FIGURE 1 from Pim Kinase Inhibitors Increase Gilteritinib Cytotoxicity in FLT3-ITD Acute Myeloid Leukemia Through GSK-3β Activation and c-Myc and Mcl-1 Proteasomal Degradation

Pim inhibitor and gilteritinib combination treatment is synergistic in cells with FLT3-ITD. A,C, Apoptosis induction. Ba/F3-ITD, MV4-11, and MOLM-14 cells were treated with the FLT3 inhibitor gilteritinib (15 nmol/L for Ba/F3-ITD and 10 nmol/L for MV4-11 and MOLM-14) and/or the Pim inhibitor AZD1208 (1 µmol/L; A) or TP-3654 (1 µmol/L; C) or DMSO control for 48 hours in triplicate experiments. Apoptosis was analyzed by Annexin V and PI staining, measured by flow cytometry. ****, P P P B,D, Cytotoxicity. Ba/F3-ITD cells seeded at 5,000 cells/well and MV4-11 and MOLM-14 cells at 10,000 cells/well in 96-well plates were treated for 48 hours with gilteritinib and/or AZD1208 (B) or TP-3654 (D) as single drugs and in combinations at the concentrations shown, in triplicate. Cytotoxicity was measured by the WST-1 assay, and drug combination effects were determined by Chou-Talalay analysis. Synergism was defined by combination index values