The use of CpG-free plasmids to mediate persistent gene expression following repeated aerosol delivery of pDNA/PEI complexes
The use of CpG-free plasmids to mediate persistent gene expression following repeated aerosol delivery of pDNA/PEI complexes
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DOI：
10.1016/j.biomaterials.2012.04.019
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发表时间：
2012-08-01
期刊：
影响因子：
14
通讯作者：
Gill, Deborah R.
中科院分区：
文献类型：
--
作者：
Davies, Lee A.;Hyde, Stephen C.;Gill, Deborah R.
气溶胶基因治疗为治疗获得性和遗传性肺部疾病提供了巨大的潜力。为了治疗慢性肺病如囊性纤维化、哮喘和肺气肿，非病毒基因疗法可能需要重复施用以维持缓慢分裂或终末分化的肺上皮细胞中的转基因表达。当与质粒DNA（pDNA）复合时，合成聚合物25 kDa支链聚乙烯亚胺（PEI）可被配制用于气雾剂递送至肺部。我们表明，pDNA/PEI气雾剂制剂可以重复施用到小鼠气道至少10次，没有可检测的毒性。有趣的是，与单次给药相比，重复递送后的峰值报告基因活性显著降低高达75%，尽管在每次后续气溶胶暴露时pDNA肺沉积相似。虽然抑制的确切机制尚不清楚，但它不依赖于小鼠品系，不涉及免疫应答，并且由PEI介导。重要的是，使用56天的给药间隔，第四代无CpG质粒的递送产生高水平的持续转基因表达，其在随后的施用中进一步加强。这些数据一起表明，pDNA/PEI气溶胶制剂提供了用于基因递送至肺的通用平台，导致适于治疗慢性肺病的持续转基因表达。(C)2012爱思唯尔有限公司保留所有权利。
Aerosol gene therapy offers great potential for treating acquired and inherited lung diseases. For treatment of chronic lung diseases such as cystic fibrosis, asthma and emphysema, non-viral gene therapy will likely require repeated administration to maintain transgene expression in slowly dividing, or terminally differentiated, lung epithelial cells. When complexed with plasmid DNA (pDNA), the synthetic polymer, 25 kDa branched Polyethylenimine (PEI), can be formulated for aerosol delivery to the lungs. We show that pDNA/PEI aerosol formulations can be repeatedly administered to airways of mice on at least 10 occasions with no detectable toxicity. Interestingly, peak reporter gene activity upon repeated delivery was significantly reduced by up to 75% compared with a single administration, despite similar pDNA lung deposition at each subsequent aerosol exposure. Although the precise mechanism of inhibition is unknown, it is independent of mouse strain, does not involve an immune response, and is mediated by PEI. Importantly, using a dosing interval of 56 days, delivery of a fourth-generation, CpG-free plasmid generated high-level, sustained transgene expression, which was further boosted at subsequent administrations. Together these data indicate that pDNA/PEI aerosol formulations offer a versatile platform for gene delivery to the lung resulting in sustained transgene expression suitable for treatment of chronic lung diseases. (C) 2012 Elsevier Ltd. All rights reserved.