{"title":"Integrated electrophysiological and payload-release assessment of neuronal responses to antibody-drug conjugates in human iPSC-derived sensory neurons: implications for <i>in vitro</i> neurotoxicity evaluation.","authors":"Taeko Kuroda, Xiaobo Han, Toshiki Mochizuki, Yuto Ishibashi, Aya Goto, Yuki Seki, Norimasa Miyamoto, Takashi Yoshinaga, Shoji Asakura, Ikuro Suzuki","doi":"10.3389/ftox.2026.1855330","DOIUrl":"10.3389/ftox.2026.1855330","url":null,"abstract":"<p><p>Antibody-drug conjugates (ADCs) enable tumor-specific drug delivery; however, target antigen-independent adverse events, including peripheral neuropathy, remain a significant clinical concern. The mechanisms underlying ADC-induced neurotoxicity, particularly the relationship between payload release and alterations in neuronal function, are not yet fully understood. In this study, we established an analytical platform that integrates microelectrode array (MEA) recordings using human induced pluripotent stem cell (iPSC)-derived sensory neurons with quantitative measurement of payload levels detected after ADC exposure under culture conditions, enabling functional evaluation of the direct effects of ADCs and their payloads on neuronal activity. The microtubule inhibitors monomethyl auristatin E (MMAE) and Lys-SMCC-DM1, when administered as free payloads, suppressed neuronal activity, as indicated by reduced firing rates, decreased spike amplitudes, together with neurite fragmentation. In contrast, their corresponding ADCs, brentuximab vedotin and trastuzumab emtansine, showed limited detectable effects on neuronal activity under the present experimental conditions. Conversely, the topoisomerase I inhibitor SN-38 enhanced neuronal activity, as evidenced by increased firing rates and spike amplitudes without marked morphological disruption. Similar functional changes were observed with sacituzumab govitecan. Furthermore, quantitative analysis revealed substantially higher measured payload levels at the 168-h endpoint following sacituzumab govitecan exposure than following brentuximab vedotin or trastuzumab emtansine exposure under the present experimental conditions. These findings suggest that the neuronal functional responses observed following ADC exposure may be associated with differences in measured payload levels and are consistent with the possibility of target antigen-independent effects. The integrated platform developed in this study enables quantitative assessment of neuronal functional responses to ADCs and payloads and may serve as a complementary tool for <i>in vitro</i> neurotoxicity evaluation during ADC design and development.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1855330"},"PeriodicalIF":5.1,"publicationDate":"2026-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13646448/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149139586","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-23eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1924250
Chanamon Maleeratana, Phattarawadee Wattanasuntorn, Saranya Poapolathep, Imourana Alassane-Kpembi, Johanna Fink-Gremmels, Isabelle P Oswald, Amnart Poapolathep
{"title":"Combined exposure to emerging <i>Fusarium</i> mycotoxins produces concentration-dependent synergistic toxicity in Jurkat T cells.","authors":"Chanamon Maleeratana, Phattarawadee Wattanasuntorn, Saranya Poapolathep, Imourana Alassane-Kpembi, Johanna Fink-Gremmels, Isabelle P Oswald, Amnart Poapolathep","doi":"10.3389/ftox.2026.1924250","DOIUrl":"10.3389/ftox.2026.1924250","url":null,"abstract":"<p><strong>Introduction: </strong>Beauvericin (BEA), enniatin B (ENN B), and moniliformin (MON), mainly produced by Fusarium species, are often described as emerging mycotoxins due to their frequent detection in cereals and cereal-based products and their current lack of regulatory limits. Despite increasing evidence regarding their individual toxicities-including immunotoxic, genotoxic, reproductive, and cardiotoxic effects-information on their combined toxicity and the underlying mechanisms remains limited. To address this gap, this study aimed to evaluate the cytotoxic effects, reactive oxygen species (ROS) generation, and interaction patterns of BEA, ENN B, and MON in human Jurkat T cells following a 24-hour exposure under individual, dual, and triple treatment conditions.