Systemic Delivery of Genes to Retina Using Adeno-Associated Viruses.
Systemic Delivery of Genes to Retina Using Adeno-Associated Viruses.
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使用腺相关病毒将基因系统性递送至视网膜。
DOI：
10.1007/978-3-030-27378-1_18
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发表时间：
2019
影响因子：
--
通讯作者：
Lobanova,EkaterinaS
中科院分区：
文献类型：
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作者：
Simpson,ChiabP;Bolch,SusanN;Zhu,Ping;Weidert,Frances;Dinculescu,Astra;Lobanova,EkaterinaS
超过80个基因的突变导致光感受器退化。虽然使用AAV载体将基因视网膜下递送至感光神经元已被证明是各种小鼠模型中的有效治疗和研究工具，但即使在健康动物中，手术程序本身也可能导致视网膜功能丧失，使实验研究的解释复杂化，并需要彻底设计的控制。一种非侵入性的方法，如通过血液系统递送带有AAV的基因，可能是工具开发的一个有希望的方向。先前的研究已经确定，AAV 9能够穿过血脑和血视网膜屏障，甚至具有有限的能力来阻断光感受器。AAV-PHP.eB是一种新的基于AAV 9的突变衣壳，其穿过血脑屏障并在成年小鼠中有效地转导中枢神经系统。在这里，我们研究了它穿过血视网膜屏障和视网膜神经元的能力。对照实验表明，这种衣壳通过玻璃体内给药对视网膜细胞的作用几乎不存在，但通过视网膜下途径对感光细胞的作用效率很高。AAV-PHP.eB在成年小鼠中的全身递送在整个视网膜中稳健地转导水平细胞，但不转导光感受器。我们的研究表明，AAV-PHP.eB穿过视网膜内血-视网膜屏障（IR-BRB），有效地转导位于IR-BRB附近的水平细胞，但进一步穿透视网膜并到达光感受器的能力非常有限。
Mutations in more than 80 genes lead to photoreceptor degeneration. Although subretinal delivery of genes to photoreceptor neurons using AAV vectors has proven itself as an efficient therapeutic and investigative tool in various mouse models, the surgical procedure itself could lead to loss of retinal function even in healthy animals, complicating the interpretation of experimental studies and requiring thoroughly designed controls. A noninvasive approach, such as a systemic delivery of genes with AAV through the bloodstream, may serve as a promising direction in tool development. Previous studies have established that AAV9 is capable of crossing the blood-brain and blood-retina barrier and even has a limited capacity to transduce photoreceptors. AAV-PHP.eB is a novel AAV9-based mutant capsid that crosses the blood-brain barrier and efficiently transduces central nervous system in the adult mice. Here, we investigated its ability to cross the blood-retina barrier and transduce retinal neurons. Control experiments demonstrated virtually nonexisting ability of this capsid to transduce retinal cells via intravitreal administration but high efficiency to transduce photoreceptors via subretinal route. Systemic delivery of AAV-PHP.eB in adult mice robustly transduced horizontal cells throughout the entire retina, but not photoreceptors. Our study suggests that AAV-PHP.eB crosses the intra-retinal blood-retinal barrier (IR-BRB), efficiently transduces horizontal cells located adjacent to IR-BRB, but has very limited ability to further penetrate retina and reach photoreceptors.