Figure 1.
Selective inhibitors of AKT positively combine with PKC412 in the presence of adherent HS-5 stroma against MOLM14-luc+ cells.
(A) Approximately two-day proliferation study performed with MOLM14-luc+ cells cultured in the presence of adherent HS-5 stroma testing the combination of PKC412 and KIN001-102 versus each agent alone. (B) MOLM14-luc+ cells cultured in the presence of adherent HS-5 stroma for approximately two days: PKC412 (40 nM)+/− Akt inhibitors (660 nM). (C) Approximately two-day PKC412 treatment of MOLM14-luc+ cells cultured in the absence and presence of human stroma. (D) Approximately two-day treatment of adherent HS-5 stroma: PKC412 (40 nM) +/− Akt inhibitors (660 nM). (E) Calcusyn combination indices derived from 4-point concentration proliferation experiments. The cut-off for nearly additive effects (C.I.: 1.1) is marked by a dashed line.
Figure 2.
Selective inhibitors of AKT positively combine with PKC412 in the presence of SCM against MOLM14-luc+ cells.
(A–D) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in the presence of HS-5 SCM. (E) Approximately two-day PKC412 treatment of MOLM14-luc+ cells cultured in the absence or presence of HS-5 SCM (n = 2). (F) Calcusyn combination indices derived from the 4-point concentration proliferation experiments shown in A-D. The cut-off for nearly additive effects (C.I.: 1.1) is marked by a dashed line.
Table 1.
Effects of PKC412 and KIN001-102, alone and combined, on MOLM14-luc+ cell cycle progression (following 24 hours of treatment) and apoptosis (following 48 hours of treatment) when cells are cultured in the presence of 50% HS-5 SCM.
Table 2.
Effects of PKC412 and KIN001-102, alone and combined, on MOLM14-luc+ cell cycle progression (following 24 hours of treatment) and apoptosis (following 48 hours of treatment) when cells are cultured in the presence of RPMI+10% FBS.
Figure 3.
Selective inhibitors of AKT positively combine with PKC412 in RPMI+10% FBS against MOLM14-luc+ cells.
(A–D) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in RPMI+10% FBS. (E) Calcusyn combination indices. The cut-off for nearly additive effects (C.I.: 1.1) is marked by a dashed line.
Figure 4.
Selective inhibitors of AKT positively combine with AC220 in RPMI+10% FBS against MOLM14-luc+ cells.
(A–D) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with AC220 in RPMI+10% FBS. (E) Calcusyn combination indices. The cut-off for nearly additive effects (C.I.: 1.1) is marked by a dashed line.
Figure 5.
Selective inhibitors of AKT positively combine with PKC412 in the absence and presence of HS-5 SCM against MOLM13-luc+ cells.
(A–B) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in the presence of RPMI+10% FBS. (C–D) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in 80% HS-5 SCM. (E) Treatment of MOLM13-luc+ cells with PKC412 in either RPMI+10% FBS or 80% HS-5 SCM.
Figure 6.
Selective inhibitors of AKT positively combine with PKC412 in RPMI+10% FBS against MV4,11 and Ba/F3-FLT3-ITD cells.
(A–C) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in RPMI+10% FBS against MV4,11 cells. (D–F) Approximately two-day proliferation studies performed with selective AKT inhibitors in combination with PKC412 in RPMI+10% FBS against Ba/F3-FLT3-ITD cells.
Table 3.
Calcusyn software-derived combination indices.
Figure 7.
Phospho-Akt mediates synergy observed between allosteric Akt inhibitor, KIN001-102, and PKC412.
Immunoblots of protein lysates prepared from MOLM14-luc+ cells treated for 2 hours with PKC412 (40 nM), KIN001-102 (165, 330, 660 nM), or a combination of the two agents in the presence of 50% SCM. Data shown are representative of two independent experiments in which similar results were achieved.
Figure 8.
Ability of Akt inhibitors to positively combine with PKC412 or AC220 against AML patient samples in the presence of cytoprotective SCM.
(A) Approximately two-day proliferation study performed with a selective Akt inhibitor in combination with PKC412 in the presence of HS-5 SCM against mutant FLT3-positive AML#2. (B) Approximately two-day combination studies: AC220 (0.4 nM) +/− selective AKT inhibitors (660 nM) against MOLM14-luc+ cells in the presence of 50% HS-5 SCM. (C) Approximately two-day combination studies: AC220 (0.4 nM) +/− selective AKT inhibitors (660 nM) against MOLM14-luc+ cells in the presence of RPMI+10% FBS. (D) Approximately two-day combination studies: PKC412 (40 nM)+/− selective AKT inhibitors (660 nM) against primary AML patient cells in the presence of 50% HS-5 SCM. (E) Approximately two-day combination studies: AC220 (0.4 nM) +/− selective AKT inhibitors (660 nM) against primary AML patient cells in the presence of 50% SCM. (F) Ability of Akt inhibitors to positively combine with PKC412 or AC220 against primary AML cells in the presence of cytoprotective SCM. Patient information is provided in Table S1.