</p><p><strong>Methods: </strong>The experimental approach evaluated cellular responses and toxicological interactions across single and combined exposures. Cytotoxicity was assessed using the MTT assay to determine cell viability, while intracellular ROS levels were measured using a fluorescence-based assay. To analyze the dual and mixture interaction patterns among the mycotoxins, the study utilized the Chou-Talalay method for quantitative assessment of synergism and antagonism.</p><p><strong>Results: </strong>The individual toxicity assessments yielded IC<sub>50</sub> values of 1.81 μM for BEA, 4.47 μM for ENN B, and 40.45 μM for MON, establishing the potency ranking of BEA > ENN B > MON. Across all treatments, cell viability decreased in a concentration-dependent manner, which was consistently accompanied by an elevation in intracellular ROS fluorescence signals. Combination Index (CI) analysis of binary mixtures (BEA + ENN B, BEA + MON, and ENN B + MON) revealed synergistic interactions at low to moderate effect levels (IC<sub>10</sub>-IC<sub>50</sub>), which shifted toward antagonism at higher effect levels (IC<sub>75</sub>-IC<sub>90</sub>). In contrast, the ternary mixture of all three mycotoxins (BEA + ENN B + MON) predominantly exhibited sustained synergism across the entire evaluated concentration range (IC<sub>10</sub>-IC<sub>90</sub>).</p><p><strong>Discussion: </strong>Overall, these findings demonstrate that co-exposure to BEA, ENN B, and MON leads to complex interactive toxicities, with the ternary mixture showing persistent synergistic effects based on Combination Index analysis. Although these cellular responses strongly correlate with elevated ROS-associated fluorescence signals, direct experimental validation is still required to confirm the specific downstream molecular mechanisms involved.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1924250"},"PeriodicalIF":5.1,"publicationDate":"2026-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13641735/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149119580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-23eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1927574
Henk J van Lingen, Paraskevi Zagana, Changlin Ke, Edoardo Saccenti, Marta Baccaro, Maria I Klapa, Zoi G Lada, Nico W van den Brink, Maria Suarez-Diez
{"title":"Integrated transcriptomic and metabolomic ecotoxicity assessment of earthworms exposed to ZnO and Mn-doped ZnO nanomaterials.","authors":"Henk J van Lingen, Paraskevi Zagana, Changlin Ke, Edoardo Saccenti, Marta Baccaro, Maria I Klapa, Zoi G Lada, Nico W van den Brink, Maria Suarez-Diez","doi":"10.3389/ftox.2026.1927574","DOIUrl":"10.3389/ftox.2026.1927574","url":null,"abstract":"<p><strong>Introduction: </strong>Zinc oxide (ZnO)-containing nanomaterials may elicit toxic responses in environmental organisms such as earthworms. In this study, we investigate the time-dependent gene expression and metabolomic response in earthworms exposed to ZnO and Mn-doped ZnO (ZnO:Mn) nanomaterials (both at 850 mg/kg soil) and MnCl<sub>2</sub>.</p><p><strong>Methods: </strong>Earthworms were exposed to ZnO nanomaterial (850 mg/kg soil) for 2, 3, 4, or 7 days, to ZnO:Mn nanomaterial (850 mg/kg soil) for 2, 4, 7, or 14 days, and to MnCl<sub>2</sub> (45 mg/kg soil) for 14 days. Time-dependent molecular responses were characterized by transcriptomic and metabolomic analyses.</p><p><strong>Results and discussion: </strong>Strong differential gene expression was observed after 4 and 2 days of exposure to ZnO and ZnO:Mn, respectively. Moderate differential gene expression was observed after 14 days of exposure to ZnO:Mn and MnCl<sub>2</sub>. Gene ontology (GO) enrichment analysis revealed that differentially expressed genes (DEGs) in earthworms exposed to ZnO were commonly associated with muscles and earthworm movements. Genes upregulated after 2 days of exposure to ZnO:Mn were linked to GO terms associated with microtubule-based structures mediating cellular motility. Downregulated genes were enriched to GO terms among which several were related to energy metabolism. For earthworms exposed to ZnO:Mn and MnCl<sub>2</sub> for 14 days, few DEGs were involved in GO terms related to diverse biological implications. Aligning with the limited number of GO terms observed, metabolomic disruptions only included increased glutamate and a tendency for increased aspartate in response to MnCl<sub>2</sub> and ZnO:Mn exposure for 14 days, respectively. Other amino acids and the oxidized-to-reduced glutathione ratio, an indicator of oxidative stress, were not affected. In summary, exposing earthworms to ZnO and ZnO:Mn nanomaterials induced a transient response in differential gene expression related to muscle biology, energy metabolism, and translation, which largely disappeared after 14 days. Combined with minimal metabolomic changes, these findings do not point to substantial long-term ecotoxic effects.