Development of preclinical models of EVI1-rearranged AML. (A) Schematic outline of the generation of a transplantation-based RUNX1/EVI1-driven mouse model allowing controllable oncogene expression. (B) Kaplan-Meier survival analysis of mice transplanted with mHSPCs expressing Nras^G12D combined with either vector control or RUNX1/EVI1 (n ≥ 5). (C) Bioluminescence imaging of mice transplanted with RUNX1/EVI1-expressing leukemia cells that received either Dox treatment (4 mg/mL) or regular drinking water. (D) Kaplan-Meier survival analysis of mice transplanted with RUNX1/EVI1-expressing leukemia cells that received Dox treatment (4 mg/mL) compared with untreated controls (n = 5). (E) Flow cytometric analyses of BM-derived leukemia cells of mice transplanted with RUNX1/EVI1-expressing leukemia cells that received Dox treatment (4 mg/mL) compared with untreated controls. Relative amounts of GFP⁺CD45.2⁺ donor cells were quantified and further characterized toward their immunophenotype. (F) BM cytospins of mice transplanted with RUNX1/EVI1-expressing leukemia cells at indicated time points upon Dox treatment (4 mg/mL) compared with untreated controls. (G) Principal component analysis of the gene expression of human EVI1-expressing AML cell lines (n = 7), human patients with AML harboring EVI1 rearrangements (n = 12) and murine leukemia models expressing RUNX1/EVI1 (n = 6).