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References (13)
-
T. Schettgen, A. Alt, D. Preim, D. Keller, T. Kraus (2012)
Biological monitoring of indoor-exposure to dioxin-like and non-dioxin-like polychlorinated biphenyls (PCB) in a public building.Toxicology letters, 213 1
-
Chunyun Zhang, Xueshu Li, S. Flor, P. Ruiz, A. Kruve, G. Ludewig, H. Lehmler (2022)
Metabolism of 3-Chlorobiphenyl (PCB 2) in a Human-Relevant Cell Line: Evidence of Dechlorinated MetabolitesEnvironmental Science & Technology, 56
-
T. Kraus, M. Gube, J. Lang, A. Esser, W. Sturm, B. Fimm, K. Willmes, J. Neulen, J. Baron, H. Merk, T. Schettgen, Kerstin Konrad, S. Deisz, L. Rink, M. Hagmann, Birgit Fillies, W. Zschiesche, J. Wittsiepe, M. Wilhelm (2012)
Surveillance Program for Former PCB-Exposed Workers of a Transformer and Capacitor Recycling Company, Family Members, Employees of Surrounding Companies, and Area Residents—Executive SummaryJournal of Toxicology and Environmental Health, Part A, 75
-
M. Tulp, W. Bruggeman, O. Hutzinger (1977)
Reductive dechlorination of chlorobiphenylols by ratsExperientia, 33
-
T. Schettgen, M. Gube, A. Esser, A. Alt, T. Kraus (2012)
Plasma Polychlorinated Biphenyls (PCB) Levels of Workers in a Transformer Recycling Company, their Family Members, and Employees of Surrounding CompaniesJournal of Toxicology and Environmental Health, Part A, 75
-
P. Espandiari, H. Glauert, H. Lehmler, Eun Lee, C. Srinivasan, L. Robertson (2003)
Polychlorinated biphenyls as initiators in liver carcinogenesis: resistant hepatocyte model.Toxicology and applied pharmacology, 186 1
-
T. Idda, C. Bonas, Jenny Hoffmann, J. Bertram, Natalia Quinete, Natalia Quinete, T. Schettgen, K. Fietkau, A. Esser, Matthias Stope, M. Leijs, Jens Baron, Thomas Kraus, Aaron Voigt, Patrick Ziegler (2020)
Metabolic activation and toxicological evaluation of polychlorinated biphenyls in Drosophila melanogasterScientific Reports, 10
-
T Vasko, J Hoffmann, S Gostek (2018)
Telomerase gene expression bioassays indicate metabolic activation og genotoxic lower chlorinated polychlorinated biphenylsSci Rep, 8
-
EQ Song, XY Ma, XG Tian (2013)
The effect of the structure of polychlorinated biphenyls on their hydroxylation, oxidation, and glutathionyl conjugation reactionsBiomed Environ Sci BES, 26
-
Fabian Grimm, D. Hu, I. Kania-Korwel, H. Lehmler, G. Ludewig, K. Hornbuckle, M. Duffel, Å. Bergman, L. Robertson (2015)
Metabolism and metabolites of polychlorinated biphenylsCritical Reviews in Toxicology, 45
-
Martin Kraft, S. Sievering, L. Grün, K. Rauchfuss (2018)
Mono‐, di‐, and trichlorinated biphenyls (PCB 1‐PCB 39) in the indoor air of office rooms and their relevance on human blood burdenIndoor Air, 28
-
M. Stiborová, R. Indra, Michaela Moserová, L. Bořek-Dohalská, P. Hodek, E. Frei, K. Kopka, H. Schmeiser, V. Arlt (2017)
Comparison of human cytochrome P450 1A1-catalysed oxidation of benzo[a]pyrene in prokaryotic and eukaryotic expression systemsMonatshefte Fur Chemie, 148
-
N. Quinete, T. Kraus, V. Belov, Christina Aretz, A. Esser, T. Schettgen (2015)
Fast determination of hydroxylated polychlorinated biphenyls in human plasma by online solid phase extraction coupled to liquid chromatography-tandem mass spectrometry.Analytica chimica acta, 888
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Abstract
Archives of Toxicology https://doi.org/10.1007/s00204-023-03621-1 TOXICOKINE TICS AND ME TABOLISM Partial dechlorination of 2,4,4′‑trichlorobiphenyl (PCB 28) mediated by recombinant human CYP1A2 1 2 3 3 1 1 Isabella Randerath · Natalia Quinete · Julian Peter Müller · Julia Stingl · Jens Bertram · Thomas Schettgen · 1 1 Thomas Kraus · Patrick Ziegler Received: 5 July 2023 / Accepted: 4 October 2023 © The Author(s) 2023 Keywords Polychlorinated biphenyls · PCB28 · Cytochrom p450 Monooxidase · Dechlorination Introduction show for the first time the partial dechlorination of PCB 28 to the dichlorinated monohydroxylated metabolite 3-OH- Polychlorinated