α-Ketoglutarate (αKG) is a key node in many important metabolic pathways. The αKG analog N-oxalylglycine (NOG) and its cell-permeable prodrug dimethyloxalylglycine (DMOG) are extensively used to inhibit αKG-dependent dioxygenases. However, whether NOG interference with other αKG-dependent processes contributes to its mode of action remains poorly understood. Here we show that, in aqueous solutions, DMOG is rapidly hydrolyzed, yielding methyloxalylglycine (MOG). MOG elicits cytotoxicity in a manner that depends on its transport by monocarboxylate transporter 2 (MCT2) and is associated with decreased glutamine-derived tricarboxylic acid–cycle flux, suppressed mitochondrial respiration and decreased ATP production. MCT2-facilitated entry of MOG into cells leads to sufficiently high concentrations of NOG to inhibit multiple enzymes in glutamine metabolism, including glutamate dehydrogenase. These findings reveal that MCT2 dictates the mode of action of NOG by determining its intracellular concentration and have important implications for the use of (D)MOG in studying αKG-dependent signaling and metabolism.
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The data that support the findings of this study are available from the corresponding author upon reasonable request. Accession codes and relevant web links can be found in the respective legends and methods sections.
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We thank all members of the laboratory of D.A. for valuable discussions and input throughout this work, particularly J. Macpherson and N. Bevan for technical help. We are grateful to L. Cantley for advice during early stages of this work. We acknowledge S. O’Callaghan (Bio21 Institute, University of Melbourne) for the algorithm to correct for natural isotope abundance in metabolomics data. We thank J. Kleinjung for advice with statistical methods, M. Howell for advice and help with cell proliferation and viability measurements, J. Cerveira for help with flow cytometry measurements and A. Gould for comments on the manuscript. We are grateful to the staff at the Medical Research Council National Biomedical NMR Centre at the Francis Crick Institute, where NMR data were obtained. This work was funded by the MRC (MC_UP_1202/1) and by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001033), the UK Medical Research Council (FC001033) and the Wellcome Trust (FC001033) to D.A.
The authors declare no competing interests.
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Supplementary Tables 1 and 2, and Supplementary Figures 1–13
Spearman’s rank correlation coefficients of gene transcripts with IC50DMOG, determined as described in Methods. Genes identified as positively or negatively correlating with sensitivity (as determined by a 5% FDR) are highlighted.
Spearman’s rank correlation coefficients of gene transcripts with IC50DMOG, using only the top quartile of SLC16A7-expressing cell lines (213 lines) used in Fig. 6e, f. Genes identified as positively or negatively correlating with sensitivity (as determined by a 5% FDR) are highlighted.
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Fets, L., Driscoll, P.C., Grimm, F. et al. MCT2 mediates concentration-dependent inhibition of glutamine metabolism by MOG. Nat Chem Biol 14, 1032–1042 (2018). https://doi.org/10.1038/s41589-018-0136-y
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