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Multiple Myeloma, Gammopathies

A novel hypoxia-selective epigenetic agent RRx-001 triggers apoptosis and overcomes drug resistance in multiple myeloma cells

Leukemia volume 30, pages 2187–2197 (2016)Cite this article

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Abstract

The hypoxic bone marrow (BM) microenvironment confers growth/survival and drug resistance in multiple myeloma (MM) cells. Novel therapies targeting the MM cell in its hypoxic BM milieu may overcome drug resistance. Recent studies led to the development of a novel molecule RRx-001 with hypoxia-selective epigenetic and nitric oxide-donating properties. Here, we demonstrate that RRx-001 decreases the viability of MM cell lines and primary patient cells, as well as overcomes drug resistance. RRx-001 inhibits MM cell growth in the presence of BM stromal cells. RRx-001-induced apoptosis is associated with: (i) activation of caspases; (ii) release of ROS and nitrogen species; (iii) induction of DNA damage via ATM/γ-H2AX; and (iv) decrease in DNA methyltransferase (DNMT) and global methylation. RNA interference study shows a predominant role of DNMT1 in MM cell survival versus DNMT3a or DNMT3b. The deubiquitylating enzyme USP7 stimulates DNMT1 activity, and conversely, USP7-siRNA reduced DNMT1 activity and decreased MM cell viability. RRx-001 plus USP7 inhibitor P5091 triggered synergistic anti-MM activity. MM xenograft studies show that RRx-001 is well tolerated, inhibits tumor growth and enhances survival. Combining RRx-001 with pomalidomide, bortezomib or SAHA induces synergistic anti-MM activity. Our results provide the rationale for translation of RRx-001, either alone or in combination, to clinical evaluation in MM.

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Acknowledgements

This investigation was supported by National Institutes of Health Specialized Programs of Research Excellence (SPORE) grant P50100707, PO1-CA078378, and RO1 CA050947. KCA is an American Cancer Society Clinical Research Professor.

Author contributions

DSD designed and performed the experiments, interpreted data and wrote the manuscript; AD helped in acquiring confocal images; AR and YS helped with animal experiments; ZT helped with viability assays, PR contributed clinical samples; BO and JS reviewed the manuscript; DC designed research, analyzed data and wrote the manuscript; KCA analyzed data and wrote 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

    D Sharma Das, A Ray, Y Song, Z Tian, P Richardson, D Chauhan & K C Anderson

  2. Program in Cellular and Molecular Medicine, Childrens Hospital, Boston, MA, USA

    A Das

  3. EpicentRx, Inc., Mountain View, CA, USA

    B Oronsky & J Scicinski

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  1. D Sharma Das
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Correspondence to D Chauhan or K C Anderson.

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Competing interests

BO and JS are employees of EpicentRx; KCA is on Advisory board of Celgene, Millenium, Gilead and Sanofi Aventis, and is a Scientific founder of Oncopep and acetylon; DC is consultant to EpicentRx Inc. The remaining authors have no conflict of interest.

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Das, D., Ray, A., Das, A. et al. A novel hypoxia-selective epigenetic agent RRx-001 triggers apoptosis and overcomes drug resistance in multiple myeloma cells. Leukemia 30, 2187–2197 (2016). https://doi.org/10.1038/leu.2016.96

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