Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100252
Nina Y. Yuan , William D. Richards , Kailyn T. Parham , Sophia G. Clark , Kaylie Greuel , Brandon Polzin , Steven W. Smith , Connie S. Lebakken
The use of iPSC-derived complex in vitro 3D cellular constructs is a promising avenue to more accurately predict human neural toxicities and reduce the use of animal models. We have generated a neural organoid model which incorporates iPSC-derived microglia and enables interrogation of neuroinflammation induced by pre-clinical drug candidates of varying modalities and chemical compounds in industrial use. Herein we describe the generation and characterization of this model system and its utility in assessing toxicity. We exposed the neuroimmune organoids to a variety of developmental neurotoxins and measured cellular damage by release of LDH, GFAP, and NF-L into the cell culture supernatants. Additionally, to determine whether the compounds led to activation of microglia-mediated inflammation, we measured IL-8 secretion and assessed microglia-specific gene transcriptional analysis using bulk RNA sequencing. Spearman correlation matrices using both differentially expressed genes in the RNA sequencing data and pathway analysis using Gene Ontology Enrichment revealed that microglia may play a role in the toxicity of these compounds which has been widely overlooked in standardized neurotoxicity tests. Treatment of the organoids with lead acetate demonstrates a dose–response curve of IL-8 secretion and alterations in the microglial morphology. Our findings suggest that both direct neurotoxicity and indirect neuroinflammatory mechanisms contribute to the potentially harmful effects of these compounds in the developing central nervous system.
{"title":"Neural organoids incorporating microglia to assess neuroinflammation and toxicities induced by known developmental neurotoxins","authors":"Nina Y. Yuan , William D. Richards , Kailyn T. Parham , Sophia G. Clark , Kaylie Greuel , Brandon Polzin , Steven W. Smith , Connie S. Lebakken","doi":"10.1016/j.crtox.2025.100252","DOIUrl":"10.1016/j.crtox.2025.100252","url":null,"abstract":"<div><div>The use of iPSC-derived complex <em>in vitro</em> 3D cellular constructs is a promising avenue to more accurately predict human neural toxicities and reduce the use of animal models. We have generated a neural organoid model which incorporates iPSC-derived microglia and enables interrogation of neuroinflammation induced by pre-clinical drug candidates of varying modalities and chemical compounds in industrial use. Herein we describe the generation and characterization of this model system and its utility in assessing toxicity. We exposed the neuroimmune organoids to a variety of developmental neurotoxins and measured cellular damage by release of LDH, GFAP, and NF-L into the cell culture supernatants. Additionally, to determine whether the compounds led to activation of microglia-mediated inflammation, we measured IL-8 secretion and assessed microglia-specific gene transcriptional analysis using bulk RNA sequencing. Spearman correlation matrices using both differentially expressed genes in the RNA sequencing data and pathway analysis using Gene Ontology Enrichment revealed that microglia may play a role in the toxicity of these compounds which has been widely overlooked in standardized neurotoxicity tests. Treatment of the organoids with lead acetate demonstrates a dose–response curve of IL-8 secretion and alterations in the microglial morphology. Our findings suggest that both direct neurotoxicity and indirect neuroinflammatory mechanisms contribute to the potentially harmful effects of these compounds in the developing central nervous system.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100252"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144766491","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2024.100209
Aakash Patel , Suruchi Poddar , Daniel Nierenberg , Stephanie Lang , Hao Wang , Camilly Pestana Pires DeMello , Julio Gamarra , Alisha Colon , Paula Kennedy , Jeffry Roles , Jules Klion , Will Bogen , Christopher Long , Xiufang Guo , Patrick Tighe , Stephan Schmidt , Michael L. Shuler , James J. Hickman
Opioids have been the primary method used to manage pain for hundreds of years, however the increasing prescription rate of these drugs in the modern world has led to a public health crisis of overdose related deaths. Naloxone is the current standard treatment for opioid overdose rescue, but it has not been fully investigated for potential off-target toxicity effects. The current methods for pharmaceutical development do not correlate well with pre-clinical animal studies compared to clinical results, creating a need for improved methods for therapeutic evaluation. Microphysiological systems (MPS) are a rapidly growing field, and the FDA has accepted this area of research to address this concern, offering a promising alternative to traditional animal models. This study establishes a novel multi-organ MPS model of acute opioid overdose and rescue to investigate the efficacy and off-target toxicity of naloxone in combination with opioids. By integrating primary human and human induced pluripotent stem cell (hiPSC)-derived cells, including preBötzinger complex neurons, liver, cardiac, and skeletal muscle components, this study establishes a novel functional multi-organ MPS model of acute opioid overdose and rescue to investigate the efficacy and off-target toxicity of naloxone in combination with opioids, with clinically relevant functional readouts of organ function. The system was able to successfully exhibit opioid overdose using methadone, as well as rescue using naloxone evidenced by the neuronal component activity. In addition to efficacy, the multi-organ platform was able to characterize potential off-target toxicity effects of naloxone, specifically in the cardiac component.
