Fig. 7. ZM-induced AML is sensitive to Brd4 blockade.
a Heatmaps showing un-supervised k-means clustering of ZM and Brd4 ChIP-seq peaks at all promoter-proximal regions (TSS + /−2.5 kb). Four clusters (labeled as a to d, same as Fig. 3e) were produced based on their distinct ZM ChIP-seq signals. TSSs on the two DNA strands are labeled as 1 and 2 (such as a1/2 and b1/2), respectively. b IGV views of normalized Brd4 ChIP-seq signals at indicated loci in ZM-transformed AML cells. The read counts were first normalized to RPGC and then normalized to input. c Proliferation (left y-axis, normalized to DMSO-treated cells) and cell death (right y-axis, measured by trypan blue staining) of AML cells transformed by ZM alone or ZM plus NrasG12D, post-treatment with the indicated concentration of I-BET151 (x-axis) for four days. n = 3 biological replicates per group and data is presented as mean ± SD. d Wright–Giemsa staining of ZM-transformed AML cells post-treatment with DMSO or I-BET151 (0.25 uM) for four days (scale bar = 10 μm). e RT-qPCR of the indicated gene in ZM-transformed AML cells post-treatment with DMSO or the indicated concentration of I-BET151 for four days. qPCR signals from three independent experiments were normalized to those of 18S RNA and then to DMSO-treated control and presented as mean ± SD. f Bioluminescent imaging of mice transplanted with ZM + NrasG12D-transformed AML cells, two weeks post-treatment with either vehicle or I-BET151 (30 mg/kg daily IP injection). g Weight of spleens in the indicated cohorts (n = 4 mice per group) at the study endpoint. The p value was calculated by two-sided Student’s t test. h Kaplan–Meier survival curve of mock- and I-BET151-treated mice bearing the ZM + NrasG12D AML (n = cohort size). Black arrow indicates the day 7 when compound administration was initiated. The p value was calculated by two-sided log-rank test. i Model illustrates the ZM-mediated proto-oncogene activation and leukemogenesis.