AYNE selectively eliminates the AML population and targets VLCAD in vivo. (A) Schematic diagram of the study highlighting the injection of patient-derived AML or normal cord blood MNC into tail veins of NSG mice. After an engraftment period during which human cells localize to BM, mice were divided into 2 groups (vehicle or AYNE) and then treated 3 times weekly for 4 or 6 weeks. Mice were then euthanized, femurs were flushed, and human CD33⁺/CD45⁺ cells were quantified via flow cytometry. Created using BioRender. Human cells were given 1 week (B-D) or 4 weeks (E) to engraft prior to starting treatment. (B) Engraftment of healthy cord cells following treatment with 300 mg/kg per week of AYNE or vehicle control for 6 weeks. Engraftment of patient-derived AML cells following treatment with vehicle or AYNE [225 mg/kg per week (C) or 300 mg/kg per week (D)] for 6 weeks. (E) Engraftment of patient-derived AML cells following treatment with vehicle or AYNE (200 mg/kg per week) for 4 weeks. (F) Schematic diagram of the study highlighting injection of patient-derived AML cells into the tail vein of NSG mice. After 8 weeks, mice were divided into 2 groups (vehicle or AYNE) and treated with a bolus dose of AYNE (150 or 200 mg/kg) or a vehicle control. Twenty-four hours later, mice were euthanized, femurs were flushed, and cells were recovered for downstream applications. The mix of human and mice cells recovered from the femurs was quantified via flow cytometry before (G) and after (H) column purification. C16-supported ETF respiration (I) and thermostabilization of VLCAD (J) were assessed in human cells recovered from mice that received vehicle or AYNE (150 or 200 mg/kg). In (B-E,I-J), data are mean ± standard deviation. *P ≤ .05, **P ≤ .002, ***P ≤ .001, Mann-Whitney U test (B-E); 1-way ANOVA with Tukey’s post hoc test (I-J).