Environmental and Molecular Mutagenesis最新文献

{"title":"Critical Evaluation of Methods for the Identification of Aneugens.","authors":"Xiaowen Sun, Stephen D Dertinger, Azeddine Elhajouji, Daniel J Roberts, Jan van Benthem, Maik Schuler, Shambhu Roy, Alexandra Taraboletti, Connie L Chen","doi":"10.1002/em.70023","DOIUrl":"https://doi.org/10.1002/em.70023","url":null,"abstract":"<p><p>The genotoxic potential of chemicals must be evaluated in regulatory safety assessment settings, including but not limited to, the development of new pharmaceuticals, industrial chemicals, food and cosmetic ingredients, and agrochemicals. Initial assessment of the chromosome-damaging potential of chemicals is often conducted in mammalian cells using the micronucleus (MN) assay, a method capable of detecting both aneugenicity and clastogenicity. When differentiation between these modes of action (MOAs) is necessary, microscopy-based analyses using fluorescent In Situ Hybridization (FISH) or CREST staining have traditionally been employed. More recently, semi-automated in vitro new approach methods (NAMs), which leverage technologies like flow cytometry and high-content imaging, have increasingly been used across sectors due to their higher throughput and faster turnaround times. A SWOT (strengths, weaknesses, opportunities, and threats) analysis was conducted to systematically evaluate the merits and limitations of widely used NAMs in industry, with a focus on the pharmaceutical sector. Data from cultured mammalian cells exposed to reference aneugens (colchicine, taxol, and AMG900) and DNA-reactive clastogens (mitomycin C and methyl methanesulfonate) across methodologies are presented to illustrate the process of distinguishing aneugens from clastogens for the different techniques described herein. Collectively, these analyses highlight the capabilities of NAMs to distinguish aneugens from clastogens. The newer, high information content, semi-automated approaches were considered preferable to traditional microscopy-based FISH and CREST techniques as they provide insight into molecular mechanisms of aneugenicity and help optimize the design of future in vivo genotoxicity studies to facilitate deriving points of departure which may contribute to margin of exposure estimates.</p>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":" ","pages":""},"PeriodicalIF":2.3,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Background Mutation Frequencies in TK6 and L5178Y Cells: Implications for Error-Corrected Sequencing","authors":"Jaime A. Miranda,&nbsp;Azra Dad,&nbsp;Xuewei Qu,&nbsp;Ji-Eun Seo,&nbsp;Xilin Li,&nbsp;Xiaoqing Guo,&nbsp;Javier R. Revollo","doi":"10.1002/em.70024","DOIUrl":"10.1002/em.70024","url":null,"abstract":"<div>\u0000 \u0000 <p>Several error-corrected sequencing (ECS) methods can detect ultralow-frequency mutations and support mutagenicity assessments. While ECS can be applied to any DNA-containing sample, spontaneous mutations that accumulate in immortalized cell cultures—likely due to DNA replication errors—may elevate background mutation frequencies (MFs) and potentially confound ECS-based mutagenicity assessments. This study identified mutations unique to individual cells in TK6 and L5178Y populations by comparing the genomes of single-cell-derived clones to their parental cultures. These mutations resulted in MFs of 9 × 10⁻⁷ and 6 × 10⁻⁷ mutations per base pair (mut/bp) in commercially available TK6 and L5178Y cell populations, respectively. Freshly derived clonal populations from single TK6 and L5178Y cells exhibited lower MFs (0.5 × 10⁻⁷ and 1 × 10⁻⁷ mut/bp, respectively). These results suggest that commercially available TK6 and L5178Y cell populations have accumulated significant levels of background mutations that could affect the interpretation of ECS experiments. To test this hypothesis, commercially available and freshly derived clonal TK6 cell populations were grown for 5 days in medium containing 0.5, 2, and 8 μg/mL of the in vitro mutagen N4-hydroxycytidine and analyzed by HiFi sequencing, an ECS method. The results showed that freshly derived clonal populations had lower background MFs and greater relative MF fold increases upon mutagen exposure than the commercially available cell population. An alternative data analysis approach, based on absolute MF changes within each cell population, yielded more comparable results for commercial and clonal populations. These findings underscore the impact of background MFs on in vitro ECS analyses.