Programmable base editing of mutated TERT promoter inhibits brain tumour growth
Programmable base editing of mutated TERT promoter inhibits brain tumour growth
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DOI：
10.1038/s41556-020-0471-6
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发表时间：
2020-02-17
影响因子：
21.3
通讯作者：
Lu, Zhimin
中科院分区：
文献类型：
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作者：
Li, Xinjian;Qian, Xu;Lu, Zhimin
Li等人证明了使用可编程碱基编辑校正突变的TERT启动子的功效，突出了其在脑肿瘤模型中诱导衰老、停滞和消退的能力。规则间隔短回文重复序列（CRISPR）、CRISPR干扰和可编程碱基编辑已经改变了用于潜在治疗应用的真核基因组的操作（1-4）。在这里，我们利用CRISPR干扰和可编程碱基编辑来确定它们在编辑TERT基因启动子激活突变中的潜力，这种突变发生在许多不同的癌症类型中，特别是胶质母细胞瘤（5-8）。使用单向导RNA（sgRNA）引导和催化受损的空肠弯曲杆菌CRISPR相关蛋白9-融合腺嘌呤碱基编辑器（CjABE）实现了-124C>T TERT启动子突变至-124C的校正。这种修饰阻断了E26转录因子家族成员与TERT启动子的结合，降低了TERT转录和TERT蛋白表达，并诱导了癌细胞衰老和增殖停滞。局部注射表达sgRNA引导的CjABE的腺相关病毒抑制了携带TERT启动子突变的胶质瘤的生长。这些临床前概念验证研究确立了基因编辑作为癌症治疗方法的可行性，并验证了激活的TERT启动子突变作为癌症特异性治疗靶点。
Li et al. demonstrate the efficacy of correcting the mutated TERT promoter using a programmable base editing, highlighting its ability to induce senescence, arrest and regression in brain tumour models.Clustered regularly interspaced short palindromic repeats (CRISPR), CRISPR interference and programmable base editing have transformed the manipulation of eukaryotic genomes for potential therapeutic applications(1-4). Here, we exploited CRISPR interference and programmable base editing to determine their potential in editing a TERT gene promoter-activating mutation, which occurs in many diverse cancer types, particularly glioblastoma(5-8). Correction of the -124C>T TERT promoter mutation to -124C was achieved using a single guide RNA (sgRNA)-guided and catalytically impaired Campylobacter jejuni CRISPR-associated protein 9-fused adenine base editor (CjABE). This modification blocked the binding of members of the E26 transcription factor family to the TERT promoter, reduced TERT transcription and TERT protein expression, and induced cancer-cell senescence and proliferative arrest. Local injection of adeno-associated viruses expressing sgRNA-guided CjABE inhibited the growth of gliomas harbouring TERT-promoter mutations. These preclinical proof-of-concept studies establish the feasibility of gene editing as a therapeutic approach for cancer and validate activated TERT-promoter mutations as a cancer-specific therapeutic target.