Synthesizing Genotoxicity Results in the MultiFlow® Assay with
Point-of-Departure Analysis and ToxPi Visualization Techniques
Abstract
In vitro genotoxicity has historically served a hazard identification
role, with simple binary outcomes provided for each of several single
endpoint assays. This will need to change, given: i) efforts to curtail
animal testing, ii) the increased use of multiplexed in vitro assays and
the ongoing development of NAMS, and iii) the desire to holistically
consider quantitative results from multiple biomarkers/endpoints that
take potency into consideration. To help facilitate more quantitative
analyses of multiple biomarkers and/or assay streams, we explored the
combined use of PROAST and Toxicological Prioritization Index (ToxPi)
software. As a proof-of-concept, this investigation employed the
MultiFlow® DNA damage assay, focusing on γH2AX and p53 biomarkers at two
time points, whereby 10 genotoxicants were evaluated in the presence and
absence of rat liver S9 metabolic activation. Whereas PROAST was used to
calculate BMD point estimates and confidence intervals (CI), ToxPi
synthesized the BMD results into visual, quantitative summaries
conveying genotoxic potency and metabolic properties. Our analyses
suggest that ToxPi’s data synthesis and visualization modules provide
useful insights into compound potency, chemical grouping, and genotoxic
mechanisms. By integrating multiple data sources, we find that ToxPi
offers a powerful complementary approach to traditional BMD CI potency
graphs, particularly for the simultaneous analysis of multiple
biomarkers enhancing chemical potency analysis of complex datasets.