Toxicology[JOURNAL]
Sun
200 papers
Automotive gasoline-induced epigenetic modifications and genotoxic effects in occupationally exposed workers.
da Silva PVB, Sarpa M, Otero UB … +6 more , da Poça KS, da Silva Christianes AL, Giardini I, Geraldino BR, Pinto LFR, Soares-Lima SC
Toxicology · 2026 May · PMID 42190885 · Publisher ↗
Occupational exposure to automotive gasoline, a complex mixture containing carcinogenic compounds such as benzene, toluene, and xylene (BTX), has been associated with both genotoxic and epigenetic effects. However, the s...
Occupational exposure to automotive gasoline, a complex mixture containing carcinogenic compounds such as benzene, toluene, and xylene (BTX), has been associated with both genotoxic and epigenetic effects. However, the specific molecular mechanisms underlying these alterations remain unclear. This study aimed to investigate global and gene-specific DNA methylation alterations in gas station workers occupationally exposed to automotive gasoline. A cross-sectional study was conducted with 217 participants from Rio de Janeiro, Brazil, divided into three groups: unexposed individuals, workers exposed via inhalation only, and workers exposed through both inhalation and dermal routes. Urinary trans,trans-muconic acid (t,t-MA), hippuric acid (HA) and methylhippuric acid (MHA) were quantified as biomarkers of benzene, toluene and xylene exposure, respectively. Global DNA methylation (LINE1 and ALU elements) and promoter methylation of DNA repair genes (MGMT, MSH3, and PARP1) were analyzed by pyrosequencing. Exposed groups showed significantly higher urinary MHA levels compared to unexposed individuals, though no significant correlation was found between exposure biomarkers and genotoxic damage. Workers exposed via inhalation only exhibited increased DNA damage. MGMT methylation levels were decreased in workers exposed through both inhalation and dermal routes, PARP1 exhibited increased methylation levels in workers exposed exclusively via inhalation, whereas MSH3 showed higher methylation in both exposed groups compared to the unexposed group. Additionally, PARP1 promoter methylation levels were directly correlated with urinary HA and MHA concentrations. Lower methylation of ALU elements was also observed upon exposure. Finally, mediation analysis showed that MGMT methylation significantly partially mediated exposure effects on DNA damage. These findings suggest the concurrent contribution of direct genotoxic mechanisms, related to the induction of DNA damage by BTX and epigenetic mechanisms, by which changes in DNA methylation may lead to the silencing of repair genes, thereby indirectly contributing to BTX-induced genomic instability. In this context, the observed methylation patterns show potential as effect biomarkers, providing relevant tools for the early detection of biological alterations, as well as for risk monitoring and prevention in occupationally exposed workers.
Identification of estrogenic and DNA damaging chemicals using a combined morphological and genotoxic biomarker assay.
Aßmann AS, Recoules C, Ertych N … +3 more , Oelgeschlaeger M, Audebert M, Fetz V
Toxicology · 2026 May · PMID 42176844 · Publisher ↗
One challenge for current chemical risk assessment is the integration of different in vitro based new approach methodologies (NAMs) to establish a concept for Next-Generation Risk Assessment (NGRA). It is particularly di...
One challenge for current chemical risk assessment is the integration of different in vitro based new approach methodologies (NAMs) to establish a concept for Next-Generation Risk Assessment (NGRA). It is particularly difficult to assess activities of chemicals associated with carcinogenicity that is not mediated by direct effects on genome integrity, known as non-genotoxic carcinogens (NGTxC). NGTxC activities may be induced through various mechanisms and cellular pathways. Thus, it is important to combine readouts, reflecting distinct mechanisms in a single test system, to limit the total number of tests needed. In this study, the previously described E-Morph screening assay (ESA), based on human MCF-7 breast cancer cells, for detection of ER-agonistic chemicals, was expanded for a simultaneous detection of DNA damage, to facilitate the distinction of genotoxic and non-genotoxic nuclear-receptor mediated carcinogens. For assessing the assay performance, we assembled a chemical library including chemicals with genotoxic or NGTxC mode of action and monitor their ER-agonistic and genotoxic activity after application of six different concentrations starting from 100 µM. The same set of chemicals was tested in a human liver cell line (HepG2) with the γH2AX/pH3 genotoxicity method. We observed a strong correlation between the two cell lines for the detection of genotoxic effects. Additionally, we observed a strong correlation between γH2AX induction and ER-agonistic activity in MCF-7 cells and aneugenicity in HepG2 cells. Consequently, the extended ESA is a promising new NAM that might play an important role in future NGRA for identifying and characterizing carcinogenic hazards.
ALDH1A3-dependent ferroptosis mediates hexafluoropropylene oxide dimer acid-induced trophoblast dysfunction.
Li S, Xu H, Li S … +3 more , Wang J, Wan B, Ju H
Toxicology · 2026 Sep · PMID 42176843 · Publisher ↗
Hexafluoropropylene oxide dimer acid (HFPO-DA), a persistent per- and polyfluoroalkyl substance, has been epidemiologically associated with adverse pregnancy outcomes; however, the molecular mechanisms underlying its pla...
