Abstract
We have administered a recombinant adenovirus vector (AdCFTR) containing the normal human CFTR cDNA to the nasal and bronchial epithelium of four individuals with cystic fibrosis (CF). We show that this vector can express the CFTR cDNA in the CF respiratory epithelium in vivo. With doses up to 2 × 109 pfu, there was no recombination/complementation or shedding of the vector or rise of neutralizing antibody titres. At 2 × 109 pfu, a transient systemic and pulmonary syndrome was observed, possibly mediated by interleukin-6. Follow-up at 6–12 months demonstrated no long term adverse effects. Thus, it is feasible to use an adenovirus vector to transfer and express the CFTR cDNA in the respiratory epithelium of individuals with CF. Correction of the CF phenotype of the airway epithelium might be achieved with this strategy.
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References
Boat, T.F., Welsh, M.J. & Beaudet, A.L. Cystic fibrosis. in The Metabolic Basis of Inherited Disease. (eds Scriver, C.R., Beaudet, A.L., Sly, W.S. & Valle, D. ) 2649–2680 (McGraw-Hill, New York, 1989).
Rommens, J.M. et al. Identification of the cystic fibrosis gene: chromosome walking and jumping. Science 245, 1059–1065 (1989).
Riordan, J.R. et al. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 245, 1066–1073 (1989).
Kerem, B.-S. et al. Identification of the cystic fibrosis gene: genetic analysis. Science 245, 1073–1080 (1989).
Gregory, R.J. et al. Expression and characterization of the cystic fibrosis transmembrane conductance regulator. Nature 347, 382–386 (1990).
Welsh, M.J. et al. Cystic fibrosis transmembrane conductance regulator: a chloride channel with novel regulation. Neuron 8, 821–829 (1992).
Cheng, S.H. et al. Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis. Cell 63, 827–834 (1990).
Collins, F.S. Cystic fibrosis: molecular biology and therapeutic implications. Science 256, 774–779 (1992).
Welsh, M.J. & Smith, A.E. Molecular mechanisms of CFTR chloride channel dysfunction in cystic fibrosis. Cell 73, 1251–1254 (1993).
Beddrossian, C.W.M., Greenberg, S.D., Singer, D.B., Hansen, J.J. & Rosenberg, H.S. The lung in cystic fibrosis: a qualitative study including prevalence of pathologic findings among different age groups. Hum. Pathol. 7, 195–204 (1976).
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).
Drumm, M.L. et al. Correction of the cystic fibrosis defect in vitro by retrovirus-mediated gene transfer. Cell 62, 1227–1233 (1990).
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).
Blaese, R.M. et al. The ADA human gene therapy clinical protocol. Hum. gene Ther. 1, 327–362 (1990).
Grossman, M. et al. Successful ex vivo gene therapy directed to liver in a patient with familial hypercholesterolaemia. Nature Genet. 6, 335–341 (1994).
Crystal, R.G. Protocol for gene therapy of the respiratory manifestations of cystic fibrosis using a replication deficient recombinant adenovirus to transfer the normal cystic fibrosis transmembrane conductance regulator cDNA to the airway epithelium. Fed. Register 58, 21737–21738 (1992).
Rosenfeld, M.A. et al. Adenovirus-mediated transfer of a recombinant α1-antitrypsin gene to the lung epithelium in vivo. Science 252, 431–434 (1991).
Mastrangeli, A. et al. Diversity of airway epithelial cell targets for in vivo recombinant adenovirus-mediated gene transfer. J. clin. Invest. 91, 225–234 (1993).
Huebner, R.J. et al. Newly recognized respiratory tract viruses. in A. Rev. Microbiol. 12, 49–76 (1958).
Straus, S.E. Adenovirus infections in humans. in The Adenoviruses. (ed. Ginsberg, H. S. ) 451–496 (Plenum Press, New York, 1984).
Rosenfeld, M.A. et al. Gene transfer to freshly isolated human respiratory epithelial cells in vitro using a replication deficient adenovirus containing the human cystic fibrosis transmembrane conductance regulator cDNA. Hum. gene Ther. 5, 331–342 (1994).
Horowitz, M.S. Adenoviridae and their replicationin Virology. (eds Fields, B.N. & Knipe, D.M.) 1679–1740 (Raven Press, New York, 1990).
Welsh, M.J. Cystic fibrosis gene therapy using an adenovirus vector: in vivo safety and efficacy in the nasal epithelium. Fed. Register 58, 21737 (1992).
Wilson, J.M. Gene therapy of cystic fibrosis lung diseases using E1 deleted adenovirus: A phase 1 trial. Fed. Register 58, 47906 (1992).
