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Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1–deleted adenoviruses

Abstract

We describe the use of a human bronchial xenograft model for studying the efficiency and biology of in vivo gene transfer into human bronchial epithelia with recombinant E1 deleted adenoviruses. All cell types in the surface epithelium except basal cells efficiently expressed the adenoviral transduced recombinant genes, lacZ and CFTR, for 3–5 weeks. Stable transgene expression was associated with high level expression of the early adenoviral gene, E2a, in a subset of transgene expressing cells and virtually undetectable expression of the late adenoviral genes encoding the structural proteins, hexon and fiber. These studies begin to address important issues that relate to safety and in vivo efficacy of recombinant adenoviruses for gene delivery into the human airway.

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References

  1. Boat, T.F., Welsh, M.J., Beaudet, A.L. Cystic Fibrosis In The Metabolic Basis of Inherited Disease. 6th edn (eds C.R. Scriver, A.L. Beaudet, W.S. Sly & D. Valle) 2649–2680 (McGraw-Hill, New York, 1989).

    Google Scholar 

  2. Rommens, J.M. et al. Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245, 1059–1065 (1989).

    Article  CAS  Google Scholar 

  3. Riordan, J.R. et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245, 1066–1072 (1989).

    Article  CAS  Google Scholar 

  4. Kerem, B.-S., Rommens et al. Identification of the cystic fibrosis gene: genetic analysis. Science 245, 1073–1080, (1989).

    Article  CAS  Google Scholar 

  5. Gregory, R.J. et al. Expression and characterization of the cystic fibrosis transmembrane conductance regulator. Nature 347, 382–285 (1990).

    Article  CAS  Google Scholar 

  6. Bear, C.D. et al. Purification and functional reconstitution of the cystic fibrosis transmembrane conductance regulator (CFTR). Cell 68, 809–818 (1992).

    Article  CAS  Google Scholar 

  7. Rich, D.P. et al. Expression of cystic fibrosis transmembrane conductance regulator corrects defective chloride channel regulation in cystic fibrosis airway epithelial cells. Nature 347, 358–363 (1990).

    Article  CAS  Google Scholar 

  8. Drumm, M.T. et al. Correction of the cystic fibrosis defect in vitro by retrovirus-mediated gene transfer. Cell 62, 1227–1233 (1990).

    Article  CAS  Google Scholar 

  9. Cotton, C.U., Shutts, M.J., Knowles, M.R., Gatzy, J.T. & Boucher, R.C. Abnormal apical cell membrane in cystic fibrosis respiratory epithelium. J. clin. Invest. 79, 80–85 (1987).

    Article  CAS  Google Scholar 

  10. Welsh, M.J. Abnormal regulation of ion channels in cystic fibrosis epithelia. FASEB 4, 2718–2725 (1990).

    Article  CAS  Google Scholar 

  11. Trapnell, B.C. et al. Expression of the cystic fibrosis transmembrane conductance regulator gene in the respiratory tract of normal individuals and individuals with cystic fibrosis. Proc. natn Acad. Sci. U.S.A. 88, 6565–6569 (1991).

    Article  CAS  Google Scholar 

  12. Sarkadi, B. et al. Biochemical characterization of the cystic fibrosis transmembrane conductance regulator in normal and cystic fibrosis epithelial cells. J. biol Chem. 267, 2087–2095 (1992).

    CAS  PubMed  Google Scholar 

  13. Zeitlin, P.L. et al. CFTR protein expression in primary and cultured epithelia. Proc. natn Acad. Sci U.S.A. 89, 344–347 (1992).

    Article  CAS  Google Scholar 

  14. Verdugo, P. Goblet cells secretion and mucogenesis. Ann. Rev. Physiol. 52, 157–176 (1990).

    Article  CAS  Google Scholar 

  15. Engelhardt, J.F. et al. Submucosal glands are the predominant site of CFTR expression in human bronchus. Nature Genet. 2, 240–248 (1992).

    Article  CAS  Google Scholar 

  16. Breeze, R.G. & Wheeldon, E.B. The cells of the pulmonary airways. Am. Rev. Resp. Dis. 116, 705–777 (1977).

    Article  CAS  Google Scholar 

  17. Ayers, M.M. & Jeffery, P.K. Proliferation and differentiation in mammalian airway epithelium. Eur. resp. J. 1, 58–80 (1988).

    CAS  Google Scholar 

  18. Blenkinsopp, W.K. Proliferation of respiratory tract epithelium in the rat. Exp. cell Res. 46, 144–154, (1967).

    Article  CAS  Google Scholar 

  19. Engelhardt, J.E., Yankaskas, J.R. & Wilson, J.M. In vivo retroviral gene transfer into human bronchial epithelia of xenografts. J. clin. Invest. 90, 2598–2607 (1992).

