Abstract
Ochratoxin A (OTA) is known to be strongly bound to serum albumin, but it remains unknown how albumin affects its metabolism and kinetics. To close this gap, we used a mouse model, where heterozygous albumin deletion reduces serum albumin to concentrations similar to hypoalbuminemic patients and completely eliminates albumin by a homozygous knockout. OTA and its potential metabolites (OTα, 4-OH-OTA, 7ʹ-OH-OTA, OTHQ, OP-OTA, OTB-GSH, OTB-NAC, OTB) were time-dependently analyzed in plasma, bile, and urine by LC–MS/MS and were compared to previously published hepatotoxicity and nephrotoxicity data. Homozygous albumin deletion strongly accelerated plasma clearance as well as biliary and urinary excretion of the parent compound and its hydroxylation products. Decreasing albumin in mice by the heterozygous and even more by the homozygous knockout leads to an increase in the parent compound in urine which corresponded to increased nephrotoxicity. The role of albumin in OTA-induced hepatotoxicity is more complex, since heterozygous but not homozygous nor wild-type mice showed a strong biliary increase in the toxic open lactone OP-OTA. Correspondingly, OTA-induced hepatotoxicity was higher in heterozygous than in wild-type and homozygous animals. We present evidence that albumin-mediated retention of OTA in hepatocytes is required for formation of the toxic OP-OTA, while complete albumin elimination leads to rapid biliary clearance of OTA from hepatocytes with less formation of OP-OTA. In conclusion, albumin has a strong influence on metabolism and toxicity of OTA. In hypoalbuminemia, the parent OTA is associated with increased nephrotoxicity and the open lactone with increased hepatotoxicity.
Similar content being viewed by others
Data availability
Data is provided within the manuscript or supplementary information files.
References
Abrunhosa L, Paterson RRM, Venâncio A (2010) Biodegradation of ochratoxin A for food and feed decontamination. Toxins (basel) 2:1078–1099. https://doi.org/10.3390/toxins2051078
Bittner A, Cramer B, Humpf HU (2013) Matrix binding of ochratoxin A during roasting. J Agric Food Chem 51:12737–12743. https://doi.org/10.1021/jf403984x
Cramer B, Königs M, Humpf H-U (2008) Identification and in vitro cytotoxicity of ochratoxin A degradation products formed during coffee roasting. J Agric Food Chem 56:5673–5681. https://doi.org/10.1021/jf801296z
Dai J, Park G, Wright MW, Adams M, Akman SA, Manderville RA (2002) Detection and characterization of a glutathione conjugate of ochratoxin A. Chem Res Toxicol 15:1581–1588. https://doi.org/10.1021/tx0255929
Dekant R, Langer M, Lupp M, Adaku Chilaka C, Mally A (2021) In vitro and in vivo analysis of ochratoxin A-derived glucuronides and mercapturic acids as biomarkers of exposure. Toxins (Basel) 13. https://doi.org/10.3390/toxins13080587
EFSA (2020) Risk assessment of ochratoxin A in food. EFSA Journal 18. https://doi.org/10.2903/j.efsa.2020.6113
Eskola M, Kos G, Elliott CT, Hajšlová J, Mayar S, Krska R (2020) Worldwide contamination of food-crops with mycotoxins: validity of the widely cited ‘FAO estimate’ of 25. Crit Rev Food Sci Nutr 60:2773–2789. https://doi.org/10.1080/10408398.2019.1658570
Faisal Z, Derdák D, Lemli B, Kunsági-Máté S, Bálint M, Hetényi C, Csepregi R, Kőszegi T, Sueck F, Cramer B, Humpf H-U, Poór M (2018) Interaction of 2’R-ochratoxin A with serum albumins: binding site, effects of site markers, thermodynamics, species differences of albumin-binding, and influence of albumin on its toxicity in MDCK cells. Toxins (basel) 10:353. https://doi.org/10.3390/toxins10090353
Gautier J, Richoz J, Welti DH, Markovic J, Gremaud E, Guengerich FP, Turesky RJ (2001a) Metabolism of ochratoxin A: absence of formation of genotoxic derivatives by human and rat enzymes. Chem Res Toxicol 14:34–45. https://doi.org/10.1021/tx000070j
