Celebrating Scholarship and Creativity Day (2018-)

Document Type

Presentation

Publication Date

4-30-2026

Disciplines

Biochemistry, Biophysics, and Structural Biology

Advisor

Claire Otteson

Abstract

Glioblastoma is the most common malignant primary brain tumor in adults, with a median survival of 15 months post-diagnosis. Resistance to standard temozolomide (TMZ) chemotherapy is common and is strongly influenced by O6-methylguanine-DNA methyltransferase (MGMT), a DNA repair enzyme that reverses TMZ induced DNA damage. Reduced MGMT expression is associated with greater TMZ sensitivity and improved survival. Although MGMT promoter cytosine-phosphate-guanine dinucleotide (CpG) methylation is linked to treatment response, the individual CpG sites responsible for MGMT silencing remain unclear. This research aims to identify which sites are necessary to suppress MGMT expression and increase TMZ sensitivity. This project will map MGMT promoter CpG methylation patterns using bisulfite sequencing, PCR, and Sanger sequencing to identify which CpG sites directly control MGMT gene silencing in glioblastoma. CRISPR mediated epigenetic editing will be used to methylate specific CpG sites and measure resulting changes in MGMT mRNA and protein expression. Finally, TMZ dose response assays will be conducted to calculate IC50 values and determine whether targeted methylation is sufficient to increase TMZ sensitivity. The novelty of this proposal lies in experimentally testing causal CpG specific methylation effects rather than relying on promoter wide correlation studies. These results may refine MGMT biomarker precision and improve prediction of patient response to TMZ therapy. Long term, identifying CpG sites that directly regulate MGMT expression could support targeted epigenetic strategies to sensitize resistant glioblastoma tumors. Future work could expand this approach to investigate other regulatory elements outside of the promoter that also influence MGMT expression.

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