2. Academic Validation
  3. Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts

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

  • NPJ Precis Oncol. 2023 May 18;7(1):44. doi: 10.1038/s41698-023-00386-9.
C Sturtzel # 1 2,  S Grissenberger # 1,  P Bozatzi 1,  E Scheuringer 1 2,  A Wenninger-Weinzierl 1 2,  Z Zajec 3,  J Dernovšek 3,  S Pascoal 1,  V Gehl 1,  A Kutsch 4,  A Granig 4,  F Rifatbegovic 1,  M Carre 5 6,  A Lang 7 8,  I Valtingojer 9,  J Moll 9 10,  D Lötsch 7 8,  F Erhart 7 8,  G Widhalm 7 8,  D Surdez 11,  O Delattre 12,  N André 5 6,  J Stampfl 4,  T Tomašič 3,  S Taschner-Mandl 13,  M Distel 14 15
Affiliations

Affiliations

  • 1 St. Anna Children's Cancer Research Institute (CCRI), Vienna, Austria.
  • 2 Zebrafish Platform Austria for Preclinical Drug Screening (ZANDR), Vienna, Austria.
  • 3 Faculty of Pharmacy, University of Ljubljana, Ljubljana, Slovenia.
  • 4 Christian Doppler Laboratory for Advanced Polymers for Biomaterials and 3D Printing, TU Wien, Vienna, Austria.
  • 5 Service d'Hématologie & Oncologie Pédiatrique, Timone Hospital, AP-HM, Marseille, France.
  • 6 Centre de Recherche en Cancérologie de Marseille (CRCM), Aix-Marseille Université, CNRS, Inserm, Institut Paoli Calmettes, Marseille, France.
  • 7 Department of Neurosurgery, Medical University of Vienna, Vienna, Austria.
  • 8 Central Nervous System Tumors Unit, Comprehensive Cancer Center, Medical University of Vienna, Vienna, Austria.
  • 9 Department of Molecular Oncology, Sanofi Research Center, Vitry-sur-Seine, France.
  • 10 Renon Biotech and Pharma Consulting, Unterinn am Ritten (Bz), Italy.
  • 11 Balgrist University Hospital, Faculty of Medicine, University of Zurich (UZH), Zurich, Switzerland.
  • 12 INSERM U830, Diversity and Plasticity of Childhood Tumors Lab, PSL Research University, SIREDO Oncology Center, Institut Curie Research Center, Paris, France.
  • 13 St. Anna Children's Cancer Research Institute (CCRI), Vienna, Austria. sabine.taschner@ccri.at.
  • 14 St. Anna Children's Cancer Research Institute (CCRI), Vienna, Austria. martin.distel@ccri.at.
  • 15 Zebrafish Platform Austria for Preclinical Drug Screening (ZANDR), Vienna, Austria. martin.distel@ccri.at.
  • # Contributed equally.
PMID: 37202469 DOI: 10.1038/s41698-023-00386-9
Abstract

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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