{"title":"Microphysiological system to address the opioid crisis: A novel multi-organ model of acute opioid overdose and recovery","authors":"Aakash Patel , Suruchi Poddar , Daniel Nierenberg , Stephanie Lang , Hao Wang , Camilly Pestana Pires DeMello , Julio Gamarra , Alisha Colon , Paula Kennedy , Jeffry Roles , Jules Klion , Will Bogen , Christopher Long , Xiufang Guo , Patrick Tighe , Stephan Schmidt , Michael L. Shuler , James J. Hickman","doi":"10.1016/j.crtox.2024.100209","DOIUrl":"10.1016/j.crtox.2024.100209","url":null,"abstract":"<div><div>Opioids have been the primary method used to manage pain for hundreds of years, however the increasing prescription rate of these drugs in the modern world has led to a public health crisis of overdose related deaths. Naloxone is the current standard treatment for opioid overdose rescue, but it has not been fully investigated for potential off-target toxicity effects. The current methods for pharmaceutical development do not correlate well with pre-clinical animal studies compared to clinical results, creating a need for improved methods for therapeutic evaluation. Microphysiological systems (MPS) are a rapidly growing field, and the FDA has accepted this area of research to address this concern, offering a promising alternative to traditional animal models. This study establishes a novel multi-organ MPS model of acute opioid overdose and rescue to investigate the efficacy and off-target toxicity of naloxone in combination with opioids. By integrating primary human and human induced pluripotent stem cell (hiPSC)-derived cells, including preBötzinger complex neurons, liver, cardiac, and skeletal muscle components, this study establishes a novel functional multi-organ MPS model of acute opioid overdose and rescue to investigate the efficacy and off-target toxicity of naloxone in combination with opioids, with clinically relevant functional readouts of organ function. The system was able to successfully exhibit opioid overdose using methadone, as well as rescue using naloxone evidenced by the neuronal component activity. In addition to efficacy, the multi-organ platform was able to characterize potential off-target toxicity effects of naloxone, specifically in the cardiac component.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100209"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11745978/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143001854","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}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100223
John T. Gamble , Chad Deisenroth
The U.S. Environmental Protection Agency (U.S. EPA) is mandated to develop new approach methods (NAMs) to detect chemicals risks to susceptible populations, including effects on pregnant women and their offspring. With limited hazard information available for current and new chemicals, NAMs can provide greater relevance to human biology, mechanistic insight, and higher testing capacity than traditional animal models. The DevTox Germ Layer Reporter (GLR) model platform was recently established for high-throughput screening and prioritization of potential developmental hazards. The model platform utilizes the RUES2-GLR pluripotent stem cell reporter line that expresses fluorescent fusion protein biomarkers SOX17 (endoderm), Brachyury (mesoderm), and SOX2 (ectoderm and pluripotency); enabling a multi-lineage readout of gastrulation lineages. The DevTox GLR-Endo assay used the model platform to evaluate chemical effects on differentiating endoderm, yielding a balanced accuracy (BA) of 72% against a training set of 43 developmental toxicants and 23 non-developmental toxicants. To assess the predictivity of additional early embryonic lineages, assays for pluripotency (DevTox GLR-Pluri), ectoderm (DevTox GLR-Ecto), and mesoderm (DevTox GLR-Meso) were developed. Chemical reference set (12 developmental toxicants and 4 non-developmental toxicants) activity for each assay revealed BAs of 92% for DevTox GLR-Endo and DevTox GLR-Pluri, 71% for DevTox GLR-Ecto, and 58% for DevTox GLR-Meso. Expanded testing of the DevTox GLR-Endo and DevTox GLR-Pluri with 63 developmental and non-developmental toxicants yielded BAs of 75% and 68%, respectively. Amongst the four DevTox GLR platform assays, the DevTox GLR-Endo assay maintained the highest degree of efficacy and overall predictive accuracy for the compound set evaluated in this study.
