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EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations

Abstract

In eukaryotes, post-translational modification of histones is critical for regulation of chromatin structure and gene expression. EZH2 is the catalytic subunit of the polycomb repressive complex 2 (PRC2) and is involved in repressing gene expression through methylation of histone H3 on lysine 27 (H3K27). EZH2 overexpression is implicated in tumorigenesis and correlates with poor prognosis in several tumour types1,2,3,4,5. Additionally, somatic heterozygous mutations of Y641 and A677 residues within the catalytic SET domain of EZH2 occur in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma6,7,8,9,10. The Y641 residue is the most frequently mutated residue, with up to 22% of germinal centre B-cell DLBCL and follicular lymphoma harbouring mutations at this site. These lymphomas have increased H3K27 tri-methylation (H3K27me3) owing to altered substrate preferences of the mutant enzymes9,11,12,13. However, it is unknown whether specific, direct inhibition of EZH2 methyltransferase activity will be effective in treating EZH2 mutant lymphomas. Here we demonstrate that GSK126, a potent, highly selective, S-adenosyl-methionine-competitive, small-molecule inhibitor of EZH2 methyltransferase activity, decreases global H3K27me3 levels and reactivates silenced PRC2 target genes. GSK126 effectively inhibits the proliferation of EZH2 mutant DLBCL cell lines and markedly inhibits the growth of EZH2 mutant DLBCL xenografts in mice. Together, these data demonstrate that pharmacological inhibition of EZH2 activity may provide a promising treatment for EZH2 mutant lymphoma.

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Figure 1: Biochemical and cellular mechanistic activity of GSK126.
Figure 2: GSK126 inhibits the proliferation of several EZH2 mutant lymphoma cell lines.
Figure 3: GSK126 induces transcriptional activation in sensitive cell lines.
Figure 4: In vivo inhibition of H3K27me3 and tumour growth response with GSK126.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

The gene expression data are accessible on GEO through accession number GSE40972 and the ChIP-seq data through accession number GSE40970.

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Acknowledgements

We acknowledge members of GlaxoSmithKline’s Platform Technology and Sciences group for reagent generation and sequencing, Ocimum Biosolutions for bioinformatic support, A. Anderson for statistical analysis, P. Hoffman for assistance with the manuscript, and all members of the Cancer Epigenetics Discovery Performance Unit for their guidance and support.

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Authors

Contributions

M.T.M., G.G., R.G.K., C.F.M., M.B., S.K.V. and C.L.C. designed studies; M.T.M., H.M.O., S.K., C.T., G.S.V.A., E.D., Y.L., A.P.G., A.D.P., L.V.L., M.M., C.D., X.T. and C.F.M. performed research; M.T.M., G.G., R.G.K., A.P.G., C.F.M., S.K.V., W.H.M., D.D., P.J.T. and C.L.C. analysed data and M.T.M., G.G., R.G.K., A.P.G. and C.L.C. wrote the paper.

Corresponding author

Correspondence to Caretha L. Creasy.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-18, Supplementary Methods, Supplementary Tables 1-5 and 8-9 – see separate files for Supplementary Tables 6 and 7. (PDF 1990 kb)

Supplementary Table 6

This file contains lists of significantly differentially exposed probe sets. (XLSX 436 kb)

Supplementary Table 7

This file contains H3K27me3 Chip-seq enriched regions. (XLSX 15514 kb)

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McCabe, M., Ott, H., Ganji, G. et al. EZH2 inhibition as a therapeutic strategy for lymphoma with EZH2-activating mutations. Nature 492, 108–112 (2012). https://doi.org/10.1038/nature11606

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