Investigating the glycolytic interactome to understand cancer metabolism

描述(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更高的葡萄糖摄取和代谢，即使在充足的氧气存在的情况下也是如此。这种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似乎效率较低的代谢途径，将葡萄糖转化为以ATP形式存在的能量。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。肿瘤细胞优先利用糖酵解的观察使许多人探索新陈代谢，以此作为治疗许多肿瘤的新窗口。在这里，我们提出了一种系统和无偏见的方法来识别糖酵解相互作用蛋白(GIPS)，以便更好地了解癌症中糖酵解的调节。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，我们已经确定了富含在细胞中的相互作用具有增强的Warburg效应，这产生了与Gips增加的相互作用驱动转化状态下的糖酵解的假设。我们将使用汇集的RNAi筛选技术来确定哪些GNAP对增殖和生存至关重要。结构性方法将被应用于对GIP监管职能的洞察，这些信息将被用于生成工具，以扰乱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对细胞的整体代谢程序做出贡献。这项提案中提出的目标将极大地扩展我们对癌症新陈代谢的了解，因此与国家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.