Investigating the role of N6-methyladenosine in the development of drug resistance in glioblastoma - PROJECT SUMMARY RNA chemical modifications are critical regulators of gene expression through regulation of RNA stability, splicing, translation, structure, and localization. Because of their important roles in several biological pathways, RNA modifications are frequently implicated in disease. N6-methyladenosine (m6A), the most prevalent internal RNA modification in eukaryotic messenger RNA (mRNA), has been extensively studied and implicated in a wide-array of human diseases, most notably cancer. Due to the established role of m6A in cancer, m6A and its associated machinery have been proposed as potential diagnostic and predictive biomarkers and therapeutic targets. Although the role of m6A in several cancer is relatively well-established, its regulatory role in glioblastoma (GBM) pathogenesis remains largely unclear. GBM, a grade IV glioma tumor, is the most common primary malignant brain tumor. GBM is a devastating disease associated with poor patient prognosis, limited therapeutic options, and frequent recurrence with acquired therapy resistance. Because m6A has been extensively demonstrated to play an important regulatory role in cancer, the goal of my proposal is to characterize molecular mechanisms by which m6A regulates GBM pathogenesis, and uncover how manipulation of m6A regulation may improve GBM treatment. The first portion of my research will examine the mechanism by which m6A post-transcriptionally regulates MGMT, a critical biomarker and predictor of chemotherapy response in GBM. To accomplish this, I will map m6A sites on MGMT mRNA, determine how disrupting m6A installation on MGMT affects MGMT expression and stability, and identify reader proteins which facilitate this effect. I will then determine the consequences of this regulatory mechanism on chemoresistance of glioma cells. In the second portion of my research, building upon preliminary RNA sequencing data, I will investigate mechanisms transcriptome-wide as to how m6A regulates the transcriptome of glioma cells in response to chronic treatment with an alkylating agent. To accomplish this, in chemoresistant glioma cells, I will (1) map m6A sites, (2) temporally sequence RNAs to measure the effect of m6A perturbation on RNA stability, and (3) sequence the interactome of canonical downstream regulators m6A-modified transcripts. Together, these complementary studies will increase our understanding of mechanisms by which m6A regulates GBM pathogenesis and reveal vulnerabilities that can be targeted with m6A-mediated therapeutics.
