Sulfur mustard resistant keratinocytes obtained elevated glutathione levels and other changes in the antioxidative defense mechanism.

Published on Nov 26, 2017in Toxicology Letters4.374
· DOI :10.1016/J.TOXLET.2017.11.024
Simone Rothmiller6
Estimated H-index: 6
 , Sarah Schröder1
Estimated H-index: 1
 + 7 AuthorsAnnette Schmidt6
Estimated H-index: 6
 (Bundeswehr University Munich)
Sources
Abstract
Abstract Background Sulfur mustard (SM) is a potent blistering chemical warfare agent, which was first used in 1917. Despite the Chemical Weapons Convention, a use was recently reported in Syria in 2015. This emphasizes the importance to develop countermeasures against chemical warfare agents. Despite intensive research, there is still no antidote or prophylaxis available against SM. Methods The newly developed SM-resistant keratinocyte cell line HaCaT/SM was used to identify new target structures for drug development, particularly the adaptations in protective measures against oxidative stress. For this purpose, glutathione (GSH) and NAD(P)H levels, the effect of glutathione S-transferase (GST) inhibition as well as activation and expression of Nrf2, GST, glutamate cysteine ligase (GCL) and glutathione-disulfide reductase (GSR) as well as multi-drug resistance (MDR) proteins 1, 3 and 5 were investigated. Results The HaCaT/SM cells showed not only a better survival after treatment with SM or cytostatic drugs, but also hydrogen peroxide (H2O2). They exhibit more GSH even after SM treatment. Nrf2 levels were significantly lower. Inhibition of GST led to significantly decreased, activation to slightly higher IC50 values after SM treatment and a lower expression of GST was observed. The cells also expressed less GCLC and GSR. Expression of MDR1, MDR3 and MDR5 was higher under control conditions, but less stimulated by SM treatment. An increased NADP+/NADPH ratio as well as higher NAD+ levels were shown. Conclusion In summary, an improved response of the resistant cell line to oxidative stress was observed. The underlying mechanisms are elevated GSH levels as well as lower expression of Nrf2 and its targets GCLC and GST as well as GSR and MDR1, MDR3 and MDR5. GST is an especially interesting target because its inhibition already induced a significant SM sensitivity. SM resistance also caused redox equivalent level differences. Taken together, these findings provide further insight into the mechanism of SM resistance and may open a window for novel therapeutic targets in SM therapy.
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Aug 18, 2017·Toxicology Letters4.37
#1Simone RothmillerH-Index: 6
#2Markus WolfH-Index: 6
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Abstract Background MicroRNAs (miRNAs) are responsible for post-transcriptional control of protein expression. Numerous miRNAs have been identified to be responsible for the resistance of tumor cells to cytostatic drugs. Possibly, the same miRNAs also play a role in the sulfur mustard (SM)-resistance of the keratinocyte cell line HaCaT/SM as alkylating cytostatics exhibit similar cytotoxic effects as SM. Methods Basal expression levels of 1920 miRNAs in total were analyzed in HaCaT/SM compared t...
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Jul 13, 2017·Nature Communications14.92
#1Xiqian Jiang (BCM: Baylor College of Medicine)H-Index: 8
#2Jianwei Chen (BCM: Baylor College of Medicine)H-Index: 19
Last. Jin Wang (BCM: Baylor College of Medicine)H-Index: 33
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Fluorescent sensors for small biomolecules are needed to shed insight into real-time cellular processes. Here the authors develop RealThiol, a sensor that can quantitatively monitor glutathione dynamics in living cells, and measure increased antioxidant capability of activated neurons and glutathione changes during ferroptosis.
