Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways
Long-term persistence of gene expression from adeno-associated virus serotype 5 in the mouse airways
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DOI：
10.1038/sj.gt.3302815
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发表时间：
2006-12-01
期刊：
影响因子：
5.1
通讯作者：
Hyde, S. C.
中科院分区：
文献类型：
--
作者：
Sumner-Jones, S. G.;Davies, L. A.;Hyde, S. C.
基于血清型2（rAAV 2）的重组腺相关病毒载体已在多种临床前和临床研究中用于将转基因递送至气道。这些研究中的基因转移受到rAAV 2受体基底外侧定位的严重限制。在这里，我们研究了从AAV 5基因组和衣壳构建的载体，其利用在气道上皮细胞的顶端表面上发现的含N-连接唾液酸的受体。我们研究了将rAAV 5/5载体递送至小鼠呼吸道后的基因转移功效和转基因表达的持续时间。在鼻内衬上皮中观察到稳健的剂量依赖性转基因表达至少32周，在肺中观察到至少52周。重要的是，在肺中，由rAAV 5/5介导的转基因表达比rAAV 2/2载体高40倍。观察到对rAAV 5/5介导的转导的不同细胞偏好，其中转基因表达主要限于鼻中嗅上皮的支持细胞和肺中肺泡II型细胞。将rAAV 5/5载体施用至鼻和肺导致rAAV 5/5中和抗体的快速发展，表明重复施用可能受到宿主免疫应答的严重阻碍。
Recombinant adeno-associated virus vectors based on serotype 2 (rAAV2) have been used to deliver transgenes to the airways in a variety of pre-clinical and clinical studies. Gene transfer in these studies has been severely restricted by the basolateral localization of rAAV2 receptors. Here, we studied vectors constructed from the AAV5 genome and capsid, which utilize N-linked sialic acid-containing receptors found on the apical surface of airway epithelial cells. We investigated gene transfer efficacy and duration of transgene expression following delivery of rAAV5/5 vectors to the mouse respiratory tract. Robust, dose-dependent transgene expression was observed in the epithelium lining the nose for at least 32 weeks, and for at least 52 weeks in the lung. Importantly, in the lung, transgene expression mediated by rAAV5/5 was 40-fold greater than by rAAV2/2 vectors. A distinct cellular preference for rAAV5/5-mediated transduction was observed, with transgene expression being predominantly restricted to sustentacular cells of the olfactory epithelium in the nose and alveolar type II cells in the lung. Administration of rAAV5/5 vectors to both the nose and lungs led to the rapid development of rAAV5/5-neutralizing antibodies, suggesting that repeated administration may be severely hampered by host immune responses.