Fig. 1

In vitro genetic and pharmacologic validation in FLT3-ITD cell lines. a Changes in reads of sgRNAs targeting BCL2 in a CRISPR knockout screen with midostaurin. Three guides are targeted to BCL2 in each of four replicate screens. b BCL2 expression was knocked down in MOLM-13 cells (BCL2 KD), confirmed via immunoblot using THP-1 as a positive control for BCL2 expression, and proliferation changes compared to parental cells (P) under treatment with 10 nM midostaurin, 15 nM gilteritinib, or control DMSO. A mixed effects model was applied to the data. Compared to DMSO, the average decrease in absorbance with midostaurin + BCL2 KD was larger than the observed decrease for midostaurin in parental cells (estimated difference = − 0.35; 95% CI − 0.49, − 0.2; p = 0.002) or with BCL2 KD alone (estimated difference: − 0.37; 95% CI − 0.4, − 0.33; p < .001). Similarly, the average decrease in absorbance for gilteritinib + BCL2 KD was larger than the decreases in absorbance for gilteritinib/parental (− 0.24; 95% CI − 0.31, − 0.17; p < .001) or BCL2 KD (− 0.42; 95% CI − 0.45, − 0.39; p < .001). c FLT3-ITD cell lines MOLM-13 and MV4-11 were treated with a range of doses of either midostaurin plus venetoclax or gilteritinib plus venetoclax for 48 h, and then, MTS reagent was added and absorption read. Highest single-agent (HSA) analysis was used to determine regions of synergy. *p < 5 × 10^–2, **p < 10^–3, ***p < 10^–4