Immunomodulatory drugs disrupt the cereblon–CD147–MCT1 axis to exert antitumor activity and teratogenicity

Abstract

Immunomodulatory drugs (IMiDs), such as thalidomide and its derivatives lenalidomide and pomalidomide, are key treatment modalities for hematologic malignancies, particularly multiple myeloma (MM) and del(5q) myelodysplastic syndrome (MDS). Cereblon (CRBN), a substrate receptor of the CRL4 ubiquitin ligase complex, is the primary target by which IMiDs mediate anticancer and teratogenic effects. Here we identify a ubiquitin-independent physiological chaperone-like function of CRBN that promotes maturation of the basigin (BSG; also known as CD147) and solute carrier family 16 member 1 (SLC16A1; also known as MCT1) proteins. This process allows for the formation and activation of the CD147–MCT1 transmembrane complex, which promotes various biological functions, including angiogenesis, proliferation, invasion and lactate export. We found that IMiDs outcompete CRBN for binding to CD147 and MCT1, leading to destabilization of the CD147–MCT1 complex. Accordingly, IMiD-sensitive MM cells lose CD147 and MCT1 expression after being exposed to IMiDs, whereas IMiD-resistant cells retain their expression. Furthermore, del(5q) MDS cells have elevated CD147 expression, which is attenuated after IMiD treatment. Finally, we show that BSG (CD147) knockdown phenocopies the teratogenic effects of thalidomide exposure in zebrafish. These findings provide a common mechanistic framework to explain both the teratogenic and pleiotropic antitumor effects of IMiDs.

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Figure 1: IMiDs compete with CRBN to destabilize CD147 and MCT1 in a ubiquitin-independent manner.
Figure 2: CRBN promotes maturation, complex assembly and membrane localization of CD147 and MCT1.
Figure 3: Lenalidomide has anti-myeloma activity via CD147 and MCT1 destabilization.
Figure 4: CD147 and MCT1 mediate tumor growth and determine the response of MM cells to IMiDs in vivo.
Figure 5: CD147 and MCT1 destabilization mediates IMiD activity in del(5q) MDS.
Figure 6: Thalidomide destabilizes CD147 to cause teratogenicity.

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Acknowledgements

We thank M. Buschbeck (Institute of Predictive and Personalized Medicine of Cancer, Barcelona) for the SKK-1 and SKM-1 cell lines, X. Bustelo (Centro de Investigacion del Cancer, Salamanca) for a EGFP–CD147 expression plasmid, Y. Cang (Life Science Institute, Zheijang University) for a CRBN expression construct, B. Ebert (Dana-Farber Cancer Institute) for an IKZF3 expression construct, A. Halestrap (School of Biochemistry, University of Bristol) for MCT1 expression constructs, H. Handa (Tokyo Institute of Technology, Japan) for an aliquot of mouse monoclonal CRBN-specific antibody, K. Kadomatsu (Department of Molecular Biology, Nagoya University) for CD147 expression constructs, W. Kaelin (Dana-Farber Cancer Institute) for CRISPR-based HEK293FT CBRN−/− cells and two clones of MM1.S CBRN−/− cells (T11, T21), D. Sabatini (Whitehead Institute) for a MCT1 and an shMCT1 construct, K. Stewart (Mayo Clinic) for a shCRBN plasmid, N. Thomä (Friedrich Miescher Institute for Biomedical Research, Basel) for a MEIS2 expression construct and purified CRL4CRBN protein, K. Tohyama (Kawasaki Medical School, Okayama) for the del(5q) MDS cell line MDSL, N. Thomä and E. Fisher for suggestions, R. Rojas for fish care and B. Nuscher for help with gel filtration chromatography. This work was supported by the Helmholtz cross-program topic 'Metabolic Dysfunction' (B.S.), a fellowship of the TU Munich (KKF # B10-13; to R.E.), grants from the José Carreras Leukemia Foundation (DJCLS R 14/16; to K.S.G.), the German Research Foundation (FOR2033 GO 713/2-1 and SFB 1243 (K.S.G.), SFB 824 (U.K.), KE 222/7-1 (U.K.), BA 2851/4-1 (F.B.) and SFB 1243 (F.B.)), the German Cancer Aid (#111051 (M.H.), #111305 (U.K.) and #111430 (F.B.)) and the Wilhelm Sander Stiftung (#2012.096.1; to F.B.). An application for a patent has been filed at the European Patent Office.

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R.E., M.H., V.F.-S. and F.B. conceived and designed the research; R.E., M.H. and V.F.-S. performed most of the experiments with crucial help from B.-S.T. and A.-M.K.; U.P., U.G. and K.S.G. provided MDS samples; C.L., S.K. and H.E. provided MM samples; K.S.G. and A.-K.G. performed FACS analyses; J.S. helped with the MM xenotransplant model; L.J. helped with the PET analysis; M.R. performed immunohistochemical analyses of tumors; F.v.B., B.S. and C.H. performed zebrafish experiments; S.L. and B.K. performed mass spectrometry; R.E., M.H., V.F.-S., F.v.B., A.-K.G., B.-S.T., S.L., U.K., C.P., B.S., C.H., K.S.G., B.K. and F.B. analyzed results; and F.B. coordinated this work and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Florian Bassermann.

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F.B. and K.S.G. received honoraria and research funding from Celgene.

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Supplementary Figures 1–11 and Supplementary Table 2 (PDF 5435 kb)

Supplementary Table 1

Strep-Flag-Tandem affinity purified-CRBN-associated proteins identified by mass spectrometry analysis. (XLSX 24 kb)

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Eichner, R., Heider, M., Fernández-Sáiz, V. et al. Immunomodulatory drugs disrupt the cereblon–CD147–MCT1 axis to exert antitumor activity and teratogenicity. Nat Med 22, 735–743 (2016). https://doi.org/10.1038/nm.4128

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