Virus-mediated transduction of murine retina with adeno-associated virus: Effects of viral capsid and genome size
Virus-mediated transduction of murine retina with adeno-associated virus: Effects of viral capsid and genome size
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DOI：
10.1128/jvi.76.15.7651-7660.2002
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发表时间：
2002-08-01
影响因子：
5.4
通讯作者：
Davidson, BL
中科院分区：
文献类型：
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作者：
Yang, GS;Schmidt, M;Davidson, BL
基于腺相关病毒（AAV）的基因治疗载体显示出治疗视网膜变性疾病的前景。在先前的工作中，视网膜下注射AAV 2、AAV 5和用AAV 5衣壳假型化的AAV 2（AAV 2/5）显示可变的视网膜色素上皮（RPE）和光感受器细胞转导，而AAV 2/1主要转导RPE。为了更彻底地比较AAV 2、AAV 3、AAV 5和AAV 6的基因转移效率，我们使用体视学方法定量AAV转导至小鼠感光细胞的动力学和效率。我们观察到持续的光感受器和RPE转导由AAV 5和AAV 2高达31周，并发现AAV 5转导的体积大于AAV 2。含有全长或半长基因组的AAV 5和AAV 2/5以相似的表达起始速率转导相当数量的感光细胞。与AAV 2相比，AAV 5在手术后5周和15周转导了显著更大数量的感光细胞（分别超过1,000倍和高达400倍）。此外，注射AAV 2/5的眼睛中的基因组拷贝数是注射AAV 2的眼睛中的30倍。比较具有半长度基因组的AAV，在5周时，AAV 5转导的感光细胞仅为AAV 2的4倍，在15周时几乎相等。在DNA水平上观察到转导的增强，注射具有短基因组的AAV的视网膜中的病毒基因组拷贝数比注射含有全长基因组的AAV的视网膜中的病毒基因组拷贝数多50倍。视网膜下注射AAV 2/6在5周和15周时仅显示RPE转导，而AAV 2/3不转导视网膜细胞。我们的结论是，不同的基因组长度和AAV衣壳可能允许改善表达和/或基因转移到视网膜中的特定细胞类型。
Gene therapy vectors based on adeno-associated viruses (AAVs) show promise for the treatment of retinal degenerative diseases. In prior work, subretinal injections of AAV2, AAV5, and AAV2 pseudotyped with AAV5 capsids (AAV2/5) showed variable retinal pigmented epithelium (RPE) and photoreceptor cell transduction, while AAV2/1 predominantly transduced the RPE. To more thoroughly compare the efficiencies of gene transfer of AAV2, AAV3, AAV5, and AAV6, we quantified, using stereological methods, the kinetics and efficiency of AAV transduction to mouse photoreceptor cells. We observed persistent photoreceptor and RPE transduction by AAV5 and AAV2 up to 31 weeks and found that AAV5 transduced a greater volume than AAV2. AAV5 containing full-length or half-length genomes and AAV2/5 transduced comparable numbers of photoreceptor cells with similar rates of onset of expression. Compared to AAV2, AAV5 transduced significantly greater numbers of photoreceptor cells at 5 and 15 weeks after surgery (greater than 1,000 times and up to 400 times more, respectively). Also, there were 30 times more genome copies in eyes injected with AAV2/5 than in eyes injected with AAV2. Comparing AAVs with half-length genomes, AAV5 transduced only four times more photoreceptor cells than AAV2 at 5 weeks and nearly equivalent numbers at 15 weeks. The enhancement of transduction was seen at the DNA level, with 50 times more viral genome copies in retinas injected with AAV having short genomes than in retinas injected with AAV containing full-length ones. Subretinal injection of AAV2/6 showed only RPE transduction at 5 and 15 weeks, while AAV2/3 did not transduce retinal cells. We conclude that varying genome length and AAV capsids may allow for improved expression and/or gene transfer to specific cell types in the retina.