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Targeting the protein ubiquitination machinery in melanoma by the NEDD8-activating enzyme inhibitor pevonedistat (MLN4924)

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Abstract

Background The neddylation pathway conjugates NEDD8 to cullin-RING ligases and controls the proteasomal degradation of specific proteins involved in essential cell processes. Pevonedistat (MLN4924) is a selective small molecule targeting the NEDD8-activating enzyme (NAE) and inhibits an early step in neddylation, resulting in DNA re-replication, cell cycle arrest and death. We investigated the anti-tumor potential of pevonedistat in preclinical models of melanoma. Methods Melanoma cell lines and patient-derived tumor xenografts (PDTX) treated with pevonedistat were assessed for viability/apoptosis and tumor growth, respectively, to identify sensitive/resistant models. Gene expression microarray and gene set enrichment analyses were performed in cell lines to determine the expression profiles and pathways of sensitivity/resistance. Pharmacodynamic changes in treated-PDTX were also characterized. Results Pevonedistat effectively inhibited cell viability (IC50 < 0.3 μM) and induced apoptosis in a subset of melanoma cell lines. Sensitive and resistant cell lines exhibited distinct gene expression profiles; sensitive models were enriched for genes involved in DNA repair, replication and cell cycle regulation, while immune response and cell adhesion pathways were upregulated in resistant models. Pevonedistat also reduced tumor growth in melanoma cell line xenografts and PDTX with variable responses. An accumulation of pevonedistat-NEDD8 adduct and CDT1 was observed in sensitive tumors consistent with its mechanism of action. Conclusions This study provided preclinical evidence that NAE inhibition by pevonedistat has anti-tumor activity in melanoma and supports the clinical benefits observed in recent Phase 1 trials of this drug in melanoma patients. Further investigations are warranted to develop rational combinations and determine predictive biomarkers of pevonedistat.

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Acknowledgments

We thank the PETT lab members for critical comments.

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Authors and Affiliations

  1. Division of Medical Oncology, Department of Medicine, University of Washington School of Medicine, Seattle, WA, USA

    Kit Man Wong

  2. Developmental Therapeutics Program, Division of Medical Oncology, Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    Lindsey N. Micel, Heather M. Selby, Aik Choon Tan, Todd M. Pitts, Stacey M. Bagby, Peter J. Klauck, John J. Tentler & S. Gail Eckhardt

  3. University of Colorado Cancer Center, Aurora, CO, USA

    Aik Choon Tan, Todd M. Pitts, John J. Tentler & S. Gail Eckhardt

  4. Department of Medical Oncology, Princess Margaret Cancer Center, Toronto, ON, Canada

    Anna Spreafico

  5. Millennium Pharmaceuticals, Inc., a wholly owned subsidiary of Takeda Pharmaceutical Company Ltd, Cambridge, MA, USA

    Stephen J. Blakemore, Peter F. Smith, Alice McDonald & Allison Berger

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  1. Kit Man Wong
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Correspondence to S. Gail Eckhardt.

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Conflict of interest

Kit Man Wong declares that she has no conflict of interest. Lindsey N. Micel declares that she has no conflict of interest. Heather M. Selby declares that she has no conflict of interest. Aik Choon Tan declares that he has no conflict of interest. Todd M. Pitts declares that he has no conflict of interest. Stacey M. Bagby declares that she has no conflict of interest. Anna Spreafico declares that she has no conflict of interest. Peter J. Klauck declares that he has no conflict of interest. Stephen J. Blakemore was employed by Millennium Pharmaceuticals Inc. at the time of conducting this research. Peter F. Smith was employed by Millennium Pharmaceuticals Inc. at the time of conducting this research. Alice McDonald was employed by Millennium Pharmaceuticals Inc. at the time of conducting this research. Allison Berger was employed by Millennium Pharmaceuticals Inc. at the time of conducting this research. John J. Tentler declares that he has no conflict of interest. S. Gail Eckhardt declares that she has no conflict of interest.

S.J. Blakemore, P.G. Smith, A. McDonald and A. Berger were employed by Millennium Pharmaceuticals Inc. at the time of conducting this research.

Funding

This work was supported by a commercial research grant (A.C. Tan, T.M. Pitts, J.J. Tentler and S.G. Eckhardt) and by grants from the University of Colorado Cancer Center and Millennium Pharmaceuticals, Inc.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All animal studies described in this study were conducted at the University of Colorado Anschutz Medical Campus in accordance with the National Institutes of Health guidelines for the care and use of laboratory animals, and animals were housed in a facility accredited by the American Association for Accreditation of Laboratory Animal Care.

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Not applicable (study does not involve human subjects).

Grant support/funding

A.C. Tan, T.M. Pitts, J.J. Tentler and S.G. Eckhardt have a commercial research grant. This work in this manuscript was supported by grants from the University of Colorado Cancer Center and Millennium Pharmaceuticals, Inc.

Additional information

Kit Man Wong and Lindsey N. Micel contributed equally to work.

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Wong, K.M., Micel, L.N., Selby, H.M. et al. Targeting the protein ubiquitination machinery in melanoma by the NEDD8-activating enzyme inhibitor pevonedistat (MLN4924). Invest New Drugs 35, 11–25 (2017). https://doi.org/10.1007/s10637-016-0398-8

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