title: Humanizing the zebrafish liver shifts drug metabolic profiles and improves pharmacokinetics of CYP3A4 substrates.
abstract: Understanding and predicting whether new drug candidates will be safe in the clinic is a critical hurdle in pharmaceutical development, that relies in part on absorption, distribution, metabolism, excretion and toxicology studies in vivo. Zebrafish is a relatively new model system for drug metabolism and toxicity studies, offering whole organism screening coupled with small size and potential for high-throughput screening. Through toxicity and absorption analyses of a number of drugs, we find that zebrafish is generally predictive of drug toxicity, although assay outcomes are influenced by drug lipophilicity which alters drug uptake. In addition, liver microsome assays reveal specific differences in metabolism of compounds between human and zebrafish livers, likely resulting from the divergence of the cytochrome P450 superfamily between species. To reflect human metabolism more accurately, we generated a transgenic "humanized" zebrafish line that expresses the major human phase I detoxifying enzyme, CYP3A4, in the liver. Here, we show that this humanized line shows an elevated metabolism of CYP3A4-specific substrates compared to wild-type zebrafish. The generation of this first described humanized zebrafish liver suggests such approaches can enhance the accuracy of the zebrafish model for toxicity prediction.
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.
authors.forename: Kar Lai
authors.initials: KL
authors.lastname: Poon
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.
authors.forename: Xingang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.
authors.forename: Ashley S
authors.initials: AS
authors.lastname: Ng
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.
authors.forename: Wei Huang
authors.initials: WH
authors.lastname: Goh
authors.affiliation: Roche Pharmaceutical Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Claudia
authors.initials: C
authors.lastname: McGinnis
authors.affiliation: Roche Pharmaceutical Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Stephen
authors.initials: S
authors.lastname: Fowler
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore. tcarney@imcb.a-star.edu.sg.|Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore, 636921, Singapore. tcarney@imcb.a-star.edu.sg.
authors.forename: Tom J
authors.identifier: http://orcid.org/0000-0003-2371-1924
authors.initials: TJ
authors.lastname: Carney
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.
authors.forename: Haishan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), Agency for Science, Technology and Research (A*STAR), 61 Biopolis Drive, Singapore, 138673, Singapore.|Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore, 636921, Singapore.
authors.forename: Phillip W
authors.initials: PW
authors.lastname: Ingham
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002851:Chromatography, High Pressure Liquid; D051544:Cytochrome P-450 CYP3A / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D013058:Mass Spectrometry; D004364:Pharmaceutical Preparations / Q000737:chemistry; D010599:Pharmacokinetics; D012995:Solubility; D018675:Toxicity Tests / Q000379:methods*; D015027:Zebrafish / Q000235:genetics*
chemical_list: D004364:Pharmaceutical Preparations; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human
keywords: CYP3A4; Humanized; Liver; Microsomes; Midazolam; Toxicity; Zebrafish