Mutagenesis最新文献

{"title":"Using error-corrected sequencing for evaluating mutagenicity of molnupiravir in humans.","authors":"Page B McKinzie, Rutu R Valapil, Emily L Heil, James E Polli, Meagan B Myers, Mason G Pearce, Vasily N Dobrovolsky","doi":"10.1093/mutage/geag002","DOIUrl":"https://doi.org/10.1093/mutage/geag002","url":null,"abstract":"<p><p>Molnupiravir (MOV) is a prodrug of N-hydroxycytidine (NHC), an analog of the endogenous ribonucleoside cytidine that can be administered orally. MOV is used for treating patients infected by SARS-CoV-2, the coronavirus responsible for COVID-19. MOV and NHC are mutagenic in bacterial and in mammalian cell cultures, yet both chemicals have been negative or equivocal in nonclinical in vivo models of mutagenesis. We designed and validated a novel error-corrected sequencing (ECS) method for detecting single basepair substitutions in tissue samples obtained from genetically heterogeneous humans. The software used for this ECS method is available for free from a public on-line repository and it is suitable for analysis of in vivo and in vitro derived samples collected in various experimental scenarios. We recruited two groups of patients having COVID-19 one to three years ago, one group that received a full course of MOV therapy and the other group (matched for age and COVID-19 diagnosis date) that did not receive MOV. Using the ECS method, we determined the frequencies of basepair substitutions in nucleated cells isolated from peripheral blood of the patients. There was no observed difference in the frequency of mutations, the types of mutations, or mutational spectra between the MOV and the control groups. Also, the spectra of mutations in the MOV group did not show any evidence of the mutational signature expected from exposures to MOV or NHC based on data from mammalian cell culture models. Within the limits of this study, the dose of MOV authorized for the treatment of COVID-19 under emergency use authorization appears to have no mutational consequences for treated patients.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":""},"PeriodicalIF":4.3,"publicationDate":"2026-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145934317","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":"Tungsten carbide-cobalt can function as a particle positive control for genotoxicity in vitro in specific cell lines.","authors":"Michael J Burgum, Stephen J Evans, Ilaria Zanoni, Magda Blosi, Gareth J Jenkins, Shareen H Doak","doi":"10.1093/mutage/geaf021","DOIUrl":"10.1093/mutage/geaf021","url":null,"abstract":"<p><p>Nanoparticle genotoxicity can be induced through several mechanisms, but there are currently no nanoparticle positive controls available for the evaluation of in vitro genotoxicity. Tungsten carbide-cobalt (WC/Co) has been proposed as one possible candidate. The aim of this study was therefore to investigate the genotoxic profile of WC/Co (Co 8% wt.) utilizing the cytokinesis-blocked micronucleus (CBMN) assay, the mammalian cell gene mutation test, and comet assay following a 24-hour exposure. This was conducted in human lymphoblast (TK6) and Chinese hamster lung fibroblast (V79-4) cells. No cytotoxicity was observed in the TK6 CBMN assay even when significant induction of micronuclei was observed at 100 μg/ml (2-fold over control). In contrast, V79-4 cells demonstrated no significant genotoxicity or cytotoxicity in the CBMN assay. In the gene mutation assay significant mutagenicity was observed in V79-4 cells at 100 μg/ml (2-fold over control). Cellular uptake of the WC/Co nanoparticles was not qualitatively detected in either cell type when investigated with transmission electron microscopy. No genotoxicity was observed in either cell type with the comet assay. The data generated indicates that WC/Co nanoparticles may be used as a positive particulate control in the CBMN assay when using TK6 cells only; whilst in the gene mutation assay it can be used as a positive control for V79-4 cells. However, its use as a particle positive control is only possible when applying the highest test concentration of 100 μg/ml.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"592-602"},"PeriodicalIF":4.3,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12720410/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145213114","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":"The use of DNA repair inhibitors and the comet assay-an overview.","authors":"E Saenz-Martinez, A López de Cerain, A Azqueta","doi":"10.1093/mutage/geaf025","DOIUrl":"10.1093/mutage/geaf025","url":null,"abstract":"<p><p>The standard comet assay detects DNA strand breaks and alkali-labile sites, but these lesions are nonspecific. They may result directly from genotoxic agents or arise as intermediates during the repair of other DNA damage, such as oxidized bases or bulky DNA adducts. Different approaches have been developed to generate or trap these repair intermediates, making them