髓母细胞瘤（MB）是儿童最常见的恶性脑肿瘤之一。目前的MB临床前体内模型系统增加了我们对调节MB发育的分子机制的理解。然而，它们可能不适合大规模研究。本研究的目的是调查基于斑马鱼的异种移植模型是否可以重现 MB 生长并实现快速药物测试。将九种不同的 MB 细胞系或患者来源的细胞移植到囊胚期斑马鱼胚胎中。然后使用实时成像监测肿瘤的发展和迁移。通过 RNA 测序来研究在神经干细胞样培养基中调节细胞后转录组的变化。此外，药物治疗在 96 孔板中进行了测试。我们在此证明，将 MB 细胞移植到斑马鱼胚胎囊胚期会导致原位肿瘤生长，移植后 24 小时内即可观察到这种生长。重要的是，通过在神经干细胞样培养基中预培养细胞，可以增强移植细胞向后脑区域的归巢和肿瘤生长的侵袭性。培养条件的变化将转录组重新连接为更具迁移性和神经元表型，包括引导分子 SEMA3A 和 EFNB1 的表达，这两者都与 MB 患者较低的总生存率相关。此外，我们强调原位斑马鱼 MB 模型具有用于快速药物测试的潜力。囊胚期斑马鱼 MB 异种移植物提供了当前 MB 小鼠异种移植模型的替代方案，能够快速评估肿瘤细胞生长、向神经性和药物疗效.© 作者 2024。由牛津大学出版社代表神经肿瘤学会出版。

Medulloblastoma (MB) is one of the most common malignant brain tumors in children. Current preclinical in vivo model systems for MB have increased our understanding of molecular mechanisms regulating MB development. However, they may not be suitable for large-scale studies. The aim of this study was to investigate if a zebrafish-based xenograft model can recapitulate MB growth and enable rapid drug testing.Nine different MB cell lines or patient-derived cells were transplanted into blastula-stage zebrafish embryos. Tumor development and migration were then monitored using live imaging. RNA sequencing was performed to investigate transcriptome changes after conditioning cells in neural stem cell-like medium. Furthermore, drug treatments were tested in a 96-well format.We demonstrate here that transplantation of MB cells into the blastula stage of zebrafish embryos leads to orthotopic tumor growth that can be observed within 24 hours after transplantation. Importantly, the homing of transplanted cells to the hindbrain region and the aggressiveness of tumor growth are enhanced by pre-culturing cells in a neural stem cell-like medium. The change in culture conditions rewires the transcriptome towards a more migratory and neuronal phenotype, including the expression of guidance molecules SEMA3A and EFNB1, both of which correlate with lower overall survival in MB patients. Furthermore, we highlight that the orthotopic zebrafish MB model has the potential to be used for rapid drug testing.Blastula-stage zebrafish MB xenografts present an alternative to current MB mouse xenograft models, enabling quick evaluation of tumor cell growth, neurotropism, and drug efficacy.© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Neuro-Oncology.