Fig. 7: Pathways enriched in progenitor cancer cells expose therapeutic opportunities.

a Bar-graph showing the proportion of viable glioma stem cells (GSCs, BT333-GSC) sorted by type followed by 7 days of HLM006474 treatment, with each cluster normalized to corresponding vehicle control. Each bar in the graph represents the average of n = 3 biological replicates treated with HLM006474 as a ratio of the average of n = 3 DMSO-treated biological replicates. Data represented as mean±SE. A one-tailed, two-sample equal variance t test was used. b Representative images of GSCs at ×10 magnification (brightfield), sorted by type and treated in HLM006474 for 7 days (images correspond to 7a). n = 2 biologically independent sphere forming experiments were performed. Scale bar: 400 μm. c Forest plot showing the odds ratio of forming a tumor sphere >65 mm following 7 days of HLM006474 treatment, calculated using a multivariate logistic regression with the astrocytic GSC type as a reference, controlled for patient cell line. There was no significant difference between the two GSC lines (p > 0.2), odds ratio with 95% confidence intervals are shown. d Bioluminescence images and e signals from representative mice treated with 20 mm HLM006474 vs DMSO with corresponding f Kaplan−Meier survival plot (n = 16, eight per group). Data are represented as mean±SE. g Box−whisker plot showing the proportion of viable unsorted GSCs (BT326-GSC) after one of the following treatments: 6 days of TMZ treatment, 6 days of HLM006474 treatment, or 3 days of HLM006474 treatment followed by 3 days of TMZ treatment, normalized to corresponding vehicle control. n = 2 biological replicates per treatment. Box plot represents the first quartile, median, and third quartile with whiskers extending to 1.5 times the interquartile range. The overlaid filled dot-plots represent the mean value per biological replicate per group. P values: TMZ/HLM, 0.004; HLM/combination, 0.01. For all plots, **p < 0.01, *p < 0.05.