Validation of NBD-coupled taurocholic acid for intravital analysis of bile acid transport in liver and kidney of mice

Authors

  • Ahmed Ghallab Department of Toxicology, Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Ardeystr. 67, 44139 Dortmund, Germany. E-mail: ghallab@ifado.de https://orcid.org/0000-0003-0695-3403
  • Sebastian Kunz Institute of Pharmacology and Toxicology, Justus Liebig University Giessen, Biomedical Research Center Seltersberg, Schubertstr. 81, 35392 Giessen, Germany https://orcid.org/0009-0000-9221-5831
  • Celine Drossel Institute of Organic Chemistry, Justus Liebig University Giessen, Heinrich-Buff-Ring 17, 35392, Giessen, Germany https://orcid.org/0009-0005-0111-2324
  • Veronica Billo Institute of Pharmacology and Toxicology, Justus Liebig University Giessen, Biomedical Research Center Seltersberg, Schubertstr. 81, 35392 Giessen, Germany https://orcid.org/0009-0007-7772-3206
  • Adrian Friebel Institute of Computer Science & Saxonian Incubator for Clinical Research (SIKT), University of Leipzig, Haertelstraße 16-18, 04107 Leipzig, Germany https://orcid.org/0000-0002-9157-4418
  • Mats Georg Institute of Organic Chemistry, Justus Liebig University Giessen, Heinrich-Buff-Ring 17, 35392, Giessen, Germany https://orcid.org/0009-0006-4631-2853
  • Richard Göttlich Institute of Organic Chemistry, Justus Liebig University Giessen, Heinrich-Buff-Ring 17, 35392, Giessen, Germany https://orcid.org/0000-0002-1424-3084
  • Zaynab Hobloss Department of Toxicology, Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Ardeystr. 67, 44139 Dortmund, Germany https://orcid.org/0009-0003-8292-867X
  • Reham Hassan Department of Toxicology, Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Ardeystr. 67, 44139 Dortmund, Germany; Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, 83523 Qena, Egypt https://orcid.org/0000-0002-6569-7676
  • Maiju Myllys Department of Toxicology, Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Ardeystr. 67, 44139 Dortmund, Germany https://orcid.org/0000-0001-9117-4572
  • Abdel-latief Seddek Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, 83523 Qena, Egypt https://orcid.org/0000-0001-7568-0352
  • Noha Abdelmageed Department of Pharmacology, Faculty of Veterinary Medicine, Sohag University, 82524 Sohag, Egypt https://orcid.org/0000-0002-0563-3144
  • Paul A. Dawson Department of Pediatrics, Division of Gastroenterology, Hepatology, and Nutrition, Emory University, Atlanta, GA 30322, USA https://orcid.org/0000-0001-9760-4422
  • Erik Lindström Albireo Pharma, Inc., Boston, MA 02109, USA https://orcid.org/0009-0009-4059-6287
  • Stefan Hoehme Institute of Computer Science & Saxonian Incubator for Clinical Research (SIKT), University of Leipzig, Haertelstraße 16-18, 04107 Leipzig, Germany https://orcid.org/0000-0002-9716-9587
  • Jan G. Hengstler Department of Toxicology, Leibniz Research Centre for Working Environment and Human Factors, Technical University Dortmund, Ardeystr. 67, 44139 Dortmund, Germany. E-mail: hengstler@ifado.de https://orcid.org/0000-0002-1427-5246
  • Joachim Geyer Institute of Pharmacology and Toxicology, Justus Liebig University Giessen, Biomedical Research Center Seltersberg, Schubertstr. 81, 35392 Giessen, Germany. E-mail: Joachim.M.Geyer@vetmed.uni-giessen.de https://orcid.org/0000-0003-2663-1858

DOI:

https://doi.org/10.17179/excli2024-7707

Keywords:

Ntcp, OATP, ASBT, cholestasis, cholemic nephropathy

Abstract

Fluorophore-coupled bile acids (BA) represent an important tool for intravital analysis of BA flux in animal models of cholestatic diseases. However, addition of a fluorophore to a BA may alter transport properties. We developed and validated a 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole-coupled taurocholic acid (3β-NBD-TCA) as a probe for intravital analysis of BA homeostasis. We compared transport of 3β-NBD-TCA to [3H]-TCA in HEK293 cells stably expressing the mouse hepatic or renal BA carriers mNtcp or mAsbt, respectively. We also studied distribution kinetics intravitally in livers and kidneys of anesthetized wildtype and mOatp1a/1b cluster knockout mice (OatpKO) with and without administration of the Ntcp inhibitor Myrcludex B and the ASBT inhibitor AS0369. In vitro, 3b-NBD-TCA and [3H]-TCA showed comparable concentration- and time-dependent transport via mNtcp and mAsbt as well as similar inhibition kinetics for Myrcludex B and AS0369. Intravital analysis in the livers of wildtype and OatpKO mice revealed contribution of both mNtcp and mOatp1a/1b in the 3β-NBD-TCA uptake from the sinusoidal blood into hepatocytes. Combined deletion of mOatp1a/1b and inhibition of mNtcp by Myrcludex B blocked the uptake of 3β-NBD-TCA from sinusoidal blood into hepatocytes. This led to an increase of 3β-NBD-TCA signal in the systemic circulation including renal capillaries, followed by strong enrichment in a subpopulation of proximal renal tubular epithelial cells (TEC). The enrichment of 3β-NBD-TCA in TEC was strongly reduced by the systemic ASBT inhibitor AS0369. NBD-coupled TCA has similar transport kinetics as [3H]-TCA and can be used as a tool to study hepatorenal BA transport.

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Published

2024-10-30

How to Cite

Ghallab, A., Kunz, S., Drossel, C., Billo, V., Friebel, A., Georg, M., … Geyer, J. (2024). Validation of NBD-coupled taurocholic acid for intravital analysis of bile acid transport in liver and kidney of mice. EXCLI Journal, 23, 1330–1352. https://doi.org/10.17179/excli2024-7707

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