Exploration of the GARDskin applicability domain: Indirectly acting haptens, hydrophobic substances and UVCBs

Main Article Content

Andy Forreryd
Robin Gradin
Charles Humfrey
Len Sweet
Henrik Johansson

Abstract

Hazard assessments of skin sensitizers are increasingly being performed using new approach methodologies (NAMs), with several in chemico, in vitro and most recently also defined approaches (DAs) being accepted for regulatory use. However, keeping track of potential limitations of each method in order to define applicability domains remains a crucial component to ensure adequate predictivity as well as facilitating the appropriate selection of method(s) for each hazard assessment task. The objective of this report is to share test results generated with the GARD™skin assay on chemicals that have traditionally been considered as difficult to test in some of the conventional in vitro and in chemico OECD Test Guidelines for skin sensitization. Such compounds may include, for example, indirectly acting haptens, hydrophobic substances, and substances of unknown variable composition or biological substances (UVCBs). Based on the results of this study, the sensitivity for prediction of skin sensitizing hazard of indirectly acting haptens was 92.4% and 87.5%, when compared with LLNA (n=25) and human data (n=8), respectively. Similarly, the sensitivity for prediction of skin sensitizing hazard of hydrophobic substances was 85.1% and 100%, when compared with LLNA (n=24) and human data (n=9), respectively. Lastly, a case study involving assessment of a set of hydrophobic UVCBs (n=7) resulted in a sensitivity of 100, as compared to available reference data. Thus, it was concluded that these data provide support for the inclusion of such chemistries in the GARD™skin applicability domain, without an increased risk of false negative classifications.

Article Details

How to Cite
Forreryd, A., Gradin, R., Humfrey, C. ., Sweet, L. and Johansson, H. (2022) “Exploration of the GARDskin applicability domain: Indirectly acting haptens, hydrophobic substances and UVCBs”, ALTEX - Alternatives to animal experimentation. doi: 10.14573/altex.2201281.
Section
Articles
References

Basketter, D. A., Alepee, N., Ashikaga, T. et al. (2014). Categorization of chemicals according to their relative human skin sensitizing potency. Dermatitis 25, 11-21. doi:10.1097/DER.0000000000000003

Belot, N., Sim, B., Longmore, C., Roscoe, L. and Treasure, C. (2017). Adaptation of the KeratinoSens™ skin sensitization test to animal-product-free cell culture. ALTEX 34, 560-564. doi:10.14573/altex.1701311

Cooper, J. A., Saracci, R. and Cole, P. (1979). Describing the validity of carcinogen screening tests. British Journal of Cancer 39,87-9. doi:10.1038/bjc.1979.10

Corsini, E., Clewell, R., Cotgreave, I., Eskes, C., Kopp-Schneider, A., Westmoreland, C., Alves, P.M., Navas, J.M. and Piersma, A. (2021). ESAC Opinion on the Scientific Validity of the GARDskin and GARDpotency Test Methods. Asturiol Bofill, D., Casati, S. and Viegas Barroso, J.F. editor(s), Publications Office of the European Union, Luxembourg, 2021. doi:10.2760/626728

Edwards, A., Roscoe, L., Longmore, C., Bailey, F., Sim, B. and Treasure, C. (2018). Adaptation of the human Cell Line Activation Test (h-CLAT) to Animal-Product-Free Conditions. ALTEX 35, 477-488. doi:10.14573/altex.1710051

EURL ECVAM - EU Reference Laboratory for alternatives to animal testing (2021). GARDskin Assay Protocol. https://tsar.jrc.ec.europa.eu/system/files/Published/GARDskin%20Assay%20Protocol%20TSAR.pdf

Forreryd, A., Zeller, K. S., Lindberg, T. et al. (2016). From genome-wide arrays to tailor-made biomarker readout - Progress towards routine analysis of skin sensitizing chemicals with GARD. Toxicol In Vitro 37, 177-88. doi:10.1016/j.tiv.2016.09.013

Gradin, R., Lindstedt, M. and Johansson, H. (2019). Batch adjustment by reference alignment (BARA): Improved prediction performance in biological test sets with batch effects. PLoS One 14, e0212669. doi:10.1371/journal.pone.0212669

Gradin, R., Forreryd, A., Mattson, U., et al. (2021). Quantitative Assessment of Sensitizing Potency using a dose-response adaptation of GARDskin. Sci Rep 176, 423-432. doi:10.1038/s41598-021-98247-7

Johansson, H., Lindstedt, M., Albrekt, A. S. and Borrebaeck, C. A. (2011). A genomic biomarker signature can predict skin sensitizers using a cell-based in vitro alternative to animal tests. BMC Genomics 12, 399. doi:10.1186/1471-2164-12-399

