title: Cell Imaging Counting as a Novel Ex Vivo Approach for Investigating Drug-Induced Hepatotoxicity in Zebrafish Larvae.
abstract: Drug-induced liver injury (DILI) is the most common reason for failures during the drug development process and for safety-related withdrawal of drugs from the pharmaceutical market. Therefore, having tools and techniques that can detect hepatotoxic properties in drug candidates at an early discovery stage is highly desirable. In this study, cell imaging counting was used to measure in a fast, straightforward, and unbiased way the effect of paracetamol and tetracycline, (compounds known to cause hepatotoxicity in humans) on the amount of DsRed-labeled hepatocytes recovered by protease digestion from Tg(fabp10a:DsRed) transgenic zebrafish. The outcome was in general comparable with the results obtained using two reference methods, i.e., visual analysis of liver morphology by fluorescence microscopy and size analysis of fluorescent 2D liver images. In addition, our study shows that administering compounds into the yolk is relevant in the framework of hepatotoxicity testing. Taken together, cell imaging counting provides a novel and rapid tool for screening hepatotoxicants in early stages of drug development. This method is also suitable for testing of other organ-related toxicities subject to the organs and tissues expressing fluorescent proteins in transgenic zebrafish lines.
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. bac.nguyenxuan@kuleuven.be.
authors.forename: Xuan-Bac
authors.initials: XB
authors.lastname: Nguyen
authors.affiliation: Division Pharmaceutical Analysis, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 923, 3000 Leuven, Belgium. stanislav.kislyuk@kuleuven.be.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Kislyuk
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. duchung.pham@kuleuven.be.
authors.forename: Duc-Hung
authors.initials: DH
authors.lastname: Pham
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. angela.kecskes@aok.pte.hu.
authors.forename: Angela
authors.initials: A
authors.lastname: Kecskés
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. jan.maes@kuleuven.be.
authors.forename: Jan
authors.initials: J
authors.lastname: Maes
authors.affiliation: Division Pharmaceutical Analysis, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 923, 3000 Leuven, Belgium. deirdre.cabooter@kuleuven.be.
authors.forename: Deirdre
authors.initials: D
authors.lastname: Cabooter
authors.affiliation: Drug Delivery and Disposition, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 921, 3000 Leuven, Belgium. pieter.annaert@kuleuven.be.
authors.forename: Pieter
authors.initials: P
authors.lastname: Annaert
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. peter.dewitte@kuleuven.be.
authors.forename: Peter
authors.initials: P
authors.lastname: De Witte
authors.affiliation: Laboratory for Molecular Biodiscovery, Department of Pharmaceutical and Pharmacological Sciences, University of Leuven, O & N II Herestraat 49-box 824, 3000 Leuven, Belgium. annelii.ny@kuleuven.be.
authors.forename: Annelii
authors.initials: A
authors.lastname: Ny
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001706:Biopsy; D002452:Cell Count; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D017930:Genes, Reporter; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D007814:Larva; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008856:Microscopy, Fluorescence / Q000379:methods; D057054:Molecular Imaging / Q000379:methods; D015027:Zebrafish
keywords: cell imaging counting; drug-induced liver injury (DILI); hepatotoxicity; pharmacokinetics; zebrafish larvae