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Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures
Published on: March 28, 2017
Metabolic profiling as a powerful tool for the analysis of cellular alterations caused by 20 mycotoxins in HepG2
Andrea Gerdemann1, Matthias Behrens1, Melanie Esselen1
1Institute of Food Chemistry, University of Münster, Corrensstraße 45, 48149, Münster, Germany.
Abstract:
Mycotoxins are secondary fungal metabolites which exhibit toxic effects in low concentrations. Several mycotoxins are described as carcinogenic or immunosuppressive, but their underlying modes of action especially on molecular level have not yet been entirely elucidated. Metabolic profiling as part of the omics methods is a powerful tool to study the toxicity and the mode of action of xenobiotics. The use of hydrophilic interaction chromatography in combination with targeted mass spectrometric detection enables the selective and sensitive analysis of more than 100 polar and ionic metabolites and allows the evaluation of metabolic alterations caused by xenobiotics such as mycotoxins. For metabolic profiling, the hepato-cellular carcinoma cell line HepG2 was treated with sub-cytotoxic concentrations of 20 mycotoxins. Moniliformin and citrinin significantly affected target elements of the citric acid cycle, but also influenced glycolytic pathways and energy metabolism. Penitrem A, zearalenone, and T2 toxin mainly interfered with the urea cycle and the amino acid homeostasis. The formation of reactive oxygen species seemed to be influenced by T2 toxin and gliotoxin. Glycolysis was altered by ochratoxin A and DNA synthesis was affected by several mycotoxins. The observed effects were not limited to these metabolic reactions as the metabolic pathways are closely interrelated. In general, metabolic profiling proved to be a highly sensitive tool for hazard identification in comparison to single-target cytotoxicity assays as metabolic alterations were already observed at sub-toxic concentrations. Metabolic profiling could therefore be a powerful tool for the overall evaluation of the toxic properties of xenobiotics.
Insights
Metabolic profiling reveals how mycotoxins impact cellular processes even at low doses. This omics approach identifies toxic effects on energy metabolism and amino acid balance, aiding hazard identification.
Area of Science:
- Toxicology
- Metabolomics
- Biochemistry
Background:
- Mycotoxins are fungal metabolites with known toxic effects, but their molecular mechanisms remain unclear.
- Understanding xenobiotic toxicity requires detailed analysis of cellular metabolic changes.
- Omics methods, particularly metabolic profiling, offer a powerful approach to investigate these effects.
Purpose of the Study:
- To elucidate the molecular mechanisms of mycotoxin toxicity using metabolic profiling.
- To identify specific metabolic pathways affected by various mycotoxins.
- To evaluate metabolic profiling as a sensitive tool for xenobiotic hazard identification.
Main Methods:
- Treatment of HepG2 cells with sub-cytotoxic concentrations of 20 different mycotoxins.
- Hydrophilic interaction chromatography coupled with targeted mass spectrometry for metabolite analysis.
- Comprehensive metabolic profiling to detect alterations in cellular metabolism.
Main Results:
- Moniliformin and citrinin disrupted the citric acid cycle, glycolysis, and energy metabolism.
- Penitrem A, zearalenone, and T2 toxin affected urea cycle and amino acid homeostasis.
- T2 toxin and gliotoxin influenced reactive oxygen species formation; ochratoxin A altered glycolysis and DNA synthesis.
Conclusions:
- Metabolic profiling effectively identifies toxicity at sub-toxic concentrations, outperforming single-target assays.
- Observed metabolic alterations highlight the interconnectedness of cellular pathways.
- Metabolic profiling is a sensitive and valuable tool for evaluating the toxic properties of xenobiotics like mycotoxins.
