Functional drug screening assay reveals potential glioma therapeutics
Christian E Badr1, Thomas Wurdinger, Bakhos A Tannous
1Neuroscience Center, Department of Neurology, Massachusetts General Hospital, Boston, USA.
Abstract:
Here we describe a novel functional screening assay based on bioluminescence monitoring of the naturally secreted Gaussia luciferase (Gluc) in the conditioned medium of cultured cells. Using this assay, we identified small-molecule drugs that sensitized brain tumor cells to the tumor necrosis factor-related apoptosis-inducing ligand-induced cell death. Human glioblastoma multiforme cells were engineered by gene transfer to express Gluc as a reporter for cell viability, which can be monitored over time by bioluminescence measurements using a plate luminometer. We have optimized the Gluc assay for screening and validated it using the National Institute of Neurological Disorders and Stroke (NINDS) custom collection II library consisting of 1,040 drugs and bioactive compounds, most of which are Food and Drug Administration-approved and are able to cross the blood-brain barrier. We found that the cardiac glycosides family sensitized glioblastoma multiforme cells to the tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis. In conclusion, the Gluc secretion assay is a robust tool for functional drug screening and can be applied to many different fields including cancer.
Insights
A novel bioluminescence assay using Gaussia luciferase (Gluc) identified drugs that enhance brain tumor cell death. Cardiac glycosides were found to sensitize glioblastoma cells to apoptosis, offering a new tool for drug discovery.
Area of Science:
- Biochemistry
- Molecular Biology
- Pharmacology
Background:
- Glioblastoma multiforme (GBM) is an aggressive brain tumor with limited treatment options.
- Developing effective drug screening assays is crucial for identifying novel GBM therapies.
- Targeting apoptosis pathways offers a promising strategy for cancer treatment.
Purpose of the Study:
- To develop and validate a novel functional screening assay for drug discovery.
- To identify small-molecule drugs that sensitize glioblastoma cells to apoptosis.
- To utilize Gaussia luciferase (Gluc) secretion as a reporter for cell viability.
Main Methods:
- Engineered human glioblastoma cells to express secreted Gluc reporter.
- Monitored Gluc activity in conditioned medium using bioluminescence.
- Validated the assay with the NINDS custom collection II library (1,040 compounds).
Main Results:
- Identified small-molecule drugs that sensitize glioblastoma cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis.
- Cardiac glycosides were found to be effective sensitizers.
- The Gluc secretion assay demonstrated robustness and efficiency in drug screening.
Conclusions:
- The Gluc secretion assay is a versatile and robust tool for functional drug screening.
- This assay can be applied to various research fields, including cancer drug discovery.
- The findings highlight the potential of cardiac glycosides in sensitizing glioblastoma to apoptosis.
