Department of Ophthalmology, Erasmus MC Cancer Institute, Erasmus MC, Rotterdam, The Netherlands.
Clinical Genetics, Erasmus MC Cancer Institute, Erasmus MC, Rotterdam, The Netherlands.
Invest Ophthalmol Vis Sci. 2024 Aug 1;65(10):26. doi: 10.1167/iovs.65.10.26.
Uveal melanoma (UM) is the most common primary intraocular malignancy with a high probability of metastatic disease. Although excellent treatment options for primary UM are available, therapy for metastatic disease remain limited. Drug discovery studies using mouse models have thus far failed to provide therapeutic solutions, highlighting the need for novel models. Here, we optimize zebrafish xenografts as a potential model for drug discovery by showcasing the behavior of multiple cell lines and novel findings on mutation-dependent compound synergism/antagonism using Z-Tada; an algorithm to objectively characterize output measurements.
Prognostic relevant primary (N = 4) and metastatic UM (N = 1) cell lines or healthy melanocytes (N = 2) were inoculated at three distinct inoculation sites. Standardized quantifications independent of inoculation site were obtained using Z-Tada; an algorithm to measure tumor burden and the number, size, and distance of disseminated tumor cells. Sequentially, we utilized this model to validate combinatorial synergism or antagonism seen in vitro.
Detailed analysis of 691 zebrafish xenografts demonstrated perivitelline space inoculation provided robust data with high probability of cell dissemination. Cell lines with more invasive behavior (SF3B1mut and BAP1mut) behaved most aggressive in this model. Combinatorial drug treatment illustrated synergism or antagonism is mutation-dependent, which were confirmed in vivo. Combinatorial treatment differed per xenograft-model, as it either inhibited overall tumor burden or cell dissemination.
Perivitelline space inoculation provides robust zebrafish xenografts with the ability for high-throughput drug screening and robust data acquisition using Z-Tada. This model demonstrates that drug discovery for uveal melanoma must take mutational subclasses into account, especially in combinatorial treatment discoveries.
葡萄膜黑色素瘤（UM）是最常见的原发性眼内恶性肿瘤，具有很高的转移疾病概率。尽管有治疗原发性 UM 的优秀治疗选择，但转移性疾病的治疗仍然有限。使用小鼠模型的药物发现研究迄今为止未能提供治疗解决方案，这凸显了对新型模型的需求。在这里，我们通过展示多种细胞系的行为以及使用 Z-Tada 对突变依赖性化合物协同作用/拮抗作用的新发现，将斑马鱼异种移植优化为一种潜在的药物发现模型；一种客观描述输出测量的算法。
将具有预后相关性的原发性（N=4）和转移性 UM（N=1）细胞系或健康黑素细胞（N=2）接种在三个不同的接种部位。使用 Z-Tada 独立于接种部位进行标准化定量；一种测量肿瘤负担以及散布的肿瘤细胞数量、大小和距离的算法。随后，我们利用该模型验证了体外观察到的组合协同作用或拮抗作用。
对 691 个斑马鱼异种移植的详细分析表明，卵黄周腔接种提供了具有高细胞扩散概率的稳健数据。侵袭性更强的行为（SF3B1mut 和 BAP1mut）的细胞系在该模型中表现出最具侵袭性。组合药物治疗表明协同作用或拮抗作用是突变依赖性的，这在体内得到了证实。组合治疗因异种移植模型而异，因为它要么抑制总体肿瘤负担，要么抑制细胞扩散。
卵黄周腔接种为斑马鱼异种移植提供了稳健的数据，具有高通量药物筛选和使用 Z-Tada 进行稳健数据采集的能力。该模型表明，葡萄膜黑色素瘤的药物发现必须考虑突变亚类，尤其是在组合治疗发现中。
