BCL2 variants have reduced activity to VEN in AML cells. (A) Structure of the BCL2 protein with venetoclax (blue) binding in the BCL2 groove.16 The positions of the 3 mutated residues are illustrated (green). P2 and P4 pockets and the azaindole side chain of venetoclax are indicated. (B) Table summarizing the binding affinities of BIM or venetoclax for WT or Val148Leu BCL2 as determined by direct binding assays in comparison with previously documented binding affinities for Asp103Glu, Phe104Leu, and Gly101Val BCL2 mutants. Data represent means ± 1 standard deviation (SD) of 3 independent experiments with curves and fits shown in supplemental Figures 2 and 3. (C-D) BCL2 WT, Asp103Glu, Phe104Leu, and Val148Leu were overexpressed in (C) OCI-AML2 and (D) MOLM-13 AML cell lines. In vitro viability assessed by flow cytometric enumeration of cells excluding propidium iodide upon exposure to indicated concentrations of venetoclax or navitoclax for 48 hours. Data represent means ± 1 SD of at least 3 independent experiments.
