Influence of cell polarity on retrovirus-mediated gene transfer to differentiated human airway epithelia.
Gene transfer with recombinant murine leukemia viruses (MuLV) provides the potential to permanently correct inherited lung diseases, such as cystic fibrosis (CF). Several problems prevent the application of MuLV-based recombinant retroviruses to lung gene therapy: (i) the lack of cell proliferation in mature pulmonary epithelia, (ii) inefficient gene transfer with a vector applied to the apical surface, and (iii) low titers of many retroviral preparations. We found that keratinocyte growth factor (KGF) stimulated proliferation of differentiated human tracheal and bronchial epithelia. Approximately 50% of epithelia divided in response to KGF as assessed by bromodeoxyuridine histochemistry. In airway epithelia stimulated to divide with KGF, high-titer ampho- and xenotropic enveloped vectors preferentially infected cells from the basal side. However, treatment with hypotonic shock or EGTA transiently increased transepithelial permeability, enhancing gene transfer with the vector applied to the mucosal surfaces of KGF-stimulated epithelia. Up to 35% of cells expressed the transgene after gene transfer. By using this approach, cells throughout the epithelial sheet, including basal cells, were targeted. Moreover, the Cl- transport defect in differentiated CF airway epithelia was corrected. These findings suggest that barriers to apical infection with MuLV can be overcome.
Wang
G
G
Departments of Pediatrics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
Davidson
B L
BL
Melchert
P
P
Slepushkin
V A
VA
van Es
H H
HH
Bodner
M
M
Jolly
D J
DJ
McCray
P B
PB
D001980:Bronchi / Q000166:cytology / Q000187:drug effects / Q000821:virology; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D002463:Cell Membrane Permeability; D016764:Cell Polarity; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism / Q000628:therapy; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000821:virology; D051526:Fibroblast Growth Factor 10; D051523:Fibroblast Growth Factor 7; D005346:Fibroblast Growth Factors; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006133:Growth Substances / Q000008:administration & dosage; D006801:Humans; D009052:Leukemia Virus, Murine / Q000235:genetics*; D028061:Phosphate Transport Proteins; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D014132:Trachea / Q000166:cytology / Q000187:drug effects / Q000821:virology
C505032:CFTR protein, human; C496453:FGF7 protein, human; D051526:Fibroblast Growth Factor 10; D006133:Growth Substances; D028061:Phosphate Transport Proteins; D011991:Receptors, Virus; D050608:Sodium-Phosphate Cotransporter Proteins; D027981:Symporters; D051523:Fibroblast Growth Factor 7; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005346:Fibroblast Growth Factors