We have adapted a zebrafish (Danio rerio) tumor xenograft model for use in the study of oncolytic virotherapy. Following implantation of mammalian cancer cells into the perivitelline space of developing zebrafish embryos, both local and intravenous oncolytic virus treatments produce a tumor-specific infection with measurable antitumor effects. Tumor cells are injected at 48 h post fertilization, with oncolytic virus treatment then being administered 24 h later to allow for an initial period of tumor development and angiogenesis. Confocal fluorescent imaging is used to quantify dynamics within the tumor environment. The natural translucency of zebrafish at the embryo stage, coupled with the availability of strains with fluorescent immune and endothelial cell reporter lines, gives the model broad potential to allow for real time, in vivo investigation of important events within tumors throughout the course of virotherapy. Zebrafish xenografts offer a system with biologic fidelity to processes in human cancer development that influence oncolytic virus efficacy, and to our knowledge this is the first demonstration of the model’s use in the context of virotherapy. Compared with other models, our protocol offers a powerful, inexpensive approach to evaluating novel oncolytic viruses and oncolytic virus-based combination therapies, with potential application to investigating the impacts of virotherapy on immune response, tumor vasculature, and metastatic disease.

我们已经适应了斑马鱼 ( Danio rerio) 用于研究溶瘤病毒疗法的肿瘤异种移植模型。将哺乳动物癌细胞植入发育中的斑马鱼胚胎的卵周空间后，局部和静脉溶瘤病毒治疗都会产生具有可测量抗肿瘤作用的肿瘤特异性感染。在受精后 48 小时注射肿瘤细胞，然后在 24 小时后给予溶瘤病毒治疗，以允许肿瘤发展和血管生成的初始阶段。共聚焦荧光成像用于量化肿瘤环境中的动态。斑马鱼在胚胎阶段的天然半透明性，加上具有荧光免疫和内皮细胞报告系的菌株的可用性，使该模型具有广泛的潜力，可以实现实时、在整个病毒治疗过程中对肿瘤内的重要事件进行体内研究。斑马鱼异种移植物为影响溶瘤病毒功效的人类癌症发展过程提供了一个具有生物保真度的系统，据我们所知，这是该模型在病毒疗法背景下使用的首次证明。与其他模型相比，我们的协议提供了一种强大、廉价的方法来评估新型溶瘤病毒和基于溶瘤病毒的联合疗法，并有可能用于研究病毒疗法对免疫反应、肿瘤脉管系统和转移性疾病的影响。据我们所知，这是该模型在病毒疗法背景下的首次展示。与其他模型相比，我们的协议提供了一种强大、廉价的方法来评估新型溶瘤病毒和基于溶瘤病毒的联合疗法，并可能应用于研究病毒疗法对免疫反应、肿瘤脉管系统和转移性疾病的影响。据我们所知，这是该模型在病毒疗法背景下的首次展示。与其他模型相比，我们的协议提供了一种强大、廉价的方法来评估新型溶瘤病毒和基于溶瘤病毒的联合疗法，并可能应用于研究病毒疗法对免疫反应、肿瘤脉管系统和转移性疾病的影响。