Abstract 6057: Mechanisms mediating the radioresistance of human glioma cells growing in the murine olfactory bulb
Authors: Degorre Charlotte M. , Sutton Ian C. , Lehman Stacey L. , Shankavaram Uma T. , Camphausen Kevin A. , Tofilon Philip J. 
Published: June 2022

We have previously reported (Int J Radiat Oncol Biol Phys. 107(1):194-201, 2020) that human GBM-stem like cells (GSCs) implanted into the right striatum of nude mice migrate/invade throughout the right hemisphere (RH) as well as into the olfactory bulb (OB). The highly invasive nature of the GSCs suggested a model system for evaluating the effects of specific brain microenvironments on glioma radioresponse and the potential for topographic heterogeneity in radioresistance. Subsequent analyses revealed that GSCs in the OB were significantly more radioresistant than those in the RH. Because the OB is highly enriched in neurons and given recent data suggesting that neuron/glioma interactions drive GBM growth, in this study we first defined contact between GSCs and neurons in the OB and RH using immunohistochemistry (IHC) and confocal microscopy. In the OB 56% of tumor cells were in direct contact with neurons as compared to 24% of tumor cells in the RH. Moreover, among the tumor cells in contact with neurons 25% had 2 or more neuronal contacts in the OB versus 2% in the RH. These data suggest a significantly greater level of glioma-neurons interaction in the OB. To further investigate the OB as a radioresistant niche, RNAseq was used to define the transcriptome of NSC11 GSCs located in the OB versus the RH. Of the greater than 15,000 genes evaluated, 1887 genes were differentially expressed between the 2 sites: 872 upregulated and 1015 down regulated in the OB versus the RH. Functional analyses (IPA and GSEA) indicated that among the genes overexpressed in the OB GSCs were a group associated with the cell cycle, specifically mitosis and the spindle assembly checkpoint (SAC). Comparison of the SAC genes overexpressed in the OB with GBM clinical outcomes (TCGA) showed a significant correlation with decreased patient survival. These analyses suggested that cell cycle regulation may play a role in the radioresistance of OB GSCs. Accordingly, radiation-induced cell cycle redistribution of GSCs in the OB and the RH was determined by IHC analysis of cell cycle phase markers and flow cytometry (propidium iodide) performed on dissociated tumor cells. Results showed a significantly higher percentage of cells in G2 in the OB at 24h and 48h after 10Gy, suggestive of prolonged G2 arrest. In addition, analysis of radiation-induced DNA DSBs using γH2AX and 53BP1 nuclear foci showed faster and more complete foci dispersal after 10Gy in the OB GSCs versus those in the RH, indicative of increased DSBs repair in the OB. These data suggest that GSC radioresistance in the OB is the result of a more effective DNA damage response. Moreover, results suggest that the murine OB may provide a model system for investigating GBM radioresistance as well as the potential role of glioma/neuron interactions.

Citation Format: Charlotte M. Degorre, Ian C. Sutton, Stacey L. Lehman, Uma T. Shankavaram, Kevin A. Camphausen, Philip J. Tofilon. Mechanisms mediating the radioresistance of human glioma cells growing in the murine olfactory bulb [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6057.