Stable Isotope-Assisted Metabolomics for Deciphering Xenobiotic Metabolism in Mammalian Cell Culture

Flasch, Mira; Bueschl, Christoph; Woelflingseder, Lydia; Schwartz-Zimmermann, Heidi E.; Adam, Gerhard; Schuhmacher, Rainer; Marko, Doris; Warth, Benedikt

doi:10.1021/acschembio.9b01016

Cited by 29 publications

(20 citation statements)

References 63 publications

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“…Therefore, it was likely to speculate that due to the observed increase of monolayer tightness after CER incubation ( Figure 1 ), DON may be taken up to a lesser extent, which was not the case ( Figure 4 ). While DON-3-S was recently reported to be produced by HepG2 liver cells [ 43 ], the present study demonstrates for the first time the formation of this metabolite in differentiated Caco-2 cells ( Figure 4 ). Our data support the hypothesis that DON-3-S is preferentially excreted to the basolateral side of the transwell system ( Figure 4 ).…”

Section: Discussionsupporting

confidence: 56%

Assessing Mixture Effects of Cereulide and Deoxynivalenol on Intestinal Barrier Integrity and Uptake in Differentiated Human Caco-2 Cells

Beisl

Varga

Braun

et al. 2021

Toxins

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The human intestine is regularly exposed to ingested food contaminants, such as fungal and bacterial toxins, which have been described to co-occur in a mixed diet. Thus, it is of utmost importance to understand possible interactions between contaminants of different origin. Hence, we investigated the single and combined effects of one of the most abundant mycotoxins, deoxynivalenol (DON; 0.1 to 10 µg/mL), and the bacterial toxin cereulide (CER; 1 to 100 ng/mL) on differentiated human Caco-2 (C2BBe1) cells cultured in a transwell system. We tested the capacity of the two toxins to alter the intestinal integrity and further investigated the uptake of both compounds and the formation of selected DON metabolites. CER alone (10 and 100 ng/mL) and in combination with DON (10 ng/mL CER with 1 µg/mL DON) was found to alter the barrier function by increasing the transepithelial electrical resistance and the expression of the tight junction protein claudin-4. For the first time, DON-3-sulfate was identified as a metabolite of human intestinal cells in vitro. Moreover, co-incubation of CER and DON led to an altered ratio between DON and DON-3-sulfate. Hence, we conclude that co-exposure to CER and DON may alter the intestinal barrier function and biotransformation of intestinal cells.

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Section: Discussionsupporting

confidence: 56%

Assessing Mixture Effects of Cereulide and Deoxynivalenol on Intestinal Barrier Integrity and Uptake in Differentiated Human Caco-2 Cells

Beisl

Varga

Braun

et al. 2021

Toxins

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“…Xenobiotic-derived metabolites are usually difficult to identify as they are: (i) most often low concentrated (xenobiotics are usually tested at low concentration, and metabolic intermediates are transient and low populated), (ii) often chemical structures absent from reference databases, and (iii) few among thousands of metabolites issued from the regular cell metabolism. A typical application of 13C labeling is the follow-up of metabolization of a 13C-labeled chemical such as a xenobiotic [ 148 , 149 ]. Both intracellular and extracellular metabolites provide complimentary pictures of xenobiotics metabolization processes.…”

Section: Addressing Cellular Response To Xenobioticsmentioning

confidence: 99%

Metabolomic Approaches to Study Chemical Exposure-Related Metabolism Alterations in Mammalian Cell Cultures

Balcerczyk

Damblon

Elena-Herrmann

et al. 2020

IJMS

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Biological organisms are constantly exposed to an immense repertoire of molecules that cover environmental or food-derived molecules and drugs, triggering a continuous flow of stimuli-dependent adaptations. The diversity of these chemicals as well as their concentrations contribute to the multiplicity of induced effects, including activation, stimulation, or inhibition of physiological processes and toxicity. Metabolism, as the foremost phenotype and manifestation of life, has proven to be immensely sensitive and highly adaptive to chemical stimuli. Therefore, studying the effect of endo- or xenobiotics over cellular metabolism delivers valuable knowledge to apprehend potential cellular activity of individual molecules and evaluate their acute or chronic benefits and toxicity. The development of modern metabolomics technologies such as mass spectrometry or nuclear magnetic resonance spectroscopy now offers unprecedented solutions for the rapid and efficient determination of metabolic profiles of cells and more complex biological systems. Combined with the availability of well-established cell culture techniques, these analytical methods appear perfectly suited to determine the biological activity and estimate the positive and negative effects of chemicals in a variety of cell types and models, even at hardly detectable concentrations. Metabolic phenotypes can be estimated from studying intracellular metabolites at homeostasis in vivo, while in vitro cell cultures provide additional access to metabolites exchanged with growth media. This article discusses analytical solutions available for metabolic phenotyping of cell culture metabolism as well as the general metabolomics workflow suitable for testing the biological activity of molecular compounds. We emphasize how metabolic profiling of cell supernatants and intracellular extracts can deliver valuable and complementary insights for evaluating the effects of xenobiotics on cellular metabolism. We note that the concepts and methods discussed primarily for xenobiotics exposure are widely applicable to drug testing in general, including endobiotics that cover active metabolites, nutrients, peptides and proteins, cytokines, hormones, vitamins, etc.

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“…4,5 The main research interest of phase II modications has been for the metabolism of drugs, nutritional components, toxins, and doping agents. [6][7][8][9][10] Phase II modications have recently also been linked to the cometabolism of the gut microbiome with their human host. Mainly sulfation but also glucuronidation are important modications for metabolites derived from the co-metabolism of the microbiome and its host.…”

Section: Introductionmentioning

confidence: 99%

Investigation of the individual human sulfatome in plasma and urine samples reveals an age-dependency

et al. 2021

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Stable Isotope-Assisted Metabolomics for Deciphering Xenobiotic Metabolism in Mammalian Cell Culture

Cited by 29 publications

References 63 publications

Assessing Mixture Effects of Cereulide and Deoxynivalenol on Intestinal Barrier Integrity and Uptake in Differentiated Human Caco-2 Cells

Assessing Mixture Effects of Cereulide and Deoxynivalenol on Intestinal Barrier Integrity and Uptake in Differentiated Human Caco-2 Cells

Metabolomic Approaches to Study Chemical Exposure-Related Metabolism Alterations in Mammalian Cell Cultures

Investigation of the individual human sulfatome in plasma and urine samples reveals an age-dependency

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