Gautier JC, Holzhaeuser D, Markovic J, Gremaud E, Schilter B, Turesky RJ (2001b) Oxidative damage and stress response from ochratoxin A exposure in rats. Free Radical Biol Med 30:1089–1098. https://doi.org/10.1016/S0891-5849(01)00507-X
Ghallab A, Myllys M, Holland CH et al (2019) Influence of liver fibrosis on lobular zonation. Cells 8:1556. https://doi.org/10.3390/cells8121556
Ghallab A, Hassan R, Myllys M et al (2021a) Subcellular spatio-temporal intravital kinetics of aflatoxin B1 and ochratoxin A in liver and kidney. Arch Toxicol 95:2163–2177. https://doi.org/10.1007/s00204-021-03073-5
Ghallab A, Hassan R, Hofmann U et al (2022) Interruption of bile acid uptake by hepatocytes after acetaminophen overdose ameliorates hepatotoxicity. J Hepatol 77:71–83. https://doi.org/10.1016/j.jhep.2022.01.020
Ghallab A, González D, Strängberg E et al (2024) Inhibition of the renal apical sodium dependent bile acid transporter prevents cholemic nephropathy in mice with obstructive cholestasis. J Hepatol 80:268–281. https://doi.org/10.1016/j.jhep.2023.10.035
Ghallab A, Myllys M, Friebel A et al (2021b) Spatio-temporal multiscale analysis of western diet-fed mice reveals a translationally relevant sequence of events during NAFLD progression. Cells 10. https://doi.org/10.3390/cells10102516
Gianmoena K, Gasparoni N, Jashari A et al (2021) Epigenomic and transcriptional profiling identifies impaired glyoxylate detoxification in NAFLD as a risk factor for hyperoxaluria. Cell Rep 36:109526. https://doi.org/10.1016/j.celrep.2021.109526
Gillman IG, Clark TN, Manderville RA (1999) Oxidation of ochratoxin A by an Fe-porphyrin system: model for enzymatic activation and DNA cleavage. Chem Res Toxicol 12:1066–1076. https://doi.org/10.1021/tx9901074
Gross-Steinmeyer K, Weymann J, Hege H-G, Metzler M (2002) Metabolism and lack of DNA reactivity of the mycotoxin ochratoxin A in cultured rat and human primary hepatocytes. J Agric Food Chem 50:938–945. https://doi.org/10.1021/jf0111817
Hadjeba-Medjdoub K, Tozlovanu M, Pfohl-Leszkowicz A, Frenette C, Paugh RJ, Manderville RA (2012) Structure-activity relationships imply different mechanisms of action for ochratoxin A-mediated cytotoxicity and genotoxicity. Chem Res Toxicol 25:181–190. https://doi.org/10.1021/tx200406c
Hagelberg S, Hult K, Fuchs R (1989) Toxicokinetics of ochratoxin A in several species and its plasma-binding properties. J Appl Toxicol 9:91–96. https://doi.org/10.1002/jat.2550090204
Han Z, Tangni EK, Di Mavungu JD, Vanhaecke L, de Saeger S, Wu A, Callebaut A (2013) In vitro glucuronidation of ochratoxin A by rat liver microsomes. Toxins (basel) 5:2671–2685. https://doi.org/10.3390/toxins5122671
Hassan R (2016) Possibilities and limitations of intravital imaging. EXCLI J 15:872–874. https://doi.org/10.17179/excli2016-863
Hassan R, Friebel A, Brackhagen L et al (2022a) Hypoalbuminemia affects the spatio-temporal tissue distribution of ochratoxin A in liver and kidneys: consequences for organ toxicity. Arch Toxicol 1–15. https://doi.org/10.1007/s00204-022-03361-8
Hassan R, González D, Hobloss Z et al (2022b) Inhibition of cytochrome P450 enhances the nephro- and hepatotoxicity of ochratoxin A. Arch Toxicol 1–13. https://doi.org/10.1007/s00204-022-03395-y
Heussner AH, Bingle LEH (2015) Comparative ochratoxin toxicity: a review of the available data. Toxins (basel) 7:4253–4282. https://doi.org/10.3390/toxins7104253
Holland CH, Ramirez Flores RO, Myllys M et al (2022) Transcriptomic cross-species analysis of chronic liver disease reveals consistent regulation between humans and mice. Hepatol Commun 6:161–177. https://doi.org/10.1002/hep4.1797
