Fasbender A, Zabner J, Chillón M, Moninger T O, Puga A P, Davidson B L, Welsh M J
Howard Hughes Medical Institute, Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
J Biol Chem. 1997 Mar 7;272(10):6479-89. doi: 10.1074/jbc.272.10.6479.
Improving the efficiency of gene transfer remains an important goal in developing new treatments for cystic fibrosis and other diseases. Adenovirus vectors and nonviral vectors each have specific advantages, but they also have limitations. Adenovirus vectors efficiently escape from the endosome and enter the nucleus, but the virus shows limited binding to airway epithelia. Nonviral cationic vectors bind efficiently to the negatively charged cell surface, but they do not catalyze subsequent steps in gene transfer. To take advantage of the unique features of the two different vector systems, we noncovalently complexed cationic molecules with recombinant adenovirus encoding a transgene. Complexes of cationic polymers and cationic lipids with adenovirus increased adenovirus uptake and transgene expression in cells that were inefficiently infected by adenovirus alone. Infection by both complexes was independent of adenovirus fiber and its receptor and occurred via a different cellular pathway than adenovirus alone. Complexes of cationic molecules and adenovirus also enhanced gene transfer to differentiated human airway epithelia in vitro and to the nasal epithelium of cystic fibrosis mice in vivo. These data show that complexes of adenovirus and cationic molecules increase the efficiency of gene transfer, which may enhance the development of gene therapy.
提高基因转移效率仍然是开发囊性纤维化和其他疾病新疗法的一个重要目标。腺病毒载体和非病毒载体各有其特定优势,但也存在局限性。腺病毒载体能有效地从内体逃逸并进入细胞核,但其与气道上皮细胞的结合有限。非病毒阳离子载体能有效地与带负电荷的细胞表面结合,但它们不能催化基因转移的后续步骤。为了利用这两种不同载体系统的独特特性,我们将阳离子分子与编码转基因的重组腺病毒进行非共价复合。阳离子聚合物和阳离子脂质与腺病毒的复合物增加了腺病毒的摄取以及在单独感染效率低下的细胞中的转基因表达。这两种复合物的感染均不依赖腺病毒纤维及其受体,且通过与单独腺病毒不同的细胞途径发生。阳离子分子与腺病毒的复合物还增强了体外向分化的人气道上皮细胞以及体内向囊性纤维化小鼠鼻上皮的基因转移。这些数据表明,腺病毒与阳离子分子的复合物提高了基因转移效率,这可能会促进基因治疗的发展。