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Blockade of deubiquitylating enzyme Rpn11 triggers apoptosis in multiple myeloma cells and overcomes bortezomib resistance

Oncogene volume 36, pages 5631–5638 (2017)Cite this article

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Abstract

Proteasome inhibition is an effective therapy for multiple myeloma (MM) patients; however, the emergence of drug resistance is common. Novel therapeutic strategies to overcome proteasome inhibitor resistance are needed. In this study, we examined whether targeting deubiquitylating (DUB) enzymes upstream of 20S proteasome overcomes proteasome inhibitor resistance. Gene expression analysis, immunohistochemical studies of MM patient bone marrow, reverse transcription–PCR and protein analysis show that Rpn11/POH1, a DUB enzyme upstream of 20S proteasome, is more highly expressed in patient MM cells than in normal plasma cells. Importantly, Rpn11 expression directly correlates with poor patient survival. Loss-of-function studies show that Rpn11-siRNA knockdown decreases MM cell viability. Pharmacological inhibition of Rpn11 with O-phenanthroline (OPA) blocks cellular proteasome function, induces apoptosis in MM cells and overcomes resistance to proteasome inhibitor bortezomib. Mechanistically, Rpn11 inhibition in MM cells activates caspase cascade and endoplasmic stress response signaling. Human MM xenograft model studies demonstrate that OPA treatment reduces progression of tumor growth and prolongs survival in mice. Finally, blockade of Rpn11 increases the cytotoxic activity of anti-MM agents lenalidomide, pomalidomide or dexamethasone. Overall, our preclinical data provide the rationale for targeting DUB enzyme Rpn11 upstream of 20S proteasome to enhance cytotoxicity and overcome proteasome inhibitor resistance in MM.

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Acknowledgements

This investigation was supported by National Institutes of Health Specialized Programs of Research Excellence (SPORE) Grants P50100707, R01CA207237 and RO1CA050947. KCA is an American Cancer Society Clinical Research Professor.

Author contributions

DC conceived the project, designed research, analyzed data and wrote the manuscript; YS designed and performed the experiments and interpreted data; SL helped in RT–PCR and animal studies; AR and DSD helped in cytotoxicity assays; JQ helped in western studies; MKS performed the GEP data set analyses; NM and Y-TT provided patient samples; RDC helped in immunohistochemistry analysis; and KCA reviewed the manuscript.

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  1. D Chauhan and K C Anderson: Joint senior authors.

Authors and Affiliations

  1. Department of Medical Oncology, LeBow Institute for Myeloma Therapeutics and Jerome Lipper Myeloma Center, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA

    Y Song, A Ray, D S Das, M K Samur, Y-T Tai, N Munshi, R D Carrasco, D Chauhan & K C Anderson

  2. Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    S Li

  3. Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA

    J Qi

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Correspondence to D Chauhan or K C Anderson.

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Competing interests

KCA is on advisory board of Celgene, Millenium, Gilead and Bristol Myers Squibb, and is a Scientific Founder of Acetylon, Oncopep and C4 Therapeutics. The other authors declare no conflict of interest.

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Song, Y., Li, S., Ray, A. et al. Blockade of deubiquitylating enzyme Rpn11 triggers apoptosis in multiple myeloma cells and overcomes bortezomib resistance. Oncogene 36, 5631–5638 (2017). https://doi.org/10.1038/onc.2017.172

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