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1927574"},"PeriodicalIF":5.1,"publicationDate":"2026-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13641736/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149119576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-22eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1947679
Lucia Pittura, Sofia Vita, Mattia Resente, Federica Mongera, Adnan Khan, Maisoor Ahmed Nafees, Saif Ud Din, Nelofar Hanif, Giuseppe d'Errico, Alessandro Nardi, Francesco Regoli, Stefania Gorbi
{"title":"Microplastics in freshwater environments across the high altitude region of Gilgit-Baltistan, Pakistan.","authors":"Lucia Pittura, Sofia Vita, Mattia Resente, Federica Mongera, Adnan Khan, Maisoor Ahmed Nafees, Saif Ud Din, Nelofar Hanif, Giuseppe d'Errico, Alessandro Nardi, Francesco Regoli, Stefania Gorbi","doi":"10.3389/ftox.2026.1947679","DOIUrl":"10.3389/ftox.2026.1947679","url":null,"abstract":"<p><p>Microplastics are increasingly recognized as pervasive contaminants in remote mountain freshwater ecosystems, yet evidence from high-altitude regions remains limited. This study investigated microplastics occurrence and characteristics in surface water, sediments, and fish from the Braldu-Shigar-Skardu freshwater system in Gilgit-Baltistan, northern Pakistan. Samples were collected from remote headwaters, tributaries, main river channels, and the Satpara Lake-Dam system. Fish originated from wild populations, farming facilities, and local markets to provide a broader assessment of microplastic exposure. Matrix-specific extraction protocols were applied, followed by stereomicroscope sorting and polymer identification by µFT-IR spectroscopy. Microplastics were detected in nearly all sites and matrices, with concentrations ranging from 1 to 17 MPs L<sup>-1</sup> in water, 55-537 MPs kg<sup>-1</sup> in sediments, and 1-11 MPs individual<sup>-1</sup> in fish gastrointestinal tracts. Notably, highest frequency of microplastics occurrence was measured in farmed individuals (78%-83%), suggesting that aquaculture-related practices may represent an additional source of microplastics. Wild fish showed the lowest occurrence (25%), whereas those obtained from local markets revealed intermediate values (62%), indicating variable exposure across different pathways. Surface waters were dominated by small fragments and polypropylene, indicative of recent inputs and origin from fragmentation. Sediments showed a broader size distribution, and greater polymer diversity, consistent with their role as long-term sinks. The highest polymer diversity was highlighted in fish, indicating multiple exposure pathways. Spatial patterns did not show a geographical downstream increase, but rather a patchy distribution shaped by local anthropogenic input, hydrological connectivity, and depositional dynamics. Glacier-fed rivers primarily act as transport corridors, whereas low-energy environments such as Satpara Lake function as sinks promoting microplastics accumulation. Overall, the study provides the first integrated multi-compartment baseline for microplastics in Gilgit-Baltistan and supports future monitoring and management actions for these high altitude critical freshwater resources.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1947679"},"PeriodicalIF":5.1,"publicationDate":"2026-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13639269/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149119595","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-22eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1901119