Boucher, R.C. & Knowles, M.R. Gene therapy for cystic fibrosis using E1 deleted adenovirus: A Phase I trial in the nasal cavity. Fed. Register 58, 53814 (1993).
Wilmott, R.W., Whitsett, J. & Trapnell, B. A phase I study of gene therapy of cystic fibrosis utilizing a replication deficient recombinant adenovirus vector to deliver the human cystic fibrosis transmembrane conductance regulator cDNA to the airways. Fed. Register 58, 21739 (1993).
Rich, D.P. et al. Development and analysis of recombinant adenoviruses for gene therapy of cystic fibrosis. Hum. Gene Ther. 4, 461–476 (1993).
Englehardt, J.F. et al. Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1-deleted adenoviruses. Nature Genet. 4, 27–34.
Zabner, J. et al. Safety and efficacy of repetitive adenovirus-mediated transfer of CFTR cDNA to airway epithelia of primates and cotton rats. Nature Genet. 6, 75–83 (1994).
Bout, A. et al. Lung gene therapy: In-vivo adenovirus-mediated gene transfer to rhesus monkey airway epithelium. Hum. Gene Ther. 5, 3–10 (1994).
Englehardt, J.F. et al. Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: biological efficacy study. Hum. gene Ther. 4, 759–769 (1993).
Brody, S.L., Metzger, M., Danel, C., Rosenfeld, M. & Crystal, R.G. Acute responses of non-human primates to airway delivery of an adenovirus vector containing the human cystic fibrosis transmembrane conductance regulator cDNA. Hum. gene Ther. 5, (1994).
Danel, C., Erzurum, S.C., Yoneyama, K., Thunnissen, F.B.J.M. & Crystal, R.G. Quantitative assessment of human airway epithelial and inflammatory cell populations in cystic fibrosis. Am. Rev. respir. Dis. 145, A689 (1992).
Zabner, J. et al. Adenovirus-mediated gene transfer transiently corrects the chloride transport defect in nasal epithelia of patients with cystic fibrosis. Cell 75, 207–216 (1993).
Graham, F.L. & Smiley, J. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. gen. Virol. 36, 59–74 (1977).
Spergel, J.M. & Selina, C.-K. lnterleukin-6 enhances a cellular activity that functionally substitutes for E1A protein in transactivation. Proc. natn. Acad. Sci. U.S.A. 88, 6472–6476 (1991).
Spergel, J.M. et al. NF-IL6, a member of the C/EBP family, regulates E1A-responsive promoters in the absence of E1A. J. Virol. 66, 1021–1030 (1992).
Couch, R.B., Cate, T.R., Fleet, W.F., Gerone, P.J. & Knight, V. Aerosol-induced adenoviral illness resembling the naturally occurring illness in military recruits. Am. Rev. respir. Dis. 93, 529–535 (1966).
Ginsberg, H.S., Badger, G.F., Dingle, J.H., Jordan, W.S. Jr., & Katz, S. Etiologic relationship of the RI-67 agent to “acute” respiratory disease (ARD). J. clin. Invest. 34, 820–831 (1955).
Fox, J.P. et al. The virus watch program: a continuing surveillance of viral infections in metropolitan New York families. VI. Observations of adenovirus infections: Virus excretion patterns, antibody response, efficiency of surveillance, patterns of Infection, and relation to illness. Am. J. Epidemiol. 89, 25 (1969).
Simon, R.H. et al. Adenovirus-mediated transfer of the CFTR gene to lung of nonhuman primates: Toxicity study. Hum. Gene Ther. 4, 771–780 (1993).
Setoguchi, Y., Jaffe, H.A., Chu, C.-S. & Crystal, R.G. Intraperitoneal In vivo gene therapy to deliver α1-antitrpysin to the systemic circulation. Am. J. respir. Cell molec. Biol. 10, 369–377 (1994).
Englehardt, J., Simon, R., Zepeda, M., Yang, Y. & Wilson, J.M. Safety and efficacy of recombinant adenoviruses for lung directed gene therapy in nonhuman primates. Pediatr. Pulmonol. 9, S16.4 (1993).
Smith, T.A.G. et al. Adenovirus mediated expression of therapeutic plasma levels of human factor IX in mice. Nature Genet. 5, 397–402 (1994).
Van Snick, J. Interteukin-6: An overview. A. Rev. Immunol. 8, 253–278 (1990).
Kishimoto, T., Shizuo, A. & Tetsuya, T. Interleukin-6 and its receptor: a paradigm for cytokines. Science 256, 593–597 (1992).