</p>\u0000 </div>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 6-7","pages":"367-376"},"PeriodicalIF":2.3,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144642125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transferability, Reproducibility and Sensitivity of Mutation Quantification by Duplex Sequencing","authors":"Shaofei Zhang,&nbsp;Barbara L. Parsons,&nbsp;Devon Fitzgerald,&nbsp;Anne Ashford,&nbsp;James Todd Auman,&nbsp;Tao Chen,&nbsp;Annette Dodge,&nbsp;Azeddine Elhajouji,&nbsp;Lena Pfaller,&nbsp;Shawn Harris,&nbsp;Jake Higgins,&nbsp;Cheryl A. Hobbs,&nbsp;Francesco Marchetti,&nbsp;Matthew J. Meier,&nbsp;Meagan B. Myers,&nbsp;Jesse Salk,&nbsp;Rebecca Sahroui,&nbsp;David Schuster,&nbsp;Raja Settivari,&nbsp;Stephanie L. Smith-Roe,&nbsp;Carole L. Yauk,&nbsp;Jian Yan,&nbsp;Andrew Williams,&nbsp;Connie L. Chen","doi":"10.1002/em.70020","DOIUrl":"10.1002/em.70020","url":null,"abstract":"<p>Duplex Sequencing (DS) is an ultra-accurate, error-corrected next generation sequencing (ecNGS) technology for mutation analysis. A working group (WG) within Health and Environmental Sciences Institute's Genetic Toxicology Technical Committee is investigating the suitability of ecNGS for regulatory mutagenicity testing, using DS as a model. Initial steps to promote acceptance require demonstrating technical reproducibility across DS-experienced and inexperienced laboratories and establishing the method's sensitivity relative to conventional tests. Thus, the WG conducted a ‘reconstruction experiment’ to evaluate the transferability, reproducibility, and sensitivity of DS. TwinStrand Biosciences first applied DS to establish mutation frequency (MF) in DNA samples extracted from the livers of an untreated Sprague Dawley rat, or rats treated with either 100 mg/kg/day benzo[a]pyrene (B[a]P) for ten days or 40 mg/kg/day N-ethyl-N-nitrosourea (ENU) for three days. Using the measured MF in these original samples, mixtures were then constructed using the B[a]P- and ENU-treated samples to create “MF standards” with target MFs 1.2-, 1.5-, and 2-fold greater than the untreated control. Aliquots of these standards were distributed to seven laboratories in North America and Europe. DS libraries were prepared by each laboratory and TwinStrand. All eight laboratories met library preparation and assay performance metrics to yield high quality sequencing data with MF in the expected ‘MF standard’ range. The measured MF and mutation spectra were nearly identical across the laboratories and a 2-fold increase in MF could readily be identified in all labs relative to the untreated controls. The results confirm the high reproducibility and sensitivity of DS for mutagenicity assessment.</p>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 6-7","pages":"311-326"},"PeriodicalIF":2.3,"publicationDate":"2025-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144526978","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of Hot and Cold Regions for Mutagenesis in the Escherichia coli tdk Gene: A Catalog of Mutational Sites","authors":"Katherine Douglas,&nbsp;Dana Sorensen,&nbsp;Arnav Saud,&nbsp;Ananya Sridharan,&nbsp;Mallika Mathew,&nbsp;Jyotsna Hiranandani,&nbsp;Ava Gonick,&nbsp;Courtney Young,&nbsp;Kelly Nguyen,&nbsp;Nhu Phi,&nbsp;Jeffrey H. Miller","doi":"10.1002/em.70021","DOIUrl":"10.1002/em.70021","url":null,"abstract":"<p>We constructed a catalog of mutational sites in the <i>tdk</i> gene of <i>Escherichia coli</i> that consists of 378 different base pair substitutions at 245 different sites (base pairs). This allows us to examine the tendency of different sub-regions of the gene to be more or less prone to mutations when compared with other sub-regions. We do this by recording the number of occurrences of close to 1100 mutations resulting from 9 different mutagens or mutators at each site within a rolling 20-site interval. We can define 5 mutationally prone regions (MPRs). Aligning the profile of MPRs in <i>tdk</i> next to those we previously described in the t<i>hyA</i> gene results in a linear map with 8 MPR regions, reinforcing the concept that certain sub-regions of genes are more mutable than others, even when other variables are accounted for. The catalog should prove useful for future studies.