Lenalidomide is a highly effective treatment for myelodysplastic syndrome (MDS) with deletion of chromosome 5q (del(5q)). Here, we demonstrate that lenalidomide induces the ubiquitination of casein kinase 1A1 (CK1α) by the E3 ubiquitin ligase CUL4–RBX1–DDB1–CRBN (known as CRL4CRBN), resulting in CK1α degradation. CK1α is encoded by a gene within the common deleted region for del(5q) MDS and haploinsufficient expression sensitizes cells to lenalidomide therapy, providing a mechanistic basis for the therapeutic window of lenalidomide in del(5q) MDS. We found that mouse cells are resistant to lenalidomide but that changing a single amino acid in mouse Crbn to the corresponding human residue enables lenalidomide-dependent degradation of CK1α. We further demonstrate that minor side chain modifications in thalidomide and a novel analogue, CC-122, can modulate the spectrum of substrates targeted by CRL4CRBN. These findings have implications for the clinical activity of lenalidomide and related compounds, and demonstrate the therapeutic potential of novel modulators of E3 ubiquitin ligases.

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We thank D. Heckl for cloning of the EGI vector and technical advice, R. Mathieu and M. Paktinat for help with FACS sorting, M. Chen for help with colony management and animal care, S. Köpff for technical support, and C. Fontanillo for proteomic analysis computational support. Patient samples were provided by the Stem Cell and Xenograft Core of the University of Pennsylvania. This work was supported by the NIH (R01HL082945 and P01CA108631), the Edward P. Evans Foundation, the Gabrielle’s Angel Foundation, and a Leukemia and Lymphoma Society Scholar Award to B.L.E.; J.K. was supported by the German Research Foundation (DFG, Emmy Noether Fellowship Kr3886/2-1, Kr3886/1-1, and SFB1074) and the Else-Kröner Fresenius Foundation. E.C.F. was supported by award T32GM007753 from the National Institute of General Medical Sciences.

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Author notes

    • Jan Krönke
    •  & Emma C. Fink

    These authors contributed equally to this work.


  1. Brigham and Women’s Hospital, Division of Hematology, Boston, Massachusetts 02115, USA

    • Jan Krönke
    • , Emma C. Fink
    • , Slater N. Hurst
    • , Rebekka K. Schneider
    • , Marie McConkey
    • , Marcus Järås
    •  & Benjamin L. Ebert
  2. University Hospital of Ulm, Department of Internal Medicine III, 89081 Ulm, Germany

    • Jan Krönke
    •  & Lars Bullinger
  3. Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA

    • Jan Krönke
    • , Emma C. Fink
    • , Namrata D. Udeshi
    • , D. R. Mani
    • , Tanya Svinkina
    • , Steven A. Carr
    •  & Benjamin L. Ebert
  4. Celgene Corporation, San Diego, California 92121, USA

    • Paul W. Hollenbach
    • , Kyle J. MacBeth
    • , Philip P. Chamberlain
    • , Hon Wah Man
    • , Anita K. Gandhi
    • , Brian E. Cathers
    •  & Rajesh Chopra
  5. Roswell Park Cancer Institute, Buffalo, New York 14263, USA

    • Elizabeth Griffiths
    •  & Meir Wetzler


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J.K., E.C.F., and B.L.E. initiated the project. J.K., E.C.F., P.W.H., K.J.M., S.N.H, M.McC., A.K.G., M.J., and R.K.S. designed and performed cell experiments and protein analysis. E.C.F. designed and performed mouse competition and patient sample experiments. N.D.U., D.R.M., and T.S. performed KG-1 proteomics and analysis. E.G. and M.W. provided patient samples. P.P.C. performed the structural analysis. H.W.M. synthesized CC-122. K.J.M. provided MDS-L proteomics. B.E.C., R.C., L.B., S.A.C., and B.L.E. supervised the work. J.K., E.C.F., and B.L.E. wrote the manuscript. All authors assisted in editing the manuscript.

Competing interests

P.W.H., K.M., P.P.C., H.W.M., A.K.G., B.E.C., and R.C. are employed by Celgene Corporation. B.L.E. has consulted for Celgene. J.K. received honoraria from Celgene. All other authors have no competing interests to declare.

Corresponding author

Correspondence to Benjamin L. Ebert.

The original mass spectra may be downloaded from MassIVE (http://massive.ucsd.edu) using the identifier: MSV000079014. The data are accessible at ftp://massive.ucsd.edu/MSV000079014.

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    Supplementary information

    This file contains Supplementary Methods (Synthesis and characterization of CC-1220); a Supplementary Note (Pharmacokinetics of lenalidomide in humansand) and Supplementary Figure 1, which shows the full uncropped scans of all Western Blots with molecular weight markers.

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  1. 1.

    Supplementary Table 1

    This file contains the full data table for KG-1 proteomics studies. Proteome tab lists the effects of 1 μM and 10 μM lenalidomide on protein levels. KGG tab lists the results of ubiquitin profiling.

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