临床测序工作不断发现新的推测致癌基因，但目前缺乏有效的策略来在体内进行功能验证并研究其在肿瘤发生中的作用。本文介绍了一种使用转基因斑马鱼系统进行融合驱动横纹肌肉瘤（RMS）体内建模的流程。该策略源自于从患者样本中发现的新型融合致癌基因，在脊椎动物系统中进行功能验证，将这些基因整合到斑马鱼基因组中，然后验证它们是否确实引发了横纹肌肉瘤的肿瘤形成。在这种情况下，将融合致癌基因的人类形式插入到斑马鱼基因组中，以了解它是否是致癌基因，以及可能的肿瘤发生机制。这种方法在我们的婴儿横纹肌肉瘤和肺泡横纹肌肉瘤模型中取得了成功，这两种疾病分别由VGLL2-NCOA2和PAX3-FOXO1融合致癌基因驱动。我们所描述的斑马鱼平台是一种迅速了解融合致癌基因活性、不同和共享的融合致癌基因生物学以及是否存在任何分析的途径汇聚可能用于临床可操作靶点的方法。© 2024. 该作者通过途径Springer Science+Business Media, LLC独占许可授予Springer Nature的使用权。

Clinical sequencing efforts continue to identify novel putative oncogenes with limited strategies to perform functional validation in vivo and study their role in tumorigenesis. Here, we present a pipeline for fusion-driven rhabdomyosarcoma (RMS) in vivo modeling using transgenic zebrafish systems. This strategy originates with novel fusion-oncogenes identified from patient samples that require functional validation in vertebrate systems, integrating these genes into the zebrafish genome, and then characterizing that they indeed drive rhabdomyosarcoma tumor formation. In this scenario, the human form of the fusion-oncogene is inserted into the zebrafish genome to understand if it is an oncogene, and if so, the underlying mechanisms of tumorigenesis. This approach has been successful in our models of infantile rhabdomyosarcoma and alveolar rhabdomyosarcoma, both driven by respective fusion-oncogenes, VGLL2-NCOA2 and PAX3-FOXO1. Our described zebrafish platform is a rapid method to understand the impact of fusion-oncogene activity, divergent and shared fusion-oncogene biology, and whether any analyzed pathways converge for potential clinically actionable targets.© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.