Transgene expression in mouse airway epithelium by aerosol gene therapy with PEI-DNA complexes
Transgene expression in mouse airway epithelium by aerosol gene therapy with PEI-DNA complexes
复制标题
DOI：
10.1006/mthe.2001.0300
复制
发表时间：
2001-04-01
影响因子：
12.4
通讯作者：
Waldrep, JC
中科院分区：
文献类型：
--
作者：
Gautam, A;Densmore, CL;Waldrep, JC
针对呼吸道上皮的基因治疗具有治疗囊性纤维化和α抗胰蛋白酶缺乏等疾病的潜力。已经利用多种方法，例如鼻内或气管内滴注和气雾剂递送，将基因靶向气道。聚乙烯亚胺（PEI）是一种线性或支化的聚阳离子聚合物，已被用于将基因递送至各种器官。在本研究中，我们使用异硫氰酸荧光素 (FITC) 标记的支链 PEI，初步检查了气雾输送至 Balb/C 小鼠后 PEI 在肺部的定位。此外，在气溶胶递送 PEI-CAT DNA 后，使用氯霉素乙酰转移酶 (CAT) 蛋白的原位免疫染色来定位肺部内的转基因表达。 CAT 的免疫组织化学以及 FITC 标记的 PEI 的定位表明，气雾剂递送后，PEI-DNA 复合物沉积并随后转染传导气道（包括外周气道）中的大部分上皮细胞。气溶胶暴露后 24 小时检测到高水平的 CAT，并且在单次气溶胶暴露后长达 28 天在肺部检测到显着的 CAT 表达。数据表明，PEI-DNA 复合物的气雾输送可有效治疗囊性纤维化和 cu-l 抗胰蛋白酶缺乏症等肺部疾病。
Gene therapy targeted at the respiratory epithelium holds therapeutic potential for diseases such as cystic fibrosis and alpha anti-trypsin deficiency. A variety of approaches such as intranasal or intratracheal instillation and aerosol delivery have been utilized to target genes to the airways. Polyethylenimine (PEI), a linear or branched polycationic polymer, has been used for delivery of genes to various organs. In this study, using fluorescein isothiocyanate (FITC)-labeled branched PEI, we initially examined the localization of PEI in the lungs after aerosol delivery to Balb/C mice. Further, after aerosol delivery of PEI-CAT DNA, in situ immunostaining for chloramphenicol acetyl transferase (CAT) protein was used to localize the transgene expression within the lungs. Immunohistochemistry for CAT, as well as localization of FITC-labeled PEI, revealed that after aerosol delivery, the PEI-DNA complexes deposit and subsequently transfect most of the epithelial cells in the conducting airways (including the peripheral airways). High levels of CAT were detected at 24 h after aerosol exposure and significant CAT expression was detected in the lungs up to 28 days after a single aerosol exposure. The data suggest that aerosol delivery of PEI-DNA complexes could be effective for the treatment of pulmonary diseases such as cystic fibrosis and cu-l anti-trypsin deficiency.