title: Editor's Highlight: Transgenic Zebrafish Reporter Lines as Alternative In Vivo Organ Toxicity Models.
abstract: Organ toxicity, particularly liver toxicity, remains one of the major reasons for the termination of drug candidates in the development pipeline as well as withdrawal or restrictions of marketed drugs. A screening-amenable alternative in vivo model such as zebrafish would, therefore, find immediate application in the early prediction of unacceptable organ toxicity. To identify highly upregulated genes as biomarkers of toxic responses in the zebrafish model, a set of well-characterized reference drugs that cause drug-induced liver injury (DILI) in the clinic were applied to zebrafish larvae and adults. Transcriptome microarray analysis was performed on whole larvae or dissected adult livers. Integration of data sets from different drug treatments at different stages identified common upregulated detoxification pathways. Within these were candidate biomarkers which recurred in multiple treatments. We prioritized 4 highly upregulated genes encoding enzymes acting in distinct phases of the drug metabolism pathway. Through promoter isolation and fosmid recombineering, eGFP reporter transgenic zebrafish lines were generated and evaluated for their response to DILI drugs. Three of the 4 generated reporter lines showed a dose and time-dependent induction in endodermal organs to reference drugs and an expanded drug set. In conclusion, through integrated transcriptomics and transgenic approaches, we have developed parallel independent zebrafish in vivo screening platforms able to predict organ toxicities of preclinical drugs.
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Kar Lai
authors.initials: KL
authors.lastname: Poon
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Xingang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Genome Institute of Singapore, A*STAR (Agency for Science, Technology and Research), 60 Biopolis Street, 138672 Singapore.
authors.forename: Serene G P
authors.initials: SG
authors.lastname: Lee
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Ashley S
authors.initials: AS
authors.lastname: Ng
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Wei Huang
authors.initials: WH
authors.lastname: Goh
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Zhonghua
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Muthafar
authors.initials: M
authors.lastname: Al-Haddawi
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.
authors.forename: Haishan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Genome Institute of Singapore, A*STAR (Agency for Science, Technology and Research), 60 Biopolis Street, 138672 Singapore.
authors.forename: Sinnakaruppan
authors.initials: S
authors.lastname: Mathavan
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.|Lee Kong Chian School of Medicine, Nanyang Technological University, 50 Nanyang Avenue, 639798 Singapore.
authors.forename: Philip W
authors.initials: PW
authors.lastname: Ingham
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.|Roche Pharmaceutical Research & Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Claudia
authors.initials: C
authors.lastname: McGinnis
authors.affiliation: Institute of Molecular and Cell Biology (IMCB), A*STAR (Agency for Science, Technology and Research), 61 Biopolis Drive, 138673 Singapore.|Lee Kong Chian School of Medicine, Nanyang Technological University, 50 Nanyang Avenue, 639798 Singapore.
authors.forename: Tom J
authors.initials: TJ
authors.lastname: Carney
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D015415:Biomarkers / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015507:Drugs, Investigational / Q000008:administration & dosage / Q000009:adverse effects*; D004707:Endoderm / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005260:Female; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D017930:Genes, Reporter / Q000187:drug effects*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D008297:Male; D038081:Organogenesis / Q000187:drug effects; D011994:Recombinant Proteins / Q000378:metabolism; D013723:Teratogens / Q000633:toxicity; D018675:Toxicity Tests / Q000379:methods*; D015262:Xenobiotics / Q000008:administration & dosage / Q000633:toxicity*; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development / Q000378:metabolism
chemical_list: D015415:Biomarkers; D015507:Drugs, Investigational; D029941:Fish Proteins; D011994:Recombinant Proteins; D013723:Teratogens; D015262:Xenobiotics; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: biomarker; hepatotoxicity; toxicology; transgenesis.; zebrafish