Weber, J. et al. Phase I trial of subcutaneous interleukin-6 in patients with advanced malignancies. J. clin. Oncol. 11, 499–506 (1993).
Xin, Z., Jordana, M., Braciak, T., Ohtoshi, T. & Gauldie, J. Lipopolysaccharide induces expression of granulocyte/macrophage colony-stimulating factor, interleukin-8, and interleukin-6 in human nasal, but not lung, fibroblasts: Evidence for heterogeneity within the respiratory tract. Am. J. resp. Cell molec. Biol. 9, 225–263 (1993).
Ruef, C., Jefferson, D.M., Schlegel-Haueter, S.E. & Suter, S. Regulation of cytokine secretion by cystic fibrosis airway epithelial cells. Eur. Respir. J. 6, 1429–1436 (1993).
Melani, C. et al. Interteukin-6 expression in human neutrophil and eosinophil peripheral blood granulocytes. Blood 81, 2744–2749 (1993).
Kronborg, G. et al. Cytokines in sputum and serum from patients with cystic fibrosis and chronic Pseudomonas aeruginosa infection as markers of destructive inflammation in the lungs. Pediatr. Pulmonol. 15, 292–297 (1993).
Bonfield, T.L., Ghnaim, H.A., Panuska, J.R., Konstan, M. & Berger, M. Cytokines in cystic fibrosis BAL. Pediatr. Pulmonol. 8, A312 (1993).
Devergne, O. et al. In vivo expression of IL-1 beta and IL-6 genes during viral infections in human. Eur. Cytokine Netw. 2, 183–194 (1991).
Ginsberg, H.S. et al. A mouse model for investigating the molecular pathogenesis of adenovirus pneumonia. Proc. natn. Acad. Sci. U.S.A. 88, 1651–1655 (1991).
Ryffel, B., Mihatsch, M.J. & Woerly, G. Pathology induced by interieukin-6. Int. Rev. exp. Path. 34A, 79–89 (1993).
Hack, C.E. et al. Increased plasma levels of interieukin-6 in sepsis. Blood 74, 1704–1710 (1989).
Fiers, W. Tumor necrosis factor: characterization at the molecular, cellular and in vivo level. FEBS Lett. 285, 199–212 (1991).
Smith, S.F. et al. Comparison of human lung surface protein profiles from central and peripheral airways sampled using two regional lavage techniques. Eur. Respir. J. 1, 792–800 (1988).
Minor modification to NIH-approved human gene transfer protocols, August 26, 1993 (Office of Recombinant DNA Activities, NIH, Bethesda, Maryland, USA).
Ginsberg, H.S. et al. Role of early region 3 (E3) in pathogenesis of adenovirus disease. Proc. natn. Acad. Sci. U.S.A. 86, 3823–3827 (1989).
Kasel, J.A. Adenovirus. in Diagonostic Procedures for Viral, Ricketsial and Chlamydial Infections. (eds Lennette, E.M. & Schmidt, N.J. ) 229–255 (American Public Health Association, Washington, DC, 1979).
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).
Eissa, N.T., Chu, C.-S., Danel, C. & Crystal, R.G. Evaluation of the respiratory epithelium of normals and individuals with cystic fibrosis for the presence of adenovirus E1a sequences relevant to the use of E1a adenovirus vectors for gene therapy for the respiratory manifestations of cystic fibrosis. Hum. gene Ther. 5, 1105–1114 (1994).
Chu, C.-S, Trapnell, B.C., Curristin, S.M., Cutting, G.R. & Crystal, R.G. Extensive posttranscriptional deletion of the coding sequences for part of nucleotide-bindlng fold 1 in respiratory epithelial mRNA transcripts of the cystic fibrosis transmembrane conductance regulator gene is not associated with the clinical manifestations of cystic fibrosis. J. clin. Invest. 90, 785–790 (1992).
Frohman, M.A., Dush, M.K. & Martin, G.R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc. natn. Acad. Sci. U.S.A. 85, 8998–9002 (1988).
Knowles, M., Gatzy, J. & Boucher, R. Increased bioelectric potential difference across respiratory epithelia in cystic fibrosis. New Engl. J. Med. 305, 1489–1495 (1981).
Saltini, C. et al. Maintenance of alveolitis in patients with chronic beryllium-specific helper T cells. New Engl. J. Med. 320, 1103–1109 (1989).
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Crystal, R., McElvaney, N., Rosenfeld, M. et al. Administration of an adenovirus containing the human CFTR cDNA to the respiratory tract of individuals with cystic fibrosis. Nat Genet 8, 42–51 (1994). https://doi.org/10.1038/ng0994-42
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DOI: https://doi.org/10.1038/ng0994-42
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