Cellular Toxicity of the Mycotoxins: From In Vitro Testing to Single Cell Analysis

A special issue of Toxins (ISSN 2072-6651). This special issue belongs to the section "Mycotoxins".

Deadline for manuscript submissions: closed (15 February 2023) | Viewed by 18706

Special Issue Editors


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Guest Editor
INRA, UMR 1331 Toxalim, Research Center in Food Toxicology, 180 Chemin de Tournefeuille, F-31027 Toulouse, France
Interests: mycotoxin; intestine; immune response; swine
Special Issues, Collections and Topics in MDPI journals

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Université de Brest, EA 3882, Laboratoire Universitaire de Biodiversité et Ecologie Microbienne, ESIAB, Technopôle Brest-Iroise, 29280 Plouzané, France
Interests: cell culture; flow cytometry; cell biology; immunology; macrophage; toxicology; dendritic cells; mycotoxins; monocytes; aflatoxin; cytotoxicity assays; acute toxicity; food contaminants; food toxicology; mycotoxin metabolites
Special Issues, Collections and Topics in MDPI journals

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Guest Editor
Faculté de médecine vétérinaire - Département de biomédecine vétérinaire, Université de Montréal, Montreal, Canada
Interests: mycotoxins; pharmacodynamics; secondary metabolites; aflatoxin; toxicity; intestinal inflammation models
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Exposure to mycotoxins contaminating food and feed as well as indoor environments can cause serious adverse effects in humans and animals. Owing to variations in cell physiology, the toxicity of mycotoxins is highly dependent on the organs and, more specifically, the types of cell encountered. Precision toxicology, built upon isolation of small groups of cells or even single cells, is a new concept to probe subtle intracellular changes in response to xenobiotics. It is of great interest to assess the toxicity of mycotoxins. Cell culture testing as well as single cell analysis provide useful tools to determine the toxicity of mycotoxins, to delineate their mechanism of action, and to characterize target pathways. They can also be used to identify solutions to control or relieve the toxicological effects of these toxins. Moreover, these in vitro approaches can be valuable tools for the screening of interactive potential in the mycotoxin and/or conjugated form mixtures that humans and animals are exposed to.

This Special Issue of Toxins will present recent studies on the toxicity of mycotoxins on targeted human and animal cells, and the associated mechanisms. Therefore, papers dealing with new cellular or/and multicellular systems as well as mycotoxin interactions effects are welcome. Papers considering -omics or descriptive approaches to understand mechanisms are invited. Both research papers and review articles proposing novelties or overviews, respectively, are welcome.

Dr. Isabelle P. Oswald
Dr. Nolwenn Hymery
Dr. Imourana Alassane-Kpembi
Guest Editors

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Keywords

  • mycotoxins
  • toxicity
  • apoptosis
  • oxidative stress
  • genotoxicity
  • in vitro
  • single-cell analysis
  • signaling pathway
  • metabolomics
  • transcriptomics
  • proteomics

Published Papers (8 papers)