{"title":"Profiling assay performance in the DevTox germ layer reporter platform","authors":"John T. Gamble , Chad Deisenroth","doi":"10.1016/j.crtox.2025.100223","DOIUrl":"10.1016/j.crtox.2025.100223","url":null,"abstract":"<div><div>The U.S. Environmental Protection Agency (U.S. EPA) is mandated to develop new approach methods (NAMs) to detect chemicals risks to susceptible populations, including effects on pregnant women and their offspring. With limited hazard information available for current and new chemicals, NAMs can provide greater relevance to human biology, mechanistic insight, and higher testing capacity than traditional animal models. The DevTox Germ Layer Reporter (GLR) model platform was recently established for high-throughput screening and prioritization of potential developmental hazards. The model platform utilizes the RUES2-GLR pluripotent stem cell reporter line that expresses fluorescent fusion protein biomarkers SOX17 (endoderm), Brachyury (mesoderm), and SOX2 (ectoderm and pluripotency); enabling a multi-lineage readout of gastrulation lineages. The DevTox GLR-Endo assay used the model platform to evaluate chemical effects on differentiating endoderm, yielding a balanced accuracy (BA) of 72% against a training set of 43 developmental toxicants and 23 non-developmental toxicants. To assess the predictivity of additional early embryonic lineages, assays for pluripotency (DevTox GLR-Pluri), ectoderm (DevTox GLR-Ecto), and mesoderm (DevTox GLR-Meso) were developed. Chemical reference set (12 developmental toxicants and 4 non-developmental toxicants) activity for each assay revealed BAs of 92% for DevTox GLR-Endo and DevTox GLR-Pluri, 71% for DevTox GLR-Ecto, and 58% for DevTox GLR-Meso. Expanded testing of the DevTox GLR-Endo and DevTox GLR-Pluri with 63 developmental and non-developmental toxicants yielded BAs of 75% and 68%, respectively. Amongst the four DevTox GLR platform assays, the DevTox GLR-Endo assay maintained the highest degree of efficacy and overall predictive accuracy for the compound set evaluated in this study.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100223"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143387663","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}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100215
Débora Dummer Meira , Victor Nogueira Da Gama Kohls , Matheus Correia Casotti , Luana Santos Louro , Gabriel Mendonça Santana , Thomas Erik Santos Louro , Adriana Madeira Alvares da Silva , Lorena Souza Castro Altoé , Raquel Reis Trabach , Sonia Groisman , Elizeu Fagundes de Carvalho , Jamila Alessandra Perini Machado , Stephanie Seneff , Iúri Drumond Louro
Brazil is the world leader in pesticide consumption, and its indiscriminate use puts farmers’ health at risk. The CYP2C9 gene encodes the CYP2C9 enzyme, which metabolizes several endogenous substrates and specific xenobiotics, especially pesticides. Our goal is to study the risk of pesticide use, especially the herbicide glyphosate, in the development of diseases and the association with two CYP2C9 polymorphisms, in farmers living in the southern region of Espírito Santo state, Brazil. The allelic frequency of CYP2C9*1, CYP2C9*2 and CYP2C9*3 was determined in blood samples from individuals exposed or not to pesticides using real-time PCR. 304 blood samples were analyzed, dividing CYP2C9 genotypes into three metabolization classes: normal, intermediate, and slow. Our results indicate that normal metabolizers may be more susceptible to conditions such as high blood pressure, cardiovascular disease, and kidney problems. Intermediate metabolizers show an association with attention deficit disorder and miscarriages, suggesting that farmers’ symptoms correlated with their CYP2C9 genotype. Insufficient data prevented conclusions about slow metabolizers (*2 and/or *3). These findings suggest that the CYP2C9 genotype may influence the way farmers exposed to pesticides respond, but more research is needed to clarify causality and investigate other possible health effects. As an introductory effort, this study provides insights into the complex relationship between genetic variations and pesticide exposure, laying the groundwork for future research. This pioneering work on associations between specific genetic variations and health risks with pesticide exposure, emphasizes the importance of personalized medicine and stricter regulation of pesticide use for public health and occupational safety.
