Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Short Communication
  • Published:

Orthotopic models of pediatric brain tumors in zebrafish

Oncogene volume 34, pages 1736–1742 (2015)Cite this article

Subjects

Abstract

High-throughput screens (HTS) of compound toxicity against cancer cells can identify thousands of potential new drug-leads. But only limited numbers of these compounds can progress to expensive and labor-intensive efficacy studies in mice, creating a ‘bottle neck’ in the drug development pipeline. Approaches that triage drug-leads for further study are greatly needed. Here we provide an intermediary platform between HTS and mice by adapting mouse models of pediatric brain tumors to grow as orthotopic xenografts in the brains of zebrafish. Freshly isolated mouse ependymoma, glioma and choroid plexus carcinoma cells expressing red fluorescence protein were conditioned to grow at 34 °C. Conditioned tumor cells were then transplanted orthotopically into the brains of zebrafish acclimatized to ambient temperatures of 34 °C. Live in vivo fluorescence imaging identified robust, quantifiable and reproducible brain tumor growth as well as spinal metastasis in zebrafish. All tumor xenografts in zebrafish retained the histological characteristics of the corresponding parent mouse tumor and efficiently recruited fish endothelial cells to form a tumor vasculature. Finally, by treating zebrafish harboring ERBB2-driven gliomas with an appropriate cytotoxic chemotherapy (5-fluorouracil) or tyrosine kinase inhibitor (erlotinib), we show that these models can effectively assess drug efficacy. Our data demonstrate, for the first time, that mouse brain tumors can grow orthotopically in fish and serve as a platform to study drug efficacy. As large cohorts of brain tumor-bearing zebrafish can be generated rapidly and inexpensively, these models may serve as a powerful tool to triage drug-leads from HTS for formal efficacy testing in mice.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

References

  1. Smith MA, Seibel NL, Altekruse SF, Ries LA, Melbert DL, O'Leary M et al. Outcomes for children and adolescents with cancer: challenges for the twenty-first century. J Clin Oncol 2010; 28: 2625–2634.

    Article  Google Scholar 

  2. Kawauchi D, Robinson G, Uziel T, Gibson P, Rehg J, Gao C et al. A mouse model of the most aggressive subgroup of human medulloblastoma. Cancer Cell 2012; 21: 168–180.

    Article  CAS  Google Scholar 

  3. Gibson P, Tong Y, Robinson G, Thompson MC, Currle DS, Eden C et al. Subtypes of medulloblastoma have distinct developmental origins. Nature 2010; 468: 1095–1099.

    Article  CAS  Google Scholar 

  4. Pei Y, Moore Colin E, Wang J, Tewari Alok K, Eroshkin A, Cho Y-J et al. An Animal Model of MYC-Driven Medulloblastoma. Cancer Cell 2012; 21: 155–167.

    Article  CAS  Google Scholar 

  5. Swartling FJ, Grimmer MR, Hackett CS, Northcott PA, Fan Q-W, Goldenberg DD et al. Pleiotropic role for MYCN in medulloblastoma. Genes Dev 2010; 24: 1059–1072.

    Article  CAS  Google Scholar 

  6. Chow Lionel ML, Endersby R, Zhu X, Rankin S, Qu C, Zhang J et al. Cooperativity within and among Pten, p53, and Rb pathways induces high-grade astrocytoma in adult brain. Cancer Cell 2011; 19: 305–316.

    Article  CAS  Google Scholar 

  7. Johnson RA, Wright KD, Poppleton H, Mohankumar KM, Finkelstein D, Pounds SB et al. Cross-species genomics matches driver mutations and cell compartments to model ependymoma. Nature 2010; 466: 632–636.

    Article  CAS  Google Scholar 

  8. Romer JT, Kimura H, Magdaleno S, Sasai K, Fuller C, Baines H et al. Suppression of the Shh pathway using a small molecule inhibitor eliminates medulloblastoma in Ptc1(+/-)p53(-/-) mice. Cancer Cell 2004; 6: 229–240.

    Article  CAS  Google Scholar 

  9. Atkinson Jennifer M, Shelat Anang A, Carcaboso Angel M, Kranenburg Tanya A, Arnold LA, Boulos N et al. An integrated in vitro and in vivo high-throughput screen identifies treatment leads for ependymoma. Cancer Cell 2011; 20: 384–399.

    Article  CAS  Google Scholar 

  10. Caponigro G, Sellers WR . Advances in the preclinical testing of cancer therapeutic hypotheses. Nat Rev Drug Discov 2011; 10: 179–187.

    Article  CAS  Google Scholar 

  11. Macarron R, Banks MN, Bojanic D, Burns DJ, Cirovic DA, Garyantes T et al. Impact of high-throughput screening in biomedical research. Nat Rev Drug Discov 2011; 10: 188–195.

    Article  CAS  Google Scholar 

  12. White R, Rose K, Zon L . Zebrafish cancer: the state of the art and the path forward. Nat Rev Cancer 2013; 13: 624–636.

    Article  CAS  Google Scholar 

  13. Stoletov K, Klemke R . Catch of the day: zebrafish as a human cancer model. Oncogene 2008; 27: 4509–4520.

    Article  CAS  Google Scholar 

  14. Herbert J, Cavallaro T, Dwork AJ . A marker for primary choroid plexus neoplasms. Am J Pathol 1990; 136: 1317–1325.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Hanahan D, Weinberg Robert A . Hallmarks of cancer: the next generation. Cell 2011; 144: 646–674.

