BACKGROUND Current experimental models using either human or mouse cell lines, are not representative of the complex features of GBM. In particular, there is no model to study patient-derived iPSCs to generate a GBM model. Overexpression of c-met gene is one of the molecular features of GBM leading to increased signaling via STAT3 phosphorylation. We generated an iPSC line from a patient with c-met mutation and we asked whether we could use it to generate neuronal-like organoids mimicking features of GBM. METHODS We have generated iPSC-aggregates differentiating towards organoids. We analyzed them by gene expression profiling, immunostaining and transmission electronic microscopy analyses (TEM). RESULTS Herein we describe that c-met-mutated iPSC aggregates spontaneously differentiate into dopaminergic neurons more rapidly than control iPSC aggregates in culture. Gene expression profiling of c-met-mutated iPSC aggregates at day +90 showed neuronal- and GBM-related genes, reproducing a genomic network described in primary human GBM. Comparative TEM analyses confirmed the enrichment of these structures in intermediate filaments and abnormal cilia, a feature described in human GBM. The c-met-mutated iPSC-derived organoids, as compared to controls expressed high levels of glial fibrillary acidic protein (GFAP), which is a typical marker of human GBM, as well as high levels of phospho-MET and phospho-STAT3. The use of temozolomide (TMZ) showed a preferential cytotoxicity of this drug in c-met-mutated neuronal-like organoids. GENERAL SIGNIFICANCE This study shows the feasibility of generating "off-the shelf" neuronal-like organoid model mimicking GBM using c-met-mutated iPSC aggregates and its potential future use in research.

中文翻译：

---

新型神经元类器官模型模仿来自诱导多能干细胞（iPSC）的胶质母细胞瘤（GBM）功能。

背景技术当前使用人或小鼠细胞系的实验模型不能代表GBM的复杂特征。特别是，没有模型可以研究患者来源的iPSC来生成GBM模型。c-met基因的过度表达是GBM的分子特征之一，可导致通过STAT3磷酸化增加信号传导。我们从一名具有c-met突变的患者中产生了一条iPSC细胞系，我们询问是否可以用它来产生模仿GBM特征的神经元样类器官。方法我们已经生成了区分类器官的iPSC聚集体。我们通过基因表达谱分析，免疫染色和透射电子显微镜分析（TEM）对它们进行了分析。结果在本文中，我们描述了c-met突变的iPSC聚集体比培养中的对照iPSC聚集体更快地自发分化为多巴胺能神经元。在第90天时，c-met突变的iPSC聚集体的基因表达谱显示神经元和GBM相关的基因，复制了原代人GBM中描述的基因组网络。比较TEM分析证实了这些结构在中间丝和异常纤毛中的富集，这是人GBM中描述的特征。与对照组相比，c-met突变的iPSC衍生的类器官表达了高水平的胶质原纤维酸性蛋白（GFAP），这是人GBM的典型标志物，以及高水平的磷酸MET和磷酸STAT3。替莫唑胺（TMZ）的使用显示该药物在c-met突变的神经元样类器官中具有优先的细胞毒性。