{"title":"Laying the groundwork: Exploring pesticide exposure and genetic factors in south-eastern Brazilian farmers","authors":"Débora Dummer Meira , Victor Nogueira Da Gama Kohls , Matheus Correia Casotti , Luana Santos Louro , Gabriel Mendonça Santana , Thomas Erik Santos Louro , Adriana Madeira Alvares da Silva , Lorena Souza Castro Altoé , Raquel Reis Trabach , Sonia Groisman , Elizeu Fagundes de Carvalho , Jamila Alessandra Perini Machado , Stephanie Seneff , Iúri Drumond Louro","doi":"10.1016/j.crtox.2025.100215","DOIUrl":"10.1016/j.crtox.2025.100215","url":null,"abstract":"<div><div>Brazil is the world leader in pesticide consumption, and its indiscriminate use puts farmers’ health at risk. The <em>CYP2C9</em> gene encodes the CYP2C9 enzyme, which metabolizes several endogenous substrates and specific xenobiotics, especially pesticides. Our goal is to study the risk of pesticide use, especially the herbicide glyphosate, in the development of diseases and the association with two <em>CYP2C9</em> polymorphisms, in farmers living in the southern region of Espírito Santo state, Brazil. The allelic frequency of <em>CYP2C9</em>*1, <em>CYP2C9</em>*2 and <em>CYP2C9</em>*3 was determined in blood samples from individuals exposed or not to pesticides using real-time PCR. 304 blood samples were analyzed, dividing <em>CYP2C9</em> genotypes into three metabolization classes: normal, intermediate, and slow. Our results indicate that normal metabolizers may be more susceptible to conditions such as high blood pressure, cardiovascular disease, and kidney problems. Intermediate metabolizers show an association with attention deficit disorder and miscarriages, suggesting that farmers’ symptoms correlated with their <em>CYP2C9</em> genotype. Insufficient data prevented conclusions about slow metabolizers (*2 and/or *3). These findings suggest that the <em>CYP2C9</em> genotype may influence the way farmers exposed to pesticides respond, but more research is needed to clarify causality and investigate other possible health effects. As an introductory effort, this study provides insights into the complex relationship between genetic variations and pesticide exposure, laying the groundwork for future research. This pioneering work on associations between specific genetic variations and health risks with pesticide exposure, emphasizes the importance of personalized medicine and stricter regulation of pesticide use for public health and occupational safety.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100215"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11783379/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143078710","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}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100229
Brian B. Silver , Rick Fannin , Kevin Gerrish , Erik J. Tokar
Environmental toxicants can contribute to the development of several neurodegenerative diseases. However, the mechanisms behind this pathology are still incompletely understood. Prompt diagnosis of impending neurodegeneration is crucial for early interventions to prevent cognitive decline. Towards this end, accurate biomarkers for early neurodegenerative processes and exposure risk are needed. Extracellular vesicles (EVs) are lipid particles released by cells which contain many bioactive molecules including miRNAs. EVs may serve both as a route of propagating neurotoxic phenotypes and as a source of biomarkers for neurological disease. However, the exact mechanisms though which EVs could spread the deleterious effects of toxicants and the full spectrum of their usage as biomarkers remain unclear. Organoid models have several advantages, including potential for use in high-throughput toxicant testing and applications in personalized medicine and disease models. However, few studies have examined EV release in brain organoids to determine if the EVs could contain useful biomarkers. We employed several technologies to characterize EVs released by human cerebral organoids and their associated miRNAs. We identified that cerebral organoids consistently release EV-associated miRNA in quantities sufficient for robust analysis with NanoString. Further, pathway analyses revealed that terms related to neurodegenerative disease and nervous system signaling are associated with the recovered miRNAs. Together, these data suggest that cerebral organoids have utility as a tool for the discovery of EV-associated miRNAs involved in neurodegenerative disease and neurotoxicity.
{"title":"Characterization of extracellular vesicles and miRNA released by cerebral organoids","authors":"Brian B. Silver , Rick Fannin , Kevin Gerrish , Erik J. Tokar","doi":"10.1016/j.crtox.2025.100229","DOIUrl":"10.1016/j.crtox.2025.100229","url":null,"abstract":"<div><div>Environmental toxicants can contribute to the development of several neurodegenerative diseases. However, the mechanisms behind this pathology are still incompletely understood. Prompt diagnosis of impending neurodegeneration is crucial for early interventions to prevent cognitive decline. Towards this end, accurate biomarkers for early neurodegenerative processes and exposure risk are needed. Extracellular vesicles (EVs) are lipid particles released by cells which contain many bioactive molecules including miRNAs. EVs may serve both as a route of propagating neurotoxic phenotypes and as a source of biomarkers for neurological disease. However, the exact mechanisms though which EVs could spread the deleterious effects of toxicants and the full spectrum of their usage as biomarkers remain unclear. Organoid models have several advantages, including potential for use in high-throughput toxicant testing and applications in personalized medicine and disease models. However, few studies have examined EV release in brain organoids to determine if the EVs could contain useful biomarkers. We employed several technologies to characterize EVs released by human cerebral organoids and their associated miRNAs. We identified that cerebral organoids consistently release EV-associated miRNA in quantities sufficient for robust analysis with NanoString. Further, pathway analyses revealed that terms related to neurodegenerative disease and nervous system signaling are associated with the recovered miRNAs. Together, these data suggest that cerebral organoids have utility as a tool for the discovery of EV-associated miRNAs involved in neurodegenerative disease and neurotoxicity.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100229"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143911682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100239
Judit Kalman , Yolanda Valcárcel-Rivera , José Luis Martínez-Guitarte
Antibiotics, like other pharmaceuticals, are continuously released into the environment as a result of human activities. Although designed to target harmful bacteria, they can also affect non-target organisms in aquatic ecosystems. Standard toxicological tests often fail to detect the subtle or long term antibiotic-induced effects, but newer methods are providing valuable insights into the molecular pathways and physiological responses they affect. Chironomus riparius, a dipteran with aquatic larvae, is widely used in toxicological testing due to its sensitivity to various toxicants. However, little is known about the molecular effects of antibiotics on this species.
