Adeno-associated virus transformation into the normal miniature pig and the normal guinea pigs cochlea via scala tympani

Abstract

Objective: To investigate the expression of the miniature pig cochlea after AAV1 transfect into the cochlea via round window membrane (RWM).

Methods: Twenty miniature pigs are equally divided into four experimental groups. Twelve miniature pigs are equally divided into four control groups. Each pig was transfected with the AAV1 in the experimental group via RWM and each pig was transduced with the artificial perilymph in the control group. The expression of green fluorescent protein (GFP) was observed at 2 weeks, 3 weeks and 4 weeks, respectively. Likewise, AAV1 was delivered into the guinea pigs cochleas using the same method, and the results were compared with that of the miniature pigs.

Results: The expression was mainly in the inner hair cells of the miniature pig. The expression of GFP began to appear at 2 weeks, reached the peak at 3 weeks. It also expressed in Hensen’s cells, inner pillar cells, outer pillar cells, spiral limbus, and spiral ligament. In the meanwhile, AAV1 was delivered into guinea pig cochlea via the same method, and AAV1 was also expressed in the inner hair cells. But the expression peaked at 2 weeks, and the efficiency of the inner hair cell transfection was higher than that of the pig.

Conclusion: AAV1 can be transformed into miniature pig cochlea via scala tympani by the RWM method efficiently.

Chinese abstract

目的: 研究AAV1通过RWM < AQ3>转染进耳蜗后, 小型猪耳蜗的表达。

方法: 将20只小型猪分为均等的4个试验组。十二只小型猪分为均等的四个对照组。试验组的每只猪通过RWM受AAV1转染；对照组中每只猪受人工外淋巴转导。在<AQ4 > 2周、3周和4周分别观察到GFP的表达。同样, 使用相同的方法将AAV1转染到豚鼠耳蜗, 并将结果与​​小型猪的结果进行比较。

结果: GFP表达主要出现在小型猪的内毛细胞中。 它开始出现在2周, 3周时达到峰值。它也在Hensen细胞、内柱细胞、外柱细胞、螺旋角膜缘和螺旋韧带中有表达。同时, 通过相同的方法将AAV1输入豚鼠耳蜗, 在内毛细胞中也有AAV1表达。但表达在2周达到峰值, 并且内毛细胞转染率高于猪。

结论: 用圆窗膜法, AAV1可以有效地通过鼓膜转化成微型猪耳蜗。

Keywords:

• AAV
• miniature pig
• inner hair cell
• gene therapy

Disclosure statement

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.

Additional information

Funding

This study was supported by National High-tech R&D Program of China (863 Program) (2014AA020510); the National Natural Science Foundation of China (NSFC #81271082, 81400472). Special Cultivating and Developing Program of Beijing Science and Technology Innovation Base (z151100001615050); Key Projects of Fujian Provincial Science and Technology Department (2014I0003); Innovation and drive power engineering project of China Association for Science and Technology (2016CXQD01).