Duplex sequencing identifies genomic features that determine susceptibility to benzo(a)pyrene-induced in vivo mutations

Exposure to environmental mutagens increases the risk of cancer and genetic disorders. We used Duplex Sequencing (DS), a high-accuracy error-corrected sequencing technology, to analyze mutation induction across twenty 2.4 kb intergenic and genic targets in the bone marrow of MutaMouse males exposed to benzo(a)pyrene (BaP), a widespread environmental pollutant. DS revealed a linear dose-related induction of mutations across all targets with low intra-group variability. Heterochromatic and intergenic regions exhibited the highest mutation frequencies (MF). C:G > A:T transversions at CCA, CCC and GCC trinucleotides were enriched in BaP-exposed mice consistent with the known etiology of BaP mutagenesis. However, GC-content had no effect on mutation susceptibility. A positive correlation was observed between DS and the gold-standard transgenic rodent gene mutation assay. Overall, we demonstrate that DS is a promising approach to study in vivo mutagenesis and yields critical insight into the genomic features governing mutation susceptibility, spectrum, and variability across the genome.

Automated summary

Exposure to environmental mutagens increases the risk of cancer and genetic disorders. Using Duplex Sequencing (DS) technology, this study analyzed mutation induction in the bone marrow of MutaMouse males exposed to benzo(a)pyrene (BaP). The results revealed a linear dose-related induction of mutations across all targets, with heterochromatic and intergenic regions exhibiting the highest mutation frequencies. The types of mutations enriched in BaP-exposed mice were consistent with the known etiology of BaP mutagenesis. Furthermore, DS showed a positive correlation with the gold-standard transgenic rodent gene mutation assay.