title: Long-term drug administration in the adult zebrafish using oral gavage for cancer preclinical studies.
abstract: Zebrafish are a major model for chemical genetics, and most studies use embryos when investigating small molecules that cause interesting phenotypes or that can rescue disease models. Limited studies have dosed adults with small molecules by means of water-borne exposure or injection techniques. Challenges in the form of drug delivery-related trauma and anesthesia-related toxicity have excluded the adult zebrafish from long-term drug efficacy studies. Here, we introduce a novel anesthetic combination of MS-222 and isoflurane to an oral gavage technique for a non-toxic, non-invasive and long-term drug administration platform. As a proof of principle, we established drug efficacy of the FDA-approved BRAF(V600E) inhibitor, Vemurafenib, in adult zebrafish harboring BRAF(V600E) melanoma tumors. In the model, adult casper zebrafish intraperitoneally transplanted with a zebrafish melanoma cell line (ZMEL1) and exposed to daily sub-lethal dosing at 100 mg/kg of Vemurafenib for 2 weeks via oral gavage resulted in an average 65% decrease in tumor burden and a 15% mortality rate. In contrast, Vemurafenib-resistant ZMEL1 cell lines, generated in culture from low-dose drug exposure for 4 months, did not respond to the oral gavage treatment regimen. Similarly, this drug treatment regimen can be applied for treatment of primary melanoma tumors in the zebrafish. Taken together, we developed an effective long-term drug treatment system that will allow the adult zebrafish to be used to identify more effective anti-melanoma combination therapies and opens up possibilities for treating adult models of other diseases.
authors.affiliation: Stem Cell Program and Division of Pediatric Hematology/Oncology, Boston Children's Hospital, Boston, MA 02115, USA Howard Hughes Medical Institute, Boston, MA 02115, USA Harvard Medical School, Boston, MA 02138, USA mdang@fas.harvard.edu zon@enders.tch.harvard.edu.
authors.forename: Michelle
authors.initials: M
authors.lastname: Dang
authors.affiliation: Stem Cell Program and Division of Pediatric Hematology/Oncology, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Henderson
authors.affiliation: Harvard Medical School, Boston, MA 02138, USA Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA 02215, USA The Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
authors.forename: Levi A
authors.initials: LA
authors.lastname: Garraway
authors.affiliation: Stem Cell Program and Division of Pediatric Hematology/Oncology, Boston Children's Hospital, Boston, MA 02115, USA Howard Hughes Medical Institute, Boston, MA 02115, USA Harvard Medical School, Boston, MA 02138, USA mdang@fas.harvard.edu zon@enders.tch.harvard.edu.
authors.forename: Leonard I
authors.identifier: 0000-0003-0860-926X
authors.initials: LI
authors.lastname: Zon
mesh_terms: D000284:Administration, Oral; D000818:Animals; D030801:Animals, Genetically Modified; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D004353:Drug Evaluation, Preclinical; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D007211:Indoles / Q000494:pharmacology / Q000627:therapeutic use; D008545:Melanoma / Q000188:drug therapy / Q000473:pathology; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology / Q000627:therapeutic use; D000077484:Vemurafenib; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D007211:Indoles; D013449:Sulfonamides; D000077484:Vemurafenib; D048493:Proto-Oncogene Proteins B-raf
keywords: Adult zebrafish; Melanoma; Oral gavage; Vemurafenib