Fig. 5. Combination of doxorubicin and KPT-330 is synergistic in vitro and in vivo.

a Heatmap of synergy scores from Bliss and ZIP drug–drug interaction models between KPT-330 with vincristine, actinomycin, or doxorubicin in two FHWT cell lines. Range blue (antagonistic <−10) to red (synergy >10). Representative of at least two biological replicates for each cell line. b Mean IC50s for WT cell lines treated with doxorubicin as compared to normal kidney cells. X axis is µM. SD shown from at least two biological replicates. *P value < 0.05 from a Student’s two-sided unpaired t test. c Heatmap showing synergy scores from Bliss and ZIP drug–drug interaction models in WT with KPT-330 and doxorubicin. Range blue (antagonistic <−10) to red (synergy >10). Representative of at least two biological replicates for each cell line. d Representative 3D landscape image of the synergy score for CCLF_PEDS_0041_T2 cells treated with doxorubicin (62.5–500 nM) and KPT-330 (1.25–10 μM). e CCLF_PEDS_0041 tumor fragments were placed subcutaneously into the hind flank of NSG (NOD-SCID IL2Rgamma null) female mice. The tumor xenografts were treated with vehicle, doxorubicin, KPT-330 or the combination of doxorubicin and KPT-330 for 28 days. Log fold change of tumor volumes at day 22 were calculated as compared to time of treatment (average 114.3 mm³). **P value < 0.005, ***P value < 0.0005, ****P value <0.00005 from a Student’s unpaired two-sided t test. f Line graph depicting the tumor volume across the study days for each mouse in different treatment groups. Tumor growth was monitored until they met endpoint or the study was terminated at day 150. g Kaplan–Meier survival curves representing the probability of overall survival in the patient-derived xenografts (n = 8 per treatment group) treated with the vehicle, KPT-330, doxorubicin, and KPT-330 + doxorubicin. **P value <0.005, ****P value <0.00005.