detectable with the comet assay. Recently, the combination of the comet assay with DNA repair inhibitors like hydroxyurea and cytosine arabinoside has been proposed to detect bulky DNA adducts. These lesions are mainly repaired through nucleotide excision repair, a process that transiently produces strand breaks when damaged oligonucleotides are excised. Normally, these intermediates are rapidly repaired by DNA resynthesis and ligation. However, by inhibiting this repair step, strand breaks persist and can be detected by the comet assay. This strategy has been applied in various fields, including genotoxicity testing, environmental toxicology, human biomonitoring, and studies on DNA repair kinetics. This review focuses specifically on the use of hydroxyurea, cytosine arabinoside, and aphidicolin in in vitro experiments to evaluate the utility and specificity of this method for detecting different types of DNA lesions. Notably, in ~70% of studies reviewed, the inclusion of DNA repair inhibitors led to a significant increase in DNA damage, highlighting the added value of this approach. However, although the method enhances sensitivity to bulky adducts, it also responds to other types of damage, such as those induced by alkylating or oxidative agents.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"577-591"},"PeriodicalIF":4.3,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12720409/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145422279","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":"Water quality assessment and genotoxicity in fishes of Karamana River, Kerala, India: an insight of microplastic pollution.","authors":"Ammu Ulakesan, Shilpa Latha Sunil, Reshma Ramarajan, Shanthala Mallikarjunaiah, Hunasanahally Puttaswamygowda Gurushankara","doi":"10.1093/mutage/geaf020","DOIUrl":"10.1093/mutage/geaf020","url":null,"abstract":"<p><p>Microplastics are emerging pollutants of global concern, and their widespread presence poses a serious threat to aquatic and terrestrial ecosystems. The current study investigated the water quality and the presence of microplastics in water and native fish samples of the Karamana River, Kerala, India. The water quality was analyzed using various physicochemical parameters, including the dissolved oxygen, biochemical oxygen demand, and chemical oxygen demand. Microplastics isolated from water and native fish samples were characterized using Fourier transform infrared (FTIR) spectroscopy. DNA damage in fish liver and gill cells was assessed using the comet assay (single-cell gel electrophoresis). The water quality assessment revealed metals in the water within the acceptable limits, reduced dissolved oxygen, and increased biochemical oxygen demand and chemical oxygen demand, which indicate a river water ecosystem in hypoxic conditions, and the higher level of the most probable number index confirmed the presence of coliforms in this river. The microplastics isolated from the water and native fish samples were in fibers, fragments, film, pellets, and foams in nature. The abundance of microplastics in the river confirmed the load of microplastic pollution, which varied among the sites. FTIR spectroscopy analysis confirmed the presence of microplastic polymers such as polyethylene, polypropylene, polystyrene, polyamide, polyoxymethylene, and polyester in the water and native fish samples of the Karamana River. The increased percentage of tail DNA in the liver and gill cells of the fish inhabitants of the Karamana River, compared with the control fish, indicated DNA damage; this could be due to the microplastics in that aquatic ecosystem.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"603-612"},"PeriodicalIF":4.3,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145186338","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":"Correction to: Re: Rather et al. Carcinogenic parasites: Insights into the epidemiology and possible mechanisms of cancer.","authors":"","doi":"10.1093/mutage/geaf022","DOIUrl":"10.1093/mutage/geaf022","url":null,"abstract":"","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"613"},"PeriodicalIF":4.3,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145232848","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":"Computational prediction of mutagenicity through comprehensive cell painting analysis.","authors":"Natacha Cerisier, Emily Truong, Taku Watanabe, Taro Oshiro, Tomohiro Takahashi, Shigeaki Ito, Olivier Taboureau","doi":"10.1093/mutage/geaf014","DOIUrl":"10.1093/mutage/geaf014","url":null,"abstract":"<p><p>The mutagenicity of chemical compounds is a key consideration in toxicology, drug development, and environmental safety. Traditional methods such as the Ames test, while reliable, are time-intensive and costly. With advances in imaging and machine learning (ML), high-content assays like cell painting offer new opportunities for predictive toxicology. Cell painting captures extensive morphological features of cells, which can correlate with chemical bioactivity. In this study, we leveraged cell painting data to develop ML models for predicting mutagenicity and compared their performance with structure-based models. We used two datasets: a Broad Institute dataset containing profiles of over 30 000 molecules and a U.S.