究表明，糖酵解中间体是支持细胞增殖所必需的生物合成途径的前体。癌细胞优先使用糖酵解的观察导致许多人探索代谢作为许多肿瘤治疗机会的新窗口。在此，我们提出了一种系统和公正的方法来鉴定糖酵解相互作用蛋白（GIPs），以便更好地了解糖酵解在癌症中的调节。我们假设癌细胞中的糖酵解调节是通过GIPS和糖酵解酶之间的相互作用发生的，并且这些相互作用有助于癌细胞的恶性表型。使用基于silac的蛋白质组学，我们已经确定了Warburg效应增加的细胞中富集的相互作用，这产生了与GIPs相互作用增加驱动转化状态下糖酵解的假设。我们将使用聚合RNAi筛选技术确定哪些GIPs对增殖和生存能力至关重要。结构方法将被应用于对GIP调控功能的深入了解，这些信息将用于生成工具来破坏这些潜在的重要相互作用。最后，我们将研究转化和糖酵解蛋白相互作用组如何促进细胞的整体代谢程序。本提案提出的目标将极大地扩展我们对癌症代谢的认识，因此与美国国立卫生研究院的使命直接相关。

DESCRIPTION (provided by applicant): Cancer cells, compared to normal tissue, display increased glucose uptake and metabolism via aerobic glycolysis, even in the presence of ample oxygen. This observation, termed the Warburg Effect, was paradoxical because it meant that cancer cells used a seemingly less efficient metabolic pathway to convert glucose to energy in the form of ATP. More recent investigations have demonstrated that glycolytic intermediates serve as precursors for biosynthetic pathways necessary to support cellular proliferation. The observation that cancer cells preferentially use glycolysis has led many to explore metabolism as a novel window of therapeutic opportunity for a number of tumors. Herein we propose a systematic and unbiased approach to identify glycolytic interacting proteins (GIPs) in order to better understand regulation of glycolysis in cancer. We hypothesize that glycolytic regulation in cancer cells occurs through interactions between GIPS and glycolytic enzymes and that these interactions contribute to the malignant phenotype of cancer cells. Using SILAC-based proteomics we have identified interactions enriched in cells with increased Warburg Effect, which has generated the hypothesis that increased interactions with GIPs drive glycolysis in the transformed state. We will determine which GIPs are essential for proliferation and viability using pooled RNAi screening technologies. Structural approaches will be applied to gain insights toward GIP regulatory function and this information will be used to generate tools to disrupt these potentially important interactions. Finally we will examine how transformation and the glycolytic protein interactome contribute to the overall metabolic program of the cell. The aims presented in this proposal will greatly expand our knowledge of cancer metabolism and are thus directly relevant to the mission of the National Institutes of Health.