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Isocitrate dehydrogenase 1 mutant R132H sensitizes glioma cells to BCNU-induced oxidative stress and cell death

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Abstract

Isocitrate dehydrogenase 1 (IDH1) decarboxylates isocitrate to α-ketoglutarate (α-KG) leading to generation of NADPH, which is required to regenerate reduced glutathione (GSH), the major cellular ROS scavenger. Mutation of R132 of IDH1 abrogates generation of α-KG and leads to conversion of α-KG to 2-hydroxyglutarate. We hypothesized that glioma cells expressing mutant IDH1 have a diminished antioxidative capacity and therefore may encounter an ensuing loss of cytoprotection under conditions of oxidative stress. Our study was performed with LN229 cells stably overexpressing IDH1 R132H and wild type IDH1 or with a lentiviral IDH1 knockdown. Quantification of GSH under basal conditions and following treatment with the glutathione reductase inhibitor BCNU revealed significantly lower GSH levels in IDH1 R132H expressing cells and IDH1 KD cells compared to their respective controls. FACS analysis of cell death and ROS production also demonstrated an increased sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to BCNU, but not to temozolomide. The sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to ROS induction and cell death was further enhanced with the transaminase inhibitor aminooxyacetic acid and under glutamine free conditions, indicating that these cells were more addicted to glutaminolysis. Increased sensitivity to BCNU-induced ROS production and cell death was confirmed in HEK293 cells inducibly expressing the IDH1 mutants R132H, R132C and R132L. Based on these findings we propose that in addition to its established pro-tumorigenic effects, mutant IDH1 may also limit the resistance of gliomas to specific death stimuli, therefore opening new perspectives for therapy.

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Acknowledgments

We thank Gabriele Köpf and Hildegard König for excellent technical assistance.

Funding

This study was supported by the Deutsche Krebshilfe (Grant 108795).

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Experimental Neurosurgery, Neuroscience Center, Goethe University Hospital, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany

    Isabelle Vanessa Mohrenz, Patrick Antonietti, Sophia Voigt, Alicia Mukrowsky & Donat Kögel

  2. Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), 69120, Heidelberg, Germany

    Stefan Pusch, David Capper & Andreas von Deimling

  3. Department of Neuropathology, Institute of Pathology, Ruprecht-Karls-University Heidelberg, INF 224, 69120, Heidelberg, Germany

    David Capper, Jörg Balss, Susanne Weissert & Andreas von Deimling

  4. Institute of Biological Chemistry and Nutrition, University of Hohenheim, 70593, Stuttgart, Germany

    Jan Frank

  5. Department of Neurosurgery, Goethe University Hospital, Theodor-Stern-Kai 7, 60590, Frankfurt, Germany

    Christian Senft & Volker Seifert

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  1. Isabelle Vanessa Mohrenz
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  2. Patrick Antonietti
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  3. Stefan Pusch
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  13. Donat Kögel
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Correspondence to Donat Kögel.

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Mohrenz, I.V., Antonietti, P., Pusch, S. et al. Isocitrate dehydrogenase 1 mutant R132H sensitizes glioma cells to BCNU-induced oxidative stress and cell death. Apoptosis 18, 1416–1425 (2013). https://doi.org/10.1007/s10495-013-0877-8

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  • DOI: https://doi.org/10.1007/s10495-013-0877-8

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