Eleonora Loi, Loredana Moi, Francesca Floris, Antonietta Marongiu, Paolo Valera, Matteo Serra, Enrico Sanjust, Cesare Zavattari, Alessandro Tommasi, Giuseppe Prencipe, Fabio Boccuni, Rocco Gibilras, Maddalena Papacchini, Marco Di Basilio, Patrizia Zavattari
{"title":"Detection of cancer epigenetic alterations in a pilot cohort of workers exposed to TiO<sub>2</sub>.","authors":"Eleonora Loi, Loredana Moi, Francesca Floris, Antonietta Marongiu, Paolo Valera, Matteo Serra, Enrico Sanjust, Cesare Zavattari, Alessandro Tommasi, Giuseppe Prencipe, Fabio Boccuni, Rocco Gibilras, Maddalena Papacchini, Marco Di Basilio, Patrizia Zavattari","doi":"10.3389/ftox.2026.1901119","DOIUrl":"https://doi.org/10.3389/ftox.2026.1901119","url":null,"abstract":"<p><strong>Introduction: </strong>The extensive use of TiO<sub>2</sub> for several applications poses the problem of its safety. Notably, TiO<sub>2</sub> has been classified as type Group 2 B carcinogen by IARC in 2010. It is widely known that DNA methylation is influenced by the exposure to environmental triggers, including occupational risk factors. The aim of this study was to evaluate the presence of a cancer DNA methylation alteration in TiO<sub>2</sub> exposed workers from a TiO<sub>2</sub> powder production plant.</p><p><strong>Methods: </strong>Due to the possible entry route, by ingestion or by inhalation, of TiO<sub>2</sub> nanoparticles in the workers of our study cohort, we analysed a DNA methylation biomarker altered in lung and colorectal cancers. The analysis was conducted by MethyLight expressing DNA methylation values as Ct (lower Ct indicates higher methylation level and viceversa), in cfDNA from plasma samples comparing workers with different duties from a plant producing TiO<sub>2</sub> pigment powder, unexposed subjects as healthy controls and colorectal cancer patients as positive controls, also considering job-related variables.</p><p><strong>Results: </strong>The obtained results showed that exposed workers displayed higher methylation compared to healthy controls approaching the methylation patterns of colorectal cancer patients, even more evident for workers of some job categories. Moreover, the methylation data would highlight the importance of personal protective equipment, as observed in workers using a gas mask exhibiting a negligible methylation value compared to the job category average methylation level.</p><p><strong>Discussion: </strong>This preliminary investigation suggests the harmful role of TiO<sub>2</sub> in altering DNA methylation patterns potentially leading to tumorigenesis and highlights the importance of monitoring workers exposed to TiO<sub>2</sub> particles.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1901119"},"PeriodicalIF":5.1,"publicationDate":"2026-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13638187/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149119552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-15eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1888799
Omar A Yanouri, Adam J Schuller, Savannah M Rocha, Aidan M Briggs, Paul A Martinez, Elizabeth Fernandez, Randy Strong, Xueshu Li, Hans-Joachim Lehmler, Jonathan A Doorn, Ronald B Tjalkens
{"title":"ALDH1/2 deficiency modulates dieldrin-induced dopaminergic neurodegeneration and reactive glial morphometry in male mice.","authors":"Omar A Yanouri, Adam J Schuller, Savannah M Rocha, Aidan M Briggs, Paul A Martinez, Elizabeth Fernandez, Randy Strong, Xueshu Li, Hans-Joachim Lehmler, Jonathan A Doorn, Ronald B Tjalkens","doi":"10.3389/ftox.2026.1888799","DOIUrl":"10.3389/ftox.2026.1888799","url":null,"abstract":"<p><p>The etiology of Parkinson's disease (PD) is associated with exposure to environmental contaminants, including the organochlorine pesticide dieldrin. This is thought to occur through induction of oxidative damage and dysregulation of dopamine homeostasis via impairment of aldehyde dehydrogenase (ALDH) enzymes, resulting in neuronal stress that may contribute to glial-mediated neurotoxicity. We postulated that sub-chronic murine dieldrin exposure would induce PD-relevant histopathologic changes in the substantia nigra (SN) which would be exacerbated in ALDH-deficient mice. To test this hypothesis, six-month-old wildtype and Aldh1a1<sup>-/-</sup> X Aldh2<sup>-/-</sup> (Aldh1/2 KO) mice were fed dieldrin in chow at doses of 0, 0.2, or 0.6ppm for 6 weeks. Slide