High-Throughput 3D Tumor Spheroid Array Platform for Evaluating Sensitivity of Proton-Drug Combinations
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Updated: Oct 6, 2025
Generation of High-Throughput Three-Dimensional Tumor Spheroids for Drug Screening
Published on: September 5, 2018
Dong Woo Lee1,2, Jung Eun Kim2, Ga-Haeng Lee3
1Department of Biomedical Engineering, Konyang University, Daejeon 35365, Korea.
Abstract:
Proton beam therapy (PBT) is a critical treatment modality for head and neck squamous cell carcinoma (HNSCC). However, not much is known about drug combinations that may improve the efficacy of PBT. This study aimed to test the feasibility of a three-dimensional (3D) tumor-spheroid-based high-throughput screening platform that could assess cellular sensitivity against PBT. Spheroids of two HNSCC cell lines-Fadu and Cal27-cultured with a mixture of Matrigel were arrayed on a 384-pillar/well plate, followed by exposure to graded doses of protons or targeted drugs including olaparib at various concentrations. Calcein staining of HNSCC spheroids revealed a dose-dependent decrease in cell viability for proton irradiation or multiple targeted drugs, and provided quantitative data that discriminated the sensitivity between the two HNSCC cell lines. The combined effect of protons and olaparib was assessed by calculating the combination index from the survival rates of 4 × 4 matrices, showing that Cal27 spheroids had greater synergy with olaparib than Fadu spheroids. In contrast, adavosertib did not synergize with protons in both spheroids. Taken together, we demonstrated that the 3D pillar/well array platform was a useful tool that provided rapid, quantitative data for evaluating sensitivity to PBT and drug combinations. Our results further supported that administration of the combination of PBT and olaparib may be an effective treatment strategy for HNSCC patients.
Insights
Proton beam therapy (PBT) combined with olaparib shows promise for head and neck squamous cell carcinoma (HNSCC). A 3D spheroid platform efficiently screened drug combinations, identifying potential synergistic treatments for HNSCC.
Area of Science:
- Oncology
- Radiation Oncology
- Drug Discovery
Background:
- Proton beam therapy (PBT) is a key treatment for head and neck squamous cell carcinoma (HNSCC).
- Optimizing PBT efficacy through drug combinations requires effective screening methods.
- Limited data exists on drug combinations enhancing PBT for HNSCC.
Purpose of the Study:
- To evaluate a 3D tumor spheroid screening platform for assessing cellular sensitivity to PBT.
- To investigate drug combinations with PBT for HNSCC treatment.
- To identify synergistic effects between PBT and targeted drugs.
Main Methods:
- Utilized a 3D tumor spheroid model with HNSCC cell lines (Fadu and Cal27) on a 384-pillar/well plate.
- Exposed spheroids to varying doses of proton irradiation and targeted drugs (olaparib, adavosertib).
- Assessed cell viability using Calcein staining and calculated combination indices for drug synergy.
Main Results:
- The 3D spheroid platform provided quantitative, dose-dependent cell viability data for PBT and drugs.
- Cal27 spheroids demonstrated greater synergy with olaparib combined with PBT compared to Fadu spheroids.
- Adavosertib did not show synergistic effects with PBT in either HNSCC cell line.
Conclusions:
- The 3D pillar/well array platform is effective for rapid, quantitative screening of PBT and drug combination sensitivity.
- Combining PBT with olaparib presents a potentially effective treatment strategy for HNSCC.
- Further research into PBT and olaparib combinations is warranted for HNSCC patients.
