Novel adeno-associated virus serotypes efficiently transduce murine Photoreceptors
Novel adeno-associated virus serotypes efficiently transduce murine Photoreceptors
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DOI：
10.1128/jvi.01327-07
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发表时间：
2007-10-01
影响因子：
5.4
通讯作者：
Auricchio, Alberto
中科院分区：
文献类型：
--
作者：
Allocca, Mariacarmela;Mussolino, Claudio;Auricchio, Alberto
严重的遗传性视网膜疾病，如视网膜色素变性和莱伯先天性黑蒙，是由光感受器中优先表达的基因突变引起的。虽然腺相关病毒（AAV）介导的基因转移可以纠正动物模型中的视网膜色素上皮（RPE）缺陷，但用于纠正光感受器特异性疾病的方法效率较低。我们评估了新型AAV血清型（AAV 2/7、AAV 2/8、AAV 29、AAV 2rh.43、AAV 2rh.64R1和AAV 2hu.29R）与组成型或光感受器特异性启动子组合改善光感受器转导（光感受器拯救中的限制性步骤）的能力。基于定性分析，在小鼠视网膜下施用后，测试的所有AAV血清型有效地抑制RPE以及视杆和视锥光感受器。有趣的是，AAV 2/9有效地转导Muller细胞。为了以定性和定量的方式比较来自不同AAV和启动子的光感受器转导，我们设计了基于使用表达增强的绿色荧光蛋白和荧光素酶的双顺反子构建体的策略。我们发现，AAV 2/8和AAV 2/7介导的体内光感受器转导水平比AAV 2/5高6至8倍，AAV 2/5被认为是迄今为止用于光感受器靶向的最有效的AAV血清型。此外，在视网膜下施用AAV后，视紫红质启动子允许比测试的其他普遍存在的或光感受器特异性启动子显著更高水平的光感受器表达。最后，我们表明，AAV 2/7，AAV 2/8，和AAV 2/9优于AAV 2/5离体转导视网膜祖细胞分化成光感受器。我们得出结论，AAV 2/7或AAV 2/8和视紫红质启动子提供了最高水平的光感受器转导体内和体外，这可能会克服迄今为止在遗传性严重光感受器疾病模型中观察到的治疗成功的局限性。
Severe inherited retinal diseases, such as retinitis pigmentosa and Leber congenital amaurosis, are caused by mutations in genes preferentially expressed in photoreceptors. While adeno-associated virus (AAV)-mediated gene transfer can correct retinal pigment epithelium (RPE) defects in animal models, approaches for the correction of photoreceptor-specific diseases are less efficient. We evaluated the ability of novel AAV serotypes (AAV2/7, AAV2/8, AAV2/9, AAV2rh.43, AAV2rh.64R1, and AAV2hu.29R) in combination with constitutive or photoreceptor-specific promoters to improve photoreceptor transduction, a limiting step in photoreceptor rescue. Based on a qualitative analysis, all AAV serotypes tested efficiently transduce the RPE as well as rod and cone photoreceptors after subretinal administration in mice. Interestingly, AAV2/9 efficiently transduces Muller cells. To compare photoreceptor transduction from different AAVs and promoters in both a qualitative and quantitative manner, we designed a strategy based on the use of a bicistronic construct expressing both enhanced green fluorescent protein and luciferase. We found that AAV2/8 and AAV2/7 mediate six-to eightfold higher levels of in vivo photoreceptor transduction than AAV2/5, considered so far the most efficient AAV serotype for photoreceptor targeting. In addition, following subretinal administration of AAV, the rhodopsin promoter allows significantly higher levels of photoreceptor expression than the other ubiquitous or photoreceptor-specific promoters tested. Finally, we show that AAV2/7, AAV2/8, and AAV2/9 outperform AAV2/5 following ex vivo transduction of retinal progenitor cells differentiated into photoreceptors. We conclude that AAV2/7 or AAV2/8 and the rhodopsin promoter provide the highest levels of photoreceptor transduction both in and ex vivo and that this may overcome the limitation to therapeutic success observed so far in models of inherited severe photoreceptor diseases.