Abstract
Reporter cell lines are a particularly useful tool to screen for the skin sensitization potential of chemicals. Current cell models based on Keap1-Nrf2 mimic induction by conducting antioxidant response element-luciferase plasmids. However, plasmid-based reporters may ignore comprehensive aspects of induction, thus affecting the accuracy of hazard identification. Herein, we developed a novel HaCaT-based reporter system, EndoSens, whereby luciferase was specifically inserted into the cassette for heme oxygenase (decycling) 1 (HMOX1, the most consistent marker induced by skin sensitizers) by CRISPR/Cas9. Testing data from 20 coded substances showed an accuracy of 90%, sensitivity of 91.7%, and specificity of 87.5%, which exceeded the OECD requirement. Among the 35 chemicals examined, predictivity was better than reported for the validated KeratinoSens™. These results indicate that the EndoSens assay could advance the predictivity of skin sensitization, thus making it a promising tool for in vitro skin sensitization testing.
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Acknowledgements
This work was funded by Guangzhou Science and Technology Major Project of Industry University Research Synergetic Innovation (201704020151), the National Training Program of Innovation and Entrepreneurship for Undergraduates (201710561169), the National Natural Science Foundation of China (31871292), and Fundamental Research Funds for the Central Universities (2014ZM0067) from South China University of Technology.
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Zhong, G., Li, H., Bai, J. et al. Advancing the predictivity of skin sensitization by applying a novel HMOX1 reporter system. Arch Toxicol 92, 3103–3115 (2018). https://doi.org/10.1007/s00204-018-2287-8
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DOI: https://doi.org/10.1007/s00204-018-2287-8