Biphenyl (PCB) congeners show a long per- CB15, which is formed by human CYP1A2 from PCB28 sistence in the human body with elimination half-lives in the in addition to its already known trichlorinated metabolites. range of 1–20 years. Elimination involves xenobiotic-metab- olizing Phase I (cytochrome P450, i.e., CYP) and conjugat- ing Phase II enzymes, which can lead to the formation and Material and methods retention of OH-PCBs, PCB sulfates, PCB glucuronides, and PCB methyl sulfones (MeSO -PCBs) (Grimm et al. 2015). Analytical standards used in this study Metabolic profiles of PCB degradation are dramatically affected by chlorine substitution patterns with implications The analytical standards of the trichlorinated PCB28 for PCB toxicity and elimination half-live. Dechlorination metabolites 2,4,4′-trichloro-5-biphenylol (5-OH-PCB28), of PCBs is considered to detoxify higher-chlorinated PCBs 2′,3′,4-tr ichloro-4′-biphenylol (4′-OH-PCB25), and and has been demonstrated in animal models and in human 2,4′,5-trichloro-4-biphenylol (4-OH-PCB31) were custom cell culture (Tulp et al. 1977) (Zhang et al. 2022). synthesized at the Max Planck Institute for Biophysical PCB28 is one of the quantitatively most important con- Chemistry, Facility for Synthetic Chemistry (Göttingen, geners of industrial PCB mixtures. Together with other Germany). The characterization was performed via NMR lower-chlorinated PCBs (e.g., PCB52 and PCB101), it can spectroscopy and mass spectrometry (data not shown). be regularly detected in contaminated indoor environments 2,4,4′-trichloro-3′-biphenylol (3′-OH-PCB28) was purchased (Schettgen et al. 2012; Kraft et al. 2018) and is found in the from Combi-Blocks (San Diego, CA, USA). 4,4′-dichloro- recycling of transformers and capacitors (Schettgen et al. 3-biphenylol (3-OH-PCB15) was purchased by Chem 2011). There is experimental evidence from PCB28 that Service (West Chester, PA, USA). For quantifications an indicates a direct genotoxic effect and thus hints to possibly internal standard of C —3′-OH-PCB28 was custom syn- a direct carcinogenic effect (Vasko et al. 2018). We here thesized and characterized via NMR spectroscopy and mass spectrometry (Göttingen, Germany). * Patrick Ziegler Incubation of PCB28 with recombinant CYP1A2 pziegler@ukaachen.de and CYP1A2 expressing transgenic HEK293 cells Institute for Occupational, Social and Environmental Medicine, Medical Faculty, RWTH Aachen University, Recombinantly expressed CYP1A2 bactosomes (coex- Pauwelsstrasse 30, 52074 Aachen, Germany pressed with CYP reductase in Escherichia coli) were Department of Chemistry and Biochemistry, Institute obtained from Tebu-Bio (Offenbach, Germany) and mixed of Environment, Florida International University, 3000 NE in the concentrations described. Incubations contained 151st Street, North Miami, Florida 33181, USA PCB 28 (20 µM), HEPES buffer (50 mM pH 7.4), MgCl Institute of Clinical Pharmacology, University Hospital (30 mM), NADPH (1 mM), and 5 pmol of bactosomes in of RWTH, 52074 Aachen, Germany Vol.:(0123456789) 1 3 Archives of Toxicology a total volume of 500 µL. Incubations were performed in (50 U/mL), and streptomycin (50 μg/ml) in a total volume of triplicates and initialized by adding NADPH after 3 min of 2.5 ml. At a confluence of 80%, PCB 28 (20 µM) was added pre-incubation at 37 °C and terminated after 60 min. Control and cells were incubated for 24 h before harvest. Control samples were run in the absence of substrate, in the absence samples were run with non-transgenic HEK293 cells. of NADPH or using heat-deactivated enzymes (Idda et al. 2020). For the incubation of PCB28 with CYP1A2 express- Analysis of PCB metabolites ing transgenic HEK293 cells, 4 × 10 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supple- Supernatants from bactosome incubations or cell culture mented with 10% (vol/vol) fetal calf serum (FCS), penicillin experiments were diluted 1:2 with 80 µL acetate buffer Fig. 1 The extracted