Targeting the TERT Promoter Mutation for Treatment of Glioblastoma
Glioblastoma (GB) is a grade IV astrocytoma responsible for nearly 17,000 deaths every year. The current median survival time for GB patients is just 14 months after surgical debulking and standard of care (Radiation + Temozolomide), suggesting that new treatment options are a clinically unmet need for GB patients. TERT is a catalytic subunit of the enzyme telomerase and is crucial to the regulation of telomere production in human cells. The TERT promoter region is mutated in 86% of brain cancer patients, and could be a great target for development of novel therapeutics for treatment of GB. Currently, the pathobiology of the TERT promoter mutation in GB progression is unknown. In this work we query eight long term established glioma cell lines (T98G, U87, U251, SF767, SNB19, U118, A172, and SF767) and two normal brain cell lines (NHA and SVG) to catalog their TERT promoter mutation status, TERT mRNA expression, and TERT protein expression. We hypothesize that the presence of the TERT promoter mutation would be directly correlated with increased mRNA and protein expression. This preliminary cataloging of TERT expression in long term established glioma cell lines and normal brain cell lines will lay the foundation for studying pathobiology of TERT promoter mutation in glioma progression. DNA, RNA, and protein were isolated from each cell line and quantified using nanodrop for quality control. Western blot and qPCR data on SVG, NHA, U87, T98G, and A172 suggest that TERT mRNA expression does not correlate with protein expression and requires further investigation to understand the functionality of TERT enzyme. We will utilize Sanger sequencing to identify TERT promoter mutation in each of the cell line model and TRAP assay to study functionality of TERT enzyme.