Pre-retinal delivery of recombinant adeno-associated virus vector significantly improves retinal transduction efficiency.
Pre-retinal delivery of recombinant adeno-associated virus vector significantly improves retinal transduction efficiency.
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DOI：
10.1016/j.omtm.2021.06.005
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发表时间：
2021-09-10
期刊：
影响因子：
--
通讯作者：
Lipinski DM
中科院分区：
文献类型：
--
作者：
Zhang H;Sajdak BS;Merriman DK;Carroll J;Lipinski DM
玻璃体内注射是目前应用最广泛的眼部基因导入技术。然而，载体扩散被玻璃体中的物理屏障和中和抗体减弱。与人类一样，13线地松鼠（13-LGS）具有比更常见的啮齿动物模型（如大鼠和小鼠）更大的相对玻璃体体积，这将进一步降低玻璃体内注射途径的转导效率。我们在这里报告一个“视网膜前”注射的方法，导致脱离的后玻璃体膜，并提供载体到玻璃体和内部视网膜之间的空间。将携带普遍表达mCherry报告基因的载体注射到成年野生型13-LGS的深层玻璃体或视网膜前间隙中。然后，用非侵入性成像、免疫荧光和流式细胞术评估腺相关病毒（AAV）介导的mCherry表达。与深层玻璃体递送相比，视网膜前施用以较低的载体剂量体积实现了全视网膜基因表达，并显著增加了转导的视锥光感受器的数量。这些结果表明，视网膜前注射是一种有前途的工具，在动物模型中的基因治疗策略的发展，是一种潜在的方法，用于人类研究，特别是在年轻的个体与完整的后玻璃体膜和稳定的玻璃体。我们证明，通过将载体递送到位于内界膜和后玻璃体膜之间的视网膜前间隙中，可以显着改善使用rAAV的视锥细胞转导。如果将这种新的递送方法转化为人类受试者，可以提高基因递送效率和手术安全性。
Intravitreal injection is the most widely used injection technique for ocular gene delivery. However, vector diffusion is attenuated by physical barriers and neutralizing antibodies in the vitreous. The 13-lined ground squirrel (13-LGS), as in humans, has a larger relative vitreous body volume than the more common rodent models such as rats and mice, which would further reduce transduction efficiency with the intravitreal injection route. We report here a “pre-retinal” injection approach that leads to detachment of the posterior hyaloid membrane and delivers vector into the space between vitreous and inner retina. Vectors carrying a ubiquitously expressing mCherry reporter were injected into the deep vitreous or pre-retinal space in adult wild-type 13-LGSs. Then, adeno-associated virus (AAV)-mediated mCherry expression was evaluated with non-invasive imaging, immunofluorescence, and flow cytometry. Compared to deep vitreous delivery, pre-retinal administration achieved pan-retinal gene expression with a lower vector dose volume and significantly increased the number of transduced cone photoreceptors. These results suggest that pre-retinal injection is a promising tool in the development of gene therapy strategies in animal models and is a potential approach for use in human research, particularly in younger individuals with an intact posterior hyaloid membrane and stable vitreous. We demonstrate that transduction of cone photoreceptors using rAAV can be significantly improved through delivery of vector into the pre-retinal space located between the inner limiting membrane and posterior hyaloid membrane. If translated to human subjects, this novel delivery approach could improve gene delivery efficiency and surgical safety.
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影响因子：
3.4
作者：
Kokona, Despina;Haner, Nathanael U.;Zinkernagel, Martin S.
通讯作者：
Zinkernagel, Martin S.
影响因子：
--
作者：
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通讯作者：
Hauswirth, William W.
影响因子：
1.8
作者：
Distler, C;Dreher, Z
通讯作者：
Dreher, Z
影响因子：
12.4
作者：
Cashman, Siobhan M.;McCullough, Laura;Kumar-Singh, Rajendra
通讯作者：
Kumar-Singh, Rajendra
影响因子：
6.9
作者：
Merriman, Dana K.;Lahvis, Garet;Schill, Kelly
通讯作者：
Schill, Kelly