This study investigated the gene expression profile of C. riparius in response to antibiotics from three classes − aminoglycosides, fluoroquinolones and penicillin. Fourth instar larvae were exposed to concentrations of 0.001, 0.1 and 10 mg/L for 24 and 72 h. The expression of genes involved in hormonal regulation, detoxification, stress response and DNA repair was analysed. The results showed that all antibiotics altered mRNA levels, with three of the four (amoxicillin, neomycin and levofloxacin) downregulating genes at 24 h and upregulating them at 72 h. Genes affected by gentamicin showed the opposite trend.
These transcriptional changes in response to different antibiotics highlight the complexity of the regulatory mechanisms involved in development, detoxification, stress response and DNA repair in aquatic insects. Further research is needed to better understand the molecular effects of antibiotics on this species.
{"title":"Molecular response of Chironomus riparius to antibiotics","authors":"Judit Kalman , Yolanda Valcárcel-Rivera , José Luis Martínez-Guitarte","doi":"10.1016/j.crtox.2025.100239","DOIUrl":"10.1016/j.crtox.2025.100239","url":null,"abstract":"<div><div>Antibiotics, like other pharmaceuticals, are continuously released into the environment as a result of human activities. Although designed to target harmful bacteria, they can also affect non-target organisms in aquatic ecosystems. Standard toxicological tests often fail to detect the subtle or long term antibiotic-induced effects, but newer methods are providing valuable insights into the molecular pathways and physiological responses they affect. <em>Chironomus riparius</em>, a dipteran with aquatic larvae, is widely used in toxicological testing due to its sensitivity to various toxicants. However, little is known about the molecular effects of antibiotics on this species.</div><div>This study investigated the gene expression profile of <em>C. riparius</em> in response to antibiotics from three classes − aminoglycosides, fluoroquinolones and penicillin. Fourth instar larvae were exposed to concentrations of 0.001, 0.1 and 10 mg/L for 24 and 72 h. The expression of genes involved in hormonal regulation, detoxification, stress response and DNA repair was analysed. The results showed that all antibiotics altered mRNA levels, with three of the four (amoxicillin, neomycin and levofloxacin) downregulating genes at 24 h and upregulating them at 72 h. Genes affected by gentamicin showed the opposite trend.</div><div>These transcriptional changes in response to different antibiotics highlight the complexity of the regulatory mechanisms involved in development, detoxification, stress response and DNA repair in aquatic insects. Further research is needed to better understand the molecular effects of antibiotics on this species.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100239"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144098464","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Studies on reproductive toxicity are conducted to assess the effects of chemicals and pharmaceuticals on the reproductive function and fetal development. However, depending on the indication or modality, Fertility and Early Embryonic Development (FEED), Embryo-Fetal Development (EFD), and Pre/Postnatal Development (PPND) studies which evaluate all reproductive stages or EFD studies in a second species may not be necessary. Therefore, based on the Common Technical Document (CTD), PMDA review reports, and other documents of drugs with new active ingredients approved in Japan, we aimed to investigate the implementation status of DART studies by classifying the studies into FEED, EFD, and PPND types, and summarizing the reasons for not conducting the studies. This survey was conducted by the Reproductive and Developmental Toxicity Team of the Japan Pharmaceutical Manufacturers Association (JPMA) to address issues related to DART studies conducted as non-clinical studies for drug development. Of the three DART studies, 35% of drugs received marketing application without conducting EFD studies in at least one species. Among the three study types, PPND studies were the second most frequently waivered, with 36% not conducted. FEED studies had the lowest implementation rate among the three types of studies, with 40% not conducted. The primary reason for waiving at least one study was compliance with ICH S5, S6, S9, and M3 guidelines. In conclusion, the necessity of DART studies varied depending on the applicable ICH guideline and the characteristics of the drug, including therapeutic indication, target, endogenous substances, low exposure, and ADA formation. This suggests that the need for DART studies may be waived because of various reasons, each of which should be justified based on scientific rationale and risk analysis.
