Specific and efficient transduction of cochlear inner hair cells with recombinant adeno-associated virus type 3 vector
Specific and efficient transduction of cochlear inner hair cells with recombinant adeno-associated virus type 3 vector
复制标题
DOI：
10.1016/j.ymthe.2005.03.021
复制
发表时间：
2005-10-01
影响因子：
12.4
通讯作者：
Ozawa, K
中科院分区：
文献类型：
--
作者：
Liu, YH;Okada, T;Ozawa, K
重组腺相关病毒（AAV）载体是感兴趣的耳蜗基因治疗，因为他们的能力，介导的有效转移和长期稳定表达的治疗基因在各种各样的有丝分裂后组织与最小的载体相关的细胞毒性。在本研究中，使用七种AAV血清型（AAV 1 -5、7、8）来构建载体。与巨细胞病毒立即早期增强子相关的鸡P-肌动蛋白启动子在耳蜗细胞中的EGFP表达表明，这些血清型中的每一种都成功地靶向不同的耳蜗细胞类型。与其他血清型相反，AAV 3载体在体内以高效率特异性转导耳蜗内毛细胞，而AAV 1、2、5、7和8载体也转导这些和其他细胞类型，包括螺旋神经节和螺旋韧带细胞。在注射AAV载体后测试的频率范围内，诱发听觉脑干反应没有耳蜗功能丧失。这些发现对于进一步研究耳蜗内毛细胞的分子生物学和基因替换策略以纠正由于这些细胞中的单基因突变而导致的隐性遗传性听力损失具有价值。
Recombinant adeno-associated virus (AAV) vectors are of interest for cochlear gene therapy because of their ability to mediate the efficient transfer and long-term stable expression of therapeutic genes in a wide variety of postmitotic tissues with minimal vector-related cytotoxicity. In the present study, seven AAV serotypes (AAV1-5, 7, 8) were used to construct vectors. The expression of EGFP by the chicken P-actin promoter associated with the cytomegalovirus immediate-early enhancer in cochlear cells showed that each of these serotypes successfully targets distinct cochlear cell types. In contrast to the other serotypes, the AAV3 vector specifically transduced cochlear inner hair cells with high efficiency in vivo, while the AAV1, 2, 5, 7, and 8 vectors also transduced these and other cell types, including spiral ganglion and spiral ligament cells. There was no loss of cochlear function with respect to evoked auditory brain-stem responses over the range of frequencies tested after the injection of AAV vectors. These findings are of value for further molecular studies of cochlear inner hair cells and for gene replacement strategies to correct recessive genetic hearing loss due to monogenic mutations in these cells.