-Environmental Protection Agency dataset with images of 1200 chemicals tested at multiple concentrations. By integrating these datasets, we aimed to improve the robustness of our models. Among three algorithms tested-Random Forest, Support Vector Machine, and Extreme Gradient Boosting-the third showed the best performance for both datasets. Notably, selecting the most relevant concentration per compound, the phenotypic altering concentration, significantly improved prediction accuracy. Our models outperformed traditional quantitative structure activity relationship (QSAR) tools such as the Virtual models for property Evaluation of chemicals within a Global Architecture (VEGA) and the CompTox Dashboard for the majority of compounds, demonstrating the utility of cell painting features. The cell painting-based models revealed morphological changes related to DNA and RNA perturbation, especially in mitochondria, endoplasmic reticulum and nuclei, aligning with mutagenicity mechanisms. Despite this, certain compounds remained challenging to predict due to inherent dataset limitations and inter-laboratory variability in cell painting technology. The findings highlight the potential of cell painting in mutagenicity prediction, offering a complementary perspective to chemical structure-based models. Future work could involve harmonizing cell painting methodologies across datasets and exploring deep learning techniques to enhance predictive accuracy. Ultimately, integrating cell painting data with QSAR descriptors in hybrid models may unlock novel insights into chemical mutagenicity.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"560-574"},"PeriodicalIF":4.3,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12534217/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144775848","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":"Influence of pH and lysis duration on DNA damage detection: a comparison of neutral and alkaline comet assays.","authors":"Ruzica Pribakovic, Julia Bornhorst, Helga Stopper, Ezgi Eyluel Bankoglu","doi":"10.1093/mutage/geaf019","DOIUrl":"10.1093/mutage/geaf019","url":null,"abstract":"<p><p>The comet assay is a widely used method for measuring DNA damage and DNA repair. When DNA strand breaks happen, the supercoiling of DNA is relaxed, and after alkaline or neutral electrophoresis, depending on the type of performed comet assay, DNA moves toward the anode, forming a comet tail. Thus, with increasing frequency of DNA strand breaks, an increase in the percentage of DNA in the tail is observed. The aim of this study was to compare systematically various steps like lysis, duration of electrophoresis, and pH of the electrophoresis solution and their effect on the comet tail with regard to sensitivity for detection and quantification of DNA damage. We treated human lymphoblastoid TK6 cells with known genotoxic substances with a different mode of action and then performed both standard and modified alkaline and neutral comet assays. The modifications included Fpg- and MspI-modified comet assays. Several aspects of this comparison are investigated for the first time here. The results obtained from these experiments showed a higher %DNA in tail in the alkaline comet assay compared to the neutral comet assay. Additionally, the lysis step was not critical in the alkaline comet assay, whereas it was essential for the neutral comet assay. Results from alkaline Fpg-modified comet assay showed higher sensitivity in detecting single strand breaks and the neutral MspI-modified comet assay was better in detecting DNA double-strand breaks. Overall, our findings provided valuable insight into the differences between alkaline and neutral electrophoresis conditions in the comet assay and indicated that the alkaline comet assay is more sensitive for measuring total DNA damage.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"535-541"},"PeriodicalIF":4.3,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12534215/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144961848","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":"Understanding the cellular uptake and genotoxic potential of industrial relevant nanomaterials utilizing electron microscopy and the ToxTracker assay in vitro.","authors":"Stephen J Evans, Nynke Moelijker, Inger Brandsma, Michael J Burgum, Rosalie Elespuru, Giel Hendriks, Shareen H Doak","doi":"10.1093/mutage/geaf013","DOIUrl":"10.1093/mutage/geaf013","url":null,"abstract":"<p><p>Evaluating the genotoxic potential of nanomaterials (NMs) presents unique challenges not associated with traditional toxicological assessment. A key question in any