Raf-1 As a Regulator of Glutamine Metabolism

项目总结 RAS蛋白在癌症中经常发生突变，以激活必要的不同信号通路 对于不受控制的细胞增殖，包括负责调节新陈代谢的途径。RAS是 能够通过它的许多效应器来协调这些许多路径。尽管有大量的 由HRAS、NRAS和KRAS突变驱动的肿瘤使该蛋白成为一个有吸引力的药物靶点，其 结构和复杂的调控限制了有效的小分子抑制剂的开发。 因为RAS本身很难靶向，所以了解它的哪些效应器对其功能至关重要 致癌的背景是必要的。为此，哈瓦里实验室此前曾进行了一次CRISPR 筛选RAS相互作用因子，以确定RAS驱动的肿瘤维持所必需的效应器。 在不同的组织类型中，我们观察到致癌的RAS驱动的癌细胞对Raf-1的依赖性。以下为- 对Raf-1的UP实验表明，致癌的RAS诱导Raf-1和Raf-1的线粒体易位 与蛋白质相互作用的新型谷氨酰胺代谢调节剂。随后的实验 证明Raf-1的缺失导致依赖RAS的突变体谷氨酰胺代谢降低 举止。这项工作表明，Raf-1是RAS驱动的肿瘤的一种特异性易感性，因为它在 直接协调代谢过程。众所周知，癌细胞会重新连接它们的新陈代谢 这会导致某些新陈代谢的脆弱性，但针对这些脆弱性仍然是一个挑战。 这项工作旨在了解Raf-1在RAS驱动的癌症中调节谷氨酰胺代谢的作用 细胞。我们假设Raf-1蛋白直接调节RAS驱动的谷氨酰胺代谢 肿瘤。在目标I中，我将通过定义Raf-1与谷氨酰胺代谢的关系来阐明Raf-1对谷氨酰胺代谢的影响 在Raf-1和谷氨酰胺代谢酶之间。在AIM II中，我将确定RAS的机制- 诱导线粒体基质定位。总体而言，这项工作将为RAS提供一种新的机制- 引起癌症新陈代谢的改变。

PROJECT SUMMARY The Ras proteins are frequently mutated in cancer to activate diverse signaling pathways necessary for uncontrolled cellular proliferation, including pathways responsible for regulating metabolism. Ras is capable of coordinating these many pathways through its many effectors. Though the large number of tumors driven by HRAS, NRAS, and KRAS mutations makes the protein an attractive drug target, its structure and complex regulation have limited the development of effective small molecule inhibitors. Because Ras itself is difficult to target, understanding which of its effectors are critical for its function in an oncogenic context is necessary. To this end, the Khavari lab had previously conducted a CRISPR screen of Ras-interactors to identify effectors that were necessary for Ras-driven tumor maintenance. Across tissue types, we observed a dependence on Raf-1 in oncogenic Ras-driven cancer cells. Follow- up experiments on Raf-1 revealed that oncogenic Ras induced mitochondrial translocation of Raf-1 and novel regulators of glutamine metabolism interacting with the protein. Subsequent experiments demonstrated that loss of Raf-1 led to decreased glutamine metabolism in a mutant Ras-dependent manner. This work suggests that Raf-1 is a specific vulnerability of Ras-driven tumors due to its role in directly coordinating metabolic processes. It is well-known that cancer cells rewire their metabolism and this leads to certain metabolic vulnerabilities, but targeting those vulnerabilities remains a challenge. This work aims to understand the role of Raf-1 in regulating glutamine metabolism in Ras-driven cancer cells. We hypothesize that the Raf-1 protein directly regulates glutamine metabolism in Ras-driven tumors. In Aim I, I will elucidate the impact of Raf-1 on glutamine metabolism by defining the relationship between Raf-1 and glutamine metabolic enzymes. In Aim II, I will determine the mechanism of Ras- induced mitochondrial matrix localization. Overall, this work will provide a novel mechanism for Ras- induced alterations to cancer metabolism.