EZH2 inhibition as a targeted therapy for H3K27M mutant pediatric gliomas [ChIP-seq]

H3K27M has been shown to inhibit Polycomb Repressive Complex 2 (PRC2) activity by binding to its catalytic subunit EZH2, and although DIPGs with H3K27M mutation show global loss of H3 with trimethylated lysine 27 (H3K27me3), several genes retain H3K27me3. Here, we describe a new mouse DIPG model in which H3K27M potentiates tumourigenesis. Using this model and primary patient-derived DIPG cell lines, we show that H3K27M expressing tumours require PRC2 for proliferation. Furthermore, we demonstrate that small molecule EZH2 inhibitors abolish tumour cell growth through a mechanism involving induction of the tumour suppressor protein p16INK4A. In agreement with this, we show that tumour cells with non-functional p16INK4A do not respond to EZH2 inhibitors. Genome-wide enrichment analyses show that several genes retain H3K27me3 even in the presence of H3K27M. Several of these genes are associated with high levels of H3K27me3 in the absence of H3K27M, whereas regions with low H3K27me3 levels tend to lose H3K27me3 upon H3K27M expression. Furthermore, we find significant overlap between genes that retain H3K27me3 in mouse and human primary DIPGs expressing H3K27M. Taken together, these results show that residual PRC2 activity is required for the proliferation of H3K27M positive DIPGs, and we propose a therapeutic strategy for these tumours using EZH2 inhibitors and expression of p16INK4A as a marker for stratifying potential responding tumours.