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Updated: Feb 17, 2026
Ortho- and Ectopic Zebrafish Xeno-Engraftment of Ocular Melanoma to Recapitulate Primary Tumor and Experimental Metastasis Development
Published on: September 4, 2021
Evaluation of (fli:GFP) Casper Zebrafish Embryos as a Model for Human Conjunctival Melanoma
Kelly Cristine de Sousa Pontes1, Arwin Groenewoud2, Jinfeng Cao1,3
1Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands.
Purpose:
Conjunctival melanoma (CM) is a rare malignant disease that can lead to recurrences and metastases. There is a lack of effective treatments for the metastases, and we set out to develop a new animal model to test potential therapies. Zebrafish are being used as a model for many diseases, and our goal was to test whether this animal could be used to study CM.
Methods:
Three human CM cell lines (CRMM-1 and CM2005.1, which both harbor a B-RAF mutation, and CRMM-2, which has an N-RAS mutation) were injected into the yolk sac, around the eye, and into the duct of Cuvier of transgenic (fli:GFP) Casper zebrafish embryos. Fluorescent and confocal images were taken to assess the phenotype and the behavior of engrafted cells and to test the effect of Vemurafenib as a treatment against CM.
Results:
While the cells that had been injected inside the yolk sac died and those injected around the eye sporadically went into the circulation, the cells that had been injected into the duct of Cuvier colonized the zebrafish: cells from all three cell lines proliferated and disseminated to the eyes, where they formed clusters, and to the tail, where we noticed extravasation and micrometastases. Vemurafenib, a potent agent for treatment of B-RAF V600E-positive melanoma, inhibited outgrowth of CRMM-1 and CM2005.1 cells in a mutation-dependent way.
Conclusions:
The (fli:GFP) Casper zebrafish embryo can be used as an efficient animal model to study metastatic behavior of human CM cells and warrants further testing of drug efficacy to aid care of CM patients.
Insights
Zebrafish embryos effectively model human conjunctival melanoma (CM) metastasis, enabling drug efficacy testing. This new model shows promise for developing treatments for rare CM cancers.
Area of Science:
- Oncology
- Genetics
- Developmental Biology
Background:
- Conjunctival melanoma (CM) is a rare cancer with limited treatment options for metastatic disease.
- Developing effective animal models is crucial for studying CM progression and testing new therapies.
Purpose of the Study:
- To establish a novel zebrafish (Danio rerio) embryo model for studying conjunctival melanoma (CM) metastasis.
- To evaluate the potential of zebrafish as a platform for preclinical drug screening against CM.
Main Methods:
- Three human CM cell lines with known mutations (B-RAF or N-RAS) were xenografted into transgenic (fli:GFP) Casper zebrafish embryos.
- Engrafted cells were monitored using fluorescent and confocal microscopy to assess tumor cell behavior and metastatic potential.
- The efficacy of Vemurafenib, a targeted therapy, was tested against CM cells harboring specific mutations.
Main Results:
- Zebrafish embryos successfully supported the colonization and dissemination of human CM cells injected into the duct of Cuvier.
- Metastases were observed in the eyes and tail, demonstrating the model's ability to replicate metastatic spread.
- Vemurafenib demonstrated mutation-dependent inhibition of CM cell outgrowth, validating the model's utility for drug testing.
Conclusions:
- The transgenic zebrafish embryo serves as an efficient and valuable model for studying conjunctival melanoma metastasis.
- This model holds significant potential for advancing the development of targeted therapies and improving patient care for CM.