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May 5, 2017·Toxicology Mechanisms and Methods2.99
#1Miroslav Pohanka (Mendel University)H-Index: 33
#2Pavla Martinkova (Central European Institute of Technology)H-Index: 7
Last. Jindrich Kynicky (Central European Institute of Technology)H-Index: 6
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AbstractSulfur mustard, in a chemical name bis(2-chloroethyl) sulfide, is a chemical warfare agent. It is cytotoxic and blister forming once spread over the skin. Though exact molecular mechanism of sulfur mustard toxic action remains unknown, inflammation and oxidative stress development are considered as the most relevant pathological consequences. Applications of either low-molecular weight antioxidants or cofactors for enzymatic antioxidants are considered as suitable ways how to ameliorate ...
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Jan 1, 2017·Archives of Toxicology5.15
#1Bernhard Stenger (LMU: Ludwig Maximilian University of Munich)H-Index: 4
#2Tanja Popp (LMU: Ludwig Maximilian University of Munich)H-Index: 15
Last. Dirk Steinritz (LMU: Ludwig Maximilian University of Munich)H-Index: 24
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Transient receptor potential family channels (TRPs) have been identified as relevant targets in many pharmacological as well as toxicological studies. TRP channels are ubiquitously expressed in different tissues and act among others as sensors for different external stimuli, such as mechanical stress or noxious impacts. Recent studies suggest that one member of this family, the transient receptor potential ankyrin 1 cation channel (TRPA1), is involved in pain, itch, and various diseases, suggest...
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Apr 1, 2016·Mutation Research-genetic Toxicology and Environmental Mutagenesis2.87
#1Eisa Tahmasbpour (BMSU: Baqiyatallah University of Medical Sciences)H-Index: 12
#2Mostafa Ghanei (BMSU: Baqiyatallah University of Medical Sciences)H-Index: 40
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Abstract Sulfur mustard (SM) is a potent alkylating agent that targets several organs, especially lung tissue. Although pathological effects of SM on mustard lung have been widely considered, molecular and cellular mechanisms for these pathologies are poorly understood. We investigated changes in expression of genes related to oxidative stress (OS) and antioxidant defense caused by SM in lung tissue of patients. We performed gene expression profiling of OS and antioxidant defense in lung tissue ...
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Feb 26, 2016·Toxicology Letters4.37
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Last. Dirk Steinritz (LMU: Ludwig Maximilian University of Munich)H-Index: 24
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Sulfur mustard (SM) is a chemical warfare agent causing blistering, inflammation and ulceration of the skin. Thiol compounds such as glutathione (GSH) and N-acetylcysteine (NAC) have been suggested as potential antidotes. We investigated SM toxicity in a human keratinocyte cell line (HaCaT) and used GSH and NAC to counteract its cytotoxic effects. Cells were treated with 1, 5 or 10 mM GSH or NAC and exposed to 30, 100 or 300 mu M SM. Different treatment regimens were applied to model extra-and i...
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Feb 26, 2016·Toxicology Letters4.37
#1Dirk Steinritz (LMU: Ludwig Maximilian University of Munich)H-Index: 24
#2Enno StrieplingH-Index: 3
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Sulfur mustard (SM) is a chemical warfare agent (CWA) that was first used in World War I and in several military conflicts afterwards. The threat by SM is still present even today due to remaining stockpiles, old and abandoned remainders all over the world as well as to its ease of synthesis. CWA are banned by the Chemical Weapons Convention (CWC) interdicting their development, production, transport, stockpiling and use and are subjected to controlled destruction. The present case report descri...
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Feb 26, 2016·Toxicology Letters4.37
#1Aswin Mangerich (University of Konstanz)H-Index: 19
#2Malgorzata Debiak (University of Konstanz)H-Index: 7
Last. Alexander Bürkle (University of Konstanz)H-Index: 67
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Abstract Mustard agents are potent DNA alkylating agents with mutagenic, cytotoxic and vesicant properties. They include bi-functional agents, such as sulfur mustard (SM) or nitrogen mustard (mustine, HN2), as well as mono-functional agents, such as “half mustard” (CEES). Whereas SM has been used as a chemical warfare agent, several nitrogen mustard derivatives, such as chlorambucil and cyclophosphamide, are being used as established chemotherapeutics. Upon induction of specific forms of genotox...
