Serotype-dependent packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice
Serotype-dependent packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice
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DOI：
10.1172/jci34316
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发表时间：
2008-05-01
影响因子：
15.9
通讯作者：
Auricchio, Alberto
中科院分区：
文献类型：
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作者：
Allocca, Mariacarmela;Doria, Monica;Auricchio, Alberto
来自腺相关病毒(AAV)的载体有望用于人类的基因治疗，包括治疗视网膜失明。作为载体的AAV的一个主要限制是AAV的货运能力被认为被限制在4.7kb。在这里，我们证明了带有AAV5衣壳的载体(即rAAV2/5)比其他6种血清型更有效地整合了长达8.9kb的基因组，与rAAV2/5生产过程的效率无关。对在常见致盲疾病中突变的大鼠ABCA4和人MYO7A和CEP290基因进行了有效包装，表明这种包装效率与包装的特定序列无关。携带大基因的rAAV2/5转导后，观察到适当大小和功能的蛋白质表达。眼内注射编码ABCA4基因的rAAV2/5后，ABCA4(-/-)小鼠视网膜的蛋白质定位于视杆外节，视网膜的形态和功能得到显著而稳定的改善。这种rAAV2/5的使用可能是一种很有前途的治疗隐性Stargardt病的策略，隐性Stargardt病是最常见的遗传性黄斑变性。在AAV中包装大基因的可能性极大地扩展了该载体系统的治疗潜力。
Vectors derived from adeno-associated virus (AAV) are promising for human gene therapy, including treatment for retinal blindness. One major limitation of AAVs as vectors is that AAV cargo capacity has been considered to be restricted to 4.7 kb. Here we demonstrate that vectors with an AAV5 capsid (i.e., rAAV2/5) incorporated up to 8.9 kb of genome more efficiently than 6 other serotypes tested, independent of the efficiency of the rAAV2/5 production process. Efficient packaging of the large murine Abca4 and human MYO7A and CEP290 genes, which are mutated in common blinding diseases, was obtained, suggesting that this packaging efficiency is independent of the specific sequence packaged. Expression of proteins of the appropriate size and function was observed following transduction with rAAV2/5 carrying large genes. Intraocular administration of rAAV2/5 encoding ABCA4 resulted in protein localization to rod outer segments and significant and stable morphological and functional improvement of the retina in Abca4(-/-) mice. This use of rAAV2/5 may be a promising therapeutic strategy for recessive Stargardt disease, the most common form of inherited macular degeneration. The possibility of packaging large genes in AAV greatly expands the therapeutic potential of this vector system.