A Murine Model for Metastatic Conjunctival Melanoma

Waard, Nadine E de; Cao, Jinfeng; McGuire, Sean P.; Kolovou, Paraskevi E.; Jordanova, Ekaterina S.; Ksander, Bruce R.; Jager, Martine J.

doi:10.1167/iovs.14-15239

Cited by 20 publications

(32 citation statements)

References 42 publications

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“…The results in vitro were expected as CRMM-1 and CM2005.1 harbor a B-RAF V600E mutation, while the CRMM-2 cell line contains an N-RAS Q61L mutation. 17,27 Vemurafenib was approved in 2011 by the Food and Drug Administration for treatment of unresectable melanoma harboring B-RAF V600E mutations 43 and is a potent agent for and P values were as follows: *P < 0.05, **P < 0.01, ***P < 0.001. For all groups: n ‡ 51. treatment of B-RAF V600E-positive melanoma.…”

Section: Discussionmentioning

confidence: 99%

“…15,16 Mice have previously been used as a model to study human CM, 17,18 but there are some limitations. The major disadvantages are a slow growth and spread of the tumor, which can take weeks to months, and the high cost for reproduction and housing.…”

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confidence: 99%

“…Our group has shown that two of three available CM cell lines, CRMM1 25 and CM2005.1, 26 harbor a B-RAF V600E mutation, while the third, CRMM2, contains an N-RAS Q61L mutation. 17,27 We injected stable red fluorescently labeled (lentiviral tdTomato-blas) CM cells via different routes into the embryonic zebrafish. Thus, we determined the most effective engraftment strategy for the establishment of CM xenograft tumors in zebrafish and we observed distinct phenotypes after implantation of the three CM cell lines.…”

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confidence: 99%

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Evaluation of (fli:GFP) Casper Zebrafish Embryos as a Model for Human Conjunctival Melanoma

Pontes

Groenewoud

Cao

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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Citation: Pontes KCS, Groenewoud A, Cao J, Ataide LMS, Snaar-Jagalska E, Jager MJ. Evaluation of (fli:GFP) Casper zebrafish embryos as a model for human conjunctival melanoma. Invest Ophthalmol Vis Sci. 2017;58:6065-6071. DOI:10.1167/iovs.17-22023 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.

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Evaluation of (fli:GFP) Casper Zebrafish Embryos as a Model for Human Conjunctival Melanoma

Pontes

Groenewoud

Cao

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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“…In our model, we are directly injecting tumor cells underneath the basement membrane, to mimic the clinicopathological situation. Similar to existing tumor models 30 in other anatomical sites of the eye, [31][32][33] this mouse model has its pros and cons. A major disadvantage might be the use of cutaneous and not primary CM cells, as it is not suitable for testing human antibodies or immune-mediated human therapies.…”

Section: Discussionmentioning

confidence: 98%

“…A similar approach for uveal melanoma is used in a mouse model developed by Lattier et al and Dithmar et al, [31][32][33] in which B16F10 cutaneous melanoma cells are injected in the posterior compartment of the eye. Another approach has been performed by De Waard et al 30 Hereby, the authors inject human CM cells into severe combined immunodeficient (SCID) mice.…”

Section: Discussionmentioning

confidence: 99%

A Novel Model of Metastatic Conjunctival Melanoma in Immune-Competent Mice

Schlereth

Iden

Mescher

et al. 2015

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. Conjunctival melanoma (CM) is an ocular surface tumor that can lead to fatal metastases. Patients developing, tumor-associated lymphangiogenesis have a significantly increased risk of metastatic disease, because tumor spread primarily occurs via lymphatic vessels to the draining lymph node. Here, we describe a novel immune-competent mouse model of CM that displays tumor-associated lymphangiogenesis with development of metastatic tumors.METHODS. C57BL/6N mice received C57BL/6N-derived dermal melanoma cells (hepatocyte growth factor [HGF] cyclin dependent kinase-4 [Cdk4]þ) or B16F10 via subconjunctival injection. A clinical score quantified primary tumor growth and metastases were identified by macroscopic examination of the draining lymph nodes, lung, and spleen. Confirmation of tumors and metastases was achieved by immunohistochemical staining for markers of pigmented cells (tyrosinase related protein-2 [TRP2]) and S-100, and of cell proliferation (Ki67). The intra-and peritumoral CD31þ blood and lymphatic vessel endothelium hyaluronan receptor-1 (LYVE-1)þ lymphatic vessels were quantified immunohistochemically.RESULTS. All mice rapidly developed aggressive TRP2þ, S100þ, and Ki67þ CM. Metastatic tumors were found in the lymph node (9%) and lung (6%) of HGF-Cdk4 R24C -treated mice and in the spleen (8%) and lung (17%) of B16F10-treated mice. The amount of peri-and intratumoral blood vessels was significantly increased compared with lymphatic vessels.CONCLUSIONS. This CM model in immune-competent animals offers new possibilities to study the pathobiology of tumor growth, invasion, and mechanisms of metastatic tumor spread, and provides a robust model to explore new immune-based and antilymphangiogenic treatment modalities of this malignancy.Keywords: conjunctival melanoma, mouse model, (lymph)-angiogenesis, ocular melanoma, B16F10, hepatocyte growth factor, metastasis C onjunctival melanoma (CM) is the second most common malignancy of the ocular surface. The current treatment of primary CM is complete surgical removal of the tumor combined with adjuvant therapy in terms of radio-, cryo-, topical chemo-, and/or immunotherapy. However, even though surgical specimens indicate intact tumor-negative margins and apparent tumor removal, CM patients still display an unusually high rate of local recurrence (36%-62% of patients [1][2][3][4][5][6] ). Tumor recurrence coincides with a high risk for the development of metastases, which occurs in 12% to 25% of patients. [1][2][3] Metastatic tumors in CM patients are typically first detected in the lymph nodes draining the tumor-containing eye, indicating the primary route of tumor cell dissemination occurs via the lymphatic vessels. In support of this, we previously demonstrated that lymphangiogenesis is associated with the transition of premalignant precursors into CM.7 Moreover, the extent of lymphatic vessels within CM coincided with an increased risk of local recurrence, lymphatic spread of tumor cells, and development of metastatic disease. [8][9][10] Melanomas ar...

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