Optimization of adeno-associated virus vector-mediated gene transfer to the respiratory tract
Optimization of adeno-associated virus vector-mediated gene transfer to the respiratory tract
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DOI：
10.1038/gt.2017.19
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发表时间：
2017-05-01
期刊：
影响因子：
5.1
通讯作者：
Kume, A.
中科院分区：
文献类型：
--
作者：
Kurosaki, F.;Uchibori, R.;Kume, A.
构建了一种高效的腺相关病毒（AAV）载体，用于治疗呼吸系统疾病。在本研究中检查了可能影响基因转移至气道细胞的效率的AAV血清型、启动子和给药途径。在体外筛选的9种AAV血清型（AAV 1 -9）和体内评价的4种血清型（AAV 1、2、6、9）中，AAV 6显示最强的转基因表达。至于启动子，巨细胞病毒（CMV）早期增强子/鸡β-肌动蛋白（CAG）启动子比CMV启动子导致更稳健的转导。关于递送途径，气管内施用导致肺中强的转基因表达，而静脉内和鼻内施用途径产生可忽略的表达。AAV 6衣壳和CAG启动子的组合导致持续的表达，并且气管内施用的AAV 6-CAG载体转导肺中的支气管细胞和周细胞。这些结果表明，AAV 6-CAG载体比先前优选的AAV 2载体更有希望用于气道转导，特别是当施用到气管中时。本研究为腺相关病毒介导的肺部疾病的基因治疗提供了一个优化的策略，如囊性纤维化和肺纤维化。
An efficient adeno-associated virus (AAV) vector was constructed for the treatment of respiratory diseases. AAV serotypes, promoters and routes of administration potentially influencing the efficiency of gene transfer to airway cells were examined in the present study. Among the nine AAV serotypes (AAV1-9) screened in vitro and four serotypes (AAV1, 2, 6, 9) evaluated in vivo, AAV6 showed the strongest transgene expression. As for promoters, the cytomegalovirus (CMV) early enhancer/chicken beta-actin (CAG) promoter resulted in more robust transduction than the CMV promoter. Regarding delivery routes, intratracheal administration resulted in strong transgene expression in the lung, whereas the intravenous and intranasal administration routes yielded negligible expression. The combination of the AAV6 capsid and CAG promoter resulted in sustained expression, and the intratracheally administered AAV6-CAG vector transduced bronchial cells and pericytes in the lung. These results suggest that AAV6-CAG vectors are more promising than the previously preferred AAV2 vectors for airway transduction, particularly when administered into the trachea. The present study offers an optimized strategy for AAV-mediated gene therapy for lung diseases, such as cystic fibrosis and pulmonary fibrosis.