Development of novel formulations that enhance adenoviral-mediated gene expression in the lung in vitro and in vivo
Development of novel formulations that enhance adenoviral-mediated gene expression in the lung in vitro and in vivo
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DOI：
10.1006/mthe.2001.0411
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发表时间：
2001-07-01
影响因子：
12.4
通讯作者：
Wilson, JM
中科院分区：
文献类型：
--
作者：
Croyle, MA;Cheng, X;Wilson, JM
尽管用于囊性纤维化治疗的病毒和非病毒基因传递载体的发展取得了显著进展，但气道上皮基因转移效率低是临床应用的主要障碍。在这里，我们开发的配方，提高细胞吸收腺病毒载体。我们从一组已知可增强药物吸收的药学上可接受的化合物中选择辅料。在体外和体内评估了每种成分存在下病毒的转导效率。甘露醇和壳聚糖分别显著提高了体外转导效率和体内表达量，分别提高了4和8个log单位。最成功的配方（蔗糖、甘露醇和Pluronic F68的混合物）在体外转导了100%的A549细胞群，并在体内大小气道中产生了强烈的基因表达区域，毒性最小。剂量反应研究还表明，当放入该配方时，病毒剂量可以降低1/2 log，同时保持较高的转基因表达水平。这种配方还增强了病毒的物理稳定性。冻干制剂在25℃下保存30天后，滴度未见明显下降。
Despite remarkable progress in the development of both viral and non-viral gene delivery vectors for cystic fibrosis therapy, low efficiency of gene transfer to the airway epithelium is a major obstacle to clinical application. Here we develop formulations that enhance cellular absorption of adenoviral vectors. We selected excipients from a panel of pharmaceutically acceptable compounds known to enhance drug absorption. Transduction efficiency of the virus in the presence of each ingredient was assessed in vitro and in vivo. Mannitol and chitosan substantially enhanced transduction efficiency in vitro and augmented expression in vivo by 4 and 8 log units, respectively. The most successful formulation (a blend of sucrose, mannitol, and Pluronic F68) transduced 100% of an A549 cell population in vitro and produced areas of intense gene expression in both large and small airways in vivo with minimal toxicity. Dose response studies also indicate that when placed in this formulation, the viral dose can be lowered by 1/2 log while maintaining superior levels of transgene expression. This formulation also enhanced the physical stability of the virus. No significant loss in titer was detected from a lyophilized formulation after storage at 25 degreesC for 30 days.