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Targeted disruption of the EZH2–EED complex inhibits EZH2-dependent cancer

Nature Chemical Biology volume 9, pages 643–650 (2013)Cite this article

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Abstract

Enhancer of zeste homolog 2 (EZH2) is the histone lysine N-methyltransferase component of the Polycomb repressive complex 2 (PRC2), which, in conjunction with embryonic ectoderm development (EED) and suppressor of zeste 12 homolog, regulates cell lineage determination and homeostasis. Enzymatic hyperactivity has been linked to aberrant repression of tumor suppressor genes in diverse cancers. Here, we report the development of stabilized α-helix of EZH2 (SAH-EZH2) peptides that selectively inhibit H3 Lys27 trimethylation by dose-responsively disrupting the EZH2–EED complex and reducing EZH2 protein levels, a mechanism distinct from that reported for small-molecule EZH2 inhibitors targeting the enzyme catalytic domain. MLL-AF9 leukemia cells, which are dependent on PRC2, undergo growth arrest and monocyte-macrophage differentiation upon treatment with SAH-EZH2, consistent with observed changes in expression of PRC2-regulated, lineage-specific marker genes. Thus, by dissociating the EZH2–EED complex, we pharmacologically modulate an epigenetic 'writer' and suppress PRC2-dependent cancer cell growth.

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Figure 1: Synthesis, EED-binding activity and cellular penetration of SAH-EZH2 peptides.
Figure 2: Sequence-dependent dissociation of EED–EZH1 and EED–EZH2 complexes and impairment of PRC2 activity by SAH-EZH2.
Figure 3: SAH-EZH2 induces cell cycle arrest and inhibits proliferation of MLL-AF9 leukemia cells.
Figure 4: SAH-EZH2 induces monocyte-macrophage differentiation of MLL-AF9 leukemia cells.
Figure 5: Cell viability effects of SAH-EZH2 and GSK126.

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Acknowledgements

We thank E. Smith for graphics assistance. This work was supported by US National Institutes of Health grant U01CA105423 to S.H.O. and 5R01GM090299 and a Leukemia and Lymphoma Society Specialized Center of Research project grant to L.D.W. S.H.O. is an Investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Division of Pediatric Hematology/Oncology, Boston Children's Hospital, Boston, Massachusetts, USA

    Woojin Kim, Guoji Guo, Marc A Kerenyi, Loren D Walensky & Stuart H Orkin

  2. Harvard Medical School, Boston, Massachusetts, USA

    Woojin Kim, Gregory H Bird, Guoji Guo, Marc A Kerenyi, Loren D Walensky & Stuart H Orkin

  3. Pediatric Oncology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Woojin Kim, Gregory H Bird, Guoji Guo, Marc A Kerenyi, Loren D Walensky & Stuart H Orkin

  4. Linde Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Gregory H Bird & Loren D Walensky

  5. Pediatric Hematology, Oncology and Bone Marrow Transplantation, University of Colorado, Aurora, Colorado, USA

    Tobias Neff

  6. Howard Hughes Medical Institute, Boston, Massachusetts, USA

    Stuart H Orkin

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Contributions

W.K., L.D.W. and S.H.O. designed the experiments and wrote the manuscript. W.K. and G.H.B. synthesized the stapled peptides. T.N. generated MLL-AF9 cells and assisted W.K. with the colony-forming assay and morphological studies. G.G. performed the high-throughput microfluidic qPCR. M.A.K. assisted W.K. with the studies on HPC5, M1 and C1498 cell lines.

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Correspondence to Loren D Walensky or Stuart H Orkin.

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Competing interests

L.D.W. is a scientific advisory board member and consultant for Aileron Therapeutics.

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Kim, W., Bird, G., Neff, T. et al. Targeted disruption of the EZH2–EED complex inhibits EZH2-dependent cancer. Nat Chem Biol 9, 643–650 (2013). https://doi.org/10.1038/nchembio.1331

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