TKI treatment leads to dynamic alterations in energy metabolism in CML progenitors. Mice with CML were treated with nilotinib or vehicle for 2 days or 2 weeks, and BM c-Kit⁺ cells were selected and metabolomic profiling was performed. (A) The relative abundance of glycolytic intermediates after 2 days of nilotinib treatment is shown (n = 3-4). (B) The relative abundance of TCA cycle intermediates after 2 days of nilotinib treatment is shown (n = 3-4). (C) The ratio of ATP/ADP and GTP/GDP after 2 days of nilotinib treatment is shown (n = 3-4). (D) The relative abundance of glycolytic intermediates after 2 weeks of nilotinib treatment is shown (n =3). (E) The relative abundance of TCA cycle intermediates after 2 weeks of nilotinib treatment is shown (n = 3). (F) The ratio of ATP/ADP and GTP/GDP after 2 weeks of nilotinib treatment is shown (n = 3). (G) BM c-Kit⁺ cells were labeled with [U-¹³C₆]glucose in vitro (n = 3). The percent labeling fraction of glycolytic end products (pyruvate, lactate, and alanine) and citric acid cycle intermediates (citrate/isocitrate, α-ketoglutarate, and malate) after 2 days and after 2 weeks of vehicle (Veh) or nilotinib (NIL) treatment is shown. The labeling fractions are corrected for ¹³C natural abundance. Significance values: ∗P < .05; ∗∗P < .01; ∗∗∗P < .001. Results represent mean ± SEM of multiple replicates. 3PG/2PG, 3-phosphoglyceric acid/2-phosphoglyceric acid; ADP, adenosine diphosphate; a-KG, α-ketoglutarate; F-1,6-BP, fructose 1,6-bisphosphate; G6P/F6P, glucose 6-phosphate/fructose 6-phosphate; GDP, guanosine diphosphate; GSH, glutathione; GTP, guanosine triphosphate; ns, not significant; PEP, phosphoenolpyruvic acid.