Raf-1 As a Regulator of Glutamine Metabolism

Raf-1 作为谷氨酰胺代谢的调节剂

• 批准号: 10361184
• 负责人: Ronald Leslie Shanderson
• 金额: $ 4.09万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-15 至 2023-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361184
• 关键词: Amino Acids; Binding; Biochemical; Biological Assay; Biophysics; Biotin; CRISPR screen; Cancer cell line; Cell Line; Cell Proliferation; Cells; Cellular Metabolic Process; Client; Complex; Dependence; Development; Drug Targeting; Enzymes; Event; Genetic Transcription; Gluconeogenesis; Glutaminase; Glutamine; KRAS2 gene; Knowledge; Label; MAP Kinase Gene; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mentorship; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Mitochondrial Matrix; Molecular Chaperones; Mutate; Mutation; Oncogenic; Outer Mitochondrial Membrane; Pathway interactions; Pharmaceutical Preparations; Phosphoenolpyruvate Carboxylase; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Precision therapeutics; Prevalence; Process; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-raf; RAF1 gene; RAS genes; Regulation; Research Personnel; Role; Signal Pathway; Signal Transduction; Structure; Succinate-CoA Ligases; Techniques; Tissues; Work; cancer cell; experimental study; follow-up; insight; mutant; novel; nucleotide metabolism; programs; protein protein interaction; raf-1 Protein; ras Proteins; small molecule inhibitor; translocase; tumor; tumor metabolism

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.

项目摘要 Ras蛋白在癌症中经常发生突变，以激活所需的多种信号通路。 不受控制的细胞增殖，包括负责调节代谢的途径。Ras是 能够通过它的许多效应器协调这些途径。虽然大量的 由HRAS、NRAS和KRAS突变驱动的肿瘤使蛋白质成为有吸引力的药物靶点， 结构和复杂的调节限制了有效的小分子抑制剂的发展。 由于Ras本身难以靶向，因此了解其效应子中的哪些对于其在细胞中的功能至关重要。 一个致癌的背景是必要的。为此，Khavari实验室此前进行了一项CRISPR 筛选Ras相互作用物以鉴定Ras驱动的肿瘤维持所必需的效应物。 在不同组织类型中，我们观察到致癌Ras驱动的癌细胞对Raf-1的依赖性。关注- 对Raf-1的实验表明，致癌Ras诱导Raf-1的线粒体易位， 与蛋白质相互作用的谷氨酰胺代谢的新调节剂。随后的实验 表明Raf-1的缺失导致突变型Ras依赖性细胞中谷氨酰胺代谢降低， 方式这项工作表明，Raf-1是Ras驱动的肿瘤的一个特定的脆弱性，因为它的作用， 直接协调代谢过程。众所周知，癌细胞会重新调整其新陈代谢， 这导致了某些代谢弱点，但针对这些弱点仍然是一个挑战。 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。 肿瘤的在目的I中，我将阐明Raf-1对谷氨酰胺代谢的影响， Raf-1和谷氨酰胺代谢酶之间的关系。在目标II中，我将确定Ras的机制- 诱导线粒体基质定位。总的来说，这项工作将提供一个新的机制，Ras- 导致癌症代谢的改变。