{"title":"Strategy for reproductive and developmental toxicity (DART) studies for marketing applications in pharmaceutical development","authors":"Yusuke Kagawa , Takuro Osawa , Naomi Koyama , Takashi Tanaharu , Daisuke Kigami","doi":"10.1016/j.crtox.2025.100258","DOIUrl":"10.1016/j.crtox.2025.100258","url":null,"abstract":"<div><div>Studies on reproductive toxicity are conducted to assess the effects of chemicals and pharmaceuticals on the reproductive function and fetal development. However, depending on the indication or modality, Fertility and Early Embryonic Development (FEED), Embryo-Fetal Development (EFD), and Pre/Postnatal Development (PPND) studies which evaluate all reproductive stages or EFD studies in a second species may not be necessary. Therefore, based on the Common Technical Document (CTD), PMDA review reports, and other documents of drugs with new active ingredients approved in Japan, we aimed to investigate the implementation status of DART studies by classifying the studies into FEED, EFD, and PPND types, and summarizing the reasons for not conducting the studies. This survey was conducted by the Reproductive and Developmental Toxicity Team of the Japan Pharmaceutical Manufacturers Association (JPMA) to address issues related to DART studies conducted as non-clinical studies for drug development. Of the three DART studies, 35% of drugs received marketing application without conducting EFD studies in at least one species. Among the three study types, PPND studies were the second most frequently waivered, with 36% not conducted. FEED studies had the lowest implementation rate among the three types of studies, with 40% not conducted. The primary reason for waiving at least one study was compliance with ICH S5, S6, S9, and M3 guidelines. In conclusion, the necessity of DART studies varied depending on the applicable ICH guideline and the characteristics of the drug, including therapeutic indication, target, endogenous substances, low exposure, and ADA formation. This suggests that the need for DART studies may be waived because of various reasons, each of which should be justified based on scientific rationale and risk analysis.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100258"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145104416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100240
Alejandra López-Lanuza , Lucila Álvarez-Barrera , Hugo López-Muñoz , Rodrigo Aníbal Mateos-Nava , Juan José Rodríguez-Mercado
Thallium (Tl) is considered hazardous to health because of its high toxicity and is an emerging contaminant with two oxidation states: Tl (I) and Tl (III). However, the toxicity of Tl and its compounds can be influenced by the oxidation state of the metal. Tl (III) is the least studied oxidation state of Tl, although it may affect cell proliferation and has genotoxic potential. Therefore, the aim of the present study was to analyze the effects Tl (III) chloride (TlCl3) on cell cycle progression, the induction of DNA damage, and oxidative stress in human lymphocytes in vitro. There were no changes in cell viability after treatment with different concentrations (0.1, 0.5, 1, 5, 10, and 50 μg/mL) of TlCl3 for different exposure durations (1, 3, and 24 h), and a reduction in the number of viable cells was observed only after treatment with high concentrations (10 and 50 μg/mL) for 72 h. In addition, cells treated with 5–50 μg/mL TlCl3 for 48 and 72 h arrested in G1 phase. Moreover, TlCl3 increased DNA damage by activating the enzyme formamidopyrimidine DNA-glycosylase (FPG), which oxidized DNA bases and increased the production of reactive oxygen species. In conclusion, TlCl3 induces oxidative stress and DNA damage by oxidizing DNA bases, which may disrupt the cell cycle.