Hexafluoropropylene oxide dimer acid (HFPO-DA), a persistent per- and polyfluoroalkyl substance, has been epidemiologically associated with adverse pregnancy outcomes; however, the molecular mechanisms underlying its placental toxicity remain poorly defined. Here, we demonstrate that acute 24-h exposure of human placental trophoblast HTR-8/SVneo and JEG-3 cells to HFPO-DA (0.03-3 μM) induces a concentration-dependent impairment in cell viability, proliferation, migration, and invasion, whereas inhibitors of apoptosis, necroptosis, and pyroptosis did not show comparable protective effects. Integrated bioinformatic analysis of HFPO-DA-associated pregnancy complication genes retrieved from the Comparative Toxicogenomics Database revealed significant enrichment of ferroptosis-related pathways. Protein-protein interaction network analysis further identified ALDH1A3 as a central hub gene, and molecular docking predicted a strong binding affinity between HFPO-DA and ALDH1A3, exceeding that observed for other candidate targets. Biolayer interferometry further confirmed a direct interaction between HFPO-DA and ALDH1A3, with an apparent equilibrium dissociation constant (KD) of 6.64 × 10 M. Consistent with these in silico findings, pharmacological inhibition experiments showed that only the ferroptosis inhibitor ferrostatin-1 effectively rescued HFPO-DA-induced cytotoxicity. HFPO-DA exposure led to hallmark ferroptotic alterations, including glutathione and superoxide dismutase depletion, malondialdehyde accumulation, intracellular Fe²⁺ overload, GPX4 suppression, and increased mitochondrial superoxide production in both trophoblast cell models. Mechanistically, HFPO-DA markedly downregulated ALDH1A3 at both mRNA and protein levels. Notably, siRNA-mediated ALDH1A3 silencing alone recapitulated ferroptosis-associated biochemical and functional defects, whereas ALDH1A3 overexpression restored redox homeostasis, attenuated lipid peroxidation and iron dysregulation, and rescued trophoblast functional impairment. Collectively, these findings identify ALDH1A3 repression as a key molecular event linking HFPO-DA exposure to trophoblast ferroptosis and dysfunction, providing mechanistic insight into HFPO-DA-associated placental pathogenesis.
Time-resolved multi-omics reveals staged mitochondrial dysfunction and neurodegeneration-related changes in a tri-culture BTX neurotoxicity model.
Wu S, Duan A, Wang W … +4 more , Zhang L, Wang H, Chen X, Li B
Toxicology · 2026 Sep · PMID 42173425 · Publisher ↗
Simultaneous benzene, toluene, and xylene (BTX) exposure is a common phenomenon in the workplace and the environment, but has not been well defined by time-resolved molecular events leading to BTX-induced neurotoxicity i...
Simultaneous benzene, toluene, and xylene (BTX) exposure is a common phenomenon in the workplace and the environment, but has not been well defined by time-resolved molecular events leading to BTX-induced neurotoxicity in multicellular settings. To address these points, we derived an in vitro tri-culture system using SH-SY5Y with a supportive glial compartment (HMC3 + U87) and combined dose-dependent phenotypic profiling with time-resolved transcriptomic, proteomic and metabolic studies after 4, 12, 24, 36 and 48 h of BTX treatment. Working concentrations (IC, IC and IC) were determined at the end of an initial 24 h dose-response step. Although BTX reduced cell viability in both monoculture and co-culture models, no significant differences in viability were observed between the two models at matched doses. Conversely, the co-culture model had increased sensitivity to sub-lethal toxic responses, which was evidenced by the higher levels of ROS and more obvious concentration-dependent responses to inflammatory, injury and the apoptosis-related markers. Transcriptional pathway dynamics were shown through time-course transcriptomics: initial enrichment of the cell cycle, DNA replication, and p53 signaling; mid-stage metabolic re-programming consisting of HIF-1 signaling, glycolysis/gluconeogenesis and pentose phosphate pathway; and later-stage enrichment of oxidative phosphorylation and Parkin pathways Time-course proteomics and metabolomics respectively indicated a temporal shift into mitochondrial energy dysfunction, proteostasis dysregulation, and neurodegeneration-associated modules. The integrative multi-omics analysis revealed oxidative phosphorylation, Parkinsonism, and thermogenesis as the convergent pathways. Additional evidence of early transcriptional compensation followed by a reduction of mitochondrial and neurofunctional proteins was obtained by time-resolved qPCR and western blot validation. Such results indicate a sequence of BTX neurotoxicity and provide a biologically meaningful multi-omics scheme to study mechanisms underlying and identify biomarkers.
Synergistic cytotoxicity of benzalkonium chloride and didecyldimethylammonium chloride mixtures revealed by stress granule formation.
Kim MJ, Kim EM, Kim KK
Toxicology · 2026 Sep · PMID 42155800 · Publisher ↗
Benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) are quaternary ammonium compound (QAC)-based biocides co-formulated into a wide range of commercial disinfectants, resulting in frequent, simultaneo...
Benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC) are quaternary ammonium compound (QAC)-based biocides co-formulated into a wide range of commercial disinfectants, resulting in frequent, simultaneous exposure in humans. Despite their ubiquitous use, the toxicity caused by combined BAC and DDAC exposure remains poorly characterized. In this study, we investigated these effects in human lung adenocarcinoma A549 cells, employing real-time stress granule (SG) formation as a sensitive early-stage biomarker of sub-lethal cell-level stress. Using A549 G3BP1-eGFP knock-in cells, the two compounds induced time- and concentration-dependent SG formation individually and eIF2α phosphorylation, accompanied by a reduction in cell viability and elevation in H₂O₂ levels. Combined exposure at sub-lethal concentrations markedly potentiated SG assembly and synergistically suppressed cell viability beyond levels predicted by the Bliss independence model. At the molecular level, co-exposure robustly amplified activation of multiple unfolded protein response (UPR) branches, including the PERK/eIF2α and IRE1α/XBP1 signaling pathways, whereas individual treatments only partially engaged UPR. Collectively, these findings demonstrate that BAC and DDAC co-exposure synergistically induced toxicity through the cooperative amplification of endoplasmic reticulum stress and oxidative stress signaling, and establish SG formation as a sensitive biomarker for detecting cell-level sub-lethal perturbations not captured by conventional assays. They underscore the need for mixture-based toxicological evaluations of co-formulated disinfectants and highlight the limitations of current frameworks for assessing the safety of single compounds.
Deoxynivalenol induces the p21-mediated G₂/M arrest and upregulation of differentiation markers in human dental pulp stem cells.
Shikhaliyeva I, Albayrak G, Kığ C … +2 more , Ercin M, Gezginci-Oktayoglu S
Toxicology · 2026 Sep · PMID 42150664 · Publisher ↗
Humans are constantly exposed to mycotoxins primarily through contaminated food. Deoxynivalenol (DON) is the most frequently occurring, and its toxicity to stem cells remains unclear. Mesenchymal stem cells (MSCs), respo...
Humans are constantly exposed to mycotoxins primarily through contaminated food. Deoxynivalenol (DON) is the most frequently occurring, and its toxicity to stem cells remains unclear. Mesenchymal stem cells (MSCs), responsible for tissue regeneration, serve as tools for regenerative therapies and valuable models for studying cellular differentiation and toxicological responses. This study investigated the cytotoxic effects of DON on the cell cycle progression and differentiation in human dental pulp stem cells (DPSCs). DPSCs, isolated from healthy embedded third-molar teeth and characterized, were treated with DON (0.25-16 µg/mL). Cell viabilities were determined via MTT assay and the inhibitory concentrations (IC and IC) of DON were calculated (0.46 μg/mL and 0.23 μg/mL). Cell death, ROS levels, cell cycle progression, and adipogenic and osteogenic differentiation of DPSCs were evaluated under DON exposure. DON did not induced cell death or oxidative stress in DPSCs. However, DON induced cell cycle arrest at G2/M phase through highly upregulated (6-fold) p21, with no increase in p53 protein levels, suggesting p53-independent p21 induction. Moreover, DON increased the expression of early differentiation markers (BMP2 and C/EBPβ) under non-induced culture conditions. This is the first study that provides direct in vitro evidence of DON on the cell cycle progression and differentiation in DPSCs.
Atrazine alters kisspeptin signaling and downstream neuroendocrine regulation following embryonic exposure in zebrafish.
Stradtman SC, Sathisaran U, Dierolf BK … +3 more , Sumner G, Tamagno WA, Freeman JL
Toxicology · 2026 Sep · PMID 42142733 · Full text
Atrazine is an herbicide used to control broadleaf and grassy weeds but is also a known endocrine disrupting chemical classified by the US EPA for its effect on the luteinizing hormone (LH) surge. The US EPA's maximum co...
Atrazine is an herbicide used to control broadleaf and grassy weeds but is also a known endocrine disrupting chemical classified by the US EPA for its effect on the luteinizing hormone (LH) surge. The US EPA's maximum contaminant level (MCL) for atrazine in drinking water is 3 parts per billion (ppb; µg/L), though concentrations may exceed this during peak crop seasons. Because drinking water is the primary exposure route, studying environmentally relevant concentrations near the MCL is critical for understanding public health impacts. Atrazine has been shown in epidemiological and toxicological studies to disrupt neuroendocrine and reproductive functions, including suppression of gonadotropin-releasing hormone (GnRH) neuron activity, leading to decreased LH and follicle-stimulating hormone (FSH) surges. Given the breadth of observed effects, this study hypothesized that atrazine targets an upstream neuroendocrine regulator-the kisspeptin signaling pathway-due to its dual role in reproductive and dopaminergic regulation. Kisspeptin expression was characterized in developing zebrafish, showing increases every 24 h from 1 to 120 h post fertilization (hpf). Zebrafish were exposed during embryogenesis (1-72 hpf) to atrazine at 0, 0.3, 3, or 30 ppb. Immunofluorescence at 120 hpf showed reduced kisspeptin expression in the habenula at 3 ppb and near-complete loss of kiss1/kiss2 expression with brain disorganization at 30 ppb. Kisspeptin, LH, and FSH levels were measured at 168 hpf and 6 months post fertilization (mpf). Age- and sex-dependent alterations were observed. Behavioral tests revealed anxiety-like phenotypes in larvae and adults. These findings indicate atrazine disrupts neuroendocrine and behavioral function through kisspeptin pathway dysfunction.
Circadian PER2 promotes T-2 toxin-induced immunosenescence by activating the hippo/MST1 signaling in macrophages.
Li J, Huang X, Wei W … +5 more , Wang X, You L, Nepovimova E, Wu Q, Kuca K
Toxicology · 2026 Sep · PMID 42140474 · Publisher ↗
T-2 toxin induces severe cytotoxicity and immunosenescence, but the underlying mechanisms, particularly the role of circadian rhythm disruption, still lack a clear understanding. This study explores the function of the c...
