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EZH2 PROTACs target EZH2- and FOXM1-associated oncogenic nodes, suppressing breast cancer cell growth

Oncogene volume 43, pages 2722–2736 (2024)Cite this article

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Abstract

Breast cancer (BC) remains the second leading cause of cancer-related mortalities in women. Resistance to hormone therapies such as tamoxifen, an estrogen receptor (ER) inhibitor, is a major hurdle in the treatment of BC. Enhancer of zeste homolog 2 (EZH2), the methyltransferase component of the Polycomb repressive complex 2 (PRC2), has been implicated in tamoxifen resistance. Evidence suggests that EZH2 often functions noncanonically, in a methyltransferase-independent manner, as a transcription coactivator through interacting with oncogenic transcription factors. Unlike methyltransferase inhibitors, proteolysis targeting chimeras (PROTAC) can suppress both activating and repressive functions of EZH2. Here, we find that EZH2 PROTACs, MS177 and MS8815, effectively inhibited the growth of BC cells, including those with acquired tamoxifen resistance, to a much greater degree when compared to methyltransferase inhibitors. Mechanistically, EZH2 associates with forkhead box M1 (FOXM1) and binds to the promoters of FOXM1 target genes. EZH2 PROTACs induce degradation of both EZH2 and FOXM1, leading to reduced expression of target genes involved in cell cycle progression and tamoxifen resistance. Together, this study supports that EZH2-targeted PROTACs represent a promising avenue of research for the future treatment of BC, including in the setting of tamoxifen resistance.

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Fig. 1: MS177 and MS8815 inhibit luminal breast cancer (BC) cell growth.
Fig. 2: MS177 reduces the expression of cell cycle regulated and tamoxifen resistance associated genes.
Fig. 3: MS177 and MS8815 inhibit growth and reduce cell cycle regulated gene sets in tamoxifen resistant breast cancer cells.
Fig. 4: EZH2 solo binding sites are associated with FOXM1 target gene promoters.
Fig. 5: EZH2 physically interacts with FOXM1 in ER+ breast cancer cells and EZH2 targeted PROTACs reduce FOXM1 protein and target gene expression compared to methyltransferase inhibitors and negative controls.
Fig. 6: EZH2 mRNA expression correlates with FOXM1, FOXM1 target genes, cell cycle regulated gene sets, and genes associated with tamoxifen resistance in luminal A BC clinical samples from the METABRIC study.

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Data availability

Raw and processed sequencing files have been submitted to the Gene Expression Omnibus (GEO) database under the accession numbers of GSE270163 and GSE270769.

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Acknowledgements

We thank all members of Cai, Jin and Wang laboratories for helpful discussions during the development of this manuscript. We thank Dr. Bryan Pan for bioinformatics support, Dr. Philip Spanheimer for sharing reagents, and John Albright for technical contributions. We thank core facilities of University of North Carolina and Duke University for their professional support in tissue culture, high-throughput sequencing and bioinformatics and data storage.

Funding

This work was supported in part through the UNC Lineberger Comprehensive Cancer Center Core Support Grant P30-CA016086. This work was also supported in part by R01CA262903 (to LC), R01CA218600 (to JJ and GGW), R01CA268519 (to GGW and JJ), R01CA211336 (to GGW) and R01CA230854 (to JJ) grants from the US National Institutes of Health (NIH).

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Authors and Affiliations

  1. Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, NC, 27710, USA

    Joshua Corbin & Gang Greg Wang

  2. Duke Cancer Institute, Duke University School of Medicine, Durham, NC, 27710, USA

    Joshua Corbin, Ling Cai & Gang Greg Wang

  3. Departments of Pharmacological Sciences and Oncological Sciences, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Xufen Yu & Jian Jin

  4. Mount Sinai Center for Therapeutics Discovery, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Xufen Yu & Jian Jin

  5. Department of Pathology, Duke University School of Medicine, Durham, NC, 27710, USA

    Ling Cai & Gang Greg Wang

  6. Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC, 27599, USA

    Ling Cai & Gang Greg Wang

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  1. Joshua Corbin
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  2. Xufen Yu
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  3. Jian Jin
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Contributions

JJ, LC and GGW designed the experiments and secured fundings. JC and XY led the biological and medicinal chemistry aspects of the study, respectively, under the supervision of GGW, LC and JJ. JC led on the manuscript writing and all authors conducted data analysis and interpretation and participated in the preparation of manuscript.

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Correspondence to Ling Cai or Gang Greg Wang.

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Competing interests

JJ is listed as a co-inventor on the patent application (WO 2018/081530 A1) filed by Icahn School of Medicine at Mount Sinai covering EZH2 degraders. JJ is a cofounder and equity shareholder in Cullgen, Inc. and a consultant for Cullgen, Inc., EpiCypher, Inc., and Accent Therapeutics, Inc. The Jin laboratory received research funds from Celgene Corporation, Levo Therapeutics, Inc., Cullgen, Inc., and Cullinan Oncology, Inc.

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Corbin, J., Yu, X., Jin, J. et al. EZH2 PROTACs target EZH2- and FOXM1-associated oncogenic nodes, suppressing breast cancer cell growth. Oncogene 43, 2722–2736 (2024). https://doi.org/10.1038/s41388-024-03119-9

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