</p>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 6-7","pages":"327-340"},"PeriodicalIF":2.3,"publicationDate":"2025-06-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144368715","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of the Genotoxic Hazard of Estuarine Sediments Using an Integrative Approach With LacZ Plasmid-Based Transgenic Mice","authors":"Miguel Pinto,&nbsp;Joana Sacadura,&nbsp;Pedro M. Costa,&nbsp;Sandra Caeiro,&nbsp;Henriqueta Louro,&nbsp;Maria J. Silva","doi":"10.1002/em.70019","DOIUrl":"10.1002/em.70019","url":null,"abstract":"<div>\u0000 \u0000 <p>Under the influence of multiple anthropogenic pressures, from industrial to agricultural activities, estuaries have long been regarded as particularly sensitive ecosystems to contamination. The present study aimed at investigating the genotoxic potential of a contaminated sediment sample from an urban and industrial area of the Sado Estuary, by combining the analysis of multiple endpoints in the <i>LacZ</i> plasmid-based transgenic mouse model exposed for 28 days to contaminated estuarine sediment extracts through drinking water. The DNA and chromosome damaging effects were monitored in peripheral blood at 7-day intervals using the standard and enzyme-modified Comet assay, as well as the micronucleus assays in peripheral blood cells. After euthanasia, DNA damage was analyzed in several mouse tissues, and <i>LacZ</i> mutant frequencies were determined in the liver. Livers were also surveyed for histopathological analysis. A time-dependent increase in micronuclei frequency was seen at all tested doses, in spite of no induction of DNA damage in any organ or mutation induction in the liver of exposed mice. The liver from mice exposed to sediment extracts did not reveal major alterations besides evidence of inflammation. Overall, the integration of the endpoints analyzed in the mice is suggestive of potential chronic, rather than acute, adverse effects in vivo, and points to the need for further research in the resident human population in the area. This experimental design can be used to assess the genotoxicity of complex environmental mixtures, understand how they work, and reduce costs and resources while speeding up data collection and interpretation.</p>\u0000 </div>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 6-7","pages":"300-310"},"PeriodicalIF":2.3,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144301394","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative Analysis of TA102 and WP2 uvrA(pKM101) Strains in Detecting Nitrosamine Mutagens in the Enhanced Ames Test","authors":"Rushikesh M. Shukla,&nbsp;Darshan T. Valani,&nbsp;Chetan K. Kajavadara,&nbsp;Satyam N. Patel,&nbsp;Rajesh J. Patel,&nbsp;Laxit K. Bhatt,&nbsp;Rajesh Sundar,&nbsp;Mukul R. Jain","doi":"10.1002/em.70018","DOIUrl":"10.1002/em.70018","url":null,"abstract":"<div>\u0000 \u0000 <p>The Ames test is a fundamental assay for evaluating chemical mutagenicity, particularly for nitrosamines, which are widespread environmental and pharmaceutical contaminants. To improve sensitivity, regulatory agencies have endorsed the enhanced Ames test (EAT), which incorporates five tester strains, a 30-min pre-incubation step, and metabolic activation using both rat and hamster liver S9 fractions. While <i>Salmonella typhimurium</i> TA102 is known for its sensitivity to oxidative mutagens, its performance under EAT conditions has not been fully characterized. This study evaluated the mutagenic response of TA102 using two nitrosamine positive controls: <i>N</i>-nitrosodimethylamine (NDMA) and 1-cyclopentyl-4-nitrosopiperazine (CPNP). <i>E. coli</i> WP2 <i>uvr</i>A(pKM101) showed consistent mutagenic responses to both NDMA and CPNP, consistent with existing EAT data. TA102 demonstrated a robust response to NDMA but not to CPNP, suggesting limited sensitivity to certain nitrosamines. These findings support the continued use of WP2 <i>uvr</i>A(pKM101) in EAT protocols and highlight the limited utility of TA102 for comprehensive nitrosamine mutagenicity assessment.