Figure 2.
Synergistic effects of combined menin-MLL and FLT3 inhibition. (A) Dose-response curves from cell-viability assays after 48 hours of treatment with various FLT3 inhibitors in FLT3-ITD+ MOLM13 and MV411 cells. (B) FLT3-ITD+ and FLT3 WT human leukemia cell lines were treated with quizartinib for 48 hours. IC50 values were graphically determined by GraphPad Prism. (C) Dose-response curves from cell-viability assays after 48 hours of treatment with various FLT3 inhibitors in murine Npm1CA/+Flt3ITD/+ cells. (D) Ponatinib IC50 concentrations in murine Npm1CA/+Flt3ITD/+ and Hoxa9-Meis1–transformed cells after 48 hours of treatment. (E) Summary of FLT3 inhibitor IC50 concentrations in the human and murine leukemia cell lines assessed in this study. (F) Dose-response curves from cell-viability assays of MV411 and MOLM13 cells, comparing MI-503 (MI; 3 days for MV411 cells and 4 days for MOLM13 cells), quizartinib (Qz; 24 hours), and combinatorial MI-503 (3 or 4 days) and quizartinib (24 hours) treatment. Dashed lines indicate IC50 values. (G) Dose-response curves from cell-viability assays of Npm1CA/+Flt3ITD/+ cells comparing MI-503 (MI, 4 days), ponatinib (Po, 24 hours, left), and gilteritinib (Gil, 24 hours; right), or their combination (4 days, MI-503; 24 hours, ponatinib and gilteritinib). Dashed lines indicate IC50 values. (H) Effect of MI-503 (2.5 µM), ponatinib (100 nM), and combinatorial treatment (2.5 µM and 100 nM) on the number of total and blast-like colonies in murine Npm1CA/+Flt3ITD/+ cells, normalized to DMSO. Micrographs were taken at ×20 amplification. (I-J) Percentage of apoptotic (annexin V) and dead (4′,6-diamidino-2-phenylindole [DAPI]-stained) cells after single and combinatorial treatment with MI-503 (2.5 µM) and quizartinib (3 nM) in human cell lines (I) or MI-503 (2.5 µM) and ponatinib (100 nM) or gilteritinib (400 nM) in murine cells (J). (K) Giemsa-stained cytospins showing human MV411 and MOLM13 cells and murine Npm1CA/+Flt3ITD/+ cells after single and combinatorial treatment with MI-503 (2.5 µM; 4 days and 3 days for MV411) and FLT3 inhibitor (quizartinib, 3 nM, 24 hours; ponatinib, 100 nM, 24 hours) or their combination (day 4/24 hours and day 3/24 hours for MV411, respectively). Micrographs were taken at ×100 amplification. Error bars represent SD of 3 independent experiments, each performed in 3 technical replicates.

Synergistic effects of combined menin-MLL and FLT3 inhibition. (A) Dose-response curves from cell-viability assays after 48 hours of treatment with various FLT3 inhibitors in FLT3-ITD+ MOLM13 and MV411 cells. (B) FLT3-ITD+ and FLT3 WT human leukemia cell lines were treated with quizartinib for 48 hours. IC50 values were graphically determined by GraphPad Prism. (C) Dose-response curves from cell-viability assays after 48 hours of treatment with various FLT3 inhibitors in murine Npm1CA/+Flt3ITD/+ cells. (D) Ponatinib IC50 concentrations in murine Npm1CA/+Flt3ITD/+ and Hoxa9-Meis1–transformed cells after 48 hours of treatment. (E) Summary of FLT3 inhibitor IC50 concentrations in the human and murine leukemia cell lines assessed in this study. (F) Dose-response curves from cell-viability assays of MV411 and MOLM13 cells, comparing MI-503 (MI; 3 days for MV411 cells and 4 days for MOLM13 cells), quizartinib (Qz; 24 hours), and combinatorial MI-503 (3 or 4 days) and quizartinib (24 hours) treatment. Dashed lines indicate IC50 values. (G) Dose-response curves from cell-viability assays of Npm1CA/+Flt3ITD/+ cells comparing MI-503 (MI, 4 days), ponatinib (Po, 24 hours, left), and gilteritinib (Gil, 24 hours; right), or their combination (4 days, MI-503; 24 hours, ponatinib and gilteritinib). Dashed lines indicate IC50 values. (H) Effect of MI-503 (2.5 µM), ponatinib (100 nM), and combinatorial treatment (2.5 µM and 100 nM) on the number of total and blast-like colonies in murine Npm1CA/+Flt3ITD/+ cells, normalized to DMSO. Micrographs were taken at ×20 amplification. (I-J) Percentage of apoptotic (annexin V) and dead (4′,6-diamidino-2-phenylindole [DAPI]-stained) cells after single and combinatorial treatment with MI-503 (2.5 µM) and quizartinib (3 nM) in human cell lines (I) or MI-503 (2.5 µM) and ponatinib (100 nM) or gilteritinib (400 nM) in murine cells (J). (K) Giemsa-stained cytospins showing human MV411 and MOLM13 cells and murine Npm1CA/+Flt3ITD/+ cells after single and combinatorial treatment with MI-503 (2.5 µM; 4 days and 3 days for MV411) and FLT3 inhibitor (quizartinib, 3 nM, 24 hours; ponatinib, 100 nM, 24 hours) or their combination (day 4/24 hours and day 3/24 hours for MV411, respectively). Micrographs were taken at ×100 amplification. Error bars represent SD of 3 independent experiments, each performed in 3 technical replicates.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal