DirectIn VivoGene Transfer to Airway Epithelium Employing Adenovirus–Polylysine–DNA Complexes

Abstract

Adenovirus-polylysine-DNA complexes were evaluated for their capacity to accomplish direct in vivo gene transfer to airway epithelium employing a rodent model. Binary complexes containing transferrin or adenovirus, or combination complexes containing both transferrin and adenovirus, were evaluated. The highest in vitro gene transfer efficiency in primary cultures of airway epithelial cells was accomplished by the combination complexes. This result was paralleled in vivo. Transient gene expression of up to 1 week was observed with localization of the transduced cells to the region of the small airways. These results establish the feasibility of this type of approach for gene therapy applications.

The effect of synthetic surfactant Exosurf on gene transfer in mouse lung in vivo

“…While naked plasmid DNA delivery avoids the safety issues associated with viral vectors, 15,16 transfection efficiency in the lung from plasmid DNA is relatively low. 3,11 However, it has been suggested that certain applications of pulmonary gene therapy might require only low levels of expression for therapeutic effect. For example, in cystic fibrosis there is evidence both in vitro 29 and in vivo 30 indicating that low levels of CFTR expression could re-establish physiological ion transport.…”

“…Animal models of bronchial gene transfer have proved crucial to the evaluation of the feasibility, efficiency and safety of various therapeutic strategies. 2 In these models, in vivo transfection of respiratory epithelium through the airways has been demonstrated after delivery of either adenovirus-polylysine-DNA complexes, 3 replication-deficient adenovirus, [4][5][6][7][8] DNA complexed with cationic lipids [9][10][11][12][13] or naked plasmid DNA under the control of CMV promoter/enhancer. 14 A major advantage of transfection with naked plasmid DNA is that it avoids the now recognized safety and inflamma-tory problems associated with either viral vectors, 15,16 or gene transfer using liposomes or cationic polymers.…”

“…[17][18][19] However, the only low levels of transfection and expression are observed in the lung as the result of the administration of naked plasmid DNA. 3,11,14 Exogenous pulmonary surfactant has been reported to enhance adenovirus-mediated luciferase gene expression in rabbits 20 and improve the uniformity of ␤-galactosidase expression of replication-deficient adenovirus in rats. 21 Natural pulmonary surfactant is a combination of lipids, proteins and carbohydrates that provide alveolar stability at low lung volumes.…”

Utilization of modified surfactant-associated protein B for delivery of DNA to airway cells in culture.

“…Successful transfection of airway cells in vitro and in vivo reported in recent literature includes the use of viruses with specific tropism for airway epithelial cells but also other compounds not indigenous to the airway. To date these include replication-defective adenoviral vectors, adenoassociated viral vectors, cationic liposomes including Lipofectin and N-[1-(2,3-dioleyloxy)propyl]-N,NN-trimethylammonium, and protein-cationic peptide complexes (1)(2)(3)(4)(5)(6)(7)(8). Recently, Curiel and associates (5) have applied the strategy of Wu and associates (4) to link poly(lysine) covalently to a cell surface receptor ligand to deliver DNA to lung cells via the receptor'endosomal pathway.…”

“…The process was enhanced by coupling the ligand-DNA complex with adenovirus through an antibody or mixing the ligand-DNA with adenovirus (7). Recently, adenovirus complexed directly with poly(lysine) was shown to deliver DNA effectively in vivo (8).…”