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Research

11 pages, 2103 KiB  
Article
The Mycotoxin De-Epoxy-Deoxynivalenol (DOM-1) Increases Endoplasmic Reticulum Stress in Ovarian Theca Cells
by Angelica D. Reyes-Perea, Hilda M. Guerrero-Netro, Europa Meza-Serrano, Anthony Estienne and Christopher A. Price
Toxins 2023, 15(3), 228; https://doi.org/10.3390/toxins15030228 - 17 Mar 2023
Viewed by 1652
Abstract
Deoxynivalenol (DON) is a major mycotoxin present in animal feed and negatively affects growth and reproduction in farm species, including pigs and cattle. The mechanism of DON action involves the ribotoxic stress response (RSR), and it acts directly on ovarian granulosa cells to [...] Read more.
Deoxynivalenol (DON) is a major mycotoxin present in animal feed and negatively affects growth and reproduction in farm species, including pigs and cattle. The mechanism of DON action involves the ribotoxic stress response (RSR), and it acts directly on ovarian granulosa cells to increase cell death. In ruminants, DON is metabolized to de-epoxy-DON (DOM-1), which cannot activate the RSR but has been shown to increase cell death in ovarian theca cells. In the present study, we determined if DOM-1 acts on bovine theca cells through endoplasmic stress using an established serum-free cell culture model and to assess whether also DON activates endoplasmic stress in granulosa cells. The results show that DOM-1 increased the cleavage of ATF6 protein, increased the phosphorylation of EIF2AK3, and increased the abundance of cleaved XBP1 mRNA. Activation of these pathways led to an increased abundance of mRNA of the ER stress target genes GRP78, GRP94, and CHOP. Although CHOP is widely associated with autophagy, inhibition of autophagy did not alter the response of theca cells to DOM-1. The addition of DON to granulosa cells partially increased ER stress pathways but failed to increase the abundance of mRNA of ER stress target genes. We conclude that the mechanism of action of DOM-1, at least in bovine theca cells, is through the activation of ER stress. Full article
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9 pages, 721 KiB  
Article
In Vitro Effects of Enniatin A on Steroidogenesis and Proliferation of Bovine Granulosa Cells
by Ilaria Chiminelli, Leon J. Spicer, Excel Rio S. Maylem and Francesca Caloni
Toxins 2022, 14(10), 714; https://doi.org/10.3390/toxins14100714 - 20 Oct 2022
Cited by 8 | Viewed by 1729
Abstract
The emerging Fusarium mycotoxins enniatins (ENNs) have been the focus of new research because of their well-documented existence in various cereal and grain products. Research findings indicate that reproductive disorders may be caused by exposure to Fusarium mycotoxins, but little work has evaluated [...] Read more.
The emerging Fusarium mycotoxins enniatins (ENNs) have been the focus of new research because of their well-documented existence in various cereal and grain products. Research findings indicate that reproductive disorders may be caused by exposure to Fusarium mycotoxins, but little work has evaluated ENNs on reproductive function. Therefore, to determine the effects of ENNA on the proliferation and steroidogenesis of granulosa cells (GC), experiments were conducted using bovine GC cultures. In vitro, ENNA (1–5 μM) inhibited (p < 0.05) hormone-induced GC progesterone and estradiol production. The inhibitory effect of ENNA on estradiol production was more pronounced in small- than large-follicle GC. In large-follicle GC, 0.3 μM ENNA had no effect (p > 0.10) whereas 1 and 3 μM ENNA inhibited GC proliferation. In small-follicle GC, ENNA (1–5 μM) dramatically decreased (p < 0.05) GC proliferation. Using cell number data, the IC50 of ENNA was estimated at 2 μM for both follicle sizes. We conclude that ENNA can directly inhibit ovarian function in cattle, decreasing the proliferation and steroid production of GC. Full article
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8 pages, 962 KiB  
Article
Analysis of the Roles of the ISLR2 Gene in Regulating the Toxicity of Zearalenone Exposure in Porcine Intestinal Epithelial Cells
by Zhenbin Bi, Xuezhu Gu, Yeyi Xiao, Yajing Zhou, Wenbin Bao, Shenglong Wu and Haifei Wang
Toxins 2022, 14(9), 639; https://doi.org/10.3390/toxins14090639 - 16 Sep 2022
Cited by 3 | Viewed by 1515
Abstract
Zearalenone (ZEN) is one of the mycotoxins that pose high risks for human and animal health, as well as food safety. However, the regulators involved in ZEN cellular toxicity remain largely unknown. Herein, we showed that cell viability of porcine intestinal epithelial cells [...] Read more.
Zearalenone (ZEN) is one of the mycotoxins that pose high risks for human and animal health, as well as food safety. However, the regulators involved in ZEN cellular toxicity remain largely unknown. Herein, we showed that cell viability of porcine intestinal epithelial cells (IPEC-J2) tended to decrease with increasing doses of ZEN by the cell counting kit-8 assay. Expression of the ISLR2 (immunoglobulin superfamily containing leucine-rich repeat 2) gene in IPEC-J2 cells was significantly downregulated upon ZEN exposure. Furthermore, we found the dose–effect of ZEN on ISLR2 expression. We then overexpressed the ISLR2 gene and observed that overexpression of ISLR2 obviously reduced the effects of ZEN on cell viability, apoptosis rate and oxidative stress level. In addition, ISLR2 overexpression significantly decreased the expression of TNF-α and IFN-α induced by ZEN. Our findings revealed the effects of ZEN on the ISLR2 gene expression and indicated the ISLR2 gene as a novel regulator of ZEN-induced cytotoxicity, which provides potential molecular targets against ZEN toxicity. Full article