</p>\u0000 </div>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 5","pages":"291-297"},"PeriodicalIF":2.3,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mitochondria-Nuclear Crosstalk: Orchestrating mtDNA Maintenance","authors":"Ghazal Darfarin,&nbsp;Janice Pluth","doi":"10.1002/em.70013","DOIUrl":"10.1002/em.70013","url":null,"abstract":"<p>The mitochondria (mt) and nucleus engage in a dynamic bidirectional communication to maintain cellular homeostasis, regulating energy production, stress response, and cell fate. Anterograde signaling directs mt function, while retrograde signaling conveys metabolic and stress-related changes from mt to the nucleus. Central to this crosstalk is mitochondrial DNA (mtDNA), which encodes key oxidative phosphorylation components. MtDNA integrity is preserved through quality control mechanisms, including fusion and fission dynamics, mitophagy, and nuclear-encoded DNA repair. Disruption in these pathways contributes to mt dysfunction, oxidative stress, and genetic instability—hallmarks of aging and diseases. Additionally, redox signaling and NAD+ homeostasis integrate mt and nuclear responses, modulating transcriptional programs that support mt biogenesis and stress adaptation. This review explores the molecular mechanisms coordinating mito-nuclear interactions, emphasizing their role in maintaining mtDNA integrity and cellular equilibrium. Understanding these processes provides insights into how mt dysfunction drives aging and disease, paving the way for targeted therapeutic strategies.</p>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 5","pages":"222-242"},"PeriodicalIF":2.3,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70013","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141638","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of Biological Modifiers to a Multiplexed, Human Cell-Based DNA Damage Assay Provides Mechanistic Information on Genotoxicity and Molecular Targets","authors":"Steven M. Bryce,&nbsp;Svetlana L. Avlasevich,&nbsp;Adam Conrad,&nbsp;Nikki E. Hall,&nbsp;Kyle Tichenor,&nbsp;Erica Briggs,&nbsp;Stephen D. Dertinger,&nbsp;Jeffrey C. Bemis","doi":"10.1002/em.70017","DOIUrl":"10.1002/em.70017","url":null,"abstract":"<div>\u0000 \u0000 <p>This laboratory has reported that the combined use of In Vitro MicroFlow and MultiFlow assays provides information regarding chemicals' genotoxic mode of action (MoA). In an effort to go beyond MoA assessments, we incorporated a panel of biological response modifiers that elicit specific effects on the assays' biomarker response profiles. This was done to pursue our hypothesis that such perturbation signatures would reveal information on clastogenic mechanisms and molecular targets. For this proof-of-concept study, we exposed TK6 cells to 20 known clastogens. Cells were exposed in 96-well plates in the presence and absence of each of four modifying agents at one optimized concentration: talazoparib (PARP inhibitor), MK-8776 (CHK1 inhibitor), AZD-7648 (DNA-PK inhibitor), or a cocktail of reactive oxygen species scavengers. In parallel, cells were also exposed to each of the test chemicals for 4 h, at which time cells were washed and allowed to recover for an additional 20 h. For each of these treatment conditions, sample processing and flow cytometric analyses were performed using standard In Vitro MicroFlow and MultiFlow procedures to measure micronuclei, γH2AX, phosphohistone-H3 (p-H3), p53 activation, and relative nuclei counts. The resulting biomarker response data were processed with PROAST benchmark dose (BMD) software, with modifying agent as a covariate. Unsupervised hierarchical clustering of the collective potency metrics for various combinations of biomarkers showed that clastogens with similar genotoxic mechanisms grouped together. Overall, this study shows that in combination with biological response modifiers, MultiFlow and In Vitro MicroFlow biomarkers can provide mechanistic insights into chemical-induced genotoxicity.