Abstract B044: Elongation control of mRNA translation drives Group 3 medulloblastoma adaptation to nutrient deprivation
Authors: Alberto Delaidelli, Fares Burwag, Jessica Oliveira de Santis, Betty Yao, Haifeng Zhang, Yue Zhou Huang, Gian Luca Negri, Christopher Hughes, Gabriel Leprivier, Poul Sorensen
Journal: Cancer Research (2024)

Abstract Background: Group 3 affiliation and MYC genetic amplification are associated with poor life expectancy and substantial morbidity in children suffering from medulloblastoma (MB). The high metabolic demand induced by MYC-driven transformation sensitizes MYC-overexpressing MB to cell death under conditions of nutrient deprivation (ND). Additionally, MYC-driven transformation is known to promote mitochondrial oxidative phosphorylation (OXPHOS). We previously reported that eukaryotic Elongation Factor Kinase 2 (eEF2K), the master regulator of mRNA translation elongation, promotes survival of MYC-overexpressing tumors under ND. Interestingly, eEF2K is overexpressed in MYC-driven MB and our preliminary proteomics data highlight large-scale alterations in OXPHOS components affecting eEF2K deficient MB cells. We therefore hypothesized that eEF2K activity is required for the selective translation of mRNAs needed for efficient OXPHOS, and for the progression of MYC-driven MB. METHODS: Multiplexed enhanced Protein Dynamic Mass Spectrometry performed in eEF2K knockdown MYC-overexpressing D425 MB cells to identify mRNAs selectively translated upon eEF2K activation. Time course experiments under ND were conducted in eEF2K knockout (KO) D425 cells to assess the presence of electron transport chain (ETC) complexes I-IV in their native state (via BN-PAGE), as well as transcript expression of individual ETC complex components (by qPCR). The effects of eEF2K inactivation on oxygen consumption, metabolic fluxes and mitochondrial membrane potential were studied with Seahorse technology and JC1/TMRE staining. The viability of eEF2K KO D425 cells was assessed by Incucyte system. Finally, MB orthotopic xenograft mouse models were used to confirm in vitro observations. RESULTS: Multiple (9 out of 10 detected) components of the mitochondrial OXPHOS pathway are selectively translated upon eEF2K activation. Inactivation of eEF2K by genetic KO leads to the disassembly of ETC complexes I-IV without affecting mRNA levels of their respective components. Consistently, eEF2K KO MB cells display decreased mitochondrial membrane potential and ~20% increased proton leak through the mitochondrial membrane. In addition, eEF2K inactivation results in increased D425 cell death under ND and doubles survival of MB bearing mice fed with calorie restricted diets (p&amp;lt;0.05). CONCLUSION: Control of mRNA translation elongation by eEF2K is critical for mitochondrial ETC complex assembly and efficient OXPHOS in MYC-overexpressing MB, likely representing an adaptive response by which MYC-driven MB cells cope with acute metabolic stress. Future therapeutic studies will aim to combine eEF2K inhibition with caloric restriction mimetic drugs as eEF2K activity appears critical under metabolic stress conditions. Citation Format: Alberto Delaidelli, Fares Burwag, Jessica Oliveira de Santis, Betty Yao, Haifeng Zhang, Yue Zhou Huang, Gian Luca Negri, Christopher Hughes, Gabriel Leprivier, Poul Sorensen. Elongation control of mRNA translation drives Group 3 medulloblastoma adaptation to nutrient deprivation [abstract]. In: Proceedings of the AACR Special Conference on Brain Cancer; 2023 Oct 19-22; Minneapolis, Minnesota. Philadelphia (PA): AACR; Cancer Res 2024;84(5 Suppl_1):Abstract nr B044.