A causative understanding of genetic factors that regulate glioblastoma pathogenesis is of central importance. Here we developed an adeno-associated virus-mediated, autochthonous genetic CRISPR screen in glioblastoma. Stereotaxic delivery of a virus library targeting genes commonly mutated in human cancers into the brains of conditional-Cas9 mice resulted in tumors that recapitulate human glioblastoma. Capture sequencing revealed diverse mutational profiles across tumors. The mutation frequencies in mice correlated with those in two independent patient cohorts. Co-mutation analysis identified co-occurring driver combinations such as B2m-Nf1, Mll3-Nf1 and Zc3h13-Rb1, which were subsequently validated using AAV minipools. Distinct from Nf1-mutant tumors, Rb1-mutant tumors are undifferentiated and aberrantly express homeobox gene clusters. The addition of Zc3h13 or Pten mutations altered the gene expression profiles of Rb1 mutants, rendering them more resistant to temozolomide. Our study provides a functional landscape of gliomagenesis suppressors in vivo.

中文翻译：

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AAV 介导的直接体内 CRISPR 筛选可识别胶质母细胞瘤中的功能抑制因子。

了解调节胶质母细胞瘤发病机制的遗传因素至关重要。在这里，我们在胶质母细胞瘤中开发了一种腺相关病毒介导的本土遗传 CRISPR 筛选。将靶向人类癌症中常见突变基因的病毒库立体定向递送到条件性 Cas9 小鼠的大脑中，会导致肿瘤重现人类胶质母细胞瘤。捕获测序揭示了跨肿瘤的不同突变谱。小鼠中的突变频率与两个独立患者队列中的突变频率相关。共突变分析确定了共同发生的驱动程序组合，例如 B2m-Nf1、Mll3-Nf1 和 Zc3h13-Rb1，随后使用 AAV 小型池进行了验证。与 Nf1 突变肿瘤不同，Rb1 突变肿瘤未分化并且异常表达同源框基因簇。Zc3h13 或 Pten 突变的添加改变了 Rb1 突变体的基因表达谱，使其对替莫唑胺更具抗性。我们的研究提供了体内胶质瘤形成抑制剂的功能景观。