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

Sturtzel, Caterina (author), Zajec, Živa (author), Dernovšek, Jaka (author), Tomašič, Tihomir (author), Taschner-Mandl, Sabine (author), Distel, Martin (author)

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.

• Language: English
• Keywords: cancer models, phenotypic screening
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: 16 str.
• Numbering: Vol. 7, art. 44
• PID: 20.500.12556/RUL-146346-31b66c38-f629-0ade-335e-3ea531c9b90b
• UDC: 615.4:54:616-006
• ISSN on article: 2397-768X
• DOI: 10.1038/s41698-023-00386-9
• COBISS.SI-ID: 153170179
• Publication date in RUL: 24.05.2023

Record is a part of a journal

• Title: npj precision oncology
• Shortened title: npj precis. oncol.
• Publisher: Springer Nature, University of Minnesota, The Hormel Institute
• ISSN: 2397-768X
• COBISS.SI-ID: 526771225

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: fenotipski pregled, cebrica, tumorske celice, presajanje zdravil, ksenografti celic, onkologija, farmacevtska kemija, tumorji