Fig. 1: Validation and refinement of RUNX1 mutational signatures in CML blast crisis patients.

A Schema of RUNX1b gene isoform with mutations found in 3 MBC and 1 LBC CML patients. Horizontal scale indicates amino acid (aa) residue positions. B GSEA heatmap in left panel shows expression of genes upregulated in the Awad RUNX1 mutational signature in RUNX1 mutant (Mut) compared to wild type patients in red for upregulation and blue for downregulation. Right panel shows enrichment plot with normalised enrichment score (NES) and false discovery rate (FDR). C GSEA heatmap in left panel and enrichment plot in right panel shows enrichment of the same gene set in wild type RUNX1 CD34 + LBC patient gene expression data compared to the equivalent in MBC. D Table shows alternative gene sets that can significantly distinguish mutant RUNX1 blast crisis samples from wild type (WT) in the top half (FDR < 0.25). The same gene sets do not enrich significantly for wild type RUNX1 CD34 + LBC samples compared to the equivalent in MBC or are depleted. Negative control RUNX2 and RUNX3 gene sets also do not enrich significantly in RUNX1 mutant compared to wild type samples in the bottom half. E GSEA heatmap in left panel shows expression of the TONKS_TARGETS_OF_RUNX1_RUNX1T1_FUSION_HSC_DN gene set [11] with enrichment in RUNX1 mutant (Mut) compared to wild type patients. Right panel shows enrichment plot with normalised enrichment score (NES) and false discovery rate (FDR). F GSEA heatmap in left panel and enrichment plot in right panel shows depletion of the same gene set in wild type RUNX1 CD34 + LBC compared to the equivalent MBC gene expression data. Black bar indicates 3 CD34 + wild type RUNX1 LBC samples while yellow and red hatched bar denotes a biphenotypic wild type RUNX1 sample with characteristics of both LBC and MBC.