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The androgen receptor is a negative regulator of eIF4E phosphorylation at S209: implications for the use of mTOR inhibitors in advanced prostate cancer

Oncogene volume 36, pages 6359–6373 (2017)Cite this article

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Abstract

The antiandrogen bicalutamide is widely used in the treatment of advanced prostate cancer (PCa) in many countries, but its effect on castration-resistant PCa (CRPC) is limited. We previously showed that resistance to bicalutamide results from activation of mechanistic target of rapamycin (mTOR). Interestingly, clinical trials testing combinations of the mTOR inhibitor RAD001 with bicalutamide were effective in bicalutamide-naïve CRPC patients, but not in bicalutamide-pretreated ones. Here we investigate causes for their difference in response. Evaluation of CRPC cell lines identified resistant vs sensitive in vitro models, and revealed that increased eIF4E(S209) phosphorylation is associated with resistance to the combination. We confirmed using a human-derived tumor xenograft mouse model that bicalutamide pre-treatment is associated with an increase in eIF4E(S209) phosphorylation. Thus, AR suppressed eukaryotic initiation factor 4E (eIF4E) phosphorylation, while the use of antiandrogens relieved this suppression, thereby triggering its increase. Additional investigation in human prostatectomy samples showed that increased eIF4E phosphorylation strongly correlated with the cell proliferation marker Ki67. Small interfering RNA-mediated knockdown (k/d) of eIF4E-sensitized CRPC cells to RAD001+bicalutamide, whereas eIF4E overexpression induced resistance. Inhibition of eIF4E phosphorylation by treatment with CGP57380 (an inhibitor of mitogen-activated protein kinase-interacting serine–threonine kinases MAP kinase-interacting kinase 1 (Mnk1/2), the eIF4E upstream kinase) or inhibitors of extracellular signal-regulated kinase 1/2 (ERK1/2), the upstream kinase-regulating Mnk1/2, also sensitized CRPC cells to RAD001+bicalutamide. Examination of downstream targets of eIF4E-mediated translation, including survivin, demonstrated that eIF4E(S209) phosphorylation increased cap-independent translation, whereas its inhibition restored cap-dependent translation, which could be inhibited by mTOR inhibitors. Thus, our results demonstrate that while combinations of AR and mTOR inhibitors were effective in suppressing tumor growth by inhibiting both AR-induced transcription and mTOR-induced cap-dependent translation, pre-treatment with AR antagonists including bicalutamide increased eIF4E phosphorylation that induced resistance to combinations of AR and mTOR inhibitors by inducing cap-independent translation. We conclude that this resistance can be overcome by inhibiting eIF4E phosphorylation with Mnk1/2 or ERK1/2 inhibitors.

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Acknowledgements

We thank Novartis Pharmaceuticals for the gift of RAD001. We also thank Stephanie Soares, Department of Urology, University of California Davis, School of Medicine (Sacramento, CA, USA), for the construction of the tissue microarrays used in this study, and Yu Wang, Department of Urology, for assistance with mice experiments. Human PSA-luciferase construct was kindly provided by Dr XuBao Shi, University of California Davis, Department of Urology. CWR-R1 cells were provided by Dr Elizabeth Wilson (University of North Carolina), pRNS1-1 cells were from Dr Johng Rhim, University of the Health Sciences (Bethesda, MD, USA), whereas PC-346C cells were from Dr WM van Weerden, Josephine Nefkens Institute (Erasmus MC, Rotterdam, Netherlands). We also thank Dr Xinbin Chen (UC Davis School of Veterinary Medicine) for the pRL-HCV-FL plasmid, and Maitreyee K Jathal and Thomas M Steele (UC Davis, Department of Urology) for samples of 22Rv1 xenograft tumors. This work was supported by a Biomedical Laboratory Research and Development (BLRD) Merit Award (I01BX000400, to PMG) from the Department of Veterans Affairs, and by Awards R01CA133209 (to PMG) and R01CA185509 (to PMG) from the National Institutes of Health.

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Authors and Affiliations

  1. VA Northern California Health Care System, University of California at Davis, Sacramento, CA, USA

    L S D'Abronzo, M E Crapuchettes, R E Beggs, S Siddiqui, F U Melgoza & P M Ghosh

  2. Department of Urology, University of California Davis School of Medicine, University of California at Davis, Sacramento, CA, USA

    L S D'Abronzo, S Bose, R L Vinall, R W deVere White & P M Ghosh

  3. California Northstate University College of Pharmacy, Elk Grove, CA, USA

    R L Vinall

  4. Department of Biochemistry and Molecular Medicine, University of California at Davis, Sacramento, CA, USA

    C G Tepper & P M Ghosh

  5. Department of Medical Microbiology and Immunology, University of California at Davis, Sacramento, CA, USA

    M Mudryj

  6. Department of Public Health, Division of Biostatistics, University of California at Davis, Sacramento, CA, USA

    B P Durbin-Johnson

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  1. L S D'Abronzo
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Correspondence to P M Ghosh.

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The work reported here does not represent the views or opinions of the Department of Veteran Affairs or the United States Government. The authors declare no conflict of interest.

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D'Abronzo, L., Bose, S., Crapuchettes, M. et al. The androgen receptor is a negative regulator of eIF4E phosphorylation at S209: implications for the use of mTOR inhibitors in advanced prostate cancer. Oncogene 36, 6359–6373 (2017). https://doi.org/10.1038/onc.2017.233

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