T-2 toxin induces severe cytotoxicity and immunosenescence, but the underlying mechanisms, particularly the role of circadian rhythm disruption, still lack a clear understanding. This study explores the function of the circadian PER2 and the Hippo/MST1 signaling in T-2 toxin-induced immunosenescence using RAW 264.7 macrophages model. We found that T-2 toxin (14 nM, 0-24 h) disrupted the oscillatory rhythm of PER2, attenuating its amplitude and altering its peak-trough pattern. This disruption was mediated by T-2 toxin-induced and sustained activation of HIF-1α, as HIF-1α inhibition significantly suppressed PER2 expression. Functionally, pharmacological inhibition of PER2 with KL044 alleviated T-2 toxin-induced senescence, as evidenced by reduced SA-β-gal activity, downregulation of SASP factors (IL-6, IL-8, CCL-2), and attenuation of G1 phase cell cycle arrest. Mechanistically, we identified that PER2 acts as a positive regulator of the Hippo/MST1 signaling. T-2 toxin activated Hippo signaling, increasing MST1 and phospho-YAP levels while reducing total YAP. PER2 inhibition blunted this activation. Crucially, direct inhibition of the Hippo kinase MST1 mirrored the protective effects of PER2 inhibition, significantly reducing cellular senescence, SASP expression, and apoptosis. Our findings demonstrate that T-2 toxin promotes immunosenescence by hijacking the circadian PER2 via a HIF-1α-dependent mechanism. The upregulated PER2, in turn, activates the Hippo/MST1 signaling, driving senescence. This study unveils the PER2/Hippo axis as a novel signaling cascade critical for T-2 toxin-induced immunosenescence.
MEHP exposure disrupts the self-renewal and differentiation of GC-1 cells by up-regulating m6A levels, leading to reproductive damage in immature rats.
Ding Y, Zhou X, Hong Y … +8 more , Chen J, Tang H, Sun B, Long C, Shen L, Wu S, Wei Y, Wei G
Toxicology · 2026 Sep · PMID 42140473 · Publisher ↗
Di-(2-ethylhexyl) phthalate (DEHP) and its active metabolite mono-(2-ethylhexyl) phthalate (MEHP) are environmental endocrine disruptors associated with male reproductive toxicity, but the underlying mechanisms remain un...
Di-(2-ethylhexyl) phthalate (DEHP) and its active metabolite mono-(2-ethylhexyl) phthalate (MEHP) are environmental endocrine disruptors associated with male reproductive toxicity, but the underlying mechanisms remain unclear. This study investigated whether N6-methyladenosine (m6A) modification is involved in DEHP/MEHP-induced reproductive damage using both an immature rat DEHP exposure model in vivo and MEHP-treated GC-1 cells in vitro. In vivo, DEHP exposure induced obvious testicular injury in immature rats, including structural disorder of seminiferous tubules and decreased seminiferous epithelium height and tubule diameter. In vitro, MEHP increased global m6A levels, altered cell cycle distribution, and disrupted the expression of self-renewal- and differentiation-related markers in GC-1 cells. MeRIP-seq further revealed increased m6A peaks and enrichment of cell cycle-related pathways after MEHP exposure. Among the examined m6A regulators, YTHDF1 showed significant upregulation after MEHP treatment. Moreover, YTHDF1 knockdown reversed the abnormal expression of self-renewal- and differentiation-related proteins induced by MEHP. These findings suggest that dysregulated m6A modification, potentially involving YTHDF1, participates in DEHP/MEHP-induced male reproductive damage.
Sustained activation of M1 microglia by oligomerized alpha-synuclein released from dopaminergic neuronal damage through CD11b/Src/Erk/NOX2 axis after Paraquat exposure.
Cai Q, Jing Q, Shi G … +4 more , Liu Y, Gong L, Chen N, Huang M
Toxicology · 2026 Sep · PMID 42140472 · Publisher ↗
Environmental factors, particularly exposure to herbicides and insecticides, are closely associated with neuroinflammation and the progression of Parkinson's disease (PD). Paraquat (PQ) is a widely used herbicide that ha...
Environmental factors, particularly exposure to herbicides and insecticides, are closely associated with neuroinflammation and the progression of Parkinson's disease (PD). Paraquat (PQ) is a widely used herbicide that has been reported to damage dopaminergic neurons and promote the abnormal aggregation of alpha-synuclein (ɑ-syn). We found that PQ exposure enhances the extracellular release of alpha-syn, which aggregates in dopaminergic neurons, subsequently triggering microglia activation and sustaining chronic neuroinflammation. However, the molecular mechanisms by which PQ-induced neuron-derived ɑ-syn mediates communication with microglia have not been fully elucidated. In this study, a co-culture model of mouse neuronal cells and microglia was utilized as an in vitro system. Experimental results revealed that treatment of HT-22 cells with PQ upregulated α-syn expression, causing its aggregation near the nucleus and extracellular space of neuronal cells. Furthermore, the co-culture model demonstrated that PQ-induced neuronal cells releasing endogenous ɑ-syn or cells supplemented with recombinant α-syn in vitro were capable of inducing M1 polarization of microglia. Mechanistically, α-syn was shown to bind to CD11b, a microglia-specific pattern recognition receptor, thereby inducing a sustained pro-inflammatory response and aggravate dopaminergic neuronal injury by activating NOX2 and promoting phosphorylation of its downstream signaling molecules, Src and Erk. Pharmacological inhibition of CD11b using an RGD peptide effectively reduced NOX2 activation, as evidenced by decreased translocation of the NOX2 cytoplasmic subunit p47 from the cytoplasmic to membrane, and attenuated NOX2-mediated reactive oxygen species (ROS) production and microglial activation. These finding highlight the pivotal role of CD11b in mediating microglia-driven neuroinflammatory responses. Collectively, this study provides novel insight into the underlying mechanism by which α-syn released from PQ-treated dopaminergic neurons induces neurodegeneration through activation of the CD11b-dependent Src/Erk/NOX2 pathway in microglia.
