A comparison of linear and branched polyethylenimine (PEI) with DCChol/DOPE liposomes for gene delivery to epithelial cells in vitro and in vivo.
Polyethylenimine (PEI), a polycation with high ionic charge density, has recently been used as a gene therapy delivery agent. We have defined the optimal conditions for PEI-based transfection of airway epithelial cells in vitro and in vivo and used these conditions to restore Cl(-) channel activity in a CF mouse model. Three forms of PEI, a linear 22 kDa (ExGen 500) form and branched 25 or 50 kDa forms were evaluated. All forms of PEI significantly increased luciferase reporter gene expression compared to the liposome DCChol/DOPE in a human bronchial epithelial cell line (16HBE) irrespective of the extent of cell confluency. With subconfluent cells, gene expression was around 1000-, 200- and 25-fold higher than liposomes using linear 22, 25 and 50 kDa PEI, respectively. The transfection efficiency was reduced in confluent and polarized epithelial cells but linear 22 kDa PEI showed the smallest decrease and gave 8000-fold better transfection in polarized cells compared to liposomes. A comparison of linear 22 or 25 kDa PEI with DCChol/DOPE for airway delivery in vivo via intranasal instillation was also performed. Linear 22 kDa PEI gave significantly better luciferase reporter gene expression of 350-fold in the lung, 180-fold in the nose and 85-fold in the trachea compared to liposome. In contrast, the 25 kDa form of PEI was no better than DCChol/DOPE. Repeat dosing with linear 22 kDa PEI failed to give reporter gene delivery comparable to the initial dose. To establish that PEI can be used to deliver a physiologically relevent gene in vivo, we used it to restore Cl(-) secretion by CFTR gene delivery in the airways of a CF mouse model.
Wiseman
J W
JW
1Department of Physiology, University of Cambridge, Cambridge, UK.
Goddard
C A
CA
McLelland
D
D
Colledge
W H
WH
D000818:Animals; D001980:Bronchi / Q000378:metabolism; D002460:Cell Line; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D019833:Genistein / Q000378:metabolism; D006801:Humans; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D010714:Phosphatidylethanolamines; D011094:Polyethyleneimine; D014132:Trachea / Q000378:metabolism; D014162:Transfection / Q000379:methods*
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; C505032:CFTR protein, human; D008081:Liposomes; D010714:Phosphatidylethanolamines; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011094:Polyethyleneimine; D019833:Genistein; D008156:Luciferases