Using zebrafish models of leukemia to streamline drug screening and discovery  Adam P. Deveau, Victoria L. Bentley, Jason N. Berman  Experimental Hematology  Volume 45, Pages 1-9 (January 2017) DOI: 10.1016/j.exphem.2016.09.012 Copyright © 2016 ISEH - International Society for Experimental Hematology Terms and Conditions

Figure 1 Using zebrafish models of leukemia to streamline drug screening and discovery. (A) Modeling acute leukemia in the zebrafish can be accomplished using transient techniques such as XT of human cells into 1–3 day postfertilization embryos or using RNA or DNA to induce expression and MO to knock down expression. Permanent strategies to alter gene expression may be used for longer-term analysis and include transgenic expression or knockout techniques such as CRISPR. Both transient and permanent strategies are of value in designing drug screens and may be complementary. (B) Thorough characterization of leukemic phenotypes in the zebrafish may use a number of techniques to validate the model for chemical screening. Establishing a phenotype in fluorescently labeled transgenic lines permits live imaging and real-time analysis. Shown are bright-field images at 5× magnification of cd41:eGFP transgenic zebrafish and pu.1:NUP98-HOXA9(tNHA9);cd41:eGFP double-transgenic zebrafish. Inset at 10× magnification demonstrates expansion of green fluorescent HSCs in the tNHA9;cd41:eGFP double transgenics. This fluorescent leukemic phenotype could serve as the basis for the discovery of novel therapies that restore normal HSC numbers (adapted from Deveau et al. [46]). (C) Small-molecule chemical screening in zebrafish embryos using a phenotype-based readout. Zebrafish-based screens are amenable to high-throughput drug discovery using small aliquots of drugs in plate formats. (D) Zebrafish are a powerful tool with which to model human disease and use in phenotype-based chemical screens. Compounds identified will then be prioritized for further validation in murine studies and subsequent clinical trials. Experimental Hematology 2017 45, 1-9DOI: (10.1016/j.exphem.2016.09.012) Copyright © 2016 ISEH - International Society for Experimental Hematology Terms and Conditions