Applicability of the U-SENS™ assay for assessing skin sensitization potential of agrochemical active ingredients and formulations.
Lenze M, Benedetti MD, García MR … +3 more , Pina JI, Wikinski S, Gutiérrez ML
Toxicology · 2026 Sep · PMID 42134487 · Publisher ↗
Skin sensitization testing is essential for the regulatory classification of agrochemicals, including active ingredients (AIs) and formulations. Although several alternative methods have been developed, their application...
Skin sensitization testing is essential for the regulatory classification of agrochemicals, including active ingredients (AIs) and formulations. Although several alternative methods have been developed, their application to agrochemicals remains limited. In particular, the performance of the U-SENS™ assay, which addresses key event three (dendritic cell activation) of the adverse outcome pathway, has not yet been evaluated for these products. This study assessed the U-SENS™ applicability to predict the sensitization potential of agrochemical formulations and their AIs. Twelve formulations and nine AIs with known in vivo data were tested following OECD TG 442E. Moreover, in silico analyses were conducted to identify protein binding alerts (PBAs) and potential (a)biotic products mechanistically support the in vitro results. The assay showed balanced accuracies of 72% for formulations and 65% for AIs, comparable to previous reports for other chemical classes. Most non-sensitizing AIs lacked PBAs and metabolites, consistent with their negative U-SENS™ results, while in vivo sensitizers exhibited PBAs in the parent compound or metabolites. These findings provide preliminary evidence that U-SENS™ may contribute to the assessment of skin sensitization potential in agrochemicals. However, further studies including a wider range of substances and additional key events are required to better define its applicability domain and improve classification reliability.
Sub-chronic exposure to nanoplastics in vitro disrupts hepatic lipid homeostasis in human hepatocytes.
Mognetti B, Mannino G, Brossa A … +4 more , Molino V, Franco F, Berta GN, Bovolin P
Toxicology · 2026 Sep · PMID 42128338 · Publisher ↗
Human exposure to micro- and nanoplastics is increasingly recognized as a global concern, yet their long-term effects on human health remain poorly understood. Defining the cellular consequences of chronic exposure is es...
Human exposure to micro- and nanoplastics is increasingly recognized as a global concern, yet their long-term effects on human health remain poorly understood. Defining the cellular consequences of chronic exposure is essential for reliable risk assessment and informed environmental and public health strategies. Here, we investigated the impact of prolonged culture in presence of nanoplastics, using an in vitro hepatocyte model (HepG2) exposed to environmentally relevant concentrations (10⁴ and 10⁶ particles/mL). After 28 days, no significant changes in morphology or cell viability were observed. Despite this apparent tolerance, prolonged exposure produced marked metabolic alterations. Cellular lipid content increased in a dose-dependent manner after 120 h, peaked at day 14 (120% ± 16 and 133% ± 12), remained stable until day 21, and declined thereafter. This response was accompanied by altered expression of key lipid metabolism regulators, including SREBP-1C, FABP1, PPAR-α, and PPAR-γ. Lipidomic analysis further revealed a shift in lipid composition from day 14 onward, with increased saturated fatty acids and reduced unsaturated lipids. Although total lipid accumulation partially resolved by day 28, suggesting adaptive responses, the lipidome continued to remodel toward a potentially pathological profile. This metabolic shift was paralleled by an inflammatory signal, as indicated by increased TNF-α expression after 7 days. Together, these findings show that prolonged exposure to low nanoplastic concentrations elicits substantial lipid remodeling, potentially predisposing hepatocytes to lipotoxic and inflammatory states.
Bisphenol A induces osteoporosis by targeting LAMA4 and OLR1: Novel insights into environmental bone toxicity.
Wang Z, Chen Y, Ma J … +7 more , Yang Y, Li K, Jiao X, Xu B, Shi G, Wang L, Qi L
Toxicology · 2026 Sep · PMID 42128337 · Publisher ↗
BACKGROUND: Bisphenol A (BPA), a prevalent endocrine-disrupting chemical, is associated with osteoporosis (OP). However, the key molecular targets and mechanisms remain unclear. METHODS: A training set of 161 public tran...
