Abstract
Here we show that glioblastoma express high levels of branched-chain amino acid transaminase 1 (BCAT1), the enzyme that initiates the catabolism of branched-chain amino acids (BCAAs). Expression of BCAT1 was exclusive to tumors carrying wild-type isocitrate dehydrogenase 1 (IDH1) and IDH2 genes and was highly correlated with methylation patterns in the BCAT1 promoter region. BCAT1 expression was dependent on the concentration of α-ketoglutarate substrate in glioma cell lines and could be suppressed by ectopic overexpression of mutant IDH1 in immortalized human astrocytes, providing a link between IDH1 function and BCAT1 expression. Suppression of BCAT1 in glioma cell lines blocked the excretion of glutamate and led to reduced proliferation and invasiveness in vitro, as well as significant decreases in tumor growth in a glioblastoma xenograft model. These findings suggest a central role for BCAT1 in glioma pathogenesis, making BCAT1 and BCAA metabolism attractive targets for the development of targeted therapeutic approaches to treat patients with glioblastoma.
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Acknowledgements
We thank G. Tödt for bioinformatic expertise, M. Kirchgäßner, K. Pfleger, E. Wieland, A.-C. Klein, S. Emmerich and the DKFZ Light Microscopy Facility for excellent technical support, C. Schmidt (DKFZ) for providing the HEY1-pDest-vectors and A. von Deimling (University of Heidelberg) for providing the IDH1-specific antibodies. This work was supported by the German Federal Ministry of Education and Research (BMBF) within the National Genome Research Network NGFNplus (01GS0883 and 01GS0884) to B.R., P.L. and G.R., the German Cancer Aid (Deutsche Krebshilfe, grant number 108456) to B.R. and P.L. and the intramural funding program of the National Tumor Center Heidelberg. Y.J.P. was supported by the Roman Herzog research fellowship from the Hertie Foundation, Germany, and the Korean Health Technology R&D Project, Ministry of Health and Welfare, Republic of Korea (A120071-1211-0000200).
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M.T. and S.B. developed the concept, designed and performed experiments, analyzed data and wrote the manuscript. Y.J.P. and A.M.L. conducted the promoter methylation analysis, chromatin immunoprecipitation analysis and HEY1 knockdown. W.W. conducted dimethyl–α-KG and hypoxia experiments. M.S. assisted with experiments. S.V.P. conducted lentiviral knockdown in NCH421k cells. A.H.C.B. assisted with microchannel experiments. D.K. conducted western blot analysis of patient samples. R.M.P. and R. König mapped metabolic pathways. J.F. performed IDH1 mutation studies and immunohistochemical analyses. D.L. performed animal experiments. I.W., V.H., S.T., T.A.C., D.S., H.W. and S.M.P. assisted with the methylation analysis. A.A. and S.M.H. performed and interpreted BCAT1 inhibition experiments. C.G.R. and R. Kemkemer synthesized the microchannel chips. B.C. and C.H.-M. analyzed tissue microarray data. K.S. conducted the MS/MS and gas chromatography–mass spectrometry. W.-E.H. performed the 1H-NMR spectroscopy. M.J. performed magnetic resonance imaging. C.P. was involved in the methylation analysis. J.G.O. analyzed the mass spectrometry data. G.R. performed histological and immunohistochemical analyses and contributed to writing the manuscript. P.L. oversaw all research phases and contributed to writing the manuscript. B.R. developed the concept, supervised the project and wrote the manuscript.
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Supplementary Figures 1–10, Supplementary Tables 1–5 and Supplementary Methods (PDF 1918 kb)
Supplementary Video 1
Migration of U-87MG cells transduced with nontarget shRNA (AVI 13511 kb)
Supplementary Video 2
Migration of U-87MG cells transduced with BCAT1 shRNAI (AVI 14761 kb)
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Tönjes, M., Barbus, S., Park, Y. et al. BCAT1 promotes cell proliferation through amino acid catabolism in gliomas carrying wild-type IDH1. Nat Med 19, 901–908 (2013). https://doi.org/10.1038/nm.3217
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DOI: https://doi.org/10.1038/nm.3217
