Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts
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 po...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-05-01
|
| Series: | npj Precision Oncology |
| Online Access: | https://doi.org/10.1038/s41698-023-00386-9 |
| _version_ | 1827607274169827328 |
|---|---|
| author | C. Sturtzel S. Grissenberger P. Bozatzi E. Scheuringer A. Wenninger-Weinzierl Z. Zajec J. Dernovšek S. Pascoal V. Gehl A. Kutsch A. Granig F. Rifatbegovic M. Carre A. Lang I. Valtingojer J. Moll D. Lötsch F. Erhart G. Widhalm D. Surdez O. Delattre N. André J. Stampfl T. Tomašič S. Taschner-Mandl M. Distel |
| author_facet | C. Sturtzel S. Grissenberger P. Bozatzi E. Scheuringer A. Wenninger-Weinzierl Z. Zajec J. Dernovšek S. Pascoal V. Gehl A. Kutsch A. Granig F. Rifatbegovic M. Carre A. Lang I. Valtingojer J. Moll D. Lötsch F. Erhart G. Widhalm D. Surdez O. Delattre N. André J. Stampfl T. Tomašič S. Taschner-Mandl M. Distel |
| author_sort | C. Sturtzel |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T06:52:07Z |
| format | Article |
| id | doaj.art-706465e903bc4c068048712533a28f34 |
| institution | Directory Open Access Journal |
| issn | 2397-768X |
| language | English |
| last_indexed | 2024-03-09T06:52:07Z |
| publishDate | 2023-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Precision Oncology |
| spelling | doaj.art-706465e903bc4c068048712533a28f342023-12-03T10:22:57ZengNature Portfolionpj Precision Oncology2397-768X2023-05-017111610.1038/s41698-023-00386-9Refined high-content imaging-based phenotypic drug screening in zebrafish xenograftsC. Sturtzel0S. Grissenberger1P. Bozatzi2E. Scheuringer3A. Wenninger-Weinzierl4Z. Zajec5J. Dernovšek6S. Pascoal7V. Gehl8A. Kutsch9A. Granig10F. Rifatbegovic11M. Carre12A. Lang13I. Valtingojer14J. Moll15D. Lötsch16F. Erhart17G. Widhalm18D. Surdez19O. Delattre20N. André21J. Stampfl22T. Tomašič23S. Taschner-Mandl24M. Distel25St. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)Faculty of Pharmacy, University of LjubljanaFaculty of Pharmacy, University of LjubljanaSt. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)Christian Doppler Laboratory for Advanced Polymers for Biomaterials and 3D Printing, TU WienChristian Doppler Laboratory for Advanced Polymers for Biomaterials and 3D Printing, TU WienSt. Anna Children’s Cancer Research Institute (CCRI)Service d’Hématologie & Oncologie Pédiatrique, Timone Hospital, AP-HMDepartment of Neurosurgery, Medical University of ViennaDepartment of Molecular Oncology, Sanofi Research CenterDepartment of Molecular Oncology, Sanofi Research CenterDepartment of Neurosurgery, Medical University of ViennaDepartment of Neurosurgery, Medical University of ViennaDepartment of Neurosurgery, Medical University of ViennaBalgrist University Hospital, Faculty of Medicine, University of Zurich (UZH)INSERM U830, Diversity and Plasticity of Childhood Tumors Lab, PSL Research University, SIREDO Oncology Center, Institut Curie Research CenterService d’Hématologie & Oncologie Pédiatrique, Timone Hospital, AP-HMChristian Doppler Laboratory for Advanced Polymers for Biomaterials and 3D Printing, TU WienFaculty of Pharmacy, University of LjubljanaSt. Anna Children’s Cancer Research Institute (CCRI)St. Anna Children’s Cancer Research Institute (CCRI)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.https://doi.org/10.1038/s41698-023-00386-9 |
| spellingShingle | C. Sturtzel S. Grissenberger P. Bozatzi E. Scheuringer A. Wenninger-Weinzierl Z. Zajec J. Dernovšek S. Pascoal V. Gehl A. Kutsch A. Granig F. Rifatbegovic M. Carre A. Lang I. Valtingojer J. Moll D. Lötsch F. Erhart G. Widhalm D. Surdez O. Delattre N. André J. Stampfl T. Tomašič S. Taschner-Mandl M. Distel Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts npj Precision Oncology |
| title | Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts |
| title_full | Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts |
| title_fullStr | Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts |
| title_full_unstemmed | Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts |
| title_short | Refined high-content imaging-based phenotypic drug screening in zebrafish xenografts |
| title_sort | refined high content imaging based phenotypic drug screening in zebrafish xenografts |
| url | https://doi.org/10.1038/s41698-023-00386-9 |
| work_keys_str_mv | AT csturtzel refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT sgrissenberger refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT pbozatzi refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT escheuringer refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT awenningerweinzierl refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT zzajec refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT jdernovsek refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT spascoal refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT vgehl refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT akutsch refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT agranig refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT frifatbegovic refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT mcarre refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT alang refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT ivaltingojer refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT jmoll refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT dlotsch refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT ferhart refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT gwidhalm refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT dsurdez refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT odelattre refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT nandre refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT jstampfl refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT ttomasic refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT staschnermandl refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts AT mdistel refinedhighcontentimagingbasedphenotypicdrugscreeninginzebrafishxenografts |