Immunomodulatory drugs (IMiDs) including lenalidomide and pomalidomide bind cereblon (CRBN) and activate the CRL4CRBN ubiquitin ligase to trigger proteasomal degradation of the essential transcription factors IKZF1 and IKZF3 and multiple myeloma (MM) cytotoxicity. We have shown that CRBN is also targeted for degradation by SCFFbxo7 ubiquitin ligase. In the current study, we explored the mechanisms underlying sensitivity of MM cells to IMiDs using genome-wide CRISPR-Cas9 screening. We validate that CSN9 signalosome complex, a deactivator of Cullin-RING ubiquitin ligase, inhibits SCFFbxo7 E3 ligase-mediated CRBN degradation, thereby conferring sensitivity to IMiDs; conversely, loss of function of CSN9 signalosome activates SCFFbxo7 complex, thereby enhancing degradation of CRBN and conferring IMiD resistance. Finally, we show that pretreatment with either proteasome inhibitors or NEDD8 activating enzyme (NAE) inhibitors can abrogate degradation and maintain levels of CRBN, thereby enhancing sensitivity to IMiDs. These studies therefore demonstrate that CSN9 signalosome complex regulates sensitivity to IMiDs by modulating CRBN expression.
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We thank X.H. Feng and Z.P. Xia for expression vectors. We thank the staff from the hematologic neoplasia core facilities at Dana-Farber Cancer Institute for technical assistance. We also thank the Genome Center of WuXi AppTec Inc. for the initial data analysis of the CRISPR screening.
This study was supported by the National Institute of Health Grant; SPORE-P50100707 (KCA), P01-CA078378 (K.C.A.), R01-CA050947 (K.C.A), and R01-CA178264 (T.H. and K.C.A.). K.C.A. is an American Cancer Society Clinical Research Professor. This study was also supported in part by funds from the National 973 Plan for Basic Research (2015CB553803), National Natural Science Foundation of China (31671334), Fundamental Research Funds for the Central Universities, and Key Construction Program of the National ‘985’ Project.
J.L., T.S., and W.Z. designed and performed the research, analyzed the data, and wrote the manuscript; L.X. and M.H. performed some experiment; S.W. performed gene expression correlation analysis; Z.P. designed research and analyzed data; Y.T. provided biological material and analyzed data; T.H. designed the research, analyzed the data, wrote the manuscript, and supervised the project; K.C.A and Y.C. conceived the project, designed the research, analyzed the data, wrote the manuscript, and supervised the project.
Conflict of interest
K.C.A. serves on advisory boards to Celgene and Millennium. All other authors declare no competing financial interests.
About this article
The future of myeloma precision medicine: integrating the compendium of known drug resistance mechanisms with emerging tumor profiling technologies
Journal of Experimental & Clinical Cancer Research (2019)
Pharmacology & Therapeutics (2019)