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P1004
Overt Toxicity Screening Assay Using Live Z-TagSM Zebrafish
Presenter Timothy Baranowski, Zygogen LLC, USA
Additional Authors: Yiting Zhang, Peter Eimon (Zygogen, LLC)
Toxicity is a major cause of drug failure that often does not become apparent until lead compounds are moved into mammalian models. Consequently, there is a compelling need to develop high-throughput safety assessment techniques that can be implemented in earlier stages of drug discovery to eliminate compounds that cause potentially undesirable side effects. Cell-based toxicity assays do not preserve the complexity of a whole organism, while traditional animal models are low throughput, costly, and labor intensive. Zygogen has designed an automated in vivo zebrafish assay that can be used in early stages of development to identify compounds that induce unacceptable toxicity. To facilitate high-throughput screening, we have implemented robotic addition of compounds to zebrafish embryos arrayed in microplates, and have demonstrated automated microplate image acquisition and analysis. The toxicity assay utilizes transgenic zebrafish embryos, which express a green fluorescent protein specifically in the blood vessels to detect overt signs of toxicity. Fluorescent blood vessels are present throughout the entire embryo, enabling automated measurement of body length and shape. Zebrafish embryos are treated with test compounds at 1 day or 5 days post fertilization and imaged at subsequent time points using the Discovery-1 high-content imaging system. An automated algorithm designed by Zygogen detects compounds, which cause significant changes in embryo length and overall body shape indicative of toxicity. This zebrafish-based assay for overt toxicity provides an efficient method for conducting high-throughput safety assessment in a complex organism. The assay may be used to eliminate unfavorable compounds from the development process prior to costly lead optimization and mammalian screening.