AYNE inhibits palmitate contribution to the TCA cycle, resulting in primary AML cell death while sparing normal cells. (A) Viability of primary bulk AML cells (#14-#18), primary CD34⁺ AML cells (#2, #3, #15, #19), and normal CD34⁺ cells (n = 3) treated for 24 hours. (B) Quantification of ATP levels in primary CD34⁺ AML and CD34⁺ normal cells after 12-hour treatment. (C) Levels of ¹³C-labeled ACs in primary AML cells (#4, #22, #23, #24, #27) after 12 hours. (D) Enrichment of ¹³C atoms into FAO and TCA metabolites following 12-hour treatment with a uniformly labeled ¹³C₁₆ palmitate tracer in primary AML cells (#4, #22, #23, #24, #27) and normal MNCs (n = 3). Created using BioRender. (E) Levels of ¹³C₃ lactate, ¹³C₃ pyruvate, ¹³C₂ citrate following 12-hour treatment with a ¹³C₆ glucose tracer in primary AML cells (#3, #15, #16, #19, #25, #26) and normal MNCs (n = 5). In all experiments, primary AML cells and normal MNCs were treated with a solvent vehicle or 50 µM AYNE. In (A-E), data are mean ± standard deviation. Summary of patient cytogenetics are shown in supplemental Figure 10. Summary of statistics for (C-E) are shown in supplemental Figure 11. *P ≤ .05, **P ≤ .002, ***P ≤ .001, 2-tailed unpaired Student t test. shRNA, short hairpin RNA.