    Article  CAS  Google Scholar 

  20. Plopper, C.G., Heidsiek, J.G., Weir, A.J., St. George, J.A. & Hyde, D.M. Tracheobronchial epithelium in the adult rhesus monkey: a quantitative histochemical and ultrastructural study. Am. J. Anat. 184, 31–40 (1989).

    Article  CAS  Google Scholar 

  21. Wilson, D.W., Plopper, C.G. & Hyde, D.M. The tracheobronchial epithelium of the bonnet monkey (Macaca radiata): a quantitative ultrastructural study. Am. J. Anat. 171, 25–40 (1984).

    Article  CAS  Google Scholar 

  22. Hazinski, T.A., Ladd, P.A. & DeMatteo, C.A. Localization and induced expression of fusion genes in the rat lung. Am. J. resp. cell. molec. Biol. 4, 206–209 (1991).

    Article  CAS  Google Scholar 

  23. Yoshimura, K. et al. Expression of the human cystic fibrosis transmembrane conductance regulator gene in the mouse lung after in vivo intratracheal plasmid-mediated gene transfer. Nucl. Acids Res. 20, 3233–3240 (1992).

    Article  CAS  Google Scholar 

  24. Graham, F.L. & Prevec, L. Manipulation of Adenovirus Vectors In Methods in Molecular Biology, Vol. 7, Gene Transfer and Expression Protocols. (ed. E.J. Murray) 109–128 (Human Press, Clifton New Jersey, 1991).

    Chapter  Google Scholar 

  25. Rosenfeld, M.A. et al. Adenovirus-mediated transfer of a recombinant alpha 1-antitrypsin gene to the lung epithelium in vivo. Science 252, 431–434 (1991).

    Article  CAS  Google Scholar 

  26. Rosenfeld, M.A. et al. In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell 68, 143–155 (1992).

    Article  CAS  Google Scholar 

  27. Horwitz, M.S. Adenoviridae and Their Replication In Virology, 2nd edn (eds B. N. Fields, et al.) 1679–1721 ((Raven Press, New York, 1990).

    Google Scholar 

  28. Reichel, R., Kovesdi, I. & Nevins, J. Jr. Developmental control of a promoter-specific factor that is also regulated by the E1A gene product. Cell 48, 501–506 (1987).

    Article  CAS  Google Scholar 

  29. Green, M. & Daesch, G.E. Biochemical studies on adenovirus multiplication: Kinetics of nucleic acid and protein synthesis in suspension cultures. Virology 13, 169–176 (1961).

    Article  CAS  Google Scholar 

  30. Stratford-Perricaudet, L.D., Makeh, I., Perricaudet, M. & Briand, P. Widespread long-term gene transfer to mouse skeletal muscles and heart. J. clin. Invest. 90, 626–630 (1992).

    Article  CAS  Google Scholar 

  31. Yankaskas, J.R., Knowles, M.R., Gatzy, J.T. & Boucher, R.C. Persistence of abnormal chloride ion permeability in cystic fibrosis nasal epithelial cells in heterologous culture. Lancet 1, 954–6 (1985).

    Article  CAS  Google Scholar 

  32. Engelhardt, J.F., Allen, E.D. & Wilson, J.M. Reconstitution of tracheal grafts with a genetically modified epithelium. Proc. natn Acad. Sci U.S.A. 88, 11192–11196 (1991).

    Article  CAS  Google Scholar 

  33. Randell, S.H., Comment, C.E., Ramaekers, F.C.S. & Nettesheim, P. Properties of rat tracheal epithelial cells separated based on expression of cell surface alpha-galactosyl end groups. Am. J. resp. cell molec. Biol. 4, 544–554 (1991).

    Article  CAS  Google Scholar 

  34. Rutten, A.A.J.J.L., Bruyntjes, J.P. & Ramaekers, F.C.S. Effect of cigarette smoke condensate and vitamin A depletion on keratin expression patterns in cultured hamster tracheal epithelium. An immunohistomorphological study using monoclonal antibodies to keratins. Virchows Archiv. B. Cell. Pathol. 56, 111–117 (1988).

    Article  CAS  Google Scholar 

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Engelhardt, J., Yang, Y., Stratford-Perricaudet, L. et al. Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1–deleted adenoviruses. Nat Genet 4, 27–34 (1993). https://doi.org/10.1038/ng0593-27

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