    Article  CAS  Google Scholar 

  16. Lawson ND, Weinstein BM . In vivo imaging of embryonic vascular development using transgenic zebrafish. Dev Biol 2002; 248: 307–318.

    Article  CAS  Google Scholar 

  17. Stoletov K, Montel V, Lester RD, Gonias SL, Klemke R . High-resolution imaging of the dynamic tumor cell–vascular interface in transparent zebrafish. Proc Natl Acad Sci USA 2007; 104: 17406–17411.

    Article  CAS  Google Scholar 

  18. Pounds S, Gao CL, Johnson RA, Wright KD, Poppleton H, Finkelstein D et al. A procedure to statistically evaluate agreement of differential expression for cross-species genomics. Bioinformatics 2011; 27: 2098–2103.

    Article  CAS  Google Scholar 

  19. Walbert T, Gilbert M, Groves M, Puduvalli V, Alfred Yung WK, Conrad C et al. Combination of 6-thioguanine, capecitabine, and celecoxib with temozolomide or lomustine for recurrent high-grade glioma. J Neurooncol 2011; 102: 273–280.

    Article  CAS  Google Scholar 

  20. Prados MD, Lamborn KR, Chang S, Burton E, Butowski N, Malec M et al. Phase 1 study of erlotinib HCl alone and combined with temozolomide in patients with stable or recurrent malignant glioma. Neuro Oncol 2006; 8: 67–78.

    Article  CAS  Google Scholar 

  21. TCGA. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 2008; 455: 1061–1068.

    Article  Google Scholar 

  22. Collymore C, Rasmussen S, Tolwani RJ . Gavaging adult zebrafish. J Vis Exp 2013; 11: e50691.

    Google Scholar 

  23. Hernan R, Fasheh R, Calabrese C, Frank AJ, Maclean KH, Allard D et al. ERBB2 up-regulates S100A4 and several other prometastatic genes in medulloblastoma. Cancer Res 2003; 63: 140–148.

    CAS  PubMed  Google Scholar 

  24. Lacy J, Saadati H, Yu JB . Complications of brain tumors and their treatment. Hematol Oncol Clin North Am 2012; 26: 779–796.

    Article  Google Scholar 

  25. Lin NU, Wefel JS, Lee EQ, Schiff D, van den Bent MJ, Soffietti R et al. Challenges relating to solid tumour brain metastases in clinical trials, part 2: neurocognitive, neurological, and quality-of-life outcomes. A report from the RANO group. Lancet Oncol 2013; 14: e407–e416.

    Article  Google Scholar 

  26. White RM, Cech J, Ratanasirintrawoot S, Lin CY, Rahl PB, Burke CJ et al. DHODH modulates transcriptional elongation in the neural crest and melanoma. Nature 2011; 471: 518–522.

    Article  CAS  Google Scholar 

  27. Hong S-K, Tsang M, Dawid IB . The mych gene is required for neural crest survival during zebrafish development. PLoS One 2008; 3: e2029.

    Article  Google Scholar 

  28. Ridges S, Heaton WL, Joshi D, Choi H, Eiring A, Batchelor L et al. Zebrafish screen identifies novel compound with selective toxicity against leukemia. Blood 2012; 119: 5621–5631.

    Article  CAS  Google Scholar 

  29. Wienholds E, Schulte-Merker S, Walderich B, Plasterk RH . Target-selected inactivation of the zebrafish rag1 gene. Science 2002; 297: 99–102.

    Article  CAS  Google Scholar 

  30. White RM, Sessa A, Burke C, Bowman T, LeBlanc J, Ceol C et al. Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2008; 2: 183–189.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Institutes of Health (R01CA129541, P01CA96832 and P30CA021765, RJG), the Collaborative Ependymoma Research Network (CERN) and by ALSAC. We are grateful to the staff of the Hartwell Center for Bioinformatics and Biotechnology, and Flow Cytometry & Cell Sorting Shared Resource at St Jude Children’s Research Hospital for technical assistance.

Author information

Authors and Affiliations

  1. Department of Developmental Neurobiology, St Jude Children’s Research Hospital, Memphis, TN, USA,

    C J Eden, M Murugesan, T N Phoenix, B Nimmervoll, Y Tong, N Boulos, J Dapper, R Thiruvenkatam & R J Gilbertson

  2. Department of Chemical Biology and Therapeutics, St Jude Children’s Research Hospital, Memphis, TN, USA,

    B Ju & M R Taylor

  3. Department of Pathology, St Jude Children’s Research Hospital, Memphis, TN, USA,

    D W Ellison

  4. Department of Computational Biology and Bioinformatics, St Jude Children’s Research Hospital, Memphis, TN, USA,

    D Finkelstein

  5. Department of Oncology, St Jude Children’s Research Hospital, Memphis, TN, USA,

    K Wright

  6. Department of Biological Sciences, University of Memphis, Memphis, TN, USA,

    C A Lessman

Authors
  1. C J Eden
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. B Ju
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M Murugesan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. T N Phoenix
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B Nimmervoll
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D W Ellison
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. D Finkelstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. N Boulos
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. J Dapper
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. R Thiruvenkatam
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. C A Lessman
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. M R Taylor
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. R J Gilbertson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to M R Taylor or R J Gilbertson.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Eden, C., Ju, B., Murugesan, M. et al. Orthotopic models of pediatric brain tumors in zebrafish. Oncogene 34, 1736–1742 (2015). https://doi.org/10.1038/onc.2014.107

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2014.107

This article is cited by

Search

Advanced search

Quick links