Article
Large-scale drug screen in patient-derived IDHmut glioma stem cells identifies several FDA-approved antineoplastic agents
Hochdurchsatz- Medikamentenscreen in IDHmut Gliom-Stammzellen identifiziert multiple FDA-zugelassene Chemotherapeutika
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Authors
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Philip Dao Trong
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
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Gerhard Jungwirth
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
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Tao Yu
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
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Stefan Pusch
- Universitätsklinikum Heidelberg, Neuropathologisches Institut, Heidelberg, Deutschland
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Christel Herold-Mende
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
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Rolf Warta
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
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Andreas W. Unterberg
- Universitätsklinikum Heidelberg, Sektion Experimentelle Neurochirurgische Forschung, Heidelberg, Deutschland
| Published: | June 26, 2020 |
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Outline
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Objective: The discovery of the Isocitrate Dehydrogenase (IDH) mutation in glioma has led to a paradigm shift on how we see glioma biology. While it is clear that IDH mutated (IDHmut) and wildtype (IDHwt) tumors have to be viewed as separate entities, the underlying biological differences are still matter of extensive research. Difficulties in cultivating IDHmut glioma stem cells (GSC) have led to a paucity of preclinical models in IDHmut glioma making the discovery of new effective chemotherapeutic agents problematic. We therefore sought to perform a repurposing drug screen in six patient-derived IDHmut GSC lines to discover potential effective antineoplastic agents, already approved by the FDA.
Methods: Patient tumor tissue was obtained in our neurosurgical department to isolate and establish IDHmut GSC lines. (D)-2-hydroxyglutarate (2HG) levels were measured in the cell culture supernatant of IDH1mut GSCs using an enzymatic diaphorase/resazurin system. Intracranial xenotransplantation was performed in NOD/SCID mice for two IDHmut GSCs. The drug libraryconsistedof 146 FDA-approved drugs. Cells were cultured as neurospheres and subjected to the test compounds for 72h in concentrations ranging from 0.1nM – 1µM. Cell viability was assessed with the CellTiterGlo assay (Promega) and apoptosis was analyzed by Annexin V FACS staining.
Results: Despite several passages NCH551b, NCH1681 showed a stable production of 2HG and a robust take rate in xenotransplanted NOD/SCID mice and were therefore consideredsuitable for the drug screen. Among the 147 FDA-approved drugs seven compounds with a half maximal inhibitory concentration (IC50) below 1µMm were identified in two IDHmut GSC lines (NCH551b, NCH1681).The cytotoxic potential could be confirmed in additional four IDHmut GSC lines (NCH612, NCH620, NCH645, NCH3763). Annexin V FACS stainings showed that the rate of cells undergoing apoptosis was significantly elevated in cells treated with the candidate drugs.
Conclusion: In this study, we present a feasible preclinical model for a high-throughput drug screen in patient-derived IDHmut GSCs and identified seven FDA-approved antineoplastic agents with a high cytotoxic potential which warrant further investigations.
