Department of Otolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, PR China.
J Gene Med. 2013 Mar-Apr;15(3-4):123-33. doi: 10.1002/jgm.2701.
Gene transfer into the inner ear is a promising approach for treating sensorineural hearing loss. The special electrochemical environment of the scala media raises a formidable challenge for effective gene delivery at the same time as keeping normal cochlear function intact. The present study aimed to define a suitable strategy for preserving hearing after viral inoculation directly into the scala media performed at various postnatal developmental stages.
We assessed transgene expression of green fluorescent protein (GFP) mediated by various types of adeno-associated virus (AAV) and lentivirus (LV) in the mouse cochlea. Auditory brainstem responses were measured 30 days after inoculation to assess effects on hearing.
Patterns of GFP expression confirmed extensive exogenous gene expression in various types of cells lining the endolymphatic space. The use of different viral vectors and promoters resulted in specific cellular GFP expression patterns. AAV2/1 with cytomegalovirus promoter apparently gave the best results for GFP expression in the supporting cells. Histological examination showed normal cochlear morphology and no hair cell loss after either AAV or LV injections. We found that hearing thresholds were not significantly changed when the injections were performed in mice younger than postnatal day 5, regardless of the type of virus tested.
Viral inoculation and expression in the inner ear for the restoration of hearing must not damage cochlear function. Using normal hearing mice as a model, we have achieved this necessary step, which is required for the treatment of many types of congenital deafness that require early intervention.
将基因转入内耳是治疗感觉神经性听力损失的一种很有前途的方法。中阶的特殊电化学环境对有效基因传递提出了巨大挑战,同时又要保持正常的耳蜗功能完整。本研究旨在确定在各种出生后发育阶段将病毒直接接种到中阶时保持听力的合适策略。
我们评估了不同类型的腺相关病毒(AAV)和慢病毒(LV)介导的绿色荧光蛋白(GFP)在小鼠耳蜗中的转基因表达。接种后 30 天测量听觉脑干反应,以评估对听力的影响。
GFP 表达模式证实了内淋巴空间各种类型细胞中的广泛外源基因表达。使用不同的病毒载体和启动子导致特定的细胞 GFP 表达模式。巨细胞病毒启动子的 AAV2/1 显然为支持细胞中的 GFP 表达提供了最佳结果。组织学检查显示,无论是 AAV 还是 LV 注射后,耳蜗形态均正常,毛细胞无丢失。我们发现,当在出生后 5 天以内的小鼠中进行注射时,无论测试的病毒类型如何,听力阈值都没有明显变化。
病毒接种和内耳表达以恢复听力不能损害耳蜗功能。使用正常听力的小鼠作为模型,我们已经实现了这一必要步骤,这是治疗许多需要早期干预的先天性耳聋所必需的。