The impact of biostatistics on hazard characterization using in vitro developmental neurotoxicity assays

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Hagen Eike Keßel , Stefan Masjosthusmann, Kristina Bartmann, Jonathan Blum, Arif Dönmez, Nils Förster, Jördis Klose, Axel Mosig, Melanie Pahl, Marcel Leist, Martin Scholze, Ellen Fritsche
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In chemical safety assessment, benchmark concentrations (BMC) and their associated uncertainty are needed for the toxicological evaluation of in vitro data sets. A BMC estimation is derived from concentration-response modelling and results from various statistical decisions, which depend on factors such as experimental design and assay endpoint features. In current data practice, the experimenter is often responsible for the data analysis and therefore relies on statistical software, often without being aware of the software default settings and how they can impact the outputs of data analysis. To provide more insight into how statistical decision-making can influence the outcomes of data analysis and interpretation, we have developed an automated platform that includes statistical methods for BMC estimation, a novel endpoint-specific hazard classification system, and routines that flag data sets that are outside the applicability domain for an automatic data evaluation. We used case studies on a large dataset produced by a developmental neurotoxicity (DNT) in vitro battery (DNT IVB). Here we focused on the BMC and its confidence interval (CI) estimation as well as on final hazard classification. We identified five crucial statistical decisions the experimenter must make during data analysis: choice of replicate averaging, response data normalization, regression modelling, BMC and CI estimation, and choice of benchmark response levels. The insights gained are intended to raise more awareness among experimenters on the importance of statistical decisions and methods but also to demonstrate how important fit-for-purpose, internationally harmonized and accepted data evaluation and analysis procedures are for objective hazard classification.

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Keßel, H. E. (2023) “The impact of biostatistics on hazard characterization using in vitro developmental neurotoxicity assays”, ALTEX - Alternatives to animal experimentation, 40(4), pp. 619–634. doi: 10.14573/altex.2210171.

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