Transcriptomic and Metabolic Alterations by an Anti-epileptic Agent Elucidate Mechanistic Efficacy in Glioblastoma Cells

Abstract ID 129319 Poster Board 260 Glioblastoma Multiforme (GBM) is a highly aggressive malignant primary brain tumor with an overall median survival of merely 15 months. GBM exhibits a poor prognosis with existing therapeutics. We targeted the metabolic reprogramming in GBM to overcome challenges such as limited brain permeation and chemotherapeutic resistance. Our investigation led to the identification of Stiripentol (STP), an FDA-approved anti-epileptic drug. Our previous study demonstrated the efficacy of STP in multiple GBM cells and GBM 3D spheroids by employing various cancer biology assays. However, STP's mechanism of action in GBM is not known. Although STP's anti-epileptic activity is known to involve GABAergic potentiation, some studies have indicated it to be a lactate dehydrogenase (LDH) inhibitor. We hypothesized that metabolic and transcriptional changes underlie the effects of STP in GBM cells. We performed a comprehensive transcriptomic analysis on U87 GBM cells to elucidate mechanistic gene regulation upon treatment with STP. Additionally, metabolic alterations by STP in GBM cells were studied using the Seahorse assay for its effect on the Extra Cellular Acidification Rate (ECAR). Our transcriptomic analysis includes significant signatures of RNA seq analysis by differential gene regulation, facilitated by Illumina DRAGEN RNA pipeline and Illumina DRAGEN Differential Gene Expression DEseq2. Further, molecular and pathway analysis was performed using Panther DB and DAVID bioinformatics software. We have identified 11 significant differentially expressed genes (DEGs) between control and STP-treated U87 cells (ALDH1A3, SERPINB2, FGF1, RN7SL2, AOX1, GRB14, RRNAD1, TG, DGKA, ITGA11, and CHD2). Our bioinformatic analysis identified genes involved in metabolic, angiogenic, and p53 feedback loop pathways. Additionally, the Seahorse assay depicted a decreased glycolytic rate in STP-treated U87 cells. In conclusion, our study revealed transcriptional and metabolic alterations in U87 GBM cells, providing significant insights into the anticancer activity of STP. Ongoing studies involving silencing and overexpression experiments and protein expression of the signaling pathways will aid in further resolving mechanistic details. Our research can unveil the true potential and mechanistic insights of the anti-epileptic drug STP for GBM therapy. This research was supported by an award from the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R16GM145557 and by funds from the College of Pharmacy and Health Sciences, St. John's University, NY.