Efficient Gene Transfer Into the Mouse Lung by Fetal Intratracheal Injection of rAAV2/6.2
Efficient Gene Transfer Into the Mouse Lung by Fetal Intratracheal Injection of rAAV2/6.2
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DOI：
10.1038/mt.2010.153
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发表时间：
2010-12-01
影响因子：
12.4
通讯作者：
Debyser, Zeger
中科院分区：
文献类型：
--
作者：
Carlon, Marianne;Toelen, Jaan;Debyser, Zeger
胎儿基因治疗是治疗高死亡率或高发病率的先天性或围产期疾病的一种可能的新策略。我们开发了一种新的递送策略，直接注射到胎鼠气管中。肠内愈合（i.t.）在胚胎第18天（E18）注射比常规羊膜内（i.a.）交付.将来自腺相关病毒血清型6.2的病毒载体注射到胎鼠气管中，所述病毒载体具有气道上皮的嗜性并且先前未在胎鼠肺中进行测试。将生物发光（BL）成像（BLI）与磁共振（MR）成像（MRI）相结合，用于体内转基因表达的无创和准确定位。β-半乳糖苷酶（β-gal）的组织学分析显示，17.5%的上皮细胞在传导气道和1.5%的肺泡细胞中转导。注射后1个月内观察到稳定的基因表达。该研究表明，在胎鼠气管中直接注射rAAV 2/6.2上级i.a.用于转导肺的递送。第二，由于稳定的基因转移检测到出生后1个月，这种方法可能是有用的，以评估胎儿肺疾病的基因治疗，如囊性纤维化，需要大量的转导细胞以及延长基因表达，以获得一个稳定的表型效应。
Fetal gene therapy is one of the possible new therapeutic strategies for congenital or perinatal diseases with high mortality or morbidity. We developed a novel delivery strategy to inject directly into the fetal mouse trachea. Intratracheal (i.t.) injection at embryonic day 18 (E18) was more efficient in targeting the fetal lung than conventional intra-amniotic (i.a.) delivery. Viral vectors derived from adeno-associated virus serotype 6.2, with tropism for the airway epithelium and not earlier tested in the fetal mouse lung, were injected into the fetal trachea. Bioluminescence (BL) imaging (BLI) was combined with magnetic resonance (MR) imaging (MRI) for noninvasive and accurate localization of transgene expression in vivo. Histological analysis for beta-galactosidase (beta-gal) revealed 17.5% of epithelial cells transduced in the conducting airways and 1.5% in the alveolar cells. Stable gene expression was observed up to 1 month after injection. This study demonstrates that direct injection of rAAV2/6.2 in the fetal mouse trachea is superior to i.a. delivery for transducing the lung. Second, as stable gene transfer was detected up to 1 postnatal month, this approach may be useful to evaluate fetal gene therapy for pulmonary diseases such as cystic fibrosis, requiring both substantial numbers of transduced cells as well as prolonged gene expression to obtain a stable phenotypic effect.