Choudhury Sourav R, Harris Anne F, Cabral Damien J, Keeler Allison M, Sapp Ellen, Ferreira Jennifer S, Gray-Edwards Heather L, Johnson Jacob A, Johnson Aime K, Su Qin, Stoica Lorelei, DiFiglia Marian, Aronin Neil, Martin Douglas R, Gao Guangping, Sena-Esteves Miguel
Department of Neurology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Gene Therapy Center, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Mol Ther. 2016 Apr;24(4):726-35. doi: 10.1038/mt.2015.231. Epub 2015 Dec 28.
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.
向中枢神经系统(CNS)进行有效的基因传递对于开发针对神经疾病的新型基因疗法至关重要。腺相关病毒(AAV)载体已成为体内基因转移的有效平台,但全身给药后源自天然AAV衣壳的载体的整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VP2衣壳蛋白的N端来提高现有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,在成年小鼠全身给药后能够在整个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比AAV9高6倍和15倍。神经元转导谱在不同脑区有所不同,但在纹状体中特别高,其中AAV-AS转导36%的纹状体神经元。在猫脑和脊髓中也记录到广泛的神经元基因转移。单次静脉注射编码靶向亨廷顿蛋白(Htt)的人工微小RNA的AAV-AS载体导致成年小鼠多个CNS结构中Htt的敲低率为33%-50%。这种新型AAV-AS载体是开发用于神经退行性疾病的新基因疗法的有前途的平台。