Fig. 7: Inhibition of Rap1 signaling enhances apoptotic priming in T-ALL.

a Bar graph of significant gene ontologies based on the top 100 differentially expressed genes in malignant cells of group 2 T-ALL patients compared to group 1 T-ALL patients (Supplementary Data 7) as analyzed by DAVID Gene ontology. P-value is derived from a Modified Fisher’s Exact Test. Immunoreg: Immunoregulatory interactions between a lymphoid and non-lymphoid cell. b Ligand-receptor interactions (NATMI³⁵) between NC monocytes and malignant cells from T-ALL group 2 patients. c, d Violin plots showing surface protein expression of CD54 in NC monocytes from healthy donors (n = 102 cells (BM), n = 168 cells (PB)), T-ALL group 1 (n = 881 cells (PB), n = 1531 cells (BM)) and T-ALL group 2 (n = 1603 cells (PB), n = 1226 cells (BM)) at diagnosis (c), and CD11a and CD18 in malignant cells from T-ALL group 1 (n = 4 (BM), n = 6 (PB)) and T-ALL group 2 (n = 1 (BM), n = 4 (PB)) patients (downsampled to 300 cells/sample) at diagnosis (d). A one-way Anova test followed by Tukey’s HSD post-hoc test (c) or an unpaired t-test (d) was used for statistical analysis. e Viability as assessed by CellTiter-Glo assay upon GGTI-298 treatment in T-ALL cell lines (Jurkat, HPB-ALL and CCRF-CEM) for three days and two primary T-ALL samples from group 2 (P8 and P14) for five days. Viability is normalized to vehicle control DMSO. Each dot represents a data point from an independent replicate. Bars and error bars represent the mean and SEM of triplicates. f Heatmap of normalized cytochrome c^- cells upon 12-hour treatment with GGTI-298 or vehicle DMSO in T-ALL cell lines Jurkat, HPB-ALL and CCRF-CEM. g Most synergistic area score (MSA) (Bliss, SynergyFinder⁷⁵) of combination treatment with Rap1 inhibitor GGTI-298 and indicated drugs in T-ALL cell lines (Jurkat, HPB-ALL and CCRF-CEM) and two primary T-ALL patients samples from group 2. MSA > 10 is considered strongly synergistic (dashed line). Each dot represents the MSA and error bars indicate 95% confidence interval. h Dose-response curves (CellTiter-Glo assay) of T-ALL cell lines (Jurkat and HPB-ALL) and primary T-ALL P14 treated with various concentrations of GGTI-298 and navitoclax (top) or venetoclax (bottom) for three or five days, respectively. Viability of GGTI-298 without addition of BCL2 inhibitors were normalized and set to 100%. Most synergistic area score (MSA) (Bliss, SynergyFinder⁷⁵) is shown (see (g)). Each dot represents the mean and error bars represent SEM of three independent replicates.