ion chro- matograms of trichlorinated (left panel) and dichlorinated (right panel) OH metabolites of PCB28. a RhCYP1A2 expressed in E.coli (bacto- somes) was incubated with PCB 28 for 1 h. b HEK293human- CYP1A2 cell line was incubated with 20 µM PCB28 for 24 h with subsequent harvest of the supernatant. c human plasma sample of a PCB28 exposed individual. d Standards. Repre- sentative images from 3 differ - ent experiments are shown. M (metabolite 1–4), DM (Dichlo- rinated Metabolite). Mass transitions are related to the following metabolites: trichlo- rinated 3′-OH-PCB28, 5-OH- PCB28, 4′-OH-PCB25, 4-OH- PCB31 m/z: 271.1- > 235.0 green, m/z: 273.2—> 237.0 red, m/z: 277.1—> 241.0 pink (the last one equalizing the internal standard C -3′-OH- PCB28); dichlorinated 3-OH- PCB15):m/z: 237.2—> 201.0 blue, m/z: 239.2—> 202.9 orange (color figure online) 1 3 Archives of Toxicology 0.1 M (pH = 5.3). 100 µL of this dilution was further incu- Functional expression of CYP1A2 could be demonstrated bated with 100 µL of ammonium acetate buffer 0.1 M by conversion of the common substrate caffeine to the spe- (pH = 5.3) and 5 µL of ß-Glucuronidase/Arylsulfatase cific paraxanthine product (Supplemental Fig. 2). Incuba- enzyme overnight in a drying oven at 37 °C for enzymatic tion of HEK293humanCYP1A2 cells with PCB28 leads to hydrolysis in order to release conjugated compounds. 50 µL the formation of all 4 known monohydroxylated metabolites −1 of a mix of internal standards (10 ng mL ) and 600 µL of of PCB28 as expected, whereas in non-transgenic parental methanol were added to the samples, then mixed by vortex- HEK293 cells metabolite formation could not be observed. ing for 1 min, and centrifuged for 10 min at 4500 rpm. The In addition, HEK293humanCYP1A2 cells formed 3-OH- individual supernatants were transferred to glass LC vials CB15 from PCB28 (Fig. 1b and Table 1). and evaporated to approximately 50 µL at 45 °C under a To find out whether metabolic formation of 3-OH-PCB15 −1 gentle stream of nitrogen. Finally, 0.1 mol L ammonium occurs also in vivo, we examined plasma samples from acetate buffer was added to a final volume of 100 µL and individuals in a cohort exposed to different PCB Aroclor then transferred to an insert for analysis. The online solid mixtures (HELPcB cohort) (Kraus et al. 2012) for the pres- phase extraction (SPE) method coupled to liquid chromatog- ence of 3-OH-CB15. Indeed, we found this metabolite in raphy-tandem mass spectrometry was carried out using an samples selected for high PCB28 concentrations (Fig. 1c API 5500 QTrap mass spectrometer (AB Sciex, Darmstadt, and Table 1). Moreover, the concentration of 3-OH-CB15 Germany) equipped with electrospray ionization (ESI) inter- correlated mainly with the concentration of the major face (Quinete et al. 2015). PCB28 metabolites 4-OH-CB31 and 3´-OH-CB28 (Fig. 2), both resulting from oxidation at the dichlorinated ring of the trichlorobiphenyl structure. In fact, there is indirect Results and discussion evidence (Song et al. 2013) that 3-OH-PCB15 is also pro- duced as a metabolite of PCB15, a potential minor compo- The metabolism of PCB28 was analyzed in vitro using nent of commercial PCB mixtures such as various types of recombinant human (rh)CYP1A2 expressed in E.coli (bacto- Aroclor (Supplemental Table 1). Thus, as composition of somes), producing several monohydroxylated trichlorinated Aroclor(s) to which the plasma donors were exposed to was metabolites identified as 5-OH-PCB28 (M1, RT 18.76 min), not reported, we cannot completely exclude the possibility 4-OH-PCB31 (M3, RT 18.28 min), and 3′-OH-PCB28/ that 3-OH-PCB15 in their blood resulted from hydroxyla- 4′-OH-PCB25 (M4/M2, RT 17.09 min) by comparison with tion of PCB15. the authentic standards (Fig. 1a and Table 1) and thereby We emphasize that––contrary to the general view that corroborating with the results found in our previous studies. dechlorination of PCBs is accompanied by their detoxifica- In