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Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts

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NPJ PRECISION ONCOLOGY
卷 7, 期 1, 页码 -

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NATURE PORTFOLIO
DOI: 10.1038/s41698-023-00386-9

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Zebrafish xenotransplantation models are increasingly used for phenotypic drug screening in precision oncology. This study presents a robust workflow for drug screening in zebrafish xenografts using high-content imaging. The authors established methods for embedding xenografts in 96-well format and provided strategies for automated imaging and analysis. The assay enables quantification of small compound efficacy in large cohorts of a vertebrate model system in vivo.
Zebrafish xenotransplantation models are increasingly applied for phenotypic drug screening to identify small compounds for precision oncology. Larval zebrafish xenografts offer the opportunity to perform drug screens at high-throughput in a complex in vivo environment. However, the full potential of the larval zebrafish xenograft model has not yet been realized and several steps of the drug screening workflow still await automation to increase throughput. Here, we present a robust workflow for drug screening in zebrafish xenografts using high-content imaging. We established embedding methods for high-content imaging of xenografts in 96-well format over consecutive days. In addition, we provide strategies for automated imaging and analysis of zebrafish xenografts including automated tumor cell detection and tumor size analysis over time. We also compared commonly used injection sites and cell labeling dyes and show specific site requirements for tumor cells from different entities. We demonstrate that our setup allows us to investigate proliferation and response to small compounds in several zebrafish xenografts ranging from pediatric sarcomas and neuroblastoma to glioblastoma and leukemia. This fast and cost-efficient assay enables the quantification of anti-tumor efficacy of small compounds in large cohorts of a vertebrate model system in vivo. Our assay may aid in prioritizing compounds or compound combinations for further preclinical and clinical investigations.

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