Evaluation of Radiotherapy Enhanced by Cherenkov Photo-Activation of Psoralen (AMT)
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PO-GePV-T-74 (Sunday, 7/10/2022) [Eastern Time (GMT-4)]
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Purpose: Radiotherapy Enhanced by Cherenkov photo-Activation (RECA) is a novel radiation treatment that seeks an enhanced anti-cancer effect through the addition of psoralen, which is photo-activated by Cherenkov radiation generated during radiotherapy. This in-vitro work investigates RECA effect with 4T1 cancer cells seeded on live tissue mounted on psoralen-doped agarose.
Methods: Extensive preparatory experiments were performed to validate a novel live tissue assay for exploring RECA effects. In the culminating experiment, three, 12-well plates containing 1 cm agar, 400 μm thick coronal slice of rat brain tissue, were given no treatment, 4.5 Gy of MV radiation of three equal fractions, or 4.5 Gy of kV radiation of three equal fractions. Plates consisted of six wells containing AMT-baked agar and six wells containing standard agar with 10,000 4T1 cells in all wells. Post-irradiation, images were taken each day for four days with a Zeiss Lumar microscope equipped with a rhodamine filter to assess luminescence from each well. A CellProfiler analysis was conducted on each image.
Results: Preparatory work validated the live tissue assay model demonstrating viable growth and psoralen loading of 4T1 cells on the live tissue assay. Measurements of the average luminescence values obtained from the Firefly Luciferase Assay and CellProfiler demonstrated slight improvement in cell proliferation after five days for MV AMT (8.40±0.98-fold growth) and MV AMT fractionated (12.22±0.79-fold growth) when compared to the MV control (13.08±0.83-fold growth). This suggests possible photo-activation of the psoralen taking place and causing a slight decrease in cell viability over time.
Conclusion: A novel live tissue assay was developed and validated for exploring RECA effects in-vitro, with realistic tissue generated Cherenkov light. Procedures developed for RECA showed promising results during preparatory experiments. A significant RECA effect on cell viability was not clearly exhibited. Further investigation is required to confirm and validate the effect.
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