Safety of adeno-associated virus as cochlear gene transfer vector: Analysis of distant spread beyond injected cochleae
Safety of adeno-associated virus as cochlear gene transfer vector: Analysis of distant spread beyond injected cochleae
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DOI：
10.1006/mthe.2000.0129
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发表时间：
2000-10-01
影响因子：
12.4
通讯作者：
Lalwani, AK
中科院分区：
文献类型：
--
作者：
Kho, ST;Pettis, RM;Lalwani, AK
腺相关病毒（AAV），接种到外淋巴，已被证明是一个有效的载体介导的脑内转基因表达。在先前的工作中，对侧耳蜗中转基因表达的意外发现引起了对病毒从靶组织传播的关注。本研究旨在评估AAV引入内耳后的传播程度。将重组AAV注射到成年雄性豚鼠的左耳中，并在2或4周时处死。包括耳蜗在内的各种器官被帽状固定以表征病毒DNA的存在和表达。通过聚合酶链反应在输注和对侧耳蜗和小脑中检测到病毒DNA，但在任何其他器官（包括皮质、心脏、肺、肝脏、脾脏和肾脏）中未检测到病毒DNA。虽然病毒存在于小脑中，但该器官中的转基因表达用Western印迹或免疫组织化学均检测不到。通过免疫组化证实，在邻近输注耳蜗和对侧耳蜗的骨髓间隙中的多核巨细胞中存在转基因表达。总的来说，这些结果表明AAV传播的潜在途径，从注入耳蜗通过耳蜗水管或通过扩展通过颞骨髓空间。这项研究强调了调查减轻病毒泄漏的因素的必要性。
The adeno-associated virus (AAV), inoculated into the perilymph, has been shown to be an effective vector for mediating intracochlear transgene expression. The unexpected finding of transgene expression in the contralateral cochlea in previous work raised concern about dissemination of the virus from the target tissue. The current study was undertaken to assess the extent of AAV dissemination following its introduction into the inner ear. Adult male guinea pigs were injected with recombinant AAV into their left ears and sacrificed at 2 or 4 weeks. Various organs including the cochleae were hat-vested to characterize the presence and expression of the viral DNA. Virus DNA was detected via polymerase chain reaction in the infused and contralateral cochlea and in the cerebellum but not in any other organs, including cortex, heart, lung, liver, spleen, and kidney. Although the viral presence was established in the cerebellum, transgene expression in this organ was undetectable with either Western blot or immunohistochemistry. Transgene expression was demonstrated via immunohistochemistry in multinucleated giant cells in the bone marrow spaces adjacent to the infused and contralateral cochleae. Collectively, these results suggest potential routes for AAV dissemination from the infused cochlea via the cochlear aqueduct or by extension through the temporal bone marrow spaces. This study reinforces the need to investigate factors that mitigate viral leakage.