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Retroviral Gene Transfer of Human Adenosine Deaminase into Hematopoietic Cells

  • D. Cournoyer
  • M. Scarpa
  • K. Moore
  • R. H. Buckley
  • J. W. Belmont
  • C. T. Caskey
Conference paper
Part of the Experimental Hematology Today—1989 book series (HEMATOLOGY, volume 1989)

Abstract

We are developing a safe and efficient system of retroviral gene transfer of human ADA into hematopoietic stem cells. Inherited deficiency of ADA results in Severe Combined Immunodeficiency (SCID) and serves as a model for gene replacement therapy attempts. The vector DNN2ADA has been successful in transducing long term expression in mice via hematopoietic stem cell infection. Additional vectors with the potential to induce tissue-specific lymphoid expression have been constructed. Results obtained with one of those vectors are described.

We have developed a new system for the detection of replication competent virus which might arise from recombinational events. It involves DNA amplification of sequences located between the pol and env genes of Moloney Murine Leukemia Virus in target cells. This allows the detection of one helper-infected cell out of 104 to 105 uninfected cells.

Virus supernatant from helper free DNN2ADA producing cell lines were used to infect human bone marrow cells (BMCs). Five to 20% of unselected colony forming progenitors contained integrated copy(ies) of the vector as detectable by DNA amplification. Low copy number of the provirus were present in supernatant cells from long term bone marrow culture up to seven weeks post-infection. Transduced ADA expression was detectable following infection of BMCs derived from two ADA deficient SCID patients.

These results indicate special retroviral vectors can successfully transfer and express human ADA into mouse and human bone marrow cells.

Keywords

Polymerase Chain Reac Severe Combine Immunodeficiency Purine Nucleoside Phosphorylase Packaging Cell Line Human Bone Marrow Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Baird PA, Anderson RW, Newcombe HB, Lowry RB (1988) Genetic disorders in children and young adults: a population study. Am J Hum Genet 42: 677–693.PubMedGoogle Scholar
  2. 2.
    Belmont JW, Caskey CT (1986) Developments leading to human gene therapy. In Kucherlapati R, ed. Gene Transfer: Plenum Publishing Corp. 411–441.Google Scholar
  3. 3.
    Anderson WF (1984) Prospects for human gene therapy. Science 226: 401–409.PubMedCrossRefGoogle Scholar
  4. 4.
    Ledley FD (1987) Somatic gene therapy for human disease: background and prospects. J Pediatr 110: 167–174.PubMedCrossRefGoogle Scholar
  5. 5.
    Kredich NM, Hershfield MS (19) Im-munodeficiency diseases caused by adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds). The Metabolic Basis of Inherited Disease, McGraw-HillGoogle Scholar
  6. 6.
    Kantoff PW, Kohn DB, Mitsuya H, Armentano D, Sieberg M (1986) Correction of adenosine deaminase deficiency in cultured human T and B cells by retrovirus-mediated gene transfer. Proc Natl Acad Sci USA 83: 6563–6567.Google Scholar
  7. 7.
    Palmer TD, Hock RA, Osborne WR, Miller AD (1987) Efficient retrovirus-media- ted transfer and expression of a human adenosine deaminase gene in diploid skin fibroblasts from an adenosine deaminase-deficient human. Proc Natl Acad Sci USA 84: 1055–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Hershfield MS, Buckley RH, Greenberg ML (1987) Treatment of adenosine deaminase deficiency with polyethylene glycol-modified adenosine deaminase. N Engl J Med 316: 589–596.Google Scholar
  9. 9.
    Cepki CL, Roberts BE, Mulligan RC (1984) Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell 37: 1053–1062.CrossRefGoogle Scholar
  10. 10.
    Keller G, Paige C, Gilboa E, Wagner EF (1985) Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature 318: 149–154.PubMedCrossRefGoogle Scholar
  11. 11.
    Armentano D, Yu SF, Kantoff PW, von Ruden T, Anderson WF,Gilboa E. (1987) Effect of internal viral sequences on the utility of retroviral vectors. J Virol 61: 1647–1650.PubMedGoogle Scholar
  12. 12.
    Belmont JW, MacGregor GR, Wager-Smith K (1988) Expression of human adenosine deaminase in murine hemato-poietic cells. Mol Cell Biol 8:5116–5125.Google Scholar
  13. 13.
    Belmont JW, Henkel-Tigges J, Chang SMW (1986) Expression of human adenosine deaminase in murine haematopoietic progenitor cells following retroviral transfer. Nature 322 (24): 385–387.Google Scholar
  14. 14.
    Mann R, Mulligan RC, Baltimore D (1983) Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell 33: 153–159.PubMedCrossRefGoogle Scholar
  15. 15.
    Markowitz, D, Goff S, Bank A (1988) A safe packaging line for gene transfer: separating viral genes on two different plasmids. J Virol 62: 1120–1124.PubMedGoogle Scholar
  16. 16.
    Moore K, Fletcher F, Villalon D, Utter A, Belmont J (1989) Human adenosine deaminase expression in mice, (submitted)Google Scholar
  17. 17.
    Sen R, Baltimore D (1986) Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell 46: 705–716.PubMedCrossRefGoogle Scholar
  18. 18.
    Wirth T, Staudt L, Baltimore D (1987) An octamer oligonucleotide upstream of a TATA motif is sufficient for lym- phoid-specific promoter activity. Nature 329: 174–178.PubMedCrossRefGoogle Scholar
  19. 19.
    Markowitz D, Goff S, Bank A (1988) Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167: 400–406.PubMedGoogle Scholar
  20. 20.
    Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R (1988) Primer-directed enzymatic amplification DNA with a thermostable DNA polymerase. Science 239: 487–491.Google Scholar
  21. 21.
    Dexter TM, Spooncer E, Simmons P, Allen TD (1984) Long-term marrow culture: an overview of techniques and experience. In: Wright DG, Greenberger JS. Long-term bone marrow culture. Alan R. Liss, Inc. 57–96.Google Scholar
  22. 22.
    Honghua L, Gyllensten UB, Cui ZF, Saiki, Erlick HA, Arnheim N (1988) Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature 335: 414–417.Google Scholar
  23. 23.
    Aitken DA, Kleijer WJ, Niermeijer MF, Herbschleb-Voogt E, Galjaard H (1980) Prenatal detection of a probable hete- rozygote for ADA deficiency and severe combined immunodeficiency disease using a microradioassay. Clinical Gen 17: 293–298.CrossRefGoogle Scholar
  24. 24.
    Bender MA, Gelinas RE, Miller AD (1989) A majority of mice show long- term expression of a human 0-globin gene after retrovirus transfer into hematopoietic stem cells. Mol Cell Biol 9: 1426–1434.PubMedGoogle Scholar
  25. 25.
    Sutherland HJ, Eaves CJ, Eaves AC, Dragowska W, Lansdrop PM (1989) Characterization and partial purifica tion of human marrow cells capable of initiating long-term hemopoiesis in vitro. Blood (in press).Google Scholar
  26. 26.
    Kamel-Reid S, Dick JE (1989) Engraftment of immune-deficient mice with human hematopoietic stem cells. Science 242: 1706–1709.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1990

Authors and Affiliations

  • D. Cournoyer
  • M. Scarpa
  • K. Moore
  • R. H. Buckley
  • J. W. Belmont
  • C. T. Caskey

There are no affiliations available

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