AccScience Publishing / JBM / Online First / DOI: 10.14440/jbm.2025.0080
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RESEARCH ARTICLE

An organotypic model for investigating drug-radiation responses in the lung

Maryam Alkadhimi1 ,  Anuradha Helen Manne1 ,  Yanyan Jiang1 ,  Marcus Green1 ,  Anderson Joseph Ryan1,2*
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1 Department of Oncology, University of Oxford, Oxford, OX3 7DQ, United Kingdom
2 FastBiopharma, Watlington, OX49 5SW, United Kingdom
JBM, e99010041 https://doi.org/10.14440/jbm.2025.0080
Submitted: 6 September 2024 | Revised: 25 October 2024 | Accepted: 5 November 2024 | Published: 28 November 2024
© 2024 by the Journal of Biological Methods published by POL Scientific. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Background: Established in vivo radiobiological models are commonly used to assess anti-tumor effects and normal tissue toxicity. However, these models have notable limitations, and additional models are necessary to gain a deeper insights into drug-radiation interactions. Objective: This study aimed to develop an organotypic ex vivo model by using precision-cut lung slices (PCLSs) to evaluate radiation-induced residual deoxyribonucleic acid (DNA) damage, both alone and in combination with a pharmacological inhibitor of DNA double-strand break (DSB) repair. Methods: Left lungs from female C57BL/6 mice were dissected, perfused with 4% low-gelling-temperature agarose, and sliced into 250 μm sections. Lung slices were then incubated ex vivo for up to 7 days. The slices were irradiated using 137Cs, either with or without a DNA-dependent protein kinase (DNA-PK) inhibitor (NU7441). Tissue sections were subsequently fixed and stained for γH2AX and 53BP1, which serve as histological markers of DNA DSBs. Results: The established conditions preserved tissue viability for up to 7 days and maintained structural integrity for 2 days. DNA damage, detected through γH2AX and 53BP1 staining, was consistent between lungs irradiated ex vivo and their counterparts irradiated in vivo. In the organotypic model, radiation alone in DNA-PK-deficient SCID mice and radiation combined with DNA-PK inhibition in C57BL/6 mice led to increased residual γH2AX and 53BP1 staining. Conclusion: This study demonstrates that residual DNA damage levels following ionizing radiation in lung tissue are comparable between in vivo and ex vivo tissue slices, suggesting that PCLSs serve as a valuable organotypic model for investigating the effects of drug-radiation combinations.

Keywords
DNA double-strand breaks
Lung
DNA-dependent protein kinase
Organotypic
Ionizing radiation
DNA repair
Funding
This work was supported by UK Medical Research Council grant MC_PC_12006. In addition, MA was supported by UK Medical Research Council award 1963111.
Conflict of interest
The authors declare that they have no competing interests.
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Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TBA, Published by POL Scientific
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