Inhibition of BCAA metabolism suppresses the PRC2 activity in human acute leukemia. (A) The schema for transcriptome assays in primary human CD34⁺ AML and ALL cells with or without gabapentin treatment. Global changes in expression profiles were analyzed by the GSEA method. (B) Enrichment plots of embryonic stem (ES) core genes (WONG_ES_CELL_CORE). (C) Enrichment plots of genes marked with H3K27me3 modification in their promoters in human ES cells (BENPORATH_ES_WITH_H3K27ME3). (D) Enrichment plots of PRC2-target genes in human ES cells. (E) Aggregation plots of H3K27me3 ChIP-seq signals centered at TSSs in the levels of ES PRC2 target genes (left panel) and HSCs-related PRC2 target genes (right panel). The blue and red lines show the control and the gabapentin-treated (40 mM for 16 hours) leukemia cells, respectively. (F) Box plots showing H3K27me3 ChIP-seq signal intensities for the whole or the ES PRC2 target genes at the TSS+5 kilobasepairs (kbp) in primary CD34⁺ AML cells (left) and CD34⁺ ALL cells (right). (G) Box plots showing H3K27me3 ChIP-seq signal intensities for all genes and ES PRC2 target genes at the TSS+5 kbp in hCD34⁺ ALL cells harvested from NSG mice treated with or without the dietary restriction of BCAA in vivo. The Wilcoxon test was used for statistical analysis. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. FDR, false discovery rate; GSEA, gene set enrichment analysis; NES, normalized enriched score; NS, not significant; TSS, transcription start sites.