ClonoScreen3D - A Novel 3-Dimensional Clonogenic Screening Platform for Identification of Radiosensitizers for
Mark R Jackson1, Amanda R Richards1, Abdul-Basit Ayoola Oladipupo1
1Wolfson Wohl Cancer Research Centre, School of Cancer Sciences, University of Glasgow, Glasgow, UK.
Purpose:
Glioblastoma (GBM) is a lethal brain tumor. Standard-of-care treatment comprising surgery, radiation, and chemotherapy results in median survival rates of 12 to 15 months. Molecular-targeted agents identified using conventional 2-dimensional (2D) in vitro models of GBM have failed to improve outcome in patients, rendering such models inadequate for therapeutic target identification. A previously developed 3D GBM in vitro model that recapitulates key GBM clinical features and responses to molecular therapies was investigated for utility for screening novel radiation-drug combinations using gold-standard clonogenic survival as readout.
Methods And Materials:
Patient-derived GBM cell lines were optimized for inclusion in a 96-well plate 3D clonogenic screening platform, ClonoScreen3D. Radiation responses of GBM cells in this system were highly reproducible and comparable to those observed in low-throughout 3D assays. The screen methodology provided quantification of candidate drug single agent activity (half maximal effective concentration or EC50) and the interaction between drug and radiation (radiation interaction ratio).
Results:
The poly(ADP-ribose) polymerase inhibitors talazoparib, rucaparib, and olaparib each showed a significant interaction with radiation by ClonoScreen3D and were subsequently confirmed as true radiosensitizers by full clonogenic assay. Screening a panel of DNA damage response inhibitors revealed the expected propensity of these compounds to interact significantly with radiation (13/15 compounds). A second screen assessed a panel of compounds targeting pathways identified by transcriptomic analysis and demonstrated single agent activity and a previously unreported interaction with radiation of dinaciclib and cytarabine (radiation interaction ratio 1.28 and 1.90, respectively). These compounds were validated as radiosensitizers in full clonogenic assays (sensitizer enhancement ratio 1.47 and 1.35, respectively).
Conclusions:
The ClonoScreen3D platform was demonstrated to be a robust method to screen for single agent and radiation-drug combination activity. Using gold-standard clonogenicity, this assay is a tool for identification of radiosensitizers. We anticipate this technology will accelerate identification of novel radiation-drug combinations with genuine translational value.
Insights
A new 3D cell model, ClonoScreen3D, effectively screens for glioblastoma (GBM) radiation-drug combinations. This platform identifies potent radiosensitizers, accelerating the development of novel GBM treatments.
Area of Science:
- Oncology
- Cancer Research
- 3D Cell Culture Models
Background:
- Glioblastoma (GBM) is an aggressive brain tumor with poor prognosis.
- Conventional 2D cell models inadequately predict therapeutic responses in GBM.
- Standard treatments offer limited survival benefits, necessitating improved therapeutic strategies.
Purpose of the Study:
- To evaluate a 3D glioblastoma (GBM) in vitro model, ClonoScreen3D, for screening novel radiation-drug combinations.
- To identify potential radiosensitizing agents for GBM treatment using clonogenic survival assays.
- To assess the utility of 3D models for predicting clinical efficacy of GBM therapies.
Main Methods:
- Patient-derived GBM cell lines were adapted to a 96-well plate 3D clonogenic screening platform (ClonoScreen3D).
- The platform quantified single-agent drug activity (EC50) and drug-radiation interactions (radiation interaction ratio).
- Radiation responses were validated using gold-standard clonogenic survival assays.
Main Results:
- ClonoScreen3D demonstrated reproducible and comparable radiation responses to traditional 3D assays.
- Poly(ADP-ribose) polymerase (PARP) inhibitors (talazoparib, rucaparib, olaparib) showed significant radiation interaction and were confirmed radiosensitizers.
- Dinaciclib and cytarabine exhibited novel radiosensitizing effects, validated by clonogenic assays.
Conclusions:
- The ClonoScreen3D platform is a robust tool for screening single-agent and radiation-drug combination therapies.
- This 3D assay effectively identifies radiosensitizers with potential clinical relevance.
- The technology is expected to expedite the discovery of novel, translationally valuable radiation-drug combinations for GBM.
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