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16 pages, 2612 KiB  
Article
Assessment of Zearalenone-Induced Cell Survival and of Global Gene Regulation in Mouse TM4 Sertoli Cells
by Christian Savard, Sadaf Gawhary, Alexandre Boyer and Younes Chorfi
Toxins 2022, 14(2), 98; https://doi.org/10.3390/toxins14020098 - 26 Jan 2022
Cited by 5 | Viewed by 2562
Abstract
Zearalenone (ZEA) is a non-steroidal xenoestrogen mycotoxin produced by many Fusarium fungal species, which are common contaminants of cereal crops destined for worldwide human and animal consumption. ZEA has been reported in various male reproduction dysfonctions, including decreased fertility potential. In this report, [...] Read more.
Zearalenone (ZEA) is a non-steroidal xenoestrogen mycotoxin produced by many Fusarium fungal species, which are common contaminants of cereal crops destined for worldwide human and animal consumption. ZEA has been reported in various male reproduction dysfonctions, including decreased fertility potential. In this report, the direct effect of ZEA on the immature Sertoli TM4 cell line was evaluated. The results show that high concentrations of ZEA increase reactive oxygen species via the activation of MAPK signaling. Transcriptome analysis was performed on the TM4 cell line treated with ZEA, and genes involved in sex differentiation (Fgfr2, Igf1, Notch1, Sox9) and extracellular matrix (ECM) formation (Ctgf, Fam20a, Fbn1, Mmp9, Postn, Sparcl1, Spp1) were identified at the center of the functional protein association network, suggesting that ZEA could be detrimental to the early steps of Sertoli cell differentiation. Full article
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17 pages, 1202 KiB  
Article
Evaluation of the Antioxidant, Anti-Inflammatory and Cytoprotective Activities of Halophyte Extracts against Mycotoxin Intoxication
by Nolwenn Hymery, Xavier Dauvergne, Halima Boussaden, Stéphane Cérantola, Dorothée Faugère and Christian Magné
Toxins 2021, 13(5), 312; https://doi.org/10.3390/toxins13050312 - 27 Apr 2021
Cited by 5 | Viewed by 2526
Abstract
Twelve halophyte species belonging to different families, widely represented along French Atlantic shoreline and commonly used in traditional medicine, were screened for protective activities against mycotoxins, in order to set out new promising sources of natural ingredients for feed applications. Selected halophytic species [...] Read more.
Twelve halophyte species belonging to different families, widely represented along French Atlantic shoreline and commonly used in traditional medicine, were screened for protective activities against mycotoxins, in order to set out new promising sources of natural ingredients for feed applications. Selected halophytic species from diverse natural habitats were examined for their in vitro anti-mycotoxin activities, through viability evaluation of Madin-Darby Bovine Kidney (MDBK) and intestinal porcine enterocyte (IPEC-J2) cell lines. Besides, the in vitro antioxidant activities of plant extracts were assessed (total antioxidant and 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging bioassays). Of the 12 species, Galium arenarium, Convolvulus soldanella and Eryngium campestre exhibited the most protective action on MDBK and IPEC-J2 cells against zearalenone (ZEN) or T2 toxin contamination (restoring about 75% of cell viability at 10 μg·mL−1) without inflammation response. They also had strong antioxidant capacities (Inhibitory concentration of 50% (IC50) < 100 μg·mL−1 for DPPH radical and total antioxidant capacity (TAC) of 100 to 200 mg Ascorbic Acid Equivalent (AAE)·g−1 Dry Weight), suggesting that cell protection against intoxication involves antioxidant action. A bio-guided study showed that fractions of G. arenarium extract protect MDBK cells against T2 or ZEN toxicity and several major compounds like chlorogenic acid and asperuloside could be involved in this protective effect. Overall, our results show that the halophytes G. arenarium, C. soldanella and E. campestre should be considered further as new sources of ingredients for livestock feed with protective action against mycotoxin intoxication. Full article
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13 pages, 2242 KiB  
Article
Effects of Deoxynivalenol and Mycotoxin Adsorbent Agents on Mitogen-Activated Protein Kinase Signaling Pathways and Inflammation-Associated Gene Expression in Porcine Intestinal Epithelial Cells
by Yu-Hsiang Yu, Yi-Han Lai, Felix Shih-Hsiang Hsiao and Yeong-Hsiang Cheng
Toxins 2021, 13(5), 301; https://doi.org/10.3390/toxins13050301 - 23 Apr 2021
Cited by 17 | Viewed by 2505
Abstract
Deoxynivalenol (DON) is the most prevalent mycotoxin in swine feedstuffs. The intestinal epithelial cells represent the first target for the DON. Here, we studied the effects of DON and mycotoxin adsorbent agents on mitogen-activated protein kinase (MAPK) signaling pathways and inflammation-associated gene expression [...] Read more.
