葡萄膜黑色素瘤（UM）是最常见的原发性眼内恶性肿瘤，转移性疾病的可能性很高。尽管原发性 UM 已有优良的治疗选择，但转移性疾病的治疗仍然有限。迄今为止，使用小鼠模型的药物发现研究未能提供治疗解决方案，这凸显了对新模型的需求。在这里，我们通过展示多种细胞系的行为以及使用 Z-Tada 的突变依赖性化合物协同/拮抗作用的新发现，优化斑马鱼异种移植物作为药物发现的潜在模型；客观表征输出测量的算法。在三个不同的接种位点接种与预后相关的原发性 (N = 4) 和转移性 UM (N = 1) 细胞系或健康黑素细胞 (N = 2)。使用 Z-Tada 获得了独立于接种部位的标准化定量；一种测量肿瘤负荷以及播散肿瘤细胞的数量、大小和距离的算法。随后，我们利用该模型来验证体外观察到的组合协同作用或拮抗作用。对 691 个斑马鱼异种移植物的详细分析表明，卵周间隙接种提供了具有高细胞传播可能性的可靠数据。具有更多侵袭行为的细胞系（SF3B1mut 和 BAP1mut）在此模型中表现得最具攻击性。组合药物治疗表明协同或拮抗作用是突变依赖性的，这在体内得到了证实。每个异种移植模型的组合治疗都不同，因为它要么抑制总体肿瘤负荷，要么抑制细胞传播。卵周间隙接种为斑马鱼异种移植提供了强大的能力，能够使用 Z-Tada 进行高通量药物筛选和强大的数据采集。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，特别是在组合治疗发现中。

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.