Figure 7. In vivo experiments with xenograft AML mouse models demonstrate that leukemic cells are dependent on SSP in fructose-rich conditions.

(A) Glucose and fructose concentration in bone marrow of xenograft leukemia mice and healthy mice measured at 30 min. after IP injection of fructose (4 g/kg).

(B) Schematic of the in vivo experiment with infusion of [U-¹³C₆] glucose or fructose. NSG mice transplanted with MOLM13 cells were grouped into three different experimental conditions; one group for glucose infusion, another for fructose infusion, and the other for fructose infusion with PHGDH inhibitor (NCT-503) treatment. GLC, glucose; FRC, fructose; PHGDHi, PHGDH inhibitor. 5 hours after injection, all the mice were sacrificed and human leukemic cells were isolated from mouse bone marrow based on hCD45 marker.

(C) Enrichment of glycine (m+2) in the cells collected in the experiment (B). n = 9 for the [U-¹³C₆] glucose infusion group, n = 8 for the [U-¹³C₆] fructose infusion group, n = 10 for the [U-¹³C₆] fructose infusion with PHGDH inhibitor treatment group.

(D) Schematic of the in vivo experiment with doxycycline-inducible PHGDH knockdown. NSG mice transplanted with MOLM13 cells with sh-Ctrl or sh2-PHGDH (Figure 5B) were grouped into three different experimental conditions; one group for serial IP injection of glucose (4 g/kg), another for fructose (4 g/kg), and the other for PBS three times a week. After four weeks of injection, all the mice were sacrificed and the engraftment of human leukemic cells was analyzed based on hCD45 marker.

(E) Engraftment level of hCD45⁺ cells in bone marrow from the mice in the experiment (D).

(F) Schematic of the in vivo experiment with a patient-derived AML mouse model (cells from Patient 5 in Figure 3I and Table S1). One group was for fructose (4 g/kg) along with vehicle treatment, and the other group was for fructose along with NCT-503 (40 mg/kg) treatment. After 6 weeks of daily injection, all the mice were sacrificed and the engraftment of human leukemic cells was analyzed based on hCD45 marker.

(G) Engraftment level of hCD45⁺ cells in bone marrow from the mice in the experiment (F). Statistical analyses were conducted with unpaired two-tailed t-test: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. GLC, glucose; FRC, fructose.