Abstract
Aberrant activity of enhancer of zeste homolog 2 (EZH2) is associated with a wide range of human cancers. The interaction of EZH2 with embryonic ectoderm development (EED) is required for EZH2's catalytic activity. Inhibition of the EZH2-EED complex thus represents a novel strategy for interfering with the oncogenic potentials of EZH2 by targeting both its catalytic and non-catalytic functions. To date, there have been no reported high-throughput screening (HTS) assays for inhibitors acting at the EZH2-EED interface. In this study, we developed a fluorescence polarization (FP)-based HTS system for the discovery of EZH2-EED interaction inhibitors. The tracer peptide sequences, positions of fluorescein labeling, and a variety of physicochemical conditions were optimized. The high Z' factors (>0.9) at a variety of DMSO concentrations suggested that this system is robust and suitable for HTS. The minimal sequence requirement for the EZH2-EED interaction was determined by using this system. A pilot screening of an in-house compound library containing 1600 FDA-approved drugs identified four compounds (apomorphine hydrochloride, oxyphenbutazone, nifedipine and ergonovine maleate) as potential EZH2-EED interaction inhibitors.
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Acknowledgements
The authors disclose the following financial support for this research and/or authorship of this article: the Ministry of Science and Technology of China (2015CB910304 to Hua-liang JIANG), the National Natural Science Foundation of China (21472208 and 81625022 to Cheng LUO, 21210003, 81230076 and 91313000 to Hua-liang JIANG), and the Fund of the State Key Laboratory of Toxicology and Medical Countermeasures, Academy of Military Medical Science (TMC201505 to Cheng LUO). We are extremely grateful to the National Centre for Protein Science Shanghai (Shanghai Science Research Center, Protein Expression and Purification System) for providing instrument support and technical assistance.
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List of compound information in the pilot screening. (DOC 1297 kb)
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Zhu, Mr., Du, Dh., Hu, Jc. et al. Development of a high-throughput fluorescence polarization assay for the discovery of EZH2-EED interaction inhibitors. Acta Pharmacol Sin 39, 302–310 (2018). https://doi.org/10.1038/aps.2017.59
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DOI: https://doi.org/10.1038/aps.2017.59
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