Widespread Central Nervous System Gene Transfer and Silencing After Systemic Delivery of Novel AAV-AS Vector
Widespread Central Nervous System Gene Transfer and Silencing After Systemic Delivery of Novel AAV-AS Vector
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DOI：
10.1038/mt.2015.231
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发表时间：
2016-04-01
影响因子：
12.4
通讯作者：
Sena-Esteves, Miguel
中科院分区：
文献类型：
--
作者：
Choudhury, Sourav R.;Harris, Anne F.;Sena-Esteves, Miguel
中枢神经系统的基因转移是神经系统疾病基因治疗的关键。腺相关病毒（AAV）载体已成为体内基因转移的有效平台，但全身施用后源自天然存在的AAV衣壳的载体的总体神经元转导效率相对较低。在这里，我们研究了通过将肽基因融合到VP 2衣壳蛋白的N-末端来改善现有AAV衣壳的CNS转导的可能性。通过插入聚丙氨酸肽产生的新型载体AAV-AS能够在成年小鼠全身给药后在整个CNS中进行广泛的基因转移。AAV-AS在脊髓和大脑中的效率分别是AAV 9的6倍和15倍。神经元转导谱在脑区域中变化，但在纹状体中特别高，其中AAV-AS转导36%的纹状体神经元。广泛的神经元基因转移也记录在猫的大脑和脊髓。编码靶向亨廷顿蛋白（Htt）的人工微小RNA的AAV-AS载体的单次静脉内注射导致成年小鼠中的多个CNS结构中Htt的33-50%敲低。这种新的AAV-AS载体是一个有前途的平台，开发新的基因治疗神经退行性疾病。
Effective gene delivery to the central nervous system (CNS) is vital for development of novel gene therapies for neurological diseases. Adeno-associated virus (AAV) vectors have emerged as an effective platform for in vivo gene transfer, but overall neuronal transduction efficiency of vectors derived from naturally occurring AAV capsids after systemic administration is relatively low. Here, we investigated the possibility of improving CNS transduction of existing AAV capsids by genetically fusing peptides to the N-terminus of VP2 capsid protein. A novel vector AAV-AS, generated by the insertion of a poly-alanine peptide, is capable of extensive gene transfer throughout the CNS after systemic administration in adult mice. AAV-AS is 6- and 15-fold more efficient than AAV9 in spinal cord and cerebrum, respectively. The neuronal transduction profile varies across brain regions but is particularly high in the striatum where AAV-AS transduces 36% of striatal neurons. Widespread neuronal gene transfer was also documented in cat brain and spinal cord. A single intravenous injection of an AAV-AS vector encoding an artificial microRNA targeting huntingtin (Htt) resulted in 33-50% knockdown of Htt across multiple CNS structures in adult mice. This novel AAV-AS vector is a promising platform to develop new gene therapies for neurodegenerative disorders.