</p>\u0000 </div>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 5","pages":"280-290"},"PeriodicalIF":2.3,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70017","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144127001","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fertilized Avian Egg Fetal Liver Assays for Assessing DNA Damaging Potential of Chemicals: A Comparative Analysis With In Vitro and In Vivo Genotoxicity Assays and Rodent Carcinogenicity","authors":"Tetyana Kobets,&nbsp;Gary M. Williams","doi":"10.1002/em.70016","DOIUrl":"10.1002/em.70016","url":null,"abstract":"<div>\u0000 \u0000 <p>The ability to produce direct DNA damage (genotoxicity), which underlies the carcinogenicity of various chemicals, is typically evaluated in a regulatory-approved battery of in vitro tests with potential in vivo follow-up. Growing concerns for animal welfare and implementation of regulations restricting the use of animal testing necessitate the introduction of New Approach Methodologies (NAMs). The avian egg-based (<i>in ovo</i>) models were developed as metabolically competent NAMs capable of bioactivation, detoxication, and elimination of xenobiotics to potentially replace short-term in vivo genotoxicity assays for chemicals that are genotoxic in vitro. These models utilize avian (chicken or turkey) fetal livers for the evaluation of endpoints indicative of DNA damage produced by either direct or indirect mechanisms, the formation of nuclear DNA adducts and strand breaks. Avian embryos have genetic and morphologic resemblance to mammals and can be used for the evaluation of other endpoints including histopathology and genomic profiling. A concordance analysis of 87 and 59 chemicals assessed in the chicken and turkey models, respectively, revealed a stronger correlation with the results from in vivo genotoxicity assays (76% and 67% sensitivity, 79% and 72% specificity for chicken and turkey, respectively) compared to in vitro assays (58% and 56% sensitivity, 45% and 63% specificity for chicken and turkey, respectively). These results demonstrate that <i>in ovo</i> models detect the genotoxic potential of a broader range of compounds compared to in vitro assays with S9 supplementation. In conclusion, fertilized avian egg fetal liver assays offer a promising alternative to traditional in vivo genotoxicity assays.</p>\u0000 </div>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 5","pages":"258-279"},"PeriodicalIF":2.3,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70016","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144109742","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Applicability of In Silico New Approach Methods for the Risk Assessment of Tattoo Ink Ingredients","authors":"Prachi Pradeep,&nbsp;Stefanie Seifert,&nbsp;Ajay Vikram Singh,&nbsp;Peter Laux,&nbsp;Ralph Pirow","doi":"10.1002/em.70010","DOIUrl":"https://doi.org/10.1002/em.70010","url":null,"abstract":"<p>Tattoo inks contain several substances, including organic and inorganic pigments, additives, and solvents, which may pose a health risk to not only the tattooed skin but also to other parts of the human body due to intradermal exposure. Substances in tattoo inks are regulated by entry 75 in Annex XVII of REACH Regulation (EC) No. 1907/2006. However, despite these legal requirements, a well-defined criterion for the safety assessment of tattoo inks remains lacking. In this context, 2021 BfR opinion titled “Tattoo inks: minimum requirements and test methods” proposed a comprehensive risk assessment of pigments using in vitro/in-chemico data in accordance with OECD Guidelines and CLP Regulations. In the absence of experimental data, new approach methodologies (NAMs) may be used for data-gap filling. Therefore, this work evaluates the applicability of in silico NAMs for data-gap filling for a list of tattoo ink ingredients identified by the Joint Research Centre (JRC) and BfR for genotoxicity assessment. Experimental in vitro genotoxicity data were acquired from the International Uniform Chemical Information Database (IUCLID) which makes non-confidential REACH Study Results publicly accessible. The specific aims of this analysis were the evaluation of in silico genotoxicity predictions from publicly available QSAR tools and structural alerts, the development and validation of new QSAR models specific to tattoo ink ingredients, and the application of in silico models for categorization and prioritization of data-poor ingredients for further screening. Based on the workflow developed in this study, 4 high priority, 18 medium priority, and 2 low priority substances were identified for further assessment.</p>","PeriodicalId":11791,"journal":{"name":"Environmental and Molecular Mutagenesis","volume":"66 4","pages":"199-209"},"PeriodicalIF":2.3,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/em.70010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144091823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}