Pediatric brain cancer medulloblastoma (MB) standard-of-care results in numerous comorbidities. MB is comprised of distinct molecular subgroups. Group 3 molecular subgroup patients have the highest relapse rates and after standard-of-care have a 20% survival. Group 3 tumors have high expression of GABRA5, which codes for the α5 subunit of the γ-aminobutyric acid type A receptor (GABA_AR). We are advancing a therapeutic approach for group 3 based on GABA_AR modulation using benzodiazepine-derivatives.

We performed analysis of GABR and MYC expression in MB tumors and used molecular, cell biological, and whole-cell electrophysiology approaches to establish presence of a functional 'druggable' GABA_AR in group 3 cells.

Analysis of expression of 763 MB tumors reveals that group 3 tumors share high subgroup-specific and correlative expression of GABR genes, which code for GABA_AR subunits α5, β3 and γ2 and 3. There are ~ 1000 functional α5-GABA_ARs per group 3 patient-derived cell that mediate a basal chloride-anion efflux of 2 × 10⁹ ions/s. Benzodiazepines, designed to prefer α5-GABA_AR, impair group 3 cell viability by enhancing chloride-anion efflux with subtle changes in their structure having significant impact on potency. A potent, non-toxic benzodiazepine ('KRM-II-08') binds to the α5-GABA_AR (0.8 µM EC₅₀) enhancing a chloride-anion efflux that induces mitochondrial membrane depolarization and in response, TP53 upregulation and p53, constitutively phosphorylated at S392, cytoplasmic localization. This correlates with pro-apoptotic Bcl-2-associated death promoter protein localization.

GABRA5 expression can serve as a diagnostic biomarker for group 3 tumors, while α5-GABA_AR is a therapeutic target for benzodiazepine binding, enhancing an ion imbalance that induces apoptosis.