Cochleovestibular gene transfer in neonatal mice by canalostomy
Cochleovestibular gene transfer in neonatal mice by canalostomy
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通过耳管造口术对新生小鼠进行耳蜗前庭基因转移
DOI：
10.1097/wnr.0000000000000827
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发表时间：
2017-08-02
期刊：
影响因子：
1.7
通讯作者：
Gong, Shu-Sheng
中科院分区：
文献类型：
--
作者：
Guo, Jing-Ying;Liu, Yu-Ying;Gong, Shu-Sheng
内耳损伤会导致人类出现感音神经性听力损失和前庭功能障碍。这些疾病中有很大一部分是先天性的，涉及听觉和前庭系统。因此，纠正缺陷的基因干预必须在发育早期阶段进行。在这项研究中，我们通过半规管开窗术评估了腺相关病毒8型（AAV8）载体在新生小鼠内耳中的基因转移情况。将携带绿色荧光蛋白（GFP）基因的AAV8通过后半规管（半规管开窗术）接种到新生小鼠的内耳中。手术后30天，对动物进行游泳测试和听性脑干反应测量。然后，对动物实施安乐死并采集颞骨进行整装制片。通过免疫组织化学评估内耳中GFP的表达和形态变化。手术后，未发现前庭功能障碍的迹象，接种AAV8的耳朵与未手术耳朵之间的听性脑干反应阈值没有显著差异。在手术耳中，在耳蜗和前庭终器中检测到广泛的GFP表达且无形态学损伤。在内毛细胞、边缘细胞、前庭毛细胞和前庭支持细胞中发现了强烈的GFP表达。总之，通过半规管开窗术将AAV8接种到新生小鼠内耳中，可使内耳中外源基因广泛过表达，且不影响听力或前庭功能。它是先天性耳蜗前庭疾病基因治疗的一种有前景的方法。版权所有（C）2017威科集团。保留所有权利。
Impairments of the inner ear result in sensorineural hearing loss and vestibular dysfunction in humans. A large proportion of these disorders are congenital, and involve both auditory and vestibular systems. Therefore, genetic interventions to correct deficits must be administered during early developmental stages. In this study, we evaluated inner ear gene transfer in neonatal mice by canalostomy using an adeno-associated virus serotype 8 (AAV8) vector. AAV8 with the green fluorescence protein (GFP) gene was inoculated into the inner ear of the neonatal mice through the posterior semicircular canal (canalostomy). At 30 days following surgery, animals were subjected to swim tests and auditory brainstem response measurements. Then, the animals were euthanized and temporal bones were harvested for whole-mount preparation. GFP expression and morphological changes in the inner ear were assessed by immunohistochemistry. After surgery, no signs of vestibular dysfunction were found, and there were no significant differences in the auditory brainstem response threshold between AAV8-inoculated ears and nonsurgery ears. In the surgery ears, extensive GFP expression and no morphological lesions were detected in the cochlear and vestibular end organs. Robust GFP expression was found in inner hair cells, marginal cells, vestibular hair cells, and vestibular supporting cells. In conclusion, AAV8 inoculation through canalostomy into the inner ears of neonatal mice led to extensive overexpression of exogenous genes in the inner ear without affecting hearing or vestibular function. It serves as a promising approach for gene therapy of congenital cochleovestibular diseases.