Protection against aminoglycoside-induced ototoxicity by regulated AAV vector-mediated GDNF gene transfer into the cochlea
Protection against aminoglycoside-induced ototoxicity by regulated AAV vector-mediated GDNF gene transfer into the cochlea
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DOI：
10.1038/sj.mt.6300379
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发表时间：
2008-03-01
影响因子：
12.4
通讯作者：
Ozawa, Keiya
中科院分区：
文献类型：
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作者：
Liu, Yuhe;Okada, Takashi;Ozawa, Keiya
由于标准的氨基糖苷类药物治疗逐渐引起听力障碍伴毛细胞变性，因此药物的全身使用受到限制。基于腺相关病毒（AAV）的载体已经引起极大的兴趣，因为它们以最小的毒性介导各种有丝分裂后细胞中的稳定转基因表达。在这项研究中，我们研究了调节AAV 1介导的胶质细胞源性神经营养因子（GDNF）在耳蜗中的表达对氨基糖苷类药物引起的损伤的影响。将编码GDNF的基于AAV 1的载体或编码具有rtTA 2s-S2泰特-开调节系统的GDNF的载体以5 × 10（10）基因组拷贝/体通过圆窗直接显微注射到大鼠耳蜗中.病毒注射后7天，皮下给予卡那霉素333 mg/kg，每日两次，连续12天。GDNF在耳蜗中的表达得到证实，并成功地由Tet-on系统调节。听觉脑干反应的监测显示，在测试的频率上GDNF转导后耳蜗功能得到改善。GDNF表达可显著减少受损的螺旋神经节细胞和毛细胞。我们的研究结果表明，AAV 1介导的GDNF的表达使用一个regulatedexpression系统在耳蜗是一个有前途的策略，以保护耳蜗免受氨基糖苷类药物诱导的损害。
Since standard aminoglycoside treatment progressively causes hearing disturbance with hair cell degeneration, systemic use of the drugs is limited. Adeno-associated virus (AAV)-based vectors have been of great interest because they mediate stable transgene expression in a variety of postmitotic cells with minimal toxicity. In this study, we investigated the effects of regulated AAV1-mediated glial cell line-derived neurotrophic factor (GDNF) expression in the cochlea on aminoglycoside-induced damage. AAV1-based vectors encoding GDNF or vectors encoding GDNF with an rtTA2s- S2 Tet- on regulation system were directly microinjected into the rat cochleae through the round window at 5 x 10(10) genome copies/body. Seven days after the virus injection, a dose of 333 mg/kg of kanamycin was subcutaneously given twice daily for 12 consecutive days. GDNF expression in the cochlea was confirmed and successfully modulated by the Tet-on system. Monitoring of the auditory brain stem response revealed an improvement of cochlear function after GDNF transduction over the frequencies tested. Damaged spiral ganglion cells and hair cells were significantly reduced by GDNF expression. Our results suggest that AAV1-mediated expression of GDNF using a regulatedexpression system in the cochlea is a promising strategy to protect the cochlea from aminoglycoside-induced damage.