scanning microscopy and deep learning-based image analysis were then applied to quantify neuronal injury and glial morphometry. Dieldrin exposure resulted in male-specific dopaminergic neurodegeneration, a dose-dependent increase in astrocytic density, and disease-associated alterations in microglial morphology in the substantia nigra. Interestingly, Aldh1/2 KO attenuated neuronal loss and distinctly modulated reactive astrocytosis and microglial morphometric changes. These results highlight a sex-dependent susceptibility to dieldrin-induced neurotoxicity and suggest a role of ALDH in modulating glial reactivity in the context of PD-like neurodegeneration.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1888799"},"PeriodicalIF":5.1,"publicationDate":"2026-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13619124/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149077026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Frontiers in toxicologyPub Date : 2026-09-14eCollection Date: 2026-01-01DOI: 10.3389/ftox.2026.1854281
John S Kendrick, David A Shaw, Marie Jourdain, Claudia Senn, Helen Beyer, Fiona R Sewell
{"title":"Animal reuse in safety assessment studies: a review of current practices, challenges and 3Rs opportunities.","authors":"John S Kendrick, David A Shaw, Marie Jourdain, Claudia Senn, Helen Beyer, Fiona R Sewell","doi":"10.3389/ftox.2026.1854281","DOIUrl":"10.3389/ftox.2026.1854281","url":null,"abstract":"<p><p>Regulatory nonclinical safety assessment in animals remains an essential part of the drug development process. Historically, naive animals are used within most nonclinical studies to avoid confounding variables such as prior drug exposure. However, opportunities exist to advance the 3Rs (Replacement, Reduction and Refinement) by reusing animals in studies when scientifically and ethically justified, without compromising scientific integrity or animal welfare or exceeding permitted cumulative physiological and psychological burden. Reuse is already standard practice in pharmacokinetic (PK) and safety pharmacology studies, yet its application in toxicology remains limited. This review explores how broader adoption of reuse as a 3Rs strategy can reduce overall animal numbers, while ensuring reuse does not compromise animal welfare or the scientific integrity of the study. Animal reuse can avoid and reduce known stressors prior to study start, such as transportation, quarantine and acclimation to new environments, by allowing animals to remain in a familiar environment for longer periods extending the time for positive reinforcement training and to establish group hierarchy. Appropriate reuse supports the sustainability and ethical use of animal models. Challenges of reuse include managing the cumulative lifetime burden of the animal, ensuring adequate washout and recovery periods, and addressing scientific concerns such as immunogenicity risks associated with developing anti-drug antibodies (ADA) against biologics or pre-existing immunity to gene therapies and the rising frequency of age-related background lesions. We propose a tiered framework to guide reuse decisions: Tier 1 includes studies where reuse is already standard and achievable; Tier 2 covers studies where reuse is generally feasible with appropriate scientific and ethical rationale; and Tier 3 includes studies where reuse is more challenging due to regulatory, scientific or welfare considerations, although enrolling non-naive animals at study start may still be possible without compromising data interpretation. Enabling wider implementation of reuse will require industry-wide collaboration, standardized decision tools, and proactive engagement with contract research organizations. When applied appropriately, reuse can reduce overall animal use while maintaining robust safety assessment. In parallel with the evolving adoption of new approach methodologies, reuse offers an additional practical avenue to support a more sustainable and humane nonclinical testing paradigm.</p>","PeriodicalId":73111,"journal":{"name":"Frontiers in toxicology","volume":"8 ","pages":"1854281"},"PeriodicalIF":5.1,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13616529/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149070672","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}