Whole genome mutagenicity evaluation using Hawk-Seq™ demonstrates high inter-laboratory reproducibility and concordance with the transgenic rodent gene mutation assay

作者：Shoji Matsumura, Sayaka Hosoi, Takako Hirose, Yuki Otsubo, Kazutoshi Saito, Masaaki Miyazawa, Akihiro Kawade, Atsushi Hakura, Dai Kakiuchi, Shoji Asakura, Naoki Koyama, Yuki Okada, Satsuki Chikura, Takafumi Kimoto, Kenichi Masumura, Takayoshi Suzuki, Kei‐ichi Sugiyama · 发表于：Genes and Environment · 年份：2025 · DOI：10.1186/s41021-025-00336-w · 被引用次数：3 · 研究领域：Carcinogens and Genotoxicity Assessment、Molecular Biology Techniques and Applications、DNA Repair Mechanisms

Abstract Background Error-corrected next-generation sequencing (ecNGS) enables the sensitive detection of chemically induced mutations. Matsumura et al. reported Hawk-Seq™, an ecNGS method, demonstrating its utility in clarifying mutagenicity both qualitatively and quantitatively. To further promote the adoption of ecNGS-based assays, it is important to evaluate their inter-laboratory transferability and reproducibility. Therefore, we evaluated the inter-laboratory reproducibility of Hawk-Seq™ and its concordance with the transgenic rodent mutation (TGR) assay. Results The Hawk-Seq™ protocol was successfully transferred from the developer’s laboratory (lab A) to two additional laboratories (labs B, C). Whole genomic mutations were analyzed independently using the same genomic DNA samples from the livers of gpt delta mice exposed to benzo[ a ]pyrene (BP), N -ethyl- N -nitrosourea (ENU), and N -methyl- N -nitrosourea (MNU). In all laboratories, clear dose-dependent increases in base substitution (BS) frequencies were observed, specific to each mutagen (e.g. G:C to T:A for BP). Statistically significant increases in overall mutation frequencies (OMFs) were identified at the same doses across all laboratories, suggesting high reproducibility in mutagenicity assessment. The correlation coefficient (r 2 ) of the six types of BS frequencies exceeded 0.97 among the three laboratories for BP- or ENU-exposed samples. Thus, Hawk-Seq™ provides qualitatively and quantitatively reproducibl...