{"title":"Thallium (III) disrupts the cell cycle and induces oxidative DNA damage in human lymphocytes in vitro","authors":"Alejandra López-Lanuza , Lucila Álvarez-Barrera , Hugo López-Muñoz , Rodrigo Aníbal Mateos-Nava , Juan José Rodríguez-Mercado","doi":"10.1016/j.crtox.2025.100240","DOIUrl":"10.1016/j.crtox.2025.100240","url":null,"abstract":"<div><div>Thallium (Tl) is considered hazardous to health because of its high toxicity and is an emerging contaminant with two oxidation states: Tl (I) and Tl (III). However, the toxicity of Tl and its compounds can be influenced by the oxidation state of the metal. Tl (III) is the least studied oxidation state of Tl, although it may affect cell proliferation and has genotoxic potential. Therefore, the aim of the present study was to analyze the effects Tl (III) chloride (TlCl<sub>3</sub>) on cell cycle progression, the induction of DNA damage, and oxidative stress in human lymphocytes <em>in vitro</em>. There were no changes in cell viability after treatment with different concentrations (0.1, 0.5, 1, 5, 10, and 50 μg/mL) of TlCl<sub>3</sub> for different exposure durations (1, 3, and 24 h), and a reduction in the number of viable cells was observed only after treatment with high concentrations (10 and 50 μg/mL) for 72 h. In addition, cells treated with 5–50 μg/mL TlCl<sub>3</sub> for 48 and 72 h arrested in G<sub>1</sub> phase. Moreover, TlCl<sub>3</sub> increased DNA damage by activating the enzyme formamidopyrimidine DNA-glycosylase (FPG), which oxidized DNA bases and increased the production of reactive oxygen species. In conclusion, TlCl<sub>3</sub> induces oxidative stress and DNA damage by oxidizing DNA bases, which may disrupt the cell cycle.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100240"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144169214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100266
Zaher Merhi
Microplastics (MPs) are now recognized as pervasive environmental contaminants with emerging evidence of bioaccumulation in the human reproductive system. Recent studies have detected MPs in human follicular fluid, oocytes, placenta, and semen, raising concerns about potential effects on fertility, gamete quality, and embryo development. Experimental data from animal models and in vitro human studies demonstrate that MPs can induce oxidative stress, mitochondrial dysfunction, meiotic spindle disruption, and apoptosis, ultimately impairing fertilization and developmental potential. These findings underscore the urgency of identifying effective strategies to reduce human exposure. Ozone-based water and wastewater treatment technologies are gaining attention as a means to mitigate environmental MPs before human ingestion or contact. Laboratory, pilot-scale, and full-scale plant studies show that ozonation—alone or in combination with granular activated carbon (GAC), coagulation–flocculation, sand filtration, or catalytic oxidation—can remove or degrade a substantial proportion of MPs (up to 96%), with smaller particles often removed more efficiently. Ozone induces surface oxidation and increases hydrophilicity, promoting aggregation and facilitating downstream filtration or sedimentation. While ozonation is less effective than some advanced oxidation processes in complete degradation, it offers the advantage of integration into existing treatment infrastructure. This review synthesizes current knowledge on MPs in ovarian health and the role of ozone-based treatments in limiting exposure. It discusses detection in the ovaries, mechanisms of reproductive toxicity, and the latest engineering strategies for MP removal, with a focus on translational implications for protecting fertility. Despite promising developments, significant gaps remain in linking environmental MP reduction directly to reproductive outcomes. Multidisciplinary collaboration between environmental engineers, reproductive biologists, and clinicians is essential to close these gaps and develop evidence-based public health interventions.
{"title":"Microplastics in ovarian function and ozone-based mitigation strategies: Emerging evidence and translational implications","authors":"Zaher Merhi","doi":"10.1016/j.crtox.2025.100266","DOIUrl":"10.1016/j.crtox.2025.100266","url":null,"abstract":"<div><div>Microplastics (MPs) are now recognized as pervasive environmental contaminants with emerging evidence of bioaccumulation in the human reproductive system. Recent studies have detected MPs in human follicular fluid, oocytes, placenta, and semen, raising concerns about potential effects on fertility, gamete quality, and embryo development. Experimental data from animal models and <em>in vitro</em> human studies demonstrate that MPs can induce oxidative stress, mitochondrial dysfunction, meiotic spindle disruption, and apoptosis, ultimately impairing fertilization and developmental potential. These findings underscore the urgency of identifying effective strategies to reduce human exposure. Ozone-based water and wastewater treatment technologies are gaining attention as a means to mitigate environmental MPs before human ingestion or contact. Laboratory, pilot-scale, and full-scale plant studies show that ozonation—alone or in combination with granular activated carbon (GAC), coagulation–flocculation, sand filtration, or catalytic oxidation—can remove or degrade a substantial proportion of MPs (up to 