Hutchison RD, Steyn PS, Thompson DL (1971) The isolation and structure of 4-hydroxyochratoxin A and 7-carboxy-3,4-dihydro-8-hydroxy-3-methylisocoumarin from Penicillium viridicatum. Tetrahedron Lett 12:4033–4036. https://doi.org/10.1016/S0040-4039(01)97353-8
IARC (1993) Some naturally occurring substances: food items and constituents, heterocyclic aromatic amines and mycotoxins: IARC Working group on the evaluation of carcinogenic risks to humans, Lyon, 9–16 June 1992. IARC monographs on the evaluation of the carcinogenic risk of chemicals to humans, vol 56. IARC, Lyon
Il’ichev YV, Perry JL, Simon JD (2002) Interaction of ochratoxin A with human serum albumin. Preferential binding of the dianion and pH effects. J Phys Chem B 106:452–459. https://doi.org/10.1021/jp012314u
Kamp HG, Eisenbrand G, Schlatter J, Wurth K, Janzowski C (2005) Ochratoxin A: induction of (oxidative) DNA damage, cytotoxicity and apoptosis in mammalian cell lines and primary cells. Toxicology 206(3):413–425. https://doi.org/10.1016/j.tox.2004.08.004
Koeppert S, Ghallab A, Peglow S et al (2021) Live imaging of calciprotein particle clearance and receptor mediated uptake: role of calciprotein monomers. Front Cell Dev Biol 9:633925. https://doi.org/10.3389/fcell.2021.633925
Kőszegi T, Poór M (2016) Ochratoxin A: molecular interactions, mechanisms of toxicity and prevention at the molecular level. Toxins (basel) 8:111. https://doi.org/10.3390/toxins8040111
Kumagai S (1985) Ochratoxin A: plasma concentration and excretion into bile and urine in albumin-deficient rats. Food Chem Toxicol 23(10):941–943. https://doi.org/10.1016/0278-6915(85)90112-7
Li S, Marquardt RR, Frohlich AA, Vitti TG, Crow G (1997) Pharmacokinetics of ochratoxin A and its metabolites in rats. Toxicol Appl Pharmacol 145:82–90. https://doi.org/10.1006/taap.1997.8155
Li S, Marquardt R, Frohlich A (2000) Identification of ochratoxins and some of their metabolites in bile and urine of rats. Food Chem Toxicol 38:141–152. https://doi.org/10.1016/S0278-6915(99)00153-2
Mally A, Dekant W (2009) Mycotoxins and the kidney: modes of action for renal tumor formation by ochratoxin A in rodents. Mol Nutr & Food Res 53:467–478. https://doi.org/10.1002/mnfr.200800149
Mally A, Pepe G, Ravoori S, Fiore M, Gupta RC, Dekant W, Mosesso P (2005) Ochratoxin A causes DNA damage and cytogenetic effects but no DNA adducts in rats. Chem Res Toxicol 18:1253–1261. https://doi.org/10.1021/tx049650x
Marin S, Ramos AJ, Cano-Sancho G, Sanchis V (2013) Mycotoxins: occurrence, toxicology, and exposure assessment. Food Chem Toxicol 60:218–237. https://doi.org/10.1016/j.fct.2013.07.047
Muñoz K, Cramer B, Dopstadt J, Humpf H-U, Degen GH (2017) Evidence of ochratoxin A conjugates in urine samples from infants and adults. Mycotoxin Res 33:39–47. https://doi.org/10.1007/s12550-016-0261-y
Pfohl-Leszkowicz A, Manderville RA (2007) Ochratoxin A: an overview on toxicity and carcinogenicity in animals and humans. Mol Nutr & Food Res 51:61–99. https://doi.org/10.1002/mnfr.200600137
Pfohl-Leszkowicz A, Grosse Y, Kane A, Creppy EE, Dirheimer G (1993) Differential DNA adduct formation and disappearance in three mouse tissues after treatment with the mycotoxin ochratoxin A. Mutat Res 289:265–273. https://doi.org/10.1016/0027-5107(93)90077-S
Remetic J, Ghallab A, Hobloss Z et al (2022) Loss of bile salt export pump aggravates lipopolysaccharide-induced liver injury in mice due to impaired hepatic endotoxin clearance. Hepatology 75(5):1095–1109. https://doi.org/10.1002/hep.32289
Ringot D, Chango A, Schneider Y-J, Larondelle Y (2006) Toxicokinetics and toxicodynamics of ochratoxin A, an update. Chem Biol Interact 159:18–46. https://doi.org/10.1016/j.cbi.2005.10.106
