Diffuse intrinsic pontine glioma (DIPG) is an aggressive brain tumor that is located in the pons and primarily affects children. Nearly 80% of DIPGs harbor mutations in histone H3 genes, wherein lysine 27 is substituted with methionine (H3K27M). H3K27M has been shown to inhibit polycomb repressive complex 2 (PRC2), a multiprotein complex responsible for the methylation of H3 at lysine 27 (H3K27me), by binding to its catalytic subunit EZH2. Although DIPGs with the H3K27M mutation show global loss of H3K27me3, several genes retain H3K27me3. Here we describe a mouse model of DIPG in which H3K27M potentiates tumorigenesis. Using this model and primary patient-derived DIPG cell lines, we show that H3K27M-expressing tumors require PRC2 for proliferation. Furthermore, we demonstrate that small-molecule EZH2 inhibitors abolish tumor cell growth through a mechanism that is dependent on the induction of the tumor-suppressor protein p16INK4A. Genome-wide enrichment analyses show that the genes that retain H3K27me3 in H3K27M cells are strong polycomb targets. Furthermore, we find a highly significant overlap between genes that retain H3K27me3 in the DIPG mouse model and in human primary DIPGs expressing H3K27M. Taken together, these results show that residual PRC2 activity is required for the proliferation of H3K27M-expressing DIPGs, and that inhibition of EZH2 is a potential therapeutic strategy for the treatment of these tumors.
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We thank members of the Helin laboratory for discussions. F.M. was supported by a postdoctoral fellowship from EMBO (874-2011). D.P. was supported by a postdoctoral fellowship from EMBO (1411-2011) and the Danish Medical Research Council. I.C. was supported by a fellowship from the Lundbeck Foundation. The work in the Helin laboratory was supported by the Danish Medical Research Council (DFF – 4004-00081), the Danish National Research Foundation (DNRF 82), and through a center grant from the Novo Nordisk Foundation (The Novo Nordisk Foundation Section for Stem Cell Biology in Human Disease). A.M.C. acknowledges support from the Xarxa de Bancs de Tumors de Catalunya (XBTC) sponsored by Pla Director d'Oncologia de Catalunya, and funding from the Fondo Alicia Pueyo, AECC Scientific Foundation, European Union Seventh Framework Programme (FP7/2007-2013) under Marie Curie International Reintegration Grant (PIRG-08-GA-2010-276998) and ISCIII-FEDER (CP13/00189). We thank S. Pollard for the gift of GNS cell lines, N. Gupta for providing the SF7761 and SF8628 cell lines and L. Uhrbom and P. Lewis for plasmids.
The authors declare no competing financial interests.
Supplementary Methods, Supplementary Figures 1–13 and Supplementary Tables 1 and 4 (PDF 11060 kb)
List of genes corresponding to the four categories and gene expression changes in H3K27M cells (XLSX 294 kb)
GO term enrichment analysis on "unchanged" genes using annotation clustering tool available on the DAVID bioinformatics resources (XLSX 73 kb)
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Mohammad, F., Weissmann, S., Leblanc, B. et al. EZH2 is a potential therapeutic target for H3K27M-mutant pediatric gliomas. Nat Med 23, 483–492 (2017). https://doi.org/10.1038/nm.4293
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