BACKGROUND: Bisphenol A (BPA), a prevalent endocrine-disrupting chemical, is associated with osteoporosis (OP). However, the key molecular targets and mechanisms remain unclear. METHODS: A training set of 161 public transcriptomic samples (75 OP, 86 Controls) was used for network toxicology and 113 algorithm combinations, with an independent validation set of 20 samples (10 OP, 10 Controls, GSE7429). Additional clinical validation included label‑free proteomic profiling of bone tissue (n = 10; 5 OP, 5 Controls) and RNA sequencing of peripheral blood mononuclear cells (PBMCs) (n = 12; 6 OP, 6 Controls). Single‑cell RNA sequencing mapped cellular expression of candidate genes. Molecular docking predicted potential binding interactions. In vitro functional assays were conducted in mouse bone marrow stromal cells (BMSCs) using a non-cytotoxic BPA concentration (10 μg/mL). RESULTS: We identified and validated a set of core dysregulated genes in OP. Proteomic profiling of bone tissue revealed significant changes in five proteins: LAMA4 (log2FC = 0.971, p = 0.011), MYH7B (log2FC = 0.902, p = 0.035), FLNA (log2FC = 0.729, p = 0.013), and LAMB1 (log2FC = 0.917, p = 0.038) were upregulated, while OLR1 (log2FC = -0.954, p = 0.027) was downregulated. Notably, all these proteins except FLNA showed consistent dysregulation trends at the mRNA level in parallel transcriptomic analysis. Single-cell analysis revealed specific enrichment of LAMA4 in BMSCs, with LAMA4⁺ BMSCs exhibiting distinct intercellular communication patterns. Molecular docking computationally predicted potential binding interactions between BPA and several core targets. Functionally, non-cytotoxic BPA exposure significantly inhibited osteogenic differentiation of BMSCs, accompanied by downregulation of osteogenic markers. Among these, only LAMA4 and OLR1 showed BPA‑responsive expression changes during osteogenic differentiation that matched the direction observed in clinical multi‑omics data. Post-hoc power analysis indicated > 80% power to detect the observed fold changes at α = 0.05. CONCLUSION: This study identifies LAMA4 and OLR1 as potential novel toxicity targets through which BPA disrupts bone homeostasis. These findings provide mechanistic insights into environmental chemical-induced bone fragility and support the inclusion of skeletal endpoints in chemical risk assessment.
Mechanistic insights into atrazine-driven endothelial dysfunction: The contribution of endoplasmic reticulum stress.
Indolfi C, Correale M, Esposito E … +6 more , Lela L, Vellecco V, Bucci M, Sorrentino R, Mitidieri E, d'Emmanuele di Villa Bianca R
Toxicology · 2026 Sep · PMID 42114589 · Publisher ↗
Atrazine (ATZ) is one of the most used herbicides worldwide. To date, its impact on vascular function and the potential role as a risk factor for cardiometabolic diseases remain poorly investigated. Here, we demonstrated...
Atrazine (ATZ) is one of the most used herbicides worldwide. To date, its impact on vascular function and the potential role as a risk factor for cardiometabolic diseases remain poorly investigated. Here, we demonstrated using mouse aorta rings and an endothelial cell line that ATZ selectively impairs endothelial function without affecting vascular smooth muscle responsiveness. In isolated mouse aorta, ATZ exposure (100 nM and 1 µM) did not alter phenylephrine-induced contraction or sodium nitroprusside-mediated relaxation, indicating preserved smooth muscle function. However, ATZ significantly reduced acetylcholine- and isoprenaline-induced relaxation, suggesting a specific disruption of NO signaling. In bovine aortic endothelial cells, short-term ATZ exposure (100 nM, 30 min) affected eNOS activity in terms of a reduced eNOS dimer/monomer ratio, accompanied by decreased NO production and increased reactive oxygen species (ROS) generation, implicating eNOS uncoupling as the primary early source of oxidative stress. Prolonged exposure (100 nM, 6 h) triggered endoplasmic reticulum (ER) stress through an increase in Nox4 and ROS levels, followed by the activation of PERK/ATF4/CHOP axis. This was coupled to an increase in IL-6 and IL-8. After 24 h, PERK activation and Nox4 upregulation persisted with a trend of increase in ATF4/CHOP, suggesting a time-dependent modulation of ER stress pathways. Additionally, sustained ROS production and elevated IL-6 levels indicate a transition toward a pro-inflammatory phenotype. Overall, these findings reveal that ATZ rapidly compromises endothelial NO bioavailability, promotes oxidative stress, and activates ER stress and inflammatory pathways, highlighting its potential role in vascular dysfunction and chronic disease development.
The impact of cannabigerol exposure on human endometrial stromal cells decidualization.
Alves P, Brighton PJ, Seng-Kong C … +6 more , Nebot MT, Amaral C, Fonseca BM, Teixeira N, Brosens JJ, Correia-da-Silva G
Toxicology · 2026 Sep · PMID 42107483 · Publisher ↗
Decidualization denotes the inflammatory reprogramming of endometrial stromal cells (EnSC) into progesterone-dependent decidual cells (DC), a process essential for embryo implantation and placenta formation. Decidualizat...
Decidualization denotes the inflammatory reprogramming of endometrial stromal cells (EnSC) into progesterone-dependent decidual cells (DC), a process essential for embryo implantation and placenta formation. Decidualization also gives rise to progesterone-resistant decidual-like senescent cells (dSC), involved in extracellular matrix (ECM) remodelling and menstruation. Cannabigerol (CBG) is a non-psychotropic phytocannabinoid present in cannabis-derived products, whose impact on endometrial function remains poorly understood. The effects of CBG on decidualization were studied using an immortalized human endometrial stromal cell line (St-T1b) and primary EnSC. Cell viability was assessed following exposure to CBG (1-10 µM) during decidualization induction. CBG (2 µM) was used to evaluate its impact on decidual marker genes expression, IL-6 secretion, and transcriptomic changes (RNA-Seq). CBG inhibited the induction of the canonical decidual marker genes PRL and IGFBP1 in differentiating St-T1b cells, while this repression was not observed in decidualizing primary EnSC. Also, in these cells, CBG did not affect progesterone-dependent regulation of SCARA5 or DIO2, nor the expression of IL1RL1 and CLU, marker genes of anti-inflammatory DC and pro-inflammatory dSC, respectively. However, IL-6 secretion was reduced during the initial pro-inflammatory phase of decidualization, suggesting alterations in the cellular reprogramming of EnSC. RNA-Seq analysis identified 34 differentially expressed genes mostly associated with mitochondrial activity, lipid biosynthesis, inflammatory response and ECM remodelling. Although CBG did not disrupt canonical decidual markers in primary cells, the modulation of DC metabolism, ECM remodelling, and inflammatory response may constitute a significant risk factor for adverse pregnancy outcome.