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Feb 26, 2016·Toxicology Letters4.37
#1Markus WolfH-Index: 6
#2Markus Siegert (LMU: Ludwig Maximilian University of Munich)H-Index: 9
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Abstract Background The cell line HaCaT/SM was derived from the human keratinocyte cell line HaCaT. HaCaT/SM cells display a high resistance against sulfur mustard (SM). Intention of the presented study was to determine the cellular and molecular differences between HaCaT/SM and HaCaT so as to evaluate which changes might be responsible for being resistant against SM. Methods Both cell lines HaCaT and HaCaT/SM were analyzed with respect to their cell growth, nuclei perimeter, clonogenicity and s...
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Feb 26, 2016·Toxicology Letters4.37
#1Annette Schmidt (German Sport University Cologne)H-Index: 29
#2Dirk Steinritz (LMU: Ludwig Maximilian University of Munich)H-Index: 24
Last. Horst ThiermannH-Index: 50
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Pairs of corresponding cytotoxic drug sensitive and resistant cell lines are powerful tools to develop treatment strategies. Developing cytotoxic drug resistant cell lines is a well-established method in cancer research. In more than fifty years of sulfur mustard (SM) resistant research such a cell pair has never been produced. Hereinafter we describe the first successful approach to develop a SM resistant keratinocyte cell line. Starting with the SM sensitive keratinocyte cell line HaCaT we use...
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Cited By6
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Summary Glutathione (GSH) is a highly dynamic, high abundance molecule regulating redox homeostasis in most mammalian cells. Traditional methods could not achieve quantification of glutathione in live cells with high spatial and temporal resolution. Here, we provide protocols on how to use reversible reaction-based ratiometric fluorescent probes, RealThiol (RT) and its derivatives, to quantify GSH globally or in specific organelles. The protocols are applicable to cultured or harvested cells thr...
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Feb 28, 2020·Experimental and Molecular Pathology3.36
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Abstract Nitrogen mustard (NM) is a highly reactive bifunctional alkylating agent that induces inflammation, edema and blistering in skin. An important mechanism mediating the action of NM and related mustards is oxidative stress. In these studies a modified murine patch-test model was used to analyze DNA damage and the antioxidant/stress response following NM exposure in isolated epidermis. NM (20 μmol) was applied to glass microfiber filters affixed to a shaved dorsal region of skin of CD-1 mi...
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Abstract Sulfur mustard (SM) is a toxicant and chemical warfare agent with strong vesicant properties. The mechanisms behind SM-induced toxicity are not fully understood and no antidote or effective therapy against SM exists. Both, the risk of SM release in asymmetric conflicts or terrorist attacks and the usage of SM-derived nitrogen mustards as cancer chemotherapeutics, render the mechanisms of mustard-induced toxicity a highly relevant research subject. Herein, we review a central role of the...
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Abstract Vesicants cause a multitude of cutaneous reactions like erythema, blisters and ulcerations. After exposure to sulfur mustard (SM) and related compounds, patients present dermal symptoms typically known for chemicals categorized as skin sensitizer (e.g. hypersensitivity and flare-up phenomena). However, although some case reports led to the assumption that SM and other alkylating compounds represent sensitizers, a comprehensive investigation of SM-triggered immunological responses has no...
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Jan 25, 2019·Chemico-Biological Interactions5.19
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Abstract Inhalation of the chemical warfare agent sulfur mustard (SM) is associated with severe acute and long-term pulmonary dysfunctions and health effects. The still not completely elucidated molecular toxicology and a missing targeted therapy emphasize the need for further research. However, appropriate human data are extremely rare. In vivo animal experiments are often regarded as gold standard in toxicology but may exhibit significant differences compared to the human pulmonary anatomy and...
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Apr 23, 2018·Nature Communications14.92
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