Genome-Wide CRISPR-Cas9 Screening Identifies NF-κB/E2F6 Responsible for EGFRvIII-Associated Temozolomide Resistance in Glioblastoma
Genome-Wide CRISPR-Cas9 Screening Identifies NF-κB/E2F6 Responsible for EGFRvIII-Associated Temozolomide Resistance in Glioblastoma
复制标题
全基因组 CRISPR-Cas9 筛选鉴定出 NF-kappa B/E2F6 负责胶质母细胞瘤中与 EGFRvIII 相关的替莫唑胺耐药性
DOI：
10.1002/advs.201900782
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发表时间：
2019-07-24
期刊：
影响因子：
15.1
通讯作者：
Kang, Chunsheng
中科院分区：
文献类型：
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作者：
Huang, Kai;Liu, Xing;Kang, Chunsheng
表皮生长因子受体（EGFR）和活性突变体EGFRvIII的扩增经常发生在胶质母细胞瘤（GBM）中，并且有助于各种癌症（尤其是GBM）中的化学/放射抗性。阐明GBM中替莫唑胺（TMZ）耐药的潜在分子机制可使癌症患者受益。在成簇的规则间隔短回文重复序列（CRISPR）-Cas9文库下进行全基因组筛选以鉴定在表达EGFRvIII的GBM细胞中赋予TMZ抗性的基因。深度sgRNA测序揭示了191个负责表达EGFRvIII的GBM细胞中TMZ抗性的候选基因。值得注意的是，E2 F6被证明驱动TMZ抗性，并且E2 F6表达由EGFRvIII/AKT/NF-κ B途径控制。此外，在原位GBM细胞系异种移植物和GBM患者来源的异种移植物模型中，E2 F6显示为TMZ抗性的有希望的治疗靶标。在将临床数据与来自134名在手术后接受TMZ治疗的GBM患者的配对的原发性复发性RNA测序数据整合后，已经揭示了E2 F6表达水平是TMZ反应的预测标志物。因此，抑制E2 F6是克服GBM中TMZ抗性的有希望的策略。
Amplification of epidermal growth factor receptor (EGFR) and active mutant EGFRvIII occurs frequently in glioblastoma (GBM) and contributes to cherno/radio-resistance in various cancers, especially in GBM. Elucidating the underlying molecular mechanism of temozolomide (TMZ) resistance in GBM could benefit cancer patients. A genome-wide screening under a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 library is conducted to identify the genes that confer resistance to TMZ in EGFRvIII-expressing GBM cells. Deep sgRNA sequencing reveals 191 candidate genes that are responsible for TMZ resistance in EGFRvIII-expressing GBM cells. Notably, E2F6 is proven to drive a TMZ resistance, and E2F6 expression is controlled by the EGFRvIII/AKT/NF-kappa B pathway. Furthermore, E2F6 is shown as a promising therapeutic target for TMZ resistance in orthotopic GBM cell line xenografts and GBM patient-derived xenografts models. After integrating clinical data with paired primary-recurrent RNA sequencing data from 134 GBM patients who received TMZ treatment after surgery, it has been revealed that the E2F6 expression level is a predictive marker for TMZ response. Therefore, the inhibition of E2F6 is a promising strategy to conquer TMZ resistance in GBM.