Glioblastoma (GBM) is the most aggressive primary brain tumor with currently no treatment with long-term efficacy. Systematic relapses and therapeutic resistance are partly associated with the presence of glioblastoma stem cells (GSC) within the tumor. Existing options to overcome GSC protection are limited. Characterizing GSC resistance mechanisms appears crucial to identify efficient targeted inhibitors. A GSC model obtained by dedifferentiation of GBM cell lines (dGSC) was previously validated for GSC characteristics. In this study, viability of dGSC after treatment with diversified cytotoxic drugs was compared to parental cell lines. GSC phenotype acquisition is associated with a greatly decreased sensitivity to most cytotoxic drugs. Proteins expression, cell cycle and iron dosage analyses demonstrated that dGSC resistance acquisition is accompanied by decreased proliferation, STAT3 and Akt activation, increased iron storage and protection against oxidative stress. BMI1 and EZH2 epigenetic regulators are associated with resistance in GBM and were expressed in both dGSC and GBM cells. Akt activation in dGSC could redirect EZH2 toward histone methylation independent functions, through S21 phosphorylation. BMI1 and EZH2 inhibition decreased viability in both GBM cells and dGSC. Notably, EZH2 inhibition through GSK343 decreased proliferation and Akt activation, without impacting EZH2-associated epigenetic modification (H3K27me3), but with the induction of a ferroptosis signature. GSK343 effects on viability, proliferation and ferroptosis were confirmed in two lines of patient-derived GSC. GSK343 overcame resistant dGSC protection to trigger cell death by a mechanism involving oxidative stress. To conclude, GSK343-mediated EZH2 inhibition efficiently eliminated resistant GSC through ferroptosis, in a H3K27me3-independent manner. • Glioblastoma stem cells phenotype acquisition leads to therapeutic resistance. • Resistance is linked to low proliferation, ferroptosis defense, Akt and STAT3. • Epigenetic regulators EZH2 and BMI1 are expressed in dGSC and GBM cell lines. • EZH2 inhibition counteracts dGSC protection to trigger oxidative stress and death. • Dedifferentiation redirects EZH2 toward histone independent roles through pS21 form.
Doualle et al. (Thu,) studied this question.