SALL4 mediates teratogenicity as a thalidomide-dependent cereblon substrate

Abstract

Targeted protein degradation via small-molecule modulation of cereblon offers vast potential for the development of new therapeutics. Cereblon-binding therapeutics carry the safety risks of thalidomide, which caused an epidemic of severe birth defects characterized by forelimb shortening or phocomelia. Here we show that thalidomide is not teratogenic in transgenic mice expressing human cereblon, indicating that binding to cereblon is not sufficient to cause birth defects. Instead, we identify SALL4 as a thalidomide-dependent cereblon neosubstrate. Human mutations in SALL4 cause Duane-radial ray, IVIC, and acro-renal-ocular syndromes with overlapping clinical presentations to thalidomide embryopathy, including phocomelia. SALL4 is degraded in rabbits but not in resistant organisms such as mice because of SALL4 sequence variations. This work expands the scope of cereblon neosubstrate activity within the formerly ‘undruggable’ C2H2 zinc finger family and offers a path toward safer therapeutics through an improved understanding of the molecular basis of thalidomide-induced teratogenicity.

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Fig. 1: Human and rabbit SALL4 zinc fingers are direct, thalidomide-dependent substrates of cereblon-CRL4 in vitro, whereas mouse SALL4 is not.
Fig. 2: SALL4 is degraded in cell lines in a cereblon- and proteasome-dependent fashion upon treatment with thalidomide.
Fig. 3: SALL4 and ZFP91 examined by immunohistochemistry in rabbit, wild-type mouse, and humanized cereblon mouse testes.
Fig. 4: SALL4 localizes to the limb bud during development in rabbit embryos, and decreases upon treatment with thalidomide.
Fig. 5: Model for thalidomide-induced SALL4 ubiquitination by cereblon-CRL4 and subsequent cellular degradation by the 26 S proteasome.

Data availability

All data generated or analyzed during this study are included in this published article (and its supplementary information files).

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Acknowledgements

Many thanks to K. Kolaja, N. Collins and the teratogenicity working group for discussions relating to this work.

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M.E.M., A.C., T.C., W.F., P.A., and M. Riley. performed biochemical studies. X.Z., G.L., and C.-C.L. performed cellular experiments. S.C., C.D., Y.R., M.W., and C.-W.L. performed IHC studies. J. Hui., K.S., and K.B. planned in vivo experiments. M.E.M., S.C., G.L., J. Hui, J. Hartke, G.K., M. Rolfe., R.V., L.G.H., and P.P.C. planned studies, and all authors analyzed the data and prepared the manuscript.

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Correspondence to Philip P. Chamberlain.

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Matyskiela, M.E., Couto, S., Zheng, X. et al. SALL4 mediates teratogenicity as a thalidomide-dependent cereblon substrate. Nat Chem Biol 14, 981–987 (2018). https://doi.org/10.1038/s41589-018-0129-x

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