Deoxynivalenol (DON) is the most prevalent mycotoxin in swine feedstuffs. The intestinal epithelial cells represent the first target for the DON. Here, we studied the effects of DON and mycotoxin adsorbent agents on mitogen-activated protein kinase (MAPK) signaling pathways and inflammation-associated gene expression in porcine intestinal epithelial cells (IPEC-J2). Results showed that phosphorylation of MAPK signaling pathways (p38, ERK, and JNK) was increased after treatment of DON or lipopolysaccharide (LPS) in IPEC-J2 cells. The phosphorylation of p38, ERK, and JNK was not further enhanced after co-treatment with DON and LPS. The inos and cox-2 mRNA expression were significantly induced at 6 h after treatment of DON. DON treatment significantly increased the claudin 3 and occludin mRNA expression at 12 h. DON in combination with LPS treatment did not further increase the inflammation and tight junction-associated gene expression. The DON-induced phosphorylation of MAPK signaling pathways was impaired by mycotoxin adsorbent agent (nanoscale silicate platelets and the mixture of montmorillonites and yeast cell walls) treatment, thereby decreasing inflammation and tight junction-associated gene expression. Taken together, these findings demonstrate that DON triggers the inflammation in IPEC-J2 cells by phosphorylation of MAPK signaling pathways and LPS does not further augment the DON-induced inflammatory responses. Mycotoxin adsorbent agents can attenuate DON-induced inflammatory responses in IPEC-J2 cells through modulation of the phosphorylation of p38, ERK, and JNK. Full article
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17 pages, 3301 KiB  
Article
Transcriptome Analysis Reveals the AhR, Smad2/3, and HIF-1α Pathways as the Mechanism of Ochratoxin A Toxicity in Kidney Cells
by Min Cheol Pyo, In-Geol Choi and Kwang-Won Lee
Toxins 2021, 13(3), 190; https://doi.org/10.3390/toxins13030190 - 06 Mar 2021
Cited by 21 | Viewed by 3092
Abstract
Ochratoxin A (OTA) is a mycotoxin occurring in foods consumed by humans. Recently, there has been growing global concern regarding OTA toxicity. The main target organ of OTA is the kidney, but the mechanism underlying renal toxicity is not well known. In this [...] Read more.
Ochratoxin A (OTA) is a mycotoxin occurring in foods consumed by humans. Recently, there has been growing global concern regarding OTA toxicity. The main target organ of OTA is the kidney, but the mechanism underlying renal toxicity is not well known. In this study, human-derived proximal tubular epithelial cells, HK-2 cells, were used for RNA-sequencing (RNA-seq) and transcriptome analysis. In total, 3193 differentially expressed genes were identified upon treatment with 200 nM OTA in HK-2 cells; of these, 2224 were upregulated and 969 were downregulated. Transcriptome analysis revealed that OTA significantly affects hypoxia, epithelial-mesenchymal transition (EMT), apoptosis, and xenobiotic metabolism pathways in kidney cells. Quantitative real-time PCR analysis showed gene expression patterns similar to RNA-seq analysis. Expression of EMT markers (E-cadherin and fibronectin), apoptosis markers (caspase-3 and Bax), and kidney injury molecule-1 (KIM-1) was suppressed by inhibiting AhR expression using siRNA, and the related transcription factors, Smad2/3, and HIF-1α were downregulated. Smad2/3 suppression with siRNA could inhibit fibronetcin, caspase-3, Bax, and KIM-1 expression. Fibronetcin, caspase-3, Bax, and KIM-1 expression could be increased with HIF-1α suppression with siRNA. Taken together, these findings suggest that OTA-mediated kidney toxicity via the AhR-Smad2/3-HIF-1α signaling pathways leads to induction of EMT, apoptosis, and kidney injury. Full article
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12 pages, 5230 KiB  
Article
Primary Impacts of the Fungal Toxin Sporidesmin on HepG2 Cells: Altered Cell Adhesion without Oxidative Stress or Cell Death
by Magalie Boucher and T. William Jordan
Toxins 2021, 13(3), 179; https://doi.org/10.3390/toxins13030179 - 28 Feb 2021
Cited by 1 | Viewed by 2033
Abstract
The fungal metabolite sporidesmin is responsible for severe necrotizing inflammation of biliary tract and liver of livestock grazing on pasture containing spores of Pithomyces chartarum that synthesizes the toxin. The toxin is secreted into bile causing the erosion of the biliary epithelium accompanied [...] Read more.
The fungal metabolite sporidesmin is responsible for severe necrotizing inflammation of biliary tract and liver of livestock grazing on pasture containing spores of Pithomyces chartarum that synthesizes the toxin. The toxin is secreted into bile causing the erosion of the biliary epithelium accompanied by inflammation and damage to surrounding tissues. Toxicity has been suggested to be due to cycles of reduction and oxidation of sporidesmin leading to oxidative damage from the formation of reactive oxygen species. The current work is the first test of the oxidative stress hypothesis using cultured cells. Oxidative stress could not be detected in HepG2 cells incubated with sporidesmin using a dichlorodihydrofluorescein diacetate assay or by use of two-dimensional electrophoresis to search for oxidized peroxiredoxins. There was also no evidence for necrosis or apoptosis, although there was a loss of cell adhesion that was accompanied by the disruption of intracellular actin microfilaments that have known roles in cell adhesion. The results are consistent with a model in which altered contact between cells in situ leads to altered permeability and subsequent inflammation and necrosis, potentially from the leakage of toxic bile into surrounding tissues. There is now a need for the further characterization of the damage processes in vivo, including the investigation of altered permeability and mechanisms of cell death in the biliary tract and other affected organs. Full article
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