Fig. 2: Mutations in both the kinase and PH domains of AKT can cause drug-specific allosteric AKT inhibitor resistance. | British Journal of Cancer

Fig. 2: Mutations in both the kinase and PH domains of AKT can cause drug-specific allosteric AKT inhibitor resistance.

From: A systematic molecular and pharmacologic evaluation of AKT inhibitors reveals new insight into their biological activity

Fig. 2

a A docking model of MK-2206 bound to AKT1. b Crystal structure of AKT1 bound to miransertib. c A homology model of AKT2 bound to miransertib. d, e Effects of the Q79K mutation on the response to AKT inhibitors. d MDA-MB-361 cells were stably transduced with wild-type AKT1 or AKT1 Q79K. Cells were then treated with the indicated doses of MK-2206, ARQ 751, miransertib, GSK690693, ipatasertib or capivasertib. The fold change in cell numbers (top) and the fraction of dead cells (bottom) following 3 days of drug treatment is shown. e MDA-MB-361 cells expressing either AKT1 or the Q79K mutant were treated with increasing doses of MK-2206 or miransertib. The fold change in cell numbers (left) and the fraction of dead cells (right) following 3 days of drug treatment is shown. f The effects of the AKT1 Q79K mutation in response to MK-2206 and miransertib was evaluated in washout experiments. MDA-MB-361 cells stably expressing wild-type AKT1 or AKT1 Q79K were treated with the indicated doses of drug for either 1 h or continuously for 72 h. The fold change in cell numbers and the fraction of dead cells (bottom) at 72 h is shown. g HCT116 AKT1/2 DKO cells were stably transduced with wild-type AKT1 or AKT1 Q79K. Cells were subjected to western blot analysis, following o/n drug treatment, with the indicated antibodies. h A mutation that disrupts the kinase-domain/PH-domain interface confers resistance to all allosteric inhibitors. MDA-MB-361 cells were stably transduced with wild-type AKT1 or the mutant variants D323H, D292A (a kinase-deficient mutant), and the double mutant D323H-D292A. Cells were treated with allosteric AKT inhibitors as shown. The fold change in cell numbers and the fraction of dead cells following 4 days of drug treatment is shown (cell death %). i HCT116 AKT1/2 DKO cells were stably reconstituted with the indicated AKT variants and treated o/n with allosteric AKT inhibitors as shown. Cells were then lysed and analysed by immunoblot with the indicated antibodies. n.s., not significant. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 and ****P ≤ 0.0001.

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