Polycomb repressive complex 2 (PRC2) is a regulator of epigenetic states required for development and homeostasis. PRC2 trimethylates histone H3 at lysine 27 (H3K27me3), which leads to gene silencing, and is dysregulated in many cancers. The embryonic ectoderm development (EED) protein is an essential subunit of PRC2 that has both a scaffolding function and an H3K27me3-binding function. Here we report the identification of A-395, a potent antagonist of the H3K27me3 binding functions of EED. Structural studies demonstrate that A-395 binds to EED in the H3K27me3-binding pocket, thereby preventing allosteric activation of the catalytic activity of PRC2. Phenotypic effects observed in vitro and in vivo are similar to those of known PRC2 enzymatic inhibitors; however, A-395 retains potent activity against cell lines resistant to the catalytic inhibitors. A-395 represents a first-in-class antagonist of PRC2 protein–protein interactions (PPI) for use as a chemical probe to investigate the roles of EED-containing protein complexes.
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The authors would like to thank P. Richardson, Y. Wang, H. Zhao, R. Clark, and Z. Ji for chemistry support. Thanks to S. Duan, S. Kakavas and R. Edalji for biochemistry and biophysics support. We would also like to thank K. Bromberg and L. Lasko for designing and helping interpret the high-content analysis assay and B. Ainsworth for help with data analysis and graphing. Finally, we would like to thank E. Nicholl, T. Shah, J. Trumbull, X. Cao, and Q. Lang for valuable assistance in TSA method development and high-throughput screening. E.L-F. is the recipient of a Canadian Institutes of Health Research Banting Postdoctoral Fellowship. Use of the IMCA-CAT beamline 17-ID at the Advanced Photon Source was supported by the companies of the Industrial Macromolecular Crystallography Association through a contract with Hauptman–Woodward Medical Research Institute. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, the Canada Foundation for Innovation, the Eshelman Institute for Innovation, Genome Canada through the Ontario Genomics Institute, Innovative Medicines Initiative (EU/EFPIA) (ULTRA-DD grant no. 115766), Janssen, Merck & Co., Novartis Pharma AG, the Ontario Ministry of Economic Development and Innovation, Pfizer, the São Paulo Research Foundation-FAPESP, Takeda, and the Wellcome Trust.
This study was sponsored by AbbVie. AbbVie contributed to the study design, research, interpretation of data, writing, reviewing, and approval of the publication. Y.H., S.S., M.L.C., C.G.J., H.Z., K.M.C., B.S., D.C., K.L.K., H.-Q.L., M.P., M.A.A., D.M., J.G., J.D., S.C.P., A.M.P., R.F.S., M.T., L.J.B., D.J.O., D.J.L., W.G., S.G., F.G.B., G.G.C., C.S., and W.N.P. were employees of AbbVie at the time of the study.
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He, Y., Selvaraju, S., Curtin, M. et al. The EED protein–protein interaction inhibitor A-395 inactivates the PRC2 complex. Nat Chem Biol 13, 389–395 (2017). https://doi.org/10.1038/nchembio.2306
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