Fig. 5
From: Clinical resistance to crenolanib in acute myeloid leukemia due to diverse molecular mechanisms
IDH1 and IDH2 mutations contribute to crenolanib resistance. a Variant allele frequencies (VAFs) of FLT3-ITD/TKD and IDH1 or IDH2 during crenolanib treatment. Notably, we did not detect the IDH1 mutation in B32 before crenolanib treatment by exome sequencing. However, a targeted gene panel detected this IDH1 mutation before crenolanib treatment. b Graph depicts mean ± SEM of colony numbers of FLT3-ITD/IDH1 WT and FLT3-ITD/IDH1 R132H expressing mouse stem cells treated with gradient concentrations of crenolanib, AG5198 or two drugs in combination. c Graph depicts cell viabilities of FLT3 D835Y, IDH1 WT/ FLT3 D835Y and IDH1 R132H/ FLT3 D835Y expressing Ba/F3 cells treated with crenolanib, or crenolanib in combination with IDH1 Inhibitor (AG5198) determined by MTS assay. d Graph depicts decreased half-maximal inhibitory concentration (IC50) of crenolanib and AG5198 in combination compared to crenolanib alone. Statistical significance was determined using two-tailed nonparametric Student’s t-tests (Mann–Whitney test) comparing each group to the non-treated group. TP53 mutations co-occur with FLT3 mutations and confer crenolanib resistance. e Graph depicts VAFs of FLT3-ITD/TKD and TP53 or PPM1D mutation during crenolanib treatment. f Representative graph depicts higher mean ± SEM of cell viability of crenolanib-treated Molm13 cells expressing CRISPR/Cas9 and single-guide RNAs (sgRNAs) targeting TP53 compared to cells expressing CRISPR/Cas9 and a non-specific targeting sgRNA (NS) control. TP53_1: sgRNA1 targeting TP53; TP53_2: sgRNA2 targeting TP53. g Graph depicts mean ± SEM of crenolanib IC50 in (b). Data shown are from three or five biological replicates. Statistical significance was assessed using one-way analysis of variance (ANOVA) together with Dunn’s multiple comparisons tests and expressed as: *p < 0.05
