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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).

Author information

Author notes

    • Martje Tönjes
    •  & Sebastian Barbus

    These authors contributed equally to this work.

Affiliations

  1. Division of Molecular Genetics, German Cancer Research Center (DKFZ), Heidelberg, Germany.

    • Martje Tönjes
    • , Sebastian Barbus
    • , Wei Wang
    • , Magdalena Schlotter
    • , Sabrina V Pleier
    • , Alfa H C Bai
    • , Irene Weibrecht
    • , Volker Hovestadt
    • , Dominik Sturm
    • , Hendrik Witt
    • , Stefan M Pfister
    • , Peter Lichter
    •  & Bernhard Radlwimmer
  2. Division of Epigenomics and Cancer Risk Factors, DKFZ, Heidelberg, Germany.

    • Yoon Jung Park
    • , Anders M Lindroth
    •  & Christoph Plass
  3. Department of Nutritional Science and Food Management, College of Health Science, Ewha Womans University, Seoul, South Korea.

    • Yoon Jung Park
  4. Division of Pediatric Neurooncology, DKFZ, Heidelberg, Germany.

    • Sabrina V Pleier
    • , Dominik Sturm
    • , Hendrik Witt
    •  & Stefan M Pfister
  5. Department of Neuropathology, Heinrich Heine University, Düsseldorf, Germany.

    • Daniela Karra
    • , Jörg Felsberg
    •  & Guido Reifenberger
  6. Department of Bioinformatics and Functional Genomics, Institute of Pharmacy and Molecular Biotechnology and Bioquant, University of Heidelberg, Heidelberg, Germany.

    • Rosario M Piro
    •  & Rainer König
  7. Division of Theoretical Bioinformatics, DKFZ, Heidelberg, Germany.

    • Rosario M Piro
    •  & Rainer König
  8. Department of Human Nutrition, Foods and Exercise, Virginia Polytechnic Institute, Blacksburg, Virginia, USA.

    • Adele Addington
    •  & Susan M Hutson
  9. Clinical Cooperation Unit Neurooncology, DKFZ, Heidelberg, Germany.

    • Dieter Lemke
  10. Department of New Materials and Biosystems, Max Planck Institute for Intelligent Systems, Stuttgart, Germany.

    • Claudio G Rolli
    •  & Ralf Kemkemer
  11. Division of Experimental Neurosurgery, University of Heidelberg, Heidelberg, Germany.

    • Benito Campos
    •  & Christel Herold-Mende
  12. Department of Neurosurgery, University of Heidelberg, Heidelberg, Germany.

    • Benito Campos
    •  & Christel Herold-Mende
  13. Human Oncology and Pathogenesis Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.

    • Sevin Turcan
    •  & Timothy A Chan
  14. Department of Pediatric Oncology, Hematology and Immunology, Heidelberg University Hospital, Heidelberg, Germany.

    • Dominik Sturm
    • , Hendrik Witt
    •  & Stefan M Pfister
  15. Reutlingen University of Applied Science, Reutlingen, Germany.

    • Ralf Kemkemer
  16. Department of General Pediatrics, Division of Inborn Metabolic Diseases, University Children's Hospital, Heidelberg, Germany.

    • Kathrin Schmidt
    •  & Jürgen G Okun
  17. Core Facility, Molecular Structural Analysis, DKFZ, Heidelberg, Germany.

    • William-Edmund Hull
  18. Core Facility, Small Animal Imaging Center, DKFZ, Heidelberg, Germany.

    • Manfred Jugold
  19. German Cancer Consortium (DKTK), DKFZ, Heidelberg, Germany.

    • Guido Reifenberger

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Contributions

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Bernhard Radlwimmer.

Supplementary information

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    Supplementary Text and Figures

    Supplementary Figures 1–10, Supplementary Tables 1–5 and Supplementary Methods

Videos

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    Supplementary Video 1

    Migration of U-87MG cells transduced with nontarget shRNA

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    Supplementary Video 2

    Migration of U-87MG cells transduced with BCAT1 shRNAI

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DOI

https://doi.org/10.1038/nm.3217

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