The future of mutagenesis testing is here

Abstract: Evaluation of a substance’s potential to cause mutagenicity is a critical component of human and environmental health risk assessment. Regulatory agencies worldwide have developed test guidelines to determine whether chemicals cause mutations. While commonly used tests under these guidelines have served the regulatory community well, they measure mutations in one reporter gene and/or require transgenic rodent models and standalone experiments. Duplex Sequencing (DS) is an error-corrected next-generation sequencing (ecNGS) technology that enables highly precise quantification of mutation frequency and characterization of spectrum in potentially any species, tissue, and cell culture model. The Marchetti lab at Health Canada has conducted foundational studies using TwinStrand DuplexSeqTM Mouse and Human Mutagenesis Assays and established laboratory and bioinformatic procedures to: (a) explore mutagenic responses by DS to confirm performance across different genotoxic modes of action and diverse tissues/models; (b) evaluate qualitative and quantitative concordance relative to conventional mutagenicity endpoints; (c) define optimal experimental designs; and (d) investigate the added-value of the mechanistic information produced by DS. The work thus far indicates a robust ability to detect mutations by DS for prototypical mutagenic exposures in numerous models and concordance of DS mutation frequencies with conventional endpoints.

Furthermore, both DuplexSeqTM Mouse and Human Mutagenesis Panels show remarkable cross-laboratory reproducibility. Altogether, these efforts set the stage for the modernization of mutagenicity testing to protect human and environmental health.