Influence of Cell Polarity on Retrovirus-Mediated Gene Transfer to Differentiated Human Airway Epithelia
| Autor: | Mordechai Bodner, Doug J. Jolly, Guoshun Wang, Paul Melchert, Helmuth H. G. van Es, Vladimir Slepushkin, Paul B. McCray, Beverly L. Davidson |
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| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
Cell Membrane Permeability
Fibroblast Growth Factor 7 Cystic Fibrosis Cellular differentiation Transgene Genetic enhancement Immunology Cystic Fibrosis Transmembrane Conductance Regulator Gene Expression Bronchi Biology Microbiology chemistry.chemical_compound Retrovirus Virology Cell polarity Gene expression Humans Phosphate Transport Proteins Growth Substances Cells Cultured FGF10 Symporters Gene Transfer Techniques Cell Polarity Cell Differentiation Epithelial Cells Sodium-Phosphate Cotransporter Proteins Gene Therapy biology.organism_classification Cell biology Fibroblast Growth Factors Leukemia Virus Murine Trachea chemistry Insect Science Receptors Virus Keratinocyte growth factor Fibroblast Growth Factor 10 Cell Division |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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