96%), with smaller particles often removed more efficiently. Ozone induces surface oxidation and increases hydrophilicity, promoting aggregation and facilitating downstream filtration or sedimentation. While ozonation is less effective than some advanced oxidation processes in complete degradation, it offers the advantage of integration into existing treatment infrastructure. This review synthesizes current knowledge on MPs in ovarian health and the role of ozone-based treatments in limiting exposure. It discusses detection in the ovaries, mechanisms of reproductive toxicity, and the latest engineering strategies for MP removal, with a focus on translational implications for protecting fertility. Despite promising developments, significant gaps remain in linking environmental MP reduction directly to reproductive outcomes. Multidisciplinary collaboration between environmental engineers, reproductive biologists, and clinicians is essential to close these gaps and develop evidence-based public health interventions.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100266"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145473658","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-01DOI: 10.1016/j.crtox.2025.100264
Chi-Wei Chen , Tsung-Teng Huang , Yan-Ru Chen , Chin-Hung Liu , Chia-Hsun Hsieh , Tai-Lin Chen , Kuo-Chu Lai
Areca nut (AN), which is commonly consumed in Southeast Asia, contains bioactive compounds that may influence cellular functions. Accumulating evidence has revealed several health impacts of AN consumption, but the toxicological effects of areca nut extract (ANE) on muscle cells remain largely unexplored. Myogenesis, a critical process for muscle development and regeneration, is closely tied to metabolic activity, which governs the differentiation and function of myocytes. This study aimed to evaluate the effects of ANE on myogenesis in murine C2C12 myoblasts and differentiated myotubes. In ANE-treated C2C12 myoblasts, we observed a significant decrease in the intracellular glutamine level that was accompanied by decreased GSH levels, decreased mTOR signaling, and increased autophagy in myoblasts but not in differentiated myotubes. ANE treatment decreased glutamine and 6-phosphogluconate levels in both myoblasts and myotubes, suggesting the widespread suppression of amino acid and redox-related metabolic pathways. Moreover, ANE significantly altered the metabolomic profile, upregulating the levels of glycolysis and TCA cycle intermediates but reducing ATP levels, indicating impaired energy metabolism in differentiated myotubes. ANE also downregulated the expression of key metabolic genes, including those involved in glycolysis (AKR1B3 and LDHA), glycerol metabolism (GPD1 and GPD2), and nitrogen metabolism (GLUD1, ARG1, GS, and GLS1), indicating that ANE disrupts critical pathways involved in muscle cell metabolism and myogenesis. This study provides new insights into the mechanisms by which AN consumption affects muscle development, emphasizing the need for further research into the dietary and environmental factors influencing myogenesis.
{"title":"Areca nut extract exposure disrupts myogenesis and metabolism in C2C12 cells","authors":"Chi-Wei Chen , Tsung-Teng Huang , Yan-Ru Chen , Chin-Hung Liu , Chia-Hsun Hsieh , Tai-Lin Chen , Kuo-Chu Lai","doi":"10.1016/j.crtox.2025.100264","DOIUrl":"10.1016/j.crtox.2025.100264","url":null,"abstract":"<div><div>Areca nut (AN), which is commonly consumed in Southeast Asia, contains bioactive compounds that may influence cellular functions. Accumulating evidence has revealed several health impacts of AN consumption, but the toxicological effects of areca nut extract (ANE) on muscle cells remain largely unexplored. Myogenesis, a critical process for muscle development and regeneration, is closely tied to metabolic activity, which governs the differentiation and function of myocytes. This study aimed to evaluate the effects of ANE on myogenesis in murine C2C12 myoblasts and differentiated myotubes. In ANE-treated C2C12 myoblasts, we observed a significant decrease in the intracellular glutamine level that was accompanied by decreased GSH levels, decreased mTOR signaling, and increased autophagy in myoblasts but not in differentiated myotubes. ANE treatment decreased glutamine and 6-phosphogluconate levels in both myoblasts and myotubes, suggesting the widespread suppression of amino acid and redox-related metabolic pathways. Moreover, ANE significantly altered the metabolomic profile, upregulating the levels of glycolysis and TCA cycle intermediates but reducing ATP levels, indicating impaired energy metabolism in differentiated myotubes. ANE also downregulated the expression of key metabolic genes, including those involved in glycolysis (AKR1B3 and LDHA), glycerol metabolism (GPD1 and GPD2), and nitrogen metabolism (GLUD1, ARG1, GS, and GLS1), indicating that ANE disrupts critical pathways involved in muscle cell metabolism and myogenesis. This study provides new insights into the mechanisms by which AN consumption affects muscle development, emphasizing the need for further research into the dietary and environmental factors influencing myogenesis.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100264"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145473663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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