addition, a prominent ion peak appeared in the chroma- tion––a more differentiated view must be taken in the case togram at a retention time of 16.25 min. A comparison with the standard compounds with the closest retention time sug- Table 1 Metabolites, experiment, and concentrations for each OH- gested that this peak resembled the hydroxylated metabolite PCB congener of 4,4′-dichlorobiphenyl (3-OH-CB15). To confirm meta- bolic origin of the above species, control experiments with Metabolite Experiment Concentration (µg/L) no NADPH, no PCB28, or with heat inactivated bactosomes 3′-OH-PCB28/ HEK293human 0 were conducted. Indeed, in the control experiments none of 4′-OH- HEK293humanCYP1A2 0.21 the metabolites were detected (Supplemental Fig. 1). These PCB25 (M4/M2) rhCYP1A2 0.11 data suggested that CYP1A2 formed a partially dechlorin- Human plasma 3.15/2.17 ated hydroxylated metabolite of PCB28 presumably via the 4-OH-PCB31 (M3) HEK293human 0 classical lower-chlorinated (LC) PCB metabolism pathway HEK293humanCYP1A2 0.07 including arene oxide intermediate formation (Fig. 2a). rhCYP1A2 0.12 Former studies had shown that bactosomes might cata- Human plasma 2.38 lyze different reactions with different metabolites generated 5-OH-PCB28 (M1) HEK293human 0 than with the corresponding enzymes expressed in eukary- HEK293humanCYP1A2 1.02 otic cell cultures (Stiborová et al. 2017). To rule out the rhCYP1A2 2.14 possibility that CYP1A2 mediated dechlorination of PCB28 Human plasma 16.41 is specific for the prokaryotic expression system, we gener - 3-OH-PCB15 (DM) HEK293human 0 ated a transgenic HEK293 cell line stably overexpressing HEK293humanCYP1A2 0.16 the human cDNA of CYP1A2. The presence of genomi- rhCYP1A2 0.10 cally integrated CYP1A2 cDNA cell clones was confirmed Human plasma 2.60 by polymerase chain reaction analysis (data not shown). 1 3 Archives of Toxicology 4-OH-PCB31(M3) 3´-OH-PCB28 (M4) 3 8 2 2 R = 0.5376 R = 0.1942 p ≤ 0.0001 p ≤ 0.0001 0 0 0123 012 3 3-OH-PCB15 3-OH-PCB15 4´-OH-PCB25 (M2) PCB28 5 200 1 2 2 R = 0.2771 R = 0.2625 p ≤ 0.0001 p ≤ 0.0001 0 0 0123 0123 3-OH-PCB15 3-OH-PCB15 5-OH-PCB28 (M1) R = 0.5305 p ≤ 0.0001 3-OH-PCB15 1 3 4´-OH-PCB25 4-OH-PCB31 5-OH-PCB28 PCB28 3´-OH-PCB28 Archives of Toxicology carcinogenesis: resistant hepatocyte model. Toxicol Appl Phar- ◂Fig. 2 a Proposed scheme of the PCB28 pathway: transition states 1 macol 186(1):55–62. https:// doi. or g/ 10. 1016/ s0041- 008x(02) and 2 lead to known trichlorinated OH metabolites M1– M4 (Quinete 00018-2 et al.), transition state 3 could lead to dechlorination of PCB28 and Grimm FA, Hu D, Kania-Korwel I et al (2015) Metabolism and metab- formation of 3-OH-PCB15. b Linear regression of plasma concentra- olites of polychlorinated biphenyls. Crit Rev Toxicol 45(3):245– tions (n = 74; µg/L) of PCB28, 5-OH-PCB28, 4-OH-PCB31, 4′-OH- 272. https:// doi. org/ 10. 3109/ 10408 444. 2014. 999365 PCB25, and 3′-OHP-PCB28 versus plasma concentrations of 3-OH- Idda T, Bonas C, Hoffmann J et al (2020) Metabolic activation and PCB15 toxicological evaluation of polychlorinated biphenyls in Dros- ophila melanogaster. Sci Rep 10(1):21587. https:// doi. org/ 10. of the WHO indicator congener PCB28. For PCB15 the 1038/ s41598- 020- 78405-z Kraft M, Sievering S, Grün L, Rauchfuss K (2018) Mono-, di- and formation of a hydroquinone (PCB15-HQ) via dihydroxy- trichlorinated biphenyls (PCB 1 - PCB 39) in the indoor air of lated intermediates was clearly demonstrated in vitro (Song office rooms and their relevance on human blood burden. Indoor et al. 2013), and PCB15 also showed a mutagenic effect both Air 28(3):441–449 in vitro and in vivo. Furthermore, PCB15 had a tumorigenic Kraus T, Gube M, Lang J et al (2012) Surveillance program for former PCB-exposed workers of a transformer and capacitor recycling effect in an initiation–promotion experiment after a single company, family members, employees of surrounding companies, administration to rats, with a 100-fold higher incidence of and area residents–executive summary. J Toxicol Environ Health preneoplastic foci/cm3 in the liver (Espandiari et al. 2003). A 75(19–20):1241–1247. https://doi. or g/10. 