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High-throughput drug screening identifies compounds and molecular strategies for targeting proteasome inhibitor-resistant multiple myeloma

Leukemia volume 28, pages 2263–2267 (2014)Cite this article

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Proteasome inhibitors (PI) are a cornerstone in the treatment of multiple myeloma (MM).1 Despite high initial response rates, nearly all MM patients relapse, marking the incurable nature of the disease and the emerging clinical challenge of combating PI resistance. This has created a need for drug discovery approaches that identify new drug cocktails of PIs, such as bortezomib/VELCADE (Btz; Millennium Pharmaceuticals, Inc., Cambridge, MA, USA), and other classes of FDA-approved or investigational new drugs. With these objectives in mind, we set out in this study to develop a high-throughput drug screening (HTS) platform to identify chemical structures that selectively kill or re-sensitize PI-resistant MM cells to PIs.

In this study, we have described and implemented a cell-based HTS platform for discovering the drugs and molecular mechanisms that specifically target treatment-resistant MM cells. The cell-based approach is advantageous for two reasons. First, given the multiple known and unknown mechanisms by which cells acquire resistance to PIs,8, 9, 10 a cell-based approach removes molecular bias. Second, the use of isogenic BzS and BzR cell models provides a relative assessment of cytotoxicity and the opportunity to identify compounds that selectively target PI-resistant populations. As proof of concept, we conducted a small-scale drug screen from which we identified VRC2, a compound with the ability to restore PI sensitivity to resistant MM cells. While VRC2 is not an ideal candidate for further development due to characteristics that predict unfavorable physiochemical properties (that is, furan and diathiazol toxicophores, a high number of hydrogen bond acceptors at 13, and predicted high clearance risk), in our study, it effectively served as a molecular probe to demonstrate the utility of coupling HTS with other approaches such as chemical genomics to pinpoint molecular mechanisms and therapies that are already approved for clinical use or are in human trials. Our investigation into the mechanism of VRC2 implicated MDM2 inhibition and p53 pathway activation as molecular strategies for targeting PI-resistant cells, findings that directed us toward the more clinically advanced MDM2 inhibitor, Nutlin3a. MDM2 inhibition has been shown by others to enhance the activity of Btz,11, 12, 13 which supports the ability of our method to identify bona fide druggable mechanisms for overcoming PI resistance. Our study further demonstrates that Nutlin3a not only augments the cytotoxic activity of Btz in Btz naive cells, but can also restore sensitivity to Btz once MM cells have acquired therapeutic resistance. It is also noteworthy that we found the combination of Nutlin3a/carfilzomib to be substantially more synergistic than Nutlin3a/Btz, findings that require further investigation but carry potentially significant translational implications. The mechanistic basis for this difference is not clear, however, it may be explained by the known differences in binding kinetics between the two PIs—inhibition of the 26S proteasome by Btz is reversible, whereas carfilzomib binds irreversibly. In summary, this proof-of-concept work validates the utility of our HTS approach for discovering lead compounds and uncovering molecular mechanisms for targeting PI-resistant MM, and provides a platform and the impetus for larger-scale screening efforts.

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Author notes

  1. N G Dolloff

    Present address: 7Current address: Department of Cell and Molecular Pharmacology & Experimental Therapeutics, Medical University of South Carolina, Charleston, SC, USA,

  2. H A F Stessman and A Lulla: These authors contributed equally to this work.

  3. B G Van Ness and N G Dolloff: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA

    H A F Stessman, A Mitra, T Harding, A Mansoor & B G Van Ness

  2. Department of Medicine, Division of Hematology and Oncology, Penn State Hershey Cancer Institute, Penn State College of Medicine, Hershey, PA, USA

    A Lulla & N G Dolloff

  3. Department of Electronic and Information Engineering, Huazhong University of Science and Technology, Wuhan, China

    T Xia

  4. Department of Computer Science and Engineering, University of Minnesota, Minneapolis, MN, USA

    C L Myers

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  1. H A F Stessman
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Corresponding authors

Correspondence to B G Van Ness or N G Dolloff.

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Competing interests

BVN receives research support from Millennium Pharmaceuticals, Inc., Cambridge, MA, and Onyx Pharmaceuticals, Inc., South San Francisco, CA, USA. The remaining authors declare no conflict of interest.

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Stessman, H., Lulla, A., Xia, T. et al. High-throughput drug screening identifies compounds and molecular strategies for targeting proteasome inhibitor-resistant multiple myeloma. Leukemia 28, 2263–2267 (2014). https://doi.org/10.1038/leu.2014.214

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