Date of Award

2014

Document Type

Honors Thesis (Open Access)

Department

Colby College. Chemistry Dept.

Advisor(s)

Kevin P. Rice

Abstract

Laromustine is an experimental sulfonylhydrazine prodrug used in late-stage clinical studies against acute myeloid leukemia (AML) and glioblastoma multiforme (GBM). Despite initial promise for both indications, clinical trials for GBM have not been as successful as those for AML. To investigate methods for improving the effectiveness of laromustine in GBM and to learn more about the mechanism of action of laromustine, a chemical genetic screen will be conducted to identify agents that sensitize GBM cells to the anti-proliferative effects of laromustine. The library, which will include approximately 450 FDA-approved drugs, will be screened using a newly optimized high throughput assay based on the Click-iT EdU Microplate Assay kit (Molecular Probes). Optimization of the assay has required determining the proper cell seed density, drug concentration and incubation time, and fluorescent substrate concentration, among other variables. It was determined that low cell seed densities allow for maximal proliferation and a high signal-to-noise ratio. Furthermore, 50 μM laromustine was found to have little inhibitory effect on the proliferation of U138 cells, while higher laromustine concentrations yielded a sharp decrease in proliferation. These results suggest that reduced proliferation of cells exposed to 50 μM laromustine in combination with library compounds is a suitable marker for sensitization to laromustine. With these optimization data, a chemical screen can now be conducted, potentially revealing new therapeutic strategies to treat GBM.

Keywords

laromustine, cloretazine, glioblastoma, chemical screen, click chemistry, cell proliferation

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Biochemistry Commons

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