The functional and structural outcome of inner ear gene transfer via the vestibular and cochlear fluids in mice
The functional and structural outcome of inner ear gene transfer via the vestibular and cochlear fluids in mice
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DOI：
10.1006/mthe.2001.0490
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发表时间：
2001-12-01
影响因子：
12.4
通讯作者：
Raphael, Y
中科院分区：
文献类型：
--
作者：
Kawamoto, K;Oh, SH;Raphael, Y
小鼠是内耳基因治疗的理想模型，因为它们的基因组正在快速测序，它们的世代时间相对较短，并且它们是人类遗传性内耳疾病的有价值的模型。然而，小鼠内耳的小尺寸给外科手术带来了特殊的挑战。我们开发了一种将病毒接种到成熟小鼠内耳的新方法，使用表达细菌基因 lacZ 的复制缺陷型腺病毒。我们通过后半规管（耳蜗造口术）将病毒注入耳蜗（耳蜗造口术）。两种方法都会导致 lacZ 在外淋巴间隙细胞中表达。 1例耳蜗造口病例显示壶腹嵴感觉细胞有基因表达，而耳蜗造口组则显示柯蒂器和球囊感觉细胞有基因表达。手术后的功能测试表明，耳管造口术保留了听力，而耳蜗造口术却没有。管造口术恢复后，任何前庭功能都会暂时丧失。我们的研究结果表明，通过半规管将腺病毒载体接种到小鼠内耳中，有可能在不损害听力的情况下有效地将转基因引入前庭系统和耳蜗。
Mice present an ideal model for inner ear gene therapy because their genome is being rapidly sequenced, their generation time is relatively short, and they serve as a valuable model for human hereditary inner ear disease. However, the small size of the mouse inner ear poses a particular challenge for surgical procedures. We have developed a new approach for viral inoculation into the mature mouse inner ear, using a replication-deficient adenovirus expressing the bacterial gene lacZ. We administered the virus through the posterior semicircular canal (canalostomy) and into the cochlea (cochleostomy). Both approaches caused lacZ to be expressed in cells lining the perilymphatic space. One canalostomy case showed gene expression in sensory cells of the crista ampullaris, whereas the cochleostomy group showed gene expression in the sensory cells in the organ of Corti and saccule. Functional tests after the surgery showed that the canalostomy preserved hearing, whereas the cochleostomy did not. Any vestibular function transiently lost after the canalostomy was recovered. Our findings indicate that inoculation of adenovirus vectors into the mouse inner ear through the semicircular canal has the potential to efficiently introduce transgenes to the vestibular system and the cochlea without compromising hearing.