Adenovirus vectors targeting distinct cell types in the retina.
Adenovirus vectors targeting distinct cell types in the retina.
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针对视网膜中不同细胞类型的腺病毒载体。
DOI：
10.1167/iovs.09-4367
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发表时间：
2010
影响因子：
4.4
通讯作者：
Kumar-Singh,Rajendra
中科院分区：
文献类型：
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作者：
Sweigard,JHarry;Cashman,SiobhanM;Kumar-Singh,Rajendra
目的：许多视网膜疾病的基因治疗需要感光细胞的有效转导。尽管腺病毒（Ad）血清型5（Ad 5）不能有效地抑制光感受器，但先前的研究已经证明通过用Ad 35（Ad 5/F35）或Ad 37（Ad 5/F37）纤维假型化的Ad 5或通过缺失Ad 5五邻体基底中的RGD结构域（Ad 5 ΔRGD）来改善光感受器转导。然而，这些构建体中的每一个都含有不同的转基因盒，这妨碍了对这些载体的相对性能的评估，这是在临床中使用这些载体之前的重要考虑因素。本研究的目的是评估这些载体在视网膜中的作用，并尝试光感受器特异性转基因表达。产生含有相同表达盒的三个基于Ad 5的载体，并将其注射到成年小鼠的视网膜下腔中。通过平片、横截面、定量RT-PCR和改良的体视学技术分析眼睛的绿色荧光蛋白表达。在光感受器特异性转基因表达的背景下分析了来自小鼠视蛋白启动子的257-bp片段。测试的每种病毒有效地转导视网膜色素上皮。作者没有发现Ad 5/F35或Ad 5/F37转导光感受器的证据。相反，他们发现Ad 5/F37转导了Müller细胞。检测到Ad 5 ΔRGD的稳健的光感受器转导。在Ad 5 ΔRGD载体的背景下，发现来自257-bp小鼠视蛋白启动子的光受体特异性转基因表达。腺病毒载体可以设计为对不同的细胞群体具有向性。在Ad 5 ΔRGD载体的背景下可以实现稳健的光感受器特异性转基因表达。
Purpose.: Gene therapy for a number of retinal diseases necessitates efficient transduction of photoreceptor cells. Whereas adenovirus (Ad) serotype 5 (Ad5) does not transduce photoreceptors efficiently, previous studies have demonstrated improved photoreceptor transduction by Ad5 pseudotyped with Ad35 (Ad5/F35) or Ad37 (Ad5/F37) fiber or by the deletion of the RGD domain in the Ad5 penton base (Ad5ΔRGD). However, each of these constructs contained a different transgene cassette, preventing the evaluation of the relative performance of these vectors, an important consideration before the use of these vectors in the clinic. The aim of this study was to evaluate these vectors in the retina and to attempt photoreceptor-specific transgene expression.Methods.: Three Ad5-based vectors containing the same expression cassette were generated and injected into the subretinal space of adult mice. Eyes were analyzed for green fluorescence protein expression in flat-mounts, cross-sections, quantitative RT-PCR, and a modified stereological technique. A 257-bp fragment derived from the mouse opsin promoter was analyzed in the context of photoreceptor-specific transgene expression.Results.: Each virus tested efficiently transduced the retinal pigment epithelium. The authors found no evidence that Ad5/F35 or Ad5/F37 transduced photoreceptors. Instead, they found that Ad5/F37 transduced Müller cells. Robust photoreceptor transduction by Ad5ΔRGD was detected. Photoreceptor-specific transgene expression from the 257-bp mouse opsin promoter in the context of Ad5ΔRGD vectors was found.Conclusions.: Adenovirus vectors may be designed with tropism to distinct cell populations. Robust photoreceptor-specific transgene expression can be achieved in the context of Ad5ΔRGD vectors.