Eastman S J, Lukason M J, Tousignant J D, Murray H, Lane M D, St George J A, Akita G Y, Cherry M, Cheng S H, Scheule R K
Genzyme Corporation, Framingham, MA 01701-9322, USA.
Hum Gene Ther. 1997 Apr 10;8(6):765-73. doi: 10.1089/hum.1997.8.6-765.
Advances in gene therapy vectors and techniques hold promise for treatment of many inherited and acquired diseases. For lung indications, especially those involving the epithelium, delivery of the gene therapy vehicle ideally will involve the use of an aerosol. Aerosol delivery of transgenes using cationic lipids is currently limited by the ability to generate highly concentrated formulations of lipid:DNA complexes that are stable and retain their activity following aerosolization. We have examined many of the variables inherent in aerosolizing cationic lipid gene delivery vehicles and have devised a new formulation that incorporates small amounts of a polyethylene glycol-containing lipid. This formulation has allowed the preparation of concentrated dispersions of cationic lipid:plasmid DNA (pDNA) complexes (> 20 mM pDNA) at approximately 10-fold higher concentrations than previously reported. Most of the pDNA in these formulations was bound to the lipid component and thereby protected from nebulizer-induced shearing; the pDNA also maintained full biological activity both in vitro and in vivo. This new formulation thus represents a significant improvement over current methods to prepare concentrated, active cationic lipid gene delivery vectors, and provides a new tool with which to test gene transfer to the lung.
基因治疗载体和技术的进展为许多遗传性和获得性疾病的治疗带来了希望。对于肺部适应症，尤其是那些涉及上皮组织的疾病，理想情况下基因治疗载体的递送将涉及使用气雾剂。目前，使用阳离子脂质进行转基因的气雾剂递送受到限制，即难以生成高浓度的脂质:DNA复合物制剂，这些复合物在雾化后要保持稳定并保留其活性。我们研究了雾化阳离子脂质基因递送载体时固有的许多变量，并设计了一种新的制剂，其中包含少量含聚乙二醇的脂质。这种制剂能够制备阳离子脂质:质粒DNA（pDNA）复合物的浓缩分散液（> 20 mM pDNA），其浓度比以前报道的高出约10倍。这些制剂中的大多数pDNA与脂质成分结合，从而免受雾化器引起的剪切作用的影响；pDNA在体外和体内也均保持了完全的生物学活性。因此，这种新制剂相对于目前制备浓缩、活性阳离子脂质基因递送载体的方法有了显著改进，并提供了一种用于测试基因向肺部转移的新工具。