1080/ 15287 394. 2012. Even though its full extent is currently unclear in humans, Quinete N, Kraus T, Belov VN, Aretz C, Esser A, Schettgen T (2015) the metabolism of PCB28 to hydroxylated PCB15 and fur- Fast determination of hydroxylated polychlorinated biphenyls in ther downstream hydroquinone metabolites should also be human plasma by online solid phase extraction coupled to liquid taken into account when assessing the potential risk of PCB chromatography-tandem mass spectrometry. Anal Chim Acta mixtures. 888:94–102. https:// doi. org/ 10. 1016/j. aca. 2015. 06. 041 Schettgen T, Alt A, Preim D, Keller D, Kraus T (2011) Biological Acknowledgements This research work was supported with funds monitoring of indoor exposure to dioxin-like and non-dioxin-like from the German Social Accident Insurance (DGUV) to PZ (grant polychlorinated biphenyls (PCBs) in a public building. Toxicol number FB295). The authors are responsible for the content of this Lett 213(1):116–121 publication. Schettgen T, Gube M, Esser A, Alt A, Kraus T (2012) Plasma polychlo- rinated biphenyls (PCB) levels of workers in a transformer recy- Funding Open Access funding enabled and organized by Projekt cling company, their family members, and employees of surround- DEAL. ing companies. J Toxicol Environ Health A 75(8–10):414–422 Song EQ, Ma XY, Tian XG et al (2013) The effect of the structure Data availability All data supporting the findings of this study are of polychlorinated biphenyls on their hydroxylation, oxidation, available within the paper and its Supplementary Information. and glutathionyl conjugation reactions. Biomed Environ Sci BES 26(2):138–147. https:// doi. org/ 10. 3967/ 0895- 3988. 2013. 02. 008 Stiborová M, Indra R, Moserová M et al (2017) Comparison of human Declarations cytochrome P450 1A1-catalysed oxidation of benzo[a]pyr- ene in prokaryotic and eukaryotic expression systems. Monat- Conflict of interest The authors declare that they have no conflict of shefte Fur Chemie 148(11):1959–1969. https:// doi. org/ 10. 1007/ interest. s00706- 017- 2002-0 Tulp MT, Bruggeman WA, Hutzinger O (1977) Reductive dechlorina- Open Access This article is licensed under a Creative Commons Attri- tion of chlorobiphenylols by rats. Experientia 33(9):1134–1136. bution 4.0 International License, which permits use, sharing, adapta- https:// doi. org/ 10. 1007/ bf019 22284 tion, distribution and reproduction in any medium or format, as long Vasko T, Hoffmann J, Gostek S et al (2018) Telomerase gene expres- as you give appropriate credit to the original author(s) and the source, sion bioassays indicate metabolic activation og genotoxic lower provide a link to the Creative Commons licence, and indicate if changes chlorinated polychlorinated biphenyls. Sci Rep 8(1):16903. were made. The images or other third party material in this article are https:// doi. org/ 10. 1083/ s41598- 018- 35043-e included in the article’s Creative Commons licence, unless indicated Zhang CY, Li X, Flor S et al (2022) Metabolism of 3-chlorobiphenyl otherwise in a credit line to the material. If material is not included in (PCB 2) in a human-relevant cell line: evidence of dechlorinated the article’s Creative Commons licence and your intended use is not metabolites. Environ Sci Technol 56(17):12460–12472. https:// permitted by statutory regulation or exceeds the permitted use, you will doi. org/ 10. 1021/ acs. est. 2c036 87 need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. References Espandiari P, Glauert HP, Lehmler HJ, Lee EY, Srinivasan C, Rob- ertson LW (2003) Polychlorinated biphenyls as initiators in liver 1 3
Journal
Archives of Toxicology – Springer Journals
Published: Jan 1, 2024
Keywords: Polychlorinated biphenyls; PCB28; Cytochrom p450 Monooxidase; Dechlorination