Recombinant adeno-associated virus type 2, 4, and 5 vectors: Transduction of variant cell types and regions in the mammalian central nervous system
Recombinant adeno-associated virus type 2, 4, and 5 vectors: Transduction of variant cell types and regions in the mammalian central nervous system
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DOI：
10.1073/pnas.050581197
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发表时间：
2000-03-28
影响因子：
11.1
通讯作者：
Chiorini, JA
中科院分区：
文献类型：
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作者：
Davidson, BL;Stein, CS;Chiorini, JA
基于血清型2的重组腺相关病毒载体（rAAV 2）可以指导中枢神经系统（CNS）中的转基因表达，但尚不知道其他rAAV血清型如何作为CNS基因转移载体。血清型4和5与rAAV 2不同，并且在其衣壳区域彼此不同，这表明它们可能指导结合和进入不同的细胞类型。在这项研究中，我们研究了由rAAV 4和rAAV 5制成的编码P-半乳糖苷酶的载体与类似设计的基于rAAV 2的载体相比的向性和转导效率。将rAAV 5 β-半乳糖苷酶（β gal）或rAAV 4 β gal注射到侧脑室中导致室管膜细胞的稳定转导，阳性细胞比注射rAAV 2 β gal的小鼠中多约10倍。三种载体之间的主要差异揭示了纹状体注射。纹状体内注射rAAV 4 β gal再次导致转基因的显著室管膜特异性表达，在实质中显著缺乏转导细胞。rAAV 2 β gal和rAAV 5 β gal纹状体内注射导致β-gal阳性实质细胞，但与rAAV 2 β gal不同，rAAV 5 β gal转导神经元和星形胶质细胞。在3周和15周时，rAAV 5 β gal注射的脑中转基因阳性细胞的数量分别是rAAV 2 β gal注射的脑中转基因阳性细胞的130倍和5，000倍。此外，在rAAV 5 β gal转导的动物中，转基因阳性细胞广泛分散在整个注射的半球中。总之，我们的数据为早期的体外工作提供了体内支持，表明rAAV 4和rAAV 5通过与rAAV 2不同的受体进入细胞。这些差异可用于改善CNS疾病的基因治疗。
Recombinant adeno-associated virus vectors based on serotype 2 (rAAV2) can direct transgene expression in the central nervous system (CNS), but it is not known how other rAAV serotypes perform as CNS gene transfer vectors. Serotypes 4 and 5 are distinct from rAAV2 and from each other in their capsid regions, suggesting that they may direct binding and entry into different cell types. In this study, we examined the tropisms and transduction efficiencies of P-galactosidase-encoding vectors made from rAAV4 and rAAV5 compared with similarly designed rAAV2-based vectors. Injection of rAAV5 beta-galactosidase (beta gal) or rAAV4 beta gal into the lateral ventricle resulted in stable transduction of ependymal cells, with approximately 10-fold more positive cells than in mice injected with rAAV2 beta gal. Major differences between the three vectors were revealed upon striatal injections. Intrastriatal injection of rAAV4 beta gal resulted again in striking ependyma-specific expression of transgene, with a notable absence of transduced cells in the parenchyma. rAAV2 beta gal and rAAV5 beta gal intrastriatal injections led to beta-gal-positive parenchymal cells, but unlike rAAV2 beta gal, rAAV5 beta gal transduced both neurons and astrocytes. The number of transgene-positive cells in rAAV5 beta gal-injected brains was 130 and 5,000 times higher than in rAAV2 beta gal-injected brains at 3 and 15 wk, respectively. Moreover, transgene-positive cells were widely dispersed throughout the injected hemisphere in rAAV5 beta gal-transduced animals. Together, our data provide in vivo support for earlier in vitro work, suggesting that rAAV4 and rAAV5 gain cell entry by means of receptors distinct from rAAV2. These differences could be exploited to improve gene therapy for CNS disorders.