Rottkord U, Röhl C, Ferse I, Schulz M-C, Rückschloss U, Gekle M, Schwerdt G, Humpf H-U (2017) Structure-activity relationship of ochratoxin A and synthesized derivatives: importance of amino acid and halogen moiety for cytotoxicity. Arch Toxicol 91:1461–1471. https://doi.org/10.1007/s00204-016-1799-3
Schwerdt G, Kopf M, Gekle M (2023) The nephrotoxin ochratoxin A impairs resilience of energy homeostasis of human proximal tubule cells. Mycotoxin Res 1–11. https://doi.org/10.1007/s12550-023-00500-7
Støren O, Holm H, Størmer FC (1982) Metabolism of ochratoxin A by rats. Appl Environ Microbiol 44:785–789. https://doi.org/10.1128/aem.44.4.785-789.1982
Størmer FC, Støren O, Hansen CE, Pedersen JI, Aasen AJ (1983) Formation of (4R)- and (4S)-4-hydroxyochratoxin A and 10-hydroxyochratoxin A from ochratoxin A by rabbit liver microsomes. Appl Environ Microbiol 45:1183–1187. https://doi.org/10.1128/aem.45.4.1183-1187.1983
Studer-Rohr I, Schlatter J, Dietrich DR (2000) Kinetic parameters and intraindividual fluctuations of ochratoxin A plasma levels in humans. Arch Toxicol 74:499–510. https://doi.org/10.1007/s002040000157
Sueck F, Specht J, Cramer B, Humpf H-U (2020) Identification of ochratoxin-N-acetyl-L-cysteine as a new ochratoxin A metabolite and potential biomarker in human urine. Mycotoxin Res 36:1–10. https://doi.org/10.1007/s12550-019-00360-0
Sueck F, Poór M, Faisal Z, Gertzen CGW, Cramer B, Lemli B, Kunsági-Máté S, Gohlke H, Humpf H-U (2018) Interaction of ochratoxin A and its thermal degradation product 2’R-ochratoxin A with human serum albumin. Toxins (Basel) 10. https://doi.org/10.3390/toxins10070256
Tozlovanu M, Canadas D, Pfohl-Leszkowicz A, Frenette C, Paugh RJ, Manderville RA (2012) Glutathione conjugates of ochratoxin A as biomarkers of exposure. Arh Hig Rada Toksikol 63:417–427. https://doi.org/10.2478/10004-1254-63-2012-2202
Turesky RJ (2005) Perspective: ochratoxin A is not a genotoxic carcinogen. Chem Res Toxicol 18:1082–1090. https://doi.org/10.1021/tx050076e
Vucur M, Ghallab A, Schneider AT et al (2023) Sublethal necroptosis signaling promotes inflammation and liver cancer. Immunity 56:1578-1595.e8. https://doi.org/10.1016/j.immuni.2023.05.017
**ao H, Madhyastha S, Marquardt RR, Li S, Vodela JK, Frohlich AA, Kemppainen BW (1996) Toxicity of ochratoxin A, its opened lactone form and several of its analogs: structure-activity relationships. Toxicol Appl Pharmacol 137:182–192. https://doi.org/10.1006/taap.1996.0071
Yang S, Zhang H, de Saeger S, de Boevre M, Sun F, Zhang S, Cao X, Wang Z (2015) In vitro and in vivo metabolism of ochratoxin A: a comparative study using ultra-performance liquid chromatography-quadrupole/time-of-flight hybrid mass spectrometry. Anal Bioanal Chem 407:3579–3589. https://doi.org/10.1007/s00216-015-8570-0
Zepnik H, Völkel W, Dekant W (2003) Toxicokinetics of the mycotoxin ochratoxin A in F 344 rats after oral administration. Toxicol Appl Pharmacol 192:36–44. https://doi.org/10.1016/S0041-008X(03)00261-8
Author information
Authors and Affiliations
Contributions
A.G., J.G.H., B.C.: Study concept and design; data analysis and interpretation, drafting the manuscript. M.K.: analysis of OTA and its metabolites; data acquisition; contributed to writing of the manuscript. R.H.: animal experiments; data acquisition; contributed to writing of the manuscript. D.G.; M.M., Z.H.: data acquisition; contributed to writing of the manuscript. G.H.D., H.H.: contributed to study concept and design; analysis and interpretation of data; critical revision of the manuscript. All authors reviewed the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Kuhn, M., Hassan, R., González, D. et al. Role of albumin in the metabolism and excretion of ochratoxin A. Mycotoxin Res (2024). https://doi.org/10.1007/s12550-024-00538-1
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s12550-024-00538-1