RUNX3 mediates Aflatoxin B1-induced hepatocyte apoptosis through modulation of the hippo signaling pathway.
Mo Y, Jiang Y, Huang S … +8 more , Yijiati A, Zheng S, Hong Y, Xian X, Yu X, Xiao Y, Wei Q, He Z
Toxicology · 2026 Sep · PMID 42107482 · Publisher ↗
Aflatoxin B1 (AFB1) is a common foodborne mycotoxin that poses a serious threat to human and animal health, particularly through hepatotoxic effects. Although apoptosis is recognized as a central event in AFB1-induced li...
Aflatoxin B1 (AFB1) is a common foodborne mycotoxin that poses a serious threat to human and animal health, particularly through hepatotoxic effects. Although apoptosis is recognized as a central event in AFB1-induced liver injury, the upstream regulatory mechanisms involved remain incompletely understood. Our prior work identified increased runt-related transcription factor 3 (RUNX3) expression as a critical early event in AFB1-induced hepatocellular malignant transformation and demonstrated its potential as a biomarker of both exposure and biological effect in AFB1-exposed populations. However, the precise mechanism by which RUNX3 regulates AFB1-induced hepatocyte apoptosis remains undefined. In this study, we investigated the role of RUNX3 in AFB1-induced hepatocyte apoptosis and explored its functional relationship with the Hippo signaling pathway in two human immortalized hepatocyte cell lines: THLE-2 and L-02. AFB1 exposure significantly increased apoptotic cell death, disrupted mitochondrial membrane potential (ΔΨ), and altered the expression of apoptosis-related proteins, including BAX, BCL2, and CASPASE-3. At the same time, AFB1 treatment increased RUNX3 expression and was accompanied by marked changes in Hippo pathway-related signaling. Silencing RUNX3 significantly attenuated AFB1-induced apoptosis and partially reversed the associated Hippo pathway alterations. Similarly, pharmacological inhibition of the pathway reduced the pro-apoptotic effects of AFB1. Collectively, these findings indicate that RUNX3 mediates AFB1-induced hepatocyte apoptosis through functional modulation of the Hippo signaling pathway. This study provides mechanistic insight into the molecular events underlying AFB1-induced hepatotoxicity and suggests that RUNX3 may serve as a potential biomarker and intervention target in AFB1-related liver injury.
Towards a novel toxicity prediction pipeline, combining PBPK models and mechanistic cellular adversity models.
Burgers EJ, Sharma RP, Danilyuk TY … +8 more , Vogel S, Wijaya LS, Leist M, Zickgraf FM, Schildknecht S, Bouwman P, van de Water B, Beltman JB
Toxicology · 2026 Sep · PMID 42097329 · Publisher ↗
There is a strong need for animal-free methods that properly predict drug adversity. Especially drug-induced liver injury (DILI) is a common adverse effect that is difficult to predict with current methodology. In this s...
There is a strong need for animal-free methods that properly predict drug adversity. Especially drug-induced liver injury (DILI) is a common adverse effect that is difficult to predict with current methodology. In this study, we developed a data-driven computational adversity model for predicting the relative number of apoptotic cells in vitro, based on stress pathway activity invoked by exposure of liver cells to nitrofurantoin, diclofenac, and ketoconazole. Next, we adapted this adversity model to represent an in vivo situation and developed physiologically-based pharmacokinetic (PBPK) models for all three drugs to simulate the concentration of the drugs in the liver of humans and rats. Coupling the PBPK, stress pathway and adversity models generated quantitative systems toxicology models for rats and humans at clinically-relevant doses. For all three drugs, low levels of toxicity were predicted in rats and humans, which is consistent with in vivo observations. A detailed analysis of modifying factors (sex, age) showed that the amount of apoptotic cells is expected to increase for females and elderly people compared to males. Coupling the different models generated a novel modelling pipeline that is an important step towards in silico DILI predictions.
Transcriptomic and metabolomic analyses reveal cobalt-induced mitochondrial metabolic alterations in human cortical organoids.
Guo X, Li H, Huang Y … +8 more , Zhao L, Wang Z, Zhu X, Zeng P, Huang Y, Gao H, Cen X, Bu Q
Toxicology · 2026 Sep · PMID 42082092 · Publisher ↗
The global transition towards carbon-neutral energy systems has prompted a substantial increase in demand for cobalt (Co), making it a crucial component of lithium-ion batteries and advanced industrial materials. The wid...
