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Diffuse Intrinsic Pontine Glioma (DIPG)

Oncohistones, mutated histones with oncogenic features, affect the global chromatin landscape and drive tumorigenesis due to altered gene expression. 78% of Diffuse Intrinsic Pontine Gliomas (DIPGs) contain a histone H3 oncohistone where a missense mutation substitutes Lys 27 with methionine, affecting its ability to be methylated and repress gene expression.

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H3K27M

H3K27M serves as an oncohistone, where mutations contribute to tumor development because Ezh2 is no longer able to methylate the histone and gene expression is aberrantly upregulated.

DIPG Figure 1

Confocal immunofluorescent analysis of mouse astrocytes, containing either a knock-in FLAG-tagged K27M mutant histone H3.3 gene (left, positive) or a knock-in FLAG-tagged wild-type histone H3.3 gene (center, negative), and HeLa cells (right, negative), using Histone H3 (K27M Mutant Specific) (D3B5T) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red).

Ezh2

Changes in methylation of H3K27 are mostly due to mislocalization of Ezh2 on the genome and can be confirmed by ChIP-seq analysis.

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Ezh2 (D2C9) XP® Rabbit mAb #5246 – WB, IP, IF, F, IHC, ChIP, ChIP-seq
DIPG Figure 2

Chromatin immunoprecipitations were performed with cross-linked chromatin from Hela cells and either Ezh2 (D2C9) XP® Rabbit mAb or Tri-Methyl-Histone H3 (Lys27) (C36B11) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5ng enriched ChIP DNA for EZH2 ChIP-seq and 50ng enriched ChIP DNA for H3K27me3 ChIP-seq using SimpleChIP® ChIP-Seq DNA Library Prep Kit for Illumina® #56795, and sequenced on the Illumina NextSeq. EZH2 and H3K27me3 are known to associate with each other on chromatin. The figure shows binding of both EZH2 and H3K27me3 across the MYT1 gene.