Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy
Vector design influences hepatic genotoxicity after adeno-associated virus gene therapy
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DOI：
10.1172/jci79213
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发表时间：
2015-02-01
影响因子：
15.9
通讯作者：
Venditti, Charles P.
中科院分区：
文献类型：
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作者：
Chandler, Randy J.;LaFave, Matthew C.;Venditti, Charles P.
腺相关病毒（AAV）作为基因治疗载体的使用最近已被批准用于临床使用，并已在越来越多的临床试验中证明了疗效。然而，AAV作为载体的安全性受到了单一研究的挑战，该研究记录了AAV基因递送小鼠后的肝细胞癌（HCC）。大多数研究没有注意到AAV介导的基因递送后的遗传毒性;因此，AAV和HCC之间存在关联的可能性是有争议的。在这里，我们进行了一个全面的研究肝癌在大量的小鼠治疗性AAV基因传递。利用高通量整合位点捕获技术和全局表达分析，我们发现AAV整合入RNA印迹并积累在核（Rian）位点，以及由此产生的近端microRNA和逆转录转座子样1（Rt/l）的过表达与HCC相关。此外，我们证明了AAV载体剂量、增强子/启动子选择和基因递送的时机都是决定AAV基因递送后HCC发病率的关键因素。总之，我们的研究结果定义了影响遗传毒性的AAV介导的基因治疗的方面，并建议这些功能应考虑更安全的AAV载体和基因治疗研究的设计。
The use of adeno-associated virus (AAV) as a gene therapy vector has been approved recently for clinical use and has demonstrated efficacy in a growing number of clinical trials. However, the safety of AAV as a vector has been challenged by a single study that documented hepatocellular carcinoma (HCC) after AAV gene delivery in mice. Most studies have not noted genotoxicity following AAV-mediated gene delivery; therefore, the possibility that there is an association between AAV and HCC is controversial. Here, we performed a comprehensive study of HCC in a large number of mice following therapeutic AAV gene delivery. Using a sensitive high-throughput integration site-capture technique and global expressional analysis, we found that AAV integration into the RNA imprinted and accumulated in nucleus (Rian) locus, and the resulting overexpression of proximal microRNAs and retrotransposon-like 1 (Rt/l) were associated with HCC. In addition, we demonstrated that the AAV vector dose, enhancer/promoter selection, and the timing of gene delivery are all critical factors for determining HCC incidence after AAV gene delivery. Together, our results define aspects of AAV-mediated gene therapy that influence genotoxicity and suggest that these features should be considered for design of both safer AAV vectors and gene therapy studies.