Fig. 4

Combination of AZD6738 with Cisplatin or Fludarabine Phosphate show synergistic efficacy ex vivo and in vivo. a, Evaluation of combination treatment efficacy and synergy of AZD6738 with cisplatin in primary glioma cultures (PC). Shown as black bar is the predicted value for additive combination effects based on the Bliss Independence Criterion as outlined in Methods. Observed measurements (purple) of combination treatments are depicted in purple. Lower values than predicted indicate a synergistic effect of the combination. Shown are means ± SD (n = 1, 3 technical replicates per sample). Representative bar graphs for one AZD6738 treatment in combination with cisplatin in each PC. b, Tumor areas of control (untreated (n = 16), DMSO (n = 16)), AZD6738 [50 µM] (n = 12), cisplatin [150 µM] (n = 19) and the combination of both (n = 12) treated wildtype zebrafish embryos xenotransplanted with LN229-GFP cells. Data was collected in two independent experiments. Tumor surface areas are measured using Imaris (version 9.2.0) after 48 h of treatment. Measurements were normalized to untreated control, means ± SD of the respective groups are indicated, each dot represents one embryo. Statistical analysis using one-way ANOVA (all to all comparison of means), Sidak correction for multiple testing, shown are only comparisons with corrected p-values < 0.5. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Right panel, exemplary pictures of zebrafish embryos of each group. Left, untreated zebrafish embryo with anatomical features “eye”, “yolk sac” and “LN229-GFP cells in midbrain region” highlighted by red arrows. Scale bars: 500 μm. c, Waterfall graph (left) and Kaplan-Meier curves (right) of untreated, control treated, AZD6738 (50 mg/kg), cisplatin (1 mg/kg), AZD6738/cisplatin-treated nude mice transplanted with LN229 cells. Median survival for each group listed below waterfall plot. Statistical analysis using log-rank (Mantel Cox) test, p-value below 0.05 considered significant, n.s. “not significant”. Combination of AZD6738 plus cisplatin significantly prolongs survival compared to control treated mice