Fig. 6: Disrupting the super-enhancers using ABBV-075 inhibits CREB5 expression, consequently preventing H3.3K27M DIPG tumor growth.

a Representative IGV tracks for RNA-seq with vehicle or JQ1 treatment at a time-course and indicated ChIP-seq in DIPG cells at CREB5 gene loci. b qPCR analysis of indicated genes with vehicle or JQ1 (300 nM) treatment for 24 hours in DIPG17 cells. c Schematic of BRD4 inhibitor screening strategy. d qPCR analysis of CREB5 expression treated with series compounds for 24 hours in DIPG17 cells. e Heatmap showing the relative cell viability of DIPG17 cells and normal PPC cells treated with indicated compounds for 48 hours (n = 3). f, g Immunoblotting analysis of the indicated proteins with ABBV-075 (100 nM) treatment at indicated time points (f) and with mentioned concentrations of ABBV-075 for 24 hours (g) in DIPG17 cells. h The overall structure of molecular docking ABBV-075 (shown in blue) in complex with BRD4 protein (PDB: 3MXF). i Bar graph depicting luciferase activity of indicated reporter constructs in 293T cells. j Line graph depicting the CREB5 promoter luciferase activity with ABBV-075 treatment at various concentrations. The insert bar graph shows the luciferase activity treated with 100 nM ABBV-075 for 24 hours. k ATAC-seq profiles from DIPG17 cells showing the DNA accessibility for super-enhancer regions with or without ABBV-075 treatment (100 nM) for 24 hours. l Bar plot showing the fold change of ATAC-seq peaks with or without ABBV-075 treatment at pre-ranked super-enhancer associated genes in DIPG17 cells. m Representative IGV tracks for ATAC-seq and RNA-seq with vehicle or ABBV-075 treatment in DIPG17 cells at CREB5 gene loci. n ATAC-seq profiles from DIPG17 cells showing the DNA accessibility for CREB5 binding promoter and enhancer regions with or without ABBV-075 treatment. Metagene plot showing the average signal for ATAC-seq at CREB5-binding promoter and enhancer regions with or without ABBV-075 treatment. Data presented as mean ± s.e.m. of three independent experiments (b, d, i, and j), statistical significance was determined by a two-tailed unpaired Student’s t-test (b, d, i, and j). Experiments were repeated three times independently with similar results (f and g). Panel c created in BioRender. Zhou, W. (2025) https://BioRender.com/q17x870. Source data are provided as a Source Data file.