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Loss of the tumor suppressor Hace1 leads to ROS-dependent glutamine addiction

Oncogene volume 34, pages 4005–4010 (2015)Cite this article

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Abstract

Cellular transformation is associated with altered glutamine (Gln) metabolism. Tumor cells utilize Gln in the tricarboxylic acid (TCA) cycle to maintain sufficient pools of biosynthetic precursors to support rapid growth and proliferation. However, Gln metabolism also generates NADPH, and Gln-derived glutamate is used for synthesis of glutathione (GSH). As both NADPH and GSH are antioxidants, Gln may also contribute to redox balance in transformed cells. The Hace1 E3 ligase is a tumor suppressor inactivated in diverse human cancers. Hace1 targets the Rac1 GTPase for degradation at Rac1-dependent NADPH oxidase complexes, blocking superoxide generation by the latter. Consequently, loss of Hace1 increases reactive oxygen species (ROS) levels in vitro and in vivo. Given the link between Hace1 loss and increased ROS, we investigated whether genetic inactivation of Hace1 alters Gln metabolism. We demonstrate that mouse embryonic fibroblasts (MEFs) derived from Hace1−/− mice are highly sensitive to Gln withdrawal, leading to enhanced cell death compared with wild-type (wt) MEFs, and Gln depletion or chemical inhibition of Gln uptake blocks soft agar colony formation by Hace1−/− MEFs. Hace1−/− MEFs exhibit increased Gln uptake and ammonia secretion, and metabolic labeling using 13C-Gln revealed that Hace1 loss increases incorporation of Gln carbons into the TCA cycle intermediates. Gln starvation markedly increases ROS levels in Hace1−/− but not in wt MEFs, and treatment with the antioxidant N-acetyl cysteine or the TCA cycle intermediate oxaloacetate efficiently rescues Gln starvation-induced ROS elevation and cell death in Hace1−/− MEFs. Finally, Gln starvation increases superoxide levels in Hace1−/− MEFs, and NADPH oxidase inhibitors block the induction of superoxide and cell death by Gln starvation. Together, these results suggest that increased ROS production due to Hace1 loss leads to Gln addiction as a mechanism to cope with increased ROS-induced oxidative stress.

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Acknowledgements

This research was supported by funds from the Canadian Institute of Health Research (MOP-123416) and British Columbia Cancer Foundation through generous donations from Team Finn and Ride to Conquer Cancer (to PHS), and from the National Institutes of Health (R01 CA157996) and Damon-Runyon Cancer Research Foundation (to RJD). NC was funded by the Canadian Institutes of Health Research, Frederick Banting and Charles Best Canada Graduate Scholarship Doctoral Award.

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Author notes

  1. M Daugaard

    Present address: 5Current address: Department of Urological Sciences, University of British Columbia, Vancouver Prostate Centre, Vancouver, British Columbia, Canada V6H3Z6.,

  2. B Rotblat

    Present address: 6Current address: Department of Life Sciences, Ben Gurion University, Beersheva, Israel.,

Authors and Affiliations

  1. Department of Molecular Oncology, BC Cancer Research Centre, Vancouver, British Columbia, Canada,

    N Cetinbas, M Daugaard, S Hajee, B Rotblat, A Lopez, A Li & P H Sorensen

  2. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada

    N Cetinbas, M Daugaard, B Rotblat & P H Sorensen

  3. Department of Pediatrics, University of Texas-Southern Medical Centre, Dallas, TX, USA,

    A R Mullen & R J DeBerardinis

  4. McDermott Center for Human Growth and Development, University of Texas-Southern Medical Center, Dallas, TX, USA

    A R Mullen & R J DeBerardinis

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  1. N Cetinbas
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Correspondence to P H Sorensen.

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Cetinbas, N., Daugaard, M., Mullen, A. et al. Loss of the tumor suppressor Hace1 leads to ROS-dependent glutamine addiction. Oncogene 34, 4005–4010 (2015). https://doi.org/10.1038/onc.2014.316

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