Widespread airway distribution and short-term phenotypic correction of cystic fibrosis pigs following aerosol delivery of piggyBac/adenovirus.
Widespread airway distribution and short-term phenotypic correction of cystic fibrosis pigs following aerosol delivery of piggyBac/adenovirus.
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DOI：
10.1093/nar/gky773
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发表时间：
2018-10-12
影响因子：
14.9
通讯作者：
Sinn PL
中科院分区：
文献类型：
--
作者：
Cooney AL;Singh BK;Loza LM;Thornell IM;Hippee CE;Powers LS;Ostedgaard LS;Meyerholz DK;Wohlford-Lenane C;Stoltz DA;B McCray P Jr;Sinn PL
囊性纤维化（CF）是一种常见的遗传性疾病，由编码囊性纤维化跨膜传导调节因子（CFTR）的基因突变引起。虽然CF影响多个器官系统，但肺部慢性细菌感染和炎症是CF患者发病和死亡的主要原因。与功能性CFTR基因互补可以修复这种缺陷，无论致病突变如何。在这项研究中，我们使用了一种基因递送系统，称为piggyBac/腺病毒（Ad），它结合了基于腺病毒的载体的递送效率与基于DNA转座子的载体的持续表达。我们将piggyBac/Ad雾化吸入猪的气道，并观察到广泛的肺部分布的载体。我们量化了气道中的区域分布，并观察了非CF猪的大气道上皮细胞和小气道上皮细胞的转导，其中约30-50%的表面上皮细胞对GFP呈阳性。我们转导了多种细胞类型，包括纤毛、非纤毛、基底和粘膜下腺细胞。此外，我们通过阴离子通道活性、气道表面液体pH值和细菌杀伤能力测定，在递送表达CFTR的piggyBac/Ad后，对CF猪进行表型校正。将整合DNA转座子与腺病毒载体递送组合是从单一载体施用实现功能性CFTR校正的有效方法。
Cystic fibrosis (CF) is a common genetic disease caused by mutations in the gene coding for cystic fibrosis transmembrane conductance regulator (CFTR). Although CF affects multiple organ systems, chronic bacterial infections and inflammation in the lung are the leading causes of morbidity and mortality in people with CF. Complementation with a functional CFTR gene repairs this defect, regardless of the disease-causing mutation. In this study, we used a gene delivery system termed piggyBac/adenovirus (Ad), which combines the delivery efficiency of an adenoviral-based vector with the persistent expression of a DNA transposon-based vector. We aerosolized piggyBac/Ad to the airways of pigs and observed widespread pulmonary distribution of vector. We quantified the regional distribution in the airways and observed transduction of large and small airway epithelial cells of non-CF pigs, with ∼30–50% of surface epithelial cells positive for GFP. We transduced multiple cell types including ciliated, non-ciliated, basal, and submucosal gland cells. In addition, we phenotypically corrected CF pigs following delivery of piggyBac/Ad expressing CFTR as measured by anion channel activity, airway surface liquid pH, and bacterial killing ability. Combining an integrating DNA transposon with adenoviral vector delivery is an efficient method for achieving functional CFTR correction from a single vector administration.
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影响因子：
8.8
作者：
Cao, Huibi;Machuca, Tiago N.;Hu, Jim
通讯作者：
Hu, Jim
影响因子：
2.6
作者：
Fraser, MJ;Coszczon, T;Bauser, C
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Bauser, C
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5.1
作者：
Cao, H.;Yang, T.;Hu, J.
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Hu, J.
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Croyle, MA;Chirmule, N;Wilson, JM
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Wilson, JM
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作者：
Drazan, KE;Olthoff, KM;Shaked, A
通讯作者：
Shaked, A