Human Gene Expression in Murine Hemopoietic Cells In Vivo

  • Frederick A. Fletcher
  • Kateri A. Moore
  • Grant R. MacGregor
  • John W. Belmont
  • C. Thomas Caskey
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 34)


Somatic gene transfer offers the possibility of a new approach in the treatment of human genetic disease. Defects affecting the blood and blood forming tissues are candidates for therapies involving transfer of new genetic information into hemopoietic stem cells. One such defect, adenosine deaminase (ADA) deficiency, is being used as a model in which hemopoietic gene transfer techniques can be developed and evaluated. In this model, gene transfer is mediated by a retroviral vector. Retroviral vectors have been used extensively to deliver information to hemopoietic cells1–14. We have previously reported delivery and expression of human ADA (hADA) sequences in murine hemopoietic progenitors in vitro15 and in vivo16, but were not able to demonstrate long term stability of expression. We describe here the construction and testing of four new vectors, representing the first demonstration of efficient transfer and long term in vivo expression of hADA in murine hemopoietic cells.


Adenosine Deaminase Human Genetic Disease Hemopoietic Stem Cell Howard Hughes Medical Institute Human Gene Expression 
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  1. 1.
    Joyner A., G. Keller, R.A. Phillips, and A. Bernstein. 1983. Retrovirus transfer of a bacterial gene into mouse haematopoietic progenitor cells. Nature 305: 556.PubMedCrossRefGoogle Scholar
  2. 2.
    Miller, A.D., R.J. Eckner, D.J. Jolly, T. Friedmann, and I.M. Verma. 1984. Expression of a Retrovirus Encoding Human HPRT in Mice. Science 225: 630.PubMedCrossRefGoogle Scholar
  3. 3.
    Williams, D.A., I.R. Lemischka, D.G. Nathan, and R.C. Mulligan. 1984. Introduction of new genetic material into pluripotent haematopoietic stem cells of the mouse. Nature 310: 476.PubMedCrossRefGoogle Scholar
  4. 4.
    Dick, J.E., M.C. Magli, D. Huszar, R.A. Phillips, and A. Bernstein. 1985. Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice. Cell 42: 71.PubMedCrossRefGoogle Scholar
  5. 5.
    Keller, G., C. Paige, E. Gilboa, and E.F. Wagner. 1985. Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent heamatopoietic precursors. Nature 318: 149.PubMedCrossRefGoogle Scholar
  6. 6.
    Eglitis, M.A., P. Kantoff, E. Gilboa, and W.F. Anderson. 1985. Gene Expression in Mice after High Efficiency Retroviral Mediated Gene Transfer. Science 230: 1395.PubMedCrossRefGoogle Scholar
  7. 7.
    Lemischka, I.R., D.H. Raulet, and R.C. Mulligan. 1986. Developmental Potential and Dynamic Behavior of Hematopoietic Stem Cells. Cell 45: 917.PubMedCrossRefGoogle Scholar
  8. 8.
    Williams, D.A., S.H. Orkin, and R.C. Mulligan. 1986. Retrovirus-mediated transfer of human adenosine deaminase gene sequences into cells in culture and into murine hematopoietic cells in vivo. Proc. Nat. Acad. Sci. USA 83: 2566.PubMedCrossRefGoogle Scholar
  9. 9.
    Hock, R.A., and A.D. Miller. 1986. Retrovirus-mediated transfer and expression of drug resistance genes in human haematopoietic progenitor cells. Nature 320: 275.PubMedCrossRefGoogle Scholar
  10. 10.
    Kwok, W.W., F. Schuening, R.B. Stead, and A.D. Miller. 1986. Retroviral transfer of genes into canine hemopoietic progenitor cells in culture: a model for human gene therapy. Proc. Natl. Acad. Sci. USA 55 83: 4552.CrossRefGoogle Scholar
  11. 11.
