Targeting the Histone H3.3-K27M Mutation for the Treatment of Diffuse Intrinsic Pontine Gliomas

Aim 1: Determine whether the H3.3-K27M mutation correlates with GSK-J4 sensitivity in vitro and in vivo. We will test the anti-tumor activity of GSK-J4 in pediatric glioma cells and human orthotopic DIPG xenografts. Intracranial (brainstem) tumor growth and response to therapy will be quantitatively measured by BLI, and efficacy will be assessed by survival analysis. Aim 2: Determine whether GSKJ4 treatment alters the expression of genes that promote cell proliferation and differentiation in H3.3-K27M mutant DIPG cells. This aim will determine the extent to which GSK-J4 alters gene expression in K27M mutant DIPG cells, and will identify critical mediators of protumorigenic effects of K27M mutant. We will perform ChIP-sequence, as well as gene expression array analysis, using K27M mutant DIPG cells before and after GSKJ4 treatment. Candidate genes that are suppressed in response to GSKJ4 treatment will be investigated as pro-tumorigenic mediators of the K27M mutation using siRNA treatments of tumor cells, with results interpreted by extent of growth inhibition. Experimental Design And Methods Aim 1: Determine whether the H3.3-K27M mutation correlates with GSK-J4 sensitivity in vitro and in vivo. Aim 2: Determine whether GSKJ4 treatment alters the expression of genes that promote cell proliferation and differentiation in H3.3-K27M mutant DIPG cells.