Abstract
Our prior study showed that inhibition of 19S proteasome-associated ubiquitin receptor Rpn13 can overcome bortezomib resistance in MM cells. Here, we performed proteomic analysis of Rpn13 inhibitor (RA190)-treated MM cells and identified an antioxidant enzyme superoxide dismutase (SOD1) as a mediator of Rpn13 signaling. SOD1 levels are higher in MM patient cells versus normal PBMCs; and importantly, SOD1 expression correlates with the progression of disease and shorter survival. Functional validation studies show that RA190-induced cytotoxicity in bortezomib-sensitive and -resistant MM cells is associated with decrease in SOD1 levels; conversely, forced expression of SOD1 inhibits RA190-induced cell death. Genetic knockdown and biochemical blockade of SOD1 with LCS-1 sensitizes bortezomib-resistant MM cells to bortezomib. SOD1 inhibitor LCS-1 decreases viability in MM cell lines and patient cells. LCS-1-induced cell death is associated with: (1) increase in superoxide and ROS levels; (2) activation of caspases, and p53/p21 signaling; (3) decrease in MCL-1, BCLxL, CDC2, cyclin-B1, and c-Myc; (4) ER stress response; and (5) inhibition of proteasome function. In animal model studies, LCS-1 inhibits xenografted bortezomib-resistant human MM cell growth and prolongs host survival. Our studies therefore show that targeting Rpn13 overcomes bortezomib resistance by decreasing cellular SOD1 levels, and provide the rationale for novel therapeutics targeting SOD1 to improve patient outcome in MM.
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Acknowledgements
The grant support for this investigation was provided by Dr Miriam and Sheldon Adelson Medical Research Foundation, as well as by the National Institutes of Health Specialized Programs of Research Excellence (SPORE) grant P50100707, R01CA207237, and RO1 CA050947. KCA is an American Cancer Society Clinical Research Professor. We are thankful to Krishan Chauhan (Undergraduate Summer Intern, WIT/DFCI) for the literature research and helpful discussion on free radical-mediated signaling cascades.
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DC conceptualized the project, designed research, analyzed data, and wrote the paper; TD performed all experiments, and analyzed the data; YS helped in animal model studies; AR helped in coculture assays; and KCA provided clinical samples and reviewed the paper.
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KCA is on Advisory Board of Millenium-Takeda, Gilead, Janssen, Bristol Myers Squibb, and Sanofi Aventis, and is a Scientific Founder of Oncopep and C4 Therapeutics. DC is consultant to Stemline Therapeutic, Inc., and Equity owner in C4 Therapeutics. The remaining authors declare no conflict of interest.
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Du, T., Song, Y., Ray, A. et al. Proteomic analysis identifies mechanism(s) of overcoming bortezomib resistance via targeting ubiquitin receptor Rpn13. Leukemia 35, 550–561 (2021). https://doi.org/10.1038/s41375-020-0865-2
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DOI: https://doi.org/10.1038/s41375-020-0865-2