JPH203 combined with Ven + Aza increased antileukemia efficacy in AML. (A) Dot plot showing reads per kilobase of transcript per million mapped reads of SLC7A5, SLC43A1, and SLC43A2 stratified based on GPRC5C^hi(GPRC5C high: ≥75th percentile) and GPRC5C^low (GPRC5C low: <75th percentile), expression from the BEAT AML and the Leucegene AML cohorts. (B) Experimental scheme to assess treatment with JPH203 + Ven + Aza in leukemia cell lines and BM samples from patients with de novo AML. (C) Absolute number of live cells, as determined by annexin V staining in specimens of patients with AML treated with JPH203 for 24 hours. Normalized to the DMSO control of each AML specimen; n = 3 technical replicates per AML specimen. (D) Maximum respiration measured by the seahorse assay of patient cells with AML treated with JPH203 for 12 hours. Normalized to the DMSO control of each AML specimen; n = 2 to 5 technical replicates per AML specimen. (E) Absolute cell count of live cells as determined by annexin V staining in patient specimens with AML treated with Ven + Aza + JPH203 and measured at 24 hours. Normalized to the DMSO control of each AML specimen. CDI was calculated for each AML specimen. CDI value < 1 (synergistic), CDI  =  1 (additive), and CDI > 1 (antagonistic); n = 3 to 5 technical replicates per AML specimen. (F) Oxygen consumption levels measured by the seahorse assay of specimens of patients with AML treated with Ven + Aza + JPH203 for 12 hours. Normalized to the DMSO control of each AML specimen; n = 5 technical replicate per AML specimen. (G) Heat map of amino acids metabolites. Z-score centering of the mean value of metabolite abundance per specimen from patients with AML; n = 3 specimens from patients with AML. (H) GSEA of LSC/prognostic signatures under treatment conditions compared with the DMSO control. GSEA was performed with BH-adjusted P values after the adaptive multilevel splitting Monte Carlo approach. GAL_LSC_DOWN,⁴⁶ GOOD_PROGNOSIS_AML,⁴⁷ EPPERT_LSC_UP,⁴⁸ and SOMERVAILLE_AML_DOWN.⁴⁹ All represented by mean ± standard deviation. (A,G) Unpaired Student t test; (C-F) 2-way analysis of variance. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. For all experiments, at least 2 independent experiments were performed.