Lack of constitutively active DNA repair sensitizes glioblastomas to Akt inhibition and induces synthetic lethality with radiation treatment in a p53-dependentmanner

Treatment refractory glioblastoma (GBM) remains a major clinical problem globally and targeted therapies in GBM have not been promising to date. TCGA integrative analysis of GBM reported the striking finding of genetic alterations in the p53 and PI3K pathways in over 80% of GBMs. Given the role of these pathways in making cell-fate decisions and responding to genotoxic stress, we investigated the reliance of these two pathways in mediating radiation-resistance. We selected a panel of GBM cell lines and glioma stem cells (GSC) with wild-type TP53 (p53-wt) and mutant TP53, mutations known to interfere with p53 functionality (p53-mt). Cell lines were treated with a brain permeable inhibitor of P-Akt (ser473), phosphatidylinositol ether lipid analogue (PIA), with and without radiation treatment. Sensitivity to treatment was measured using Annexin-V/PI flow cytometry and western blot analysis for the markers of apoptotic signaling, alkaline COMET assay. All results were verified in p53 isogenic cell lines. p53-mt cell lines were selectively radiosensitized by PIA. This radiosensitization effect corresponded with an increase in DNA damage and a decrease in DNA-PKcs levels. TP53 silencing in p53-wt cells showed a similar response as the p53-mt cells. Additionally, the radiosensitization effects of Akt inhibition were not observed in normal human astrocytes suggesting this treatment strategy could have limited off-target effects. We demonstrate that the inhibition of the PI3K/Akt pathway by PIA radiosensitizes p53-mt cells by antagonizing DNA repair. In principle, this strategy could provide a large therapeutic window for the treatment of TP53 mutant tumors.

http://ift.tt/2vu3nCU