Event Title
Optimization of a Chemical Genetic Screen for Sensitization to Laromustine
Location
Diamond 145
Start Date
1-5-2014 1:00 PM
End Date
1-5-2014 2:30 PM
Project Type
Presentation- Restricted to Campus Access
Description
Laromustine is an experimental chemotherapeutic prodrug that breaks down into two active components; a chloroethylating component that causes DNA interstrand crosslinks and a carbamoylating component whose biological activity is less understood. Laromustine has been used in clinical trials to treat acute myeloid leukemia and glioblastoma multiforme (GBM). Unfortunately, the GBM clinical trials have had limited success despite the ability of laromustine to cross the blood-brain barrier and its low non-specific toxicity. GBM is the most common type of malignant brain tumor, and patients currently have a mean survival time of only 13 months. As such, finding a way to increase the potency of laromustine against GBM cells would be invaluable. To learn more about the mechanism of laromustine and investigate potential partner compounds that will sensitize GBM cells to laromustine, a chemical genetic screen will be conducted. The screen will evaluate the ability of small molecules in a library of approximately 450 FDA-approved compounds to enhance the anti-proliferative effects of laromustine in U138 GBM cells. It will be conducted using a high-throughput proliferation assay with a fluorescent readout performed in 384-well plates. Optimization of the assay has included determining the proper cell seed density, drug concentration and incubation time, and fluorescent substrate concentration, among other variables. These optimization data will allow for the facile and accurate identification of partner candidates for laromustine when the screen is conducted.
Faculty Sponsor
Julie Millard
Sponsoring Department
Colby College. Chemistry Dept.
CLAS Field of Study
Natural Sciences
Event Website
http://www.colby.edu/clas
ID
447
Optimization of a Chemical Genetic Screen for Sensitization to Laromustine
Diamond 145
Laromustine is an experimental chemotherapeutic prodrug that breaks down into two active components; a chloroethylating component that causes DNA interstrand crosslinks and a carbamoylating component whose biological activity is less understood. Laromustine has been used in clinical trials to treat acute myeloid leukemia and glioblastoma multiforme (GBM). Unfortunately, the GBM clinical trials have had limited success despite the ability of laromustine to cross the blood-brain barrier and its low non-specific toxicity. GBM is the most common type of malignant brain tumor, and patients currently have a mean survival time of only 13 months. As such, finding a way to increase the potency of laromustine against GBM cells would be invaluable. To learn more about the mechanism of laromustine and investigate potential partner compounds that will sensitize GBM cells to laromustine, a chemical genetic screen will be conducted. The screen will evaluate the ability of small molecules in a library of approximately 450 FDA-approved compounds to enhance the anti-proliferative effects of laromustine in U138 GBM cells. It will be conducted using a high-throughput proliferation assay with a fluorescent readout performed in 384-well plates. Optimization of the assay has included determining the proper cell seed density, drug concentration and incubation time, and fluorescent substrate concentration, among other variables. These optimization data will allow for the facile and accurate identification of partner candidates for laromustine when the screen is conducted.
https://digitalcommons.colby.edu/clas/2014/program/217