ETMM-07. HYPOXIC REGULATION OF METABOLIC AND STRUCTURAL GENES IN T98 GLIOBLASTOMA MULTIFORME CELLS BY RNA SEQUENCING

Glioblastoma multiforme (GBM) is the most common primary brain cancer and carries a very poor prognosis. The GBM tumor microenvironment is characterized by regions of profound hypoxia, which are associated with a variety of alterations in gene expression that confer survival, proliferation, and resistance to therapy. Multiple mechanisms have been implicated in hypoxia-associated GBM behavior including upregulation of pathways involved in angiogenesis, immunosuppression, and glucose metabolism. Our study aimed to identify changes in gene expression induced by hypoxia among T98G cells via total RNA sequencing. Human T98 GBM cell lines were cultured in a humidified incubator at 37° C and 5% CO(2) and were grown in normoxia (21% O(2)) or hypoxia (95% N(2), 5% C0(2)) for 72 hours. Total RNA was harvested, and global gene expression was evaluated via total RNA sequencing. Standard bioinformatics analysis was performed to identify changes in expression associated with hypoxia. Hypoxia in T98 cells led to significant upregulation of genes implicated in canonical glycolysis, focal adhesion, extracellular matrix reorganization, and endoplasmic reticulum-associated protein processing. We document 690 genes and 11 associated KEGG pathways that demonstrated significant enrichment (p ≤0.01 with Bonferroni, Benjamini, and False Discovery Rate corrections) induced by hypoxia. Notably, upregulation of the IRE1-mediated unfolded protein response was observed. DrugBank database analysis identified four molecules targeting genes upregulated in hypoxic T98G cells: tenecteplase (p = 0.013, 5 gene targets), succinic acid (p = 0.02, 7 targets), artenimol (p = 0.013, 13 targets), and copper (p = 0.0015, 22 targets). We document 733 genes and 6 associated KEGG pathways significantly downregulated (p ≤0.01) in hypoxia, including genes associated with DNA replication and repair, mitotic processes, and spliceosome function. Total RNA sequencing showed hypoxic upregulation of genes involved in various pathways associated with neoplastic GBM behavior and identified multiple candidate molecules which may hold therapeutic potential.