Assessment of Adeno-Associated Virus Serotype Tropism in Human Retinal Explants
Assessment of Adeno-Associated Virus Serotype Tropism in Human Retinal Explants
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DOI：
10.1089/hum.2017.179
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发表时间：
2018-02-23
影响因子：
4.2
通讯作者：
Mullins, Robert F.
中科院分区：
文献类型：
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作者：
Wiley, Luke A.;Burnight, Erin R.;Mullins, Robert F.
单基因视网膜疾病病因发现的进展，结合向视网膜递送DNA的技术，为治疗以前无法治疗的致盲疾病提供了巨大的机会。然而，为了使基因扩增最有效，需要具有正确细胞类型特异性的载体。虽然动物模型非常有用，但与人类视网膜相比，它们通常表现出视网膜细胞表面受体的差异。本研究评估了使用离体器官型外植体系统来测试人视网膜和视网膜色素上皮-脉络膜中七种不同的2型腺相关病毒（AAV 2）血清型的转导效率和嗜性-AAV 2/1、AAV 2/2、AAV 2/4、AAV 2/5、AAV 2/6、AAV 2/8和AAV 2/9-所有这些都在巨细胞病毒启动子的控制下驱动GFP的表达。培养7天后，发现AAV 2/4和AAV 2/5在转导感光细胞方面特别有效，并且AAV 2/5对外核层具有高度特异性，而AAV 2/8对光感受器的转导始终较低。为了验证器官型培养系统的真实性，还在猪模型中比较了同一组AAV的转导，其中将体内视网膜下注射与离体培养和转导的器官型培养物进行了比较。这项研究显示了不同的AAV血清型在人类视网膜中的行为，并为进一步研究这些血清型中的每一种用于遗传性视网膜变性的基于基因扩增的治疗提供了见解。
Advances in the discovery of the causes of monogenic retinal disorders, combined with technologies for the delivery of DNA to the retina, offer enormous opportunities for the treatment of previously untreatable blinding diseases. However, for gene augmentation to be most effective, vectors that have the correct cell-type specificity are needed. While animal models are very useful, they often exhibit differences in retinal cell surface receptors compared to the human retina. This study evaluated the use of an ex vivo organotypic explant system to test the transduction efficiency and tropism of seven different adeno-associated virus type 2 (AAV2) serotypes in the human retina and retinal pigment epithelium-choroid-AAV2/1, AAV2/2, AAV2/4, AAV2/5, AAV2/6, AAV2/8, and AAV2/9-all driving expression of GFP under control of the cytomegalovirus promoter. After 7 days in culture, it was found that AAV2/4 and AAV2/5 were particularly efficient at transducing photoreceptor cells and that AAV2/5 was highly specific to the outer nuclear layer, whereas AAV2/8 displayed consistently low transduction of photoreceptors. To validate the authenticity of the organotypic culture system, the transduction of the same set of AAVs was also compared in a pig model, in which sub-retinal injections in vivo were compared to cultured and transduced organotypic cultures ex vivo. This study shows how different AAV serotypes behave in the human retina and provides insight for further investigation of each of these serotypes for gene augmentation-based treatment of inherited retinal degeneration.