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Glutamate release by primary brain tumors induces epileptic activity

Abstract

Epileptic seizures are a common and poorly understood comorbidity for individuals with primary brain tumors. To investigate peritumoral seizure etiology, we implanted human-derived glioma cells into severe combined immunodeficient mice. Within 14–18 d, glioma-bearing mice developed spontaneous and recurring abnormal electroencephalogram events consistent with progressive epileptic activity. Acute brain slices from these mice showed marked glutamate release from the tumor mediated by the system xc cystine-glutamate transporter (encoded by Slc7a11). Biophysical and optical recordings showed glutamatergic epileptiform hyperexcitability that spread into adjacent brain tissue. We inhibited glutamate release from the tumor and the ensuing hyperexcitability by sulfasalazine (SAS), a US Food and Drug Administration–approved drug that blocks system xc. We found that acute administration of SAS at concentrations equivalent to those used to treat Crohn's disease in humans reduced epileptic event frequency in tumor-bearing mice compared with untreated controls. SAS should be considered as an adjuvant treatment to ameliorate peritumoral seizures associated with glioma in humans.

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Figure 1: Tumor-bearing mice show abnormal spontaneous EEG events indicative of epileptic activity.
Figure 2: Acute cortical slices from tumor-bearing mice show spontaneous epileptiform activity.
Figure 3: Acute cortical slices from tumor-bearing mice are hyperexcitable.
Figure 4: Cortical slices from glioma-bearing mice show increased cortical network activity and hyperexcitable layer 2 and 3 peritumoral pyramidal cells.
Figure 5: SAS application reduces epileptiform activity in cortical slices from glioma-bearing mice.
Figure 6: Sulfasalazine reduces frequency of epileptic activity in tumor-bearing mice.

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Acknowledgements

The authors would like to thank J. Hablitz and A. Albertson for technical assistance with diode array recordings; A. Margolies for help with histology; C. Langford for orthotopic xenografts; V. Cuddapah for editorial advice; M. McFerrin for technical support, and E. Dudek and K. Wilson at the University of Utah for training in EEG acquisition. We conducted glutamate measurements at the University of Alabama at Birmingham Targeted Metabolomics and Proteomics Facility (funded by the National Center for Research Resources grant S10 RR19231 and US National Institutes of Health grants U54 CA 100949, P50 AT00477, P30 DK079337 and P30 AR50948). We obtained bioluminescence imaging and some field electrode recordings in the Neuroscience Blueprint Core facility (Neuroscience Blueprint Core Grant NS57098). We obtained U251-MG cells from Y. Gillespie (University of Alabama at Birmingham) and U251-MGffluc cells from M. Jensen (City of Hope National Medical Center); GBM12 and GBM22 tumors were obtained from J. Sarkaria (Mayo Clinic) and provided by the University of Alabama at Birmingham Brain Tumor Animal Models Core (UAB SPORE P50-CA097247). This work was supported by US National Institutes of Health grants 2R01-NS052634, 5R01-NS036692 and 5T32NS048039-03.

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Contributions

S.C.B. and S.L.C. acquired the majority of the data presented. B.R.H. was instrumental in mouse surgeries and statistical analyses of data. V.M. (supported by an American Brain Tumor Association Basic Research Fellowship) carried out glutamate release assays. S.R. assisted in electrophysiological recordings. T.O. did western blotting and glutamate uptake assays. H.S. designed experiments, supervised all research and co-wrote the manuscript.

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Correspondence to Harald Sontheimer.

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Buckingham, S., Campbell, S., Haas, B. et al. Glutamate release by primary brain tumors induces epileptic activity. Nat Med 17, 1269–1274 (2011). https://doi.org/10.1038/nm.2453

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