NM focused toxicity study is whether the material has reached the target cell and what its subsequent subcellular localization is. This current study aimed to assess the potential of a panel of industrially relevant NMs; TiO2-NM102, TiO2-NM105, TiO2-E171, silica, polyethylene, polystyrene, carbon black, gold nanorods, tungsten carbide/cobalt, and tungsten carbide, to undergo cellular uptake in mouse embryonic stem cells, which are applied in the ToxTracker genotoxicity assay. Ultrastructural cellular analysis by transmission electron microscopy was undertaken following 100 μg/ml treatment with the test NMs for 24 h; any observed uptake was confirmed by energy-dispersive X-ray spectroscopy. Induction of DNA damage, cytotoxicity, p53 activation, protein stress, and oxidative stress was evaluated by the ToxTracker assay following 24-h treatment with the test NMs (0-100 μg/ml) in the absence of S9. TiO2-NM105, silica, polystyrene, carbon black, and tungsten carbide were all shown to undergo cellular uptake, localized in membrane-bound vesicles within the cytoplasm. None of the internalized NMs promoted a genotoxic response in ToxTracker, and similarly, no DNA damage was observed by the materials not internalized. Interestingly, of the internalized NMs, only polystyrene caused a slight cytotoxic response at 100 μg/ml treatment (10% loss in cell viability). Of the NMs not internalized, cytotoxicity was observed in mES cells treated with 100 μg/ml TiO2-NM102 (15%), polyethylene (15%), gold nanorods (35%), and tungsten carbide/cobalt (45%). In summary, this study demonstrated that TiO2-NM105, silica, polystyrene, carbon black, and tungsten carbide are non-genotoxic in vitro despite undergoing cell uptake in the ToxTracker cells. A continued focus is needed to supplement NM genotoxicity studies with cellular uptake analysis.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"550-559"},"PeriodicalIF":4.3,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12534216/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144649874","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":"Re: Rather et al. Carcinogenic parasites: insights into the epidemiology and possible mechanisms of cancer.","authors":"Armen Nersesyan","doi":"10.1093/mutage/geaf016","DOIUrl":"10.1093/mutage/geaf016","url":null,"abstract":"","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"533-534"},"PeriodicalIF":4.3,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144775849","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":"Genotoxic effects of aristolochic acid I on functional human-induced hepatocyte-like cells.","authors":"Yushi Hu, Yuyang Lei, Zhenna Gao, Yiyi Cao, Jing Xi, Yuning Ma, Qinwen Gao, Jian Fu, Xinyu Zhang, Yang Luan","doi":"10.1093/mutage/geaf012","DOIUrl":"10.1093/mutage/geaf012","url":null,"abstract":"<p><p>Aristolochic acid I (AAI) is a carcinogen associated with various human cancers. However, its causal relationship with hepatocellular carcinoma remains controversial, and inconsistent results from rodent studies have suggested species-specific differences. Here we evaluated AAI genotoxicity using functional human-induced hepatocyte-like cells (hiHep cells), a model that closely mimics primary human hepatocytes in gene expression and function, thereby shedding light on its potential hepatocarcinogenic risk in humans. First, we assessed AAI genotoxicity by evaluating AAI-DNA adducts and micronucleus frequency. In hiHep cells, AAI (0.7-2.5 µM) induced up to 105 adducts per 108 nucleotides, indicating high metabolic activation of AAI. A concentration-dependent increase in micronucleus frequency indicated a significant increase in chromosomal aberrations in hiHep cells. Considering the evidence of AAI inducing oxidative stress, we assessed 8-hydroxy-2'-deoxyguanosine and reactive oxygen species levels to evaluate DNA oxidative damage. For both indicators, significantly elevated levels were observed. A mechanism involving oxidative damage was further supported by observations of mitochondrial dysfunction, including changes in mitochondrial membrane potential and mitochondrial complex activity. Ascorbate treatment decreased AAI-induced oxidative DNA damage and DNA adduct formation, providing direct cellular evidence for free radical intermediates in AAI metabolic activation-a mechanism previously hypothesized but not experimentally validated in a human-relevant hepatocyte model. Our study findings revealed the genotoxic effects of AAI on hiHep cells and implicated oxidative stress as the key mechanism. These findings strengthen the association between AAI exposure and liver disease and highlight the potential role of antioxidant therapies in mitigating AAI-associated carcinogenesis.</p>","PeriodicalId":18889,"journal":{"name":"Mutagenesis","volume":" ","pages":"542-549"},"PeriodicalIF":4.3,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144143200","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}