Johansson, H., Gradin, R., Forreryd, A., Agemark, M. et al. (2017). Evaluation of the GARD assay in a blind Cosmetics Europe study. ALTEX 34, 515-523. doi:10.14573/altex.1701121

Johansson, H., Gradin, R., Johansson, A., et al. (2019). Validation of the GARDskin Assay for Assessment of Chemical Skin Sensitizers: Ring Trial Results of Predictive Performance and Reproducibility. Toxicol Sci 170, 374-381. doi:10.1093/toxsci/kfz108

Mehling, A., E. Adriaens, S. Casati, B. et al. (2019). In vitro RHE skin sensitisation assays: Applicability to challenging substances." Regul Toxicol Pharmacol 108, 104473. doi:10.1016/j.yrtph.2019.104473

OECD - Organisation for Economic Cooperation and Development (2014). The Adverse Outcome Pathway for Skin Sensitisation Initiated by Covalent Binding to Proteins. Part 1: Scientific Evidence. Series on Testing and Assessment No. 168. Available at: doi:10.1787/9789264221444-en

OECD - Organisation for Economic Cooperation and Development (2015a). OECD Guideline for the Testing of Chemicals No. 442C: In Chemico Skin Sensitisation: Direct Peptide Reactivity Assay (DPRA). Paris, France: Organisation for Economic Cooperation and Development. doi:10.1787/9789264229709-en

OECD - Organisation for Economic Cooperation and Development (2015b). OECD Guideline for the Testing of Chemicals No. 442D: In Vitro Skin Sensitisation: ARE-Nrf2 Luciferase Test Method. Paris, France: Organisation for Economic Cooperation and Development. doi:10.1787/9789264229822-en

OECD - Organisation for Economic Cooperation and Development (2015c). Performance standards for assessment of proposed similar or modified in vitro skin sensitisation ARE-NRF2 luciferase test methods. Series on Testing and Assessment, No. 213. Paris, France: Organisation for Economic Cooperation and Development. doi:10.1787/9789264229822-en

OECD - Organisation for Economic Cooperation and Development (2017a). OECD Guideline for the Testing of Chemicals No. 442E: In Vitro Skin Sensitisation assays addressing the AOP Key Event on Activation of Dendritic Cells. Organisation for Economic Cooperation and Development, Paris. doi:10.1787/9789264264359-en

OECD - Organisation for Economic Cooperation and Development (2017b). Revised performance standards for the assessment of proposed similar or modified in vitro Reconstructed human Cornea-like Epithelium (RhCE) test methods for eye hazard. Series on Testing and Assessment, No. 216. Paris, France: Organisation for Economic Co-operation and Development. Available at: https://www.oecd.org/officialdocuments/publicdisplaydocumentpdf/?cote=ENV/JM/MONO(2015)23&doclanguage=en

OECD - Organisation for Economic Cooperation and Development (2021a). OECD Guideline for the Testing of Chemicals No. 497: Defined Approaches on Skin Sensitisation. Organisation for Economic Cooperation and Development, Paris. doi:10.1787/b92879a4-en

OECD - Organisation for Economic Cooperation and Development (2021b). Series on Testing and Assessment No. 336: Annex 2 of the Supporting document to the Guideline (GL) on Defined Approaches (DAs) for Skin Sensitisation. Organisation for Economic Cooperation and Development, Paris. Available at: http://www.oecd.org/env/testguidelines

Patlewicz, G., S. Casati, D. A. Basketter, D. et al. (2016). Can currently available non-animal methods detect pre and pro-haptens relevant for skin sensitization? Regul Toxicol Pharmacol 82, 147-155. doi:10.1016/j.yrtph.2016.08.007

Takenouchi, O., Miyazawa, M., Saito, K., et al. (2013). Predictive performance of the human Cell Line Activation Test (h-CLAT) for lipophilic chemicals with high octanol-water partition coefficients. J Toxicol Sci 38, 599-609. doi:10.2131/jts.38.599

UN GHS - United Nations, Globally Harmonized System of Classification and Labelling of Chemicals 2015. Sixth revised edition. New York & Geneva: United Nations Publications. doi:10.18356/591dabf9-en

Urbisch, D., M. Becker, N. Honarvar, S. N. et al. (2016). Assessment of Pre- and Pro-haptens Using Nonanimal Test Methods for Skin Sensitization. Chem Res Toxicol 29, 901-13. doi:10.1021/acs.chemrestox.6b00055.

Most read articles by the same author(s)