Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish

doi:10.1073/pnas.0611302104

Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish

*Stem Cell Program and Division of Hematology/Oncology, Children's Hospital, Boston, MA 02115;
^†Dana–Farber Cancer Institute, Boston, MA 02115;
^‡Howard Hughes Medical Institute, Cambridge, MA 02138;
^§Harvard Medical School, Boston, MA 02115; and
^¶Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115

Edited by Mark T. Groudine, Fred Hutchinson Cancer Research Center, Seattle, WA, and approved April 17, 2007 (received for review December 19, 2006)

Abstract

RAS family members are among the most frequently mutated oncogenes in human cancers. Given the utility of zebrafish in both chemical and genetic screens, developing RAS-induced cancer models will make large-scale screens possible to understand further the molecular mechanisms underlying malignancy. We developed a heat shock-inducible Cre/Lox-mediated transgenic approach in which activated human kRASG12D can be conditionally induced within transgenic animals by heat shock treatment. Specifically, double transgenic fish Tg(B-actin-LoxP-EGFP-LoxP-kRASG12D; hsp70-Cre) developed four types of tumors and hyperplasia after heat shock of whole zebrafish embryos, including rhabdomyosarcoma, myeloproliferative disorder, intestinal hyperplasia, and malignant peripheral nerve sheath tumor. Using ex vivo heat shock and transplantation of whole kidney marrow cells from double transgenic animals, we were able to generate specifically kRASG12D-induced myeloproliferative disorder in recipient fish. This heat shock-inducible recombination approach allowed for the generation of multiple types of RAS-induced tumors and hyperplasia without characterizing tissue-specific promoters. Moreover, these tumors and hyperplasia closely resemble human diseases at both the morphologic and molecular levels.

Footnotes

^‖To whom correspondence should be addressed at:
HHMI/Children's Hospital, 300 Longwood Avenue, Karp 7, Boston, MA 02115.
E-mail: zon{at}enders.tch.harvard.edu
Author contributions: X.L. and D.M.L. contributed equally to this work; X.L., D.M.L., and L.I.Z. designed research; X.L., D.M.L., and M.D.K. performed research; X.L., D.M.L., J.L.K., D.S.N., and L.I.Z. analyzed data; and X.L., D.M.L., and L.I.Z. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/cgi/content/full/0611302104/DC1.
Abbreviations:

LGL-RAS,

B-actin-LoxP-EGFP-LoxP-kRASG12D;

MPD,

myeloproliferative disorder;

MPNST,

malignant peripheral nerve sheath tumor;

RMS,

rhabdomyosarcoma;

dpf,

days postfertilization;

hpf,

hours postfertilization.