Expression profiles in wild-type and IDH1 R132H/WT human astroglial cells [RNA-seq]. Homo sapiens

Mutations in the isocitrate dehydrogenase 1 (IDH1) gene are critical to oncogenesis. The exact mechanism by which mutant IDH1 drives cell transformation is still not fully understood, partially due to the difficulty of maintaining cells with endogenously mutated IDH1. We employed a “single base editing” technique and efficiently introduced the monoallelic point mutation of IDH1 R132H (IDH1R132H/WT) into non-neoplastic human astroglial cells. Characterization of our cellular models revealed that IDH1R132H/WT inhibited cell proliferation and promoted cell migration via mechanisms mediated by its oncometabolite 2-HG. Global gene expression and epigenetic analysis identified novel molecular targets of IDH1R132H/WT, namely the Hippo pathway effector, Yes-associated protein (YAP), and its downstream signaling pathway Notch. In summary, the “single base editing” strategy introduces a new paradigm that recapitulates the biological function of IDH1 R132H/WT and its oncometabolite 2-HG, which can be easily applied to other cell models. Our study provides a valuable model for novel discoveries of molecular mechanisms during IDH1 R132H/WT-driven pre-cancerous events. Overall design: We measured gene expression profiles of a wild-type and two IDH1R132H/WT clones from human astroglial cells

异柠檬酸脱氢酶1（isocitrate dehydrogenase 1, IDH1）基因的突变在肿瘤发生过程中发挥关键作用。目前，突变型IDH1驱动细胞转化的确切机制仍未完全阐明，这一困境部分源于维持内源性携带IDH1突变的细胞存在技术难度。本研究采用单碱基编辑（single base editing）技术，将IDH1 R132H的单等位基因点突变（IDH1R132H/WT）高效导入非肿瘤性人类星形胶质细胞中。对该细胞模型的表征分析显示，IDH1R132H/WT可通过其促癌代谢物2-羟基戊二酸（2-hydroxyglutarate, 2-HG）介导的信号通路，抑制细胞增殖并促进细胞迁移。全基因表达与表观遗传分析鉴定出IDH1R132H/WT的新型分子靶点：Hippo通路效应分子Yes相关蛋白（Yes-associated protein, YAP）及其下游Notch信号通路。综上，单碱基编辑策略为复刻IDH1R132H/WT及其促癌代谢物2-HG的生物学功能提供了全新研究范式，该方法可便捷地推广应用至其他细胞模型体系。本研究为解析IDH1R132H/WT驱动的癌前病变分子机制提供了极具价值的实验模型。总体实验设计：本研究检测了人类星形胶质细胞中1株野生型细胞与2株IDH1R132H/WT突变型细胞的基因表达谱。