    Chang, S.M., K. Wager Smith, T.Y. Tsao, J. Henkel Tigges., S. Vaishnav, and C.T. Caskey. 1987. Construction of a defective retrovirus containing the human hypoxanthine phosphoribosyl-transferase cDNA and its expression in cultured cells and mouse bone marrow. Mol. Cell. Biol. 7: 854.PubMedGoogle Scholar
  12. 12.
    Hawley, R.G., L. Covarrubias, T. Hawley, and B. Mintz. 2987. Handicapped retroviral vectors efficiently transduce foreign genes into hematopoietic stem cells. Proc. Natl. Acad. Sci. USA. 84: 206.Google Scholar
  13. 13.
    Kantoff, P.W., A.P. Gillio, J.R. McLachlin, C. Bordignon, M.A. Eglitis, N.A. Kernan, R.C. Moen, D.B. Kohn, S.F. Yu, and E. Karson. 1987. Expression of human adenosine deaminase in nonhuman primates after retrovirus-mediated gene transfer. J. Exp. Med. 166: 219.PubMedCrossRefGoogle Scholar
  14. 14.
    Magli, M-C., J.E. Dick, D. Huszar, A. Bernstein, and R.A. Phillips. 1987. Modulation of Gene Expression in Multiple hematopoietic cell lineages following retroviral gene transfer. Proc. Natl. Acad. Sci. USA 84: 789.PubMedCrossRefGoogle Scholar
  15. 15.
    Belmont, J.W., J. Henkel Tigges, S.M. Chang, K. Wager Smith, R.E. Kellems, J.E. Dick, M.C. Magli, R.A. Phillips, A. Bernstein, and C.T. Caskey. 1986. Expression of human adenosine deaminase in murine haematopoietic progenitor cells following retroviral transfer. Nature 322: 385.PubMedCrossRefGoogle Scholar
  16. 16.
    Belmont, J.W., J. Henkel Tigges, K. Wager Smith, SM-W. Chang, and C.T. Caskey. 1987. Transfer and Expression of Human Adenosine Deaminase Gene in Murine Bone Marrow Cells. In R.P. Gale, R. Champlin (eds): “Progress in Bone Marrow Transplantation,” New York: Alan R. Liss, p 963.Google Scholar
  17. 17.
    Manuscript in preparation.Google Scholar
  18. 18.
    Bender, M.A., T.D. Palmer, R.E. Gelinas, and A.D. Miller. 1987. Evidence that the packaging signal of Moloney murine leukemia virus extends into the gag region. J. Virol. 61: 1639.PubMedGoogle Scholar
  19. 19.
    Deschamps, J., F. Meijlink, and I. Verma. 1985. Identification of a transcriptional enhancer element upstream from the proto oncogen fos. Science 230: 1174.PubMedCrossRefGoogle Scholar
  20. 20.
    Hogan, B., F. Constantini, and E. Lacy. In Manipulating the Mouse Embryo. New York: Cold Spring Harbor Laboratory Press.Google Scholar
  21. 21.
    Bannerman, R.M. 1983. Hematology. In Foster, H.L., Small, J.D., Fox, J.G. (eds): “The Mouse in Biomedical Research:Volume III”. New York Academic Press, p 293.Google Scholar
  22. 22.
    This work was supported by the Howard Hughes Medical Institute (HHMI) and by grants from the NIH (HD21452) and from the Cystic Fibrosis Foundation (R004 7-0351). GRM is the recipient of an Arthritis Foundation post-doctoral fellowship; JWB is an Assistant Investigator and CTC is an Investigator in HHMI. The authors wish to thank Jenny Henkel-Tigges, Dianne Houston-Hawkins, Michelle Rives, Deborah Villalon, and Karen Wager-Smith for technical assistance and Elsa Perez for help in preparation of the manuscript.Google Scholar

Copyright information

© Plenum Press, New York 1988

Authors and Affiliations

  • Frederick A. Fletcher
    • 1
    • 2
  • Kateri A. Moore
    • 1
    • 2
  • Grant R. MacGregor
    • 1
    • 2
  • John W. Belmont
    • 1
    • 2
  • C. Thomas Caskey
    • 1
    • 2
  1. 1.Institute for Molecular GeneticsBaylor College of MedicineHoustonUSA
  2. 2.Howard Hughes Medical InstituteBaylor College of MedicineHoustonUSA

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