High-risk blastemal Wilms tumor can be modeled by 3D spheroid cultures in vitro
High-risk blastemal Wilms tumor can be modeled by 3D spheroid cultures in vitro
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DOI：
10.1038/s41388-019-1027-8
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发表时间：
2020-01-01
期刊：
影响因子：
8
通讯作者：
Gessler, Manfred
中科院分区：
文献类型：
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作者：
Wegert, Jenny;Zauter, Lisa;Gessler, Manfred
体外模型是癌症研究中研究肿瘤生物学和评估新治疗方案的重要工具。不幸的是，没有有效的临床前模型可用于代表肾母细胞瘤(WT) -最常见的儿童肾脏肿瘤。尤其是胚质WT亚型的高危性，目前尚未建立起少数的原代细胞系。在这里，我们描述了一种新的用于体外培养胚质WT细胞的3D方法，其中原代培养物在悬浮中生长为球状体，可以长期繁殖。除了胚质培养外，我们还可以生成代表上皮和间质WT的球体。球体培养物与相应的肿瘤切片进行免疫组织化学分析，并通过RNA测序进一步表征。球体的组织学外观与原肿瘤相似，表达了早期肾脏发育特征的标记基因和胚质WT元件。这些培养物易于遗传操作，形成了类似于人类原发肿瘤的异种移植物肿瘤。携带不同遗传驱动的WT球体的集合形成了长期寻求的药物测试和体外建模工具。
In vitro models represent a critical tool in cancer research to study tumor biology and to evaluate new treatment options. Unfortunately, there are no effective preclinical models available that represent Wilms tumor (WT) - the most common pediatric renal tumor. Especially the high-risk blastemal WT subtype is not represented by the few primary cell lines established until now. Here, we describe a new 3D approach for in vitro cultivation of blastemal WT cells, where primary cultures grown in suspension as spheroids could be propagated long-term. Besides blastemal cultures, we could generate spheroids representing epithelial and stromal WT. Spheroid cultures were analyzed by immunohistochemistry in comparison to corresponding tumor sections and were further characterized by RNA sequencing. Histological appearance of spheroids resembled the original tumor and they expressed marker genes characteristic of early renal development and blastemal WT elements. The cultures were amenable to genetic manipulation and they formed xenograft tumors, which resemble the primary human tumor. This collection of WT spheroids that carry different genetic drivers forms a long-sought tool for drug testing and in vitro modeling.