Gilteritinib Resistant MOLM-14 and MV4;11 Whole Exome Sequencing

In this study, we utilized a two-step approach to reveal temporal evolution of gilteritinib resistance. We cultured AML cells with exogenous protective factors that are normally secreted by marrow stromal cells to recapitulate microenvironmental/early resistance. Removal of these protective factors led to the development and outgrowth of intrinsic mutations that delineate late resistance. Using multiple orthogonal approaches--whole exome sequencing, genome-wide CRISPR resensitization screens, global/phospho-proteomics, metabolomics, and small-molecule inhibitor screening--we determined the critical pathways that are altered in early and late gilteritinib resistance.