Efficient Gene Delivery to Pig Airway Epithelia and Submucosal Glands Using Helper-Dependent Adenoviral Vectors
Efficient Gene Delivery to Pig Airway Epithelia and Submucosal Glands Using Helper-Dependent Adenoviral Vectors
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DOI：
10.1038/mtna.2013.55
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发表时间：
2013-10-01
影响因子：
8.8
通讯作者：
Hu, Jim
中科院分区：
文献类型：
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作者：
Cao, Huibi;Machuca, Tiago N.;Hu, Jim
气道基因递送是治疗囊性纤维化（CF）等危及生命的肺部疾病患者的一种有前途的策略。然而，这种策略必须在大型动物临床前研究中进行评估，以便将其转化为人类应用。由于人类和猪肺在解剖学和生理学上的相似性，我们利用猪作为一个大型动物模型来研究辅助依赖性腺病毒载体的气道基因递送的安全性和有效性。在支气管镜引导下，将携带人CFTR或报告基因LacZ的辅助依赖性载体雾化吸入猪体内。我们发现LacZ报告基因和hCFTR转基因产物在肺气道上皮细胞中有效表达。具有这种递送的转基因载体也可以到达粘膜下腺体。此外，hCFTR转基因蛋白定位于纤毛和非纤毛上皮细胞的顶膜，反映了野生型CF跨膜传导调节因子（CFTR）的位置。根据试验猪的有限数量，猪对气雾剂给药程序耐受良好，未显示全身毒性。这些结果为开发人CF肺基因治疗的临床策略提供了重要的见解。
Airway gene delivery is a promising strategy to treat patients with life-threatening lung diseases such as cystic fibrosis (CF). However, this strategy has to be evaluated in large animal preclinical studies in order to translate it to human applications. Because of anatomic and physiological similarities between the human and pig lungs, we utilized pig as a large animal model to examine the safety and efficiency of airway gene delivery with helper-dependent adenoviral vectors. Helper-dependent vectors carrying human CFTR or reporter gene LacZ were aerosolized intratracheally into pigs under bronchoscopic guidance. We found that the LacZ reporter and hCFTR transgene products were efficiently expressed in lung airway epithelial cells. The transgene vectors with this delivery can also reach to submucosal glands. Moreover, the hCFTR transgene protein localized to the apical membrane of both ciliated and nonciliated epithelial cells, mirroring the location of wild-type CF transmembrane conductance regulator (CFTR). Aerosol delivery procedure was well tolerated by pigs without showing systemic toxicity based on the limited number of pigs tested. These results provide important insights into developing clinical strategies for human CF lung gene therapy.