The global transition towards carbon-neutral energy systems has prompted a substantial increase in demand for cobalt (Co), making it a crucial component of lithium-ion batteries and advanced industrial materials. The widespread use and limited biological elimination of Co raises growing concerns regarding potential neurodevelopmental toxicity, yet the underlying mechanisms remain largely unresolved. Here, cortical organoids obtained from human induced pluripotent stem cells (iPSCs) were subjected to 5 - 20 μM CoCl for 28 days, with the integration of whole-transcriptomics, targeted-metabolomics, and microelectrode array (MEA). The present study has shown that Co exposure induced increased dose-dependent neural apoptosis and HIF-1α stabilization, while inhibiting neuronal generation. Multi-omics integration revealed disruption of tricarboxylic acid (TCA) cycle flux and the assembly of the mitochondrial energy metabolism respiratory chain, which may be mediated by long non-coding RNA (lncRNA)-mRNA interaction networks. These molecular and metabolic disturbances were accompanied by impaired oxidative phosphorylation (OXPHOS), reduced mitochondrial functional status, and ATP depletion. Synaptic immunostaining and MEA analysis further demonstrated that there was impaired formation of excitatory synapses and compromised electrophysiological function. Collectively, these findings provide a mechanistic basis for assessing the developmental neurotoxicity risks associated with Co and underscore the significance of epigenetic regulation of metabolic pathways as a plausible contributing factor from a human-relevant model.
In vitro assessment of respiratory sensitization potential and allergic asthma: A miRNA-based approach.
Passoni FC, Iulini M, Melzi G … +4 more , Liviero F, Marinovich M, Corsini E, Galbiati V
Toxicology · 2026 Sep · PMID 42069290 · Publisher ↗
Occupational exposure to respiratory sensitizers is a major cause of allergic asthma. Although miRNAs are increasingly linked to asthma pathogenesis, their specific role in allergic asthma remains poorly defined. This st...
Occupational exposure to respiratory sensitizers is a major cause of allergic asthma. Although miRNAs are increasingly linked to asthma pathogenesis, their specific role in allergic asthma remains poorly defined. This study aims to assess a human-based 3D in vitro model to investigate the possible role of miRNAs in allergic asthma. Calu-3 cells cultured in ALI were exposed to three respiratory sensitizers: hexamethylen diisocyanate (HDI), ammonium hexachloroplatinate (HClPt), and trimellitic anhydride (TMA) using VITROCELL® Cloud Alpha 6 System. In addition, 2,4-dinitrochlorobenzene (DNCB) as skin sensitizer and the irritant sodium dodecyl sulphate (SDS) were tested. Cytotoxicity was assessed by MTT and LDH assays, while epithelial barrier integrity was evaluated by TEER. MiRNA profiling was performed, followed by targeted analysis. Cytokine release (IFN-γ, IL-4, IL-5, IL-6, IL-8, IL-10, IL-17, IL-18, IL-33, MCP-1, MIP-1α, TGF-β and TSLP) was quantified, and miRNAs involvement was investigated using miRNA mimics and inhibitors. HMOX gene expression was assessed. MiRNA profiling revealed several miRNAs consistently downregulated across all respiratory sensitizers, including miR-26b-5p, miR-34a-5p, miR-125b-5p, miR-148a-3p, and miR-200a-3p. HClPt induced a significant increase in IL-6 release and HMOX expression, whereas HDI and TMA caused a significant reduction. To understand the possible link, specific miRNA mimic and inhibitor conditions were used. Results indicate that miR-18b-5p, miR-135b-5p and let-7a-5p regulated IL-6 release and HMOX expression. Respiratory sensitizers from different chemical classes were able to induce distinct miRNA-dependent regulatory mechanisms affecting IL-6 and HMOX expression. These findings support the relevance of human cell-based in vitro models for investigating allergic asthma.
Comparative transcriptomics reveals shared stress and repair-associated pathway modules in proximal tubule cells exposed to platinum chemotherapeutics.
Barnes DA, Redegeld FA, Verheijen M … +1 more , Masereeuw R
Toxicology · 2026 Sep · PMID 42067180 · Publisher ↗
Platinum-based chemotherapeutics, including cisplatin, carboplatin, and oxaliplatin, are essential for treating solid malignancies but are limited by dose-dependent nephrotoxicity. The transcriptional programs shared acr...
Platinum-based chemotherapeutics, including cisplatin, carboplatin, and oxaliplatin, are essential for treating solid malignancies but are limited by dose-dependent nephrotoxicity. The transcriptional programs shared across clinically used platinum agents in renal proximal tubule cells remain incompletely characterised, limiting the development of robust in vitro signatures for hazard identification and AOP development. Here, we performed comparative transcriptomic profiling and pathway enrichment analysis in conditionally immortalised proximal tubule epithelial cells exposed to low, subtoxic doses of cisplatin, carboplatin, or oxaliplatin. RNA-seq identified 1261 differentially expressed genes shared among all three compounds. Pathway enrichment analysis based on Reactome organised these shared genes into nine pathway clusters spanning genotoxic stress responses, proteostasis, cell cycle regulation, extracellular matrix (ECM) organisation, immune signalling, and tissue remodelling/repair-associated programmes. Several clusters, including proteasome-related processes, mitochondrial translation, ECM organisation, and Hedgehog/Wnt-planar cell polarity signalling, emerged as prominent shared features in this in vitro model and are prioritised here as candidate modules for follow-up testing. Collectively, these data provide a systems-level view of common transcriptional responses to three platinum agents in proximal tubule cells and nominate AOP-relevant candidate key-event-aligned transcriptional modules and shared transcriptomic signatures of proximal tubule stress/repair responses for subsequent dose-response, temporal, and functional validation.
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