Metabolic profiling as a powerful tool for the analysis of cellular alterations caused by 20 mycotoxins in HepG2 cells

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.

Dose‐dependent expression of GFI1 alters metabolism in the haematopoietic progenitors and MLL::AF9‐induced leukaemic cells

“…Intrigued by our in‐vitro and ex‐vivo observations, we next sought to gain a further understanding of the metabolic alterations. To this end, we employed HILIC‐MS metabolic profiling to quantify the intracellular levels of the intermediary metabolites of pathways such as glycolysis, TCA cycle, glutamine and fatty acid metabolism in primary MA9 GFI1‐KI and ‐KD leukaemic cells (KIT+GFP+) using a published method with instrumental modifications 18 . Leukaemic cells with low expression levels of GFI1 showed significantly increased amounts of metabolites of the glycolysis pathway (Figure 6).…”

Glycolytic interference blocks influenza A virus propagation by impairing viral polymerase-driven synthesis of genomic vRNA

“…Moreover, we were interested in metabolic changes aside from glycolysis. Via hydrophilic interaction liquid chromatography (HILIC) coupled to tandem mass spectrometry (MS/MS), as described previously [36], we analyzed major alterations of the metabolic profile of A549 cells, induced by IAV infection and/or the treatment with 2-DG after 24 h ( Fig 6 ).…”

“…The dish was washed with additional 800 µL ACN/water (4+1, v/v) and pooled with the respective cell sample. Further preparation of samples as well as chromatographic and mass spectrometric analysis were performed as described previously [36]. S1.…”

High Throughput Metabolomics In vitro Platform for The Identification of Hepatotoxicity Modes of Action

“…Therefore, even though we increased the LC spectrum signi cantly, the detection of polar metabolites was limited. Our current method could potentially be improved by optimizing the HILIC protocol as shown by (Gerdemann et al 2022), by adding pre-column derivatization steps (Walvekar et al 2018) or by implementing an additional method for energy metabolism metabolites (Balcke et al 2011).…”