A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung
A novel cationic lipid greatly enhances plasmid DNA delivery and expression in mouse lung
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DOI：
10.1073/pnas.93.21.11454
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发表时间：
1996-10-15
影响因子：
11.1
通讯作者：
Cheng, SH
中科院分区：
文献类型：
--
作者：
Wheeler, CJ;Felgner, PL;Cheng, SH
有效的肺组织基因治疗需要使用高效的载体将感兴趣的基因输送到肺细胞中。将编码氯霉素乙酰转移酶(CAT)的DNA载体滴鼻给BALB/c小鼠，可检测到小鼠肺组织中CAT活性。与单独使用裸露DNA相比，用最优配方的商用转染剂运送的质粒DNA在肺中表达的CAT活性高达10倍。由(+/-)-N-(3-氨基丙基)-N，N-二甲基-2，3-二(十二烷氧基)-1-丙胺(GAP-DLRIE)和中性脂质体二油酰磷脂酰乙醇胺(DOPE)组成的脂质体与单独使用DNA相比，CAT的表达增加了100倍以上。将GAP-DLRIE脂质体-CAT DNA复合体单次注入小鼠肺内，在感染后第1-4天达到表达高峰，然后在给药后第21天逐渐恢复到基线水平。在第21天再次给予GAP-DLRIE脂质体CAT复合体导致报告基因表达的另一个瞬时高峰。GAP-DLRIE复合β-半乳糖苷酶DNA处理的肺组织学检查显示，肺泡上皮细胞是主要的表达部位，高达1%的肺泡上皮细胞含有表达转基因的上皮细胞。
Effective gene therapy for lung tissue requires the use of efficient vehicles to deliver the gene of interest into lung cells. When plasmid DNA encoding chloramphenicol acetyltransferase (CAT) was administered intranasally to BALB/c mice without carrier lipids, CAT activity was detected in mouse lung extracts. Plasmid DNA delivered with optimally formulated commercially available transfection reagents expressed up to 10-fold more CAT activity in lung than observed with naked DNA alone. Liposome formulations consisting of (+/-)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis(dodecyloxy)-1- propanaminium bromide (GAP-DLRIE) plus the neutral colipid dioleoylphosphatidylethanolamine (DOPE) enhanced CAT expression by more than 100-fold relative to plasmid DNA alone. A single administration of GAP-DLRIE liposome-CAT DNA complexes to mouse lung elicited peak expression at days 1-4 posttransfection, followed by a gradual return to baseline by day 21 postadministration. Readministration of GAP-DLRIE liposome CAT complexes at day 21 led to another transient peak of reporter gene expression. Histological examination of lungs treated with GAP-DLRIE complexed beta-galactosidase DNA revealed that alveolar epithelial cells were the primary locus of expression and that up to 1% of all alveoli contained epithelial cells expressing the transgene.