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Bone marrow stromal cells as a vehicle for gene transfer

Gene Therapy volume 6, pages 1611–1616 (1999)Cite this article

Abstract

Adoptive transfer of genetically modified somatic cells is playing an increasingly important role in the management of a wide spectrum of human diseases. Hematopoietic stem cells and lymphocytes have been used to transfer a variety of genes, however, they have limitations. In this study, the feasibility of retroviral gene transduction of bone marrow stromal cells, and the engraftment characteristics of these cells following infusion, was investigated in a murine transplantation model. Stromal cells derived from Balb/c mouse bone marrow were transduced with a replication-defective retrovirus containing the LacZ gene. Following three rounds of transduction, between 5 and 40% of the cells were positive for the LacZ gene. A total of 2 × 106 cells were infused into the same mouse strain. After the infusion, the LacZ gene was detected by PCR in the bone marrow, spleen, liver, kidney and lung; however, only the spleen and bone marrow samples were strongly positive. Quantitative PCR demonstrated that between 3 and 5% of spleen and bone marrow cells, and 1% of liver cells contained the LacZ gene at 3 weeks after infusion; <0.2% transduced cells were found in other organs. no difference was noted in engraftment between mice with or without irradiation before transplantation, suggesting that engraftment occurred without myeloablation. the infused transduced cells persisted for up to 24 weeks. self-renewal of transplanted stromal cells was demonstrated in secondary transplant studies. ease of culture and gene transduction and tissue specificity to hematopoietic organs (bone marrow, spleen, liver) is demonstrated, indicating that stromal cells may be an ideal vehicle for gene transfer.

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References

  1. Mulligan RC . The basic science of gene therapy Science 1993 260: 926–932

    Article  CAS  Google Scholar 

  2. Ali M, Lemoine NR, Ring CJA . The use of DNA virus as vectors for gene therapy Gene Therapy 1994 1: 367–384

    CAS  PubMed  Google Scholar 

  3. Mitani K, Wakamiya M, Caskey CT . Long term expression of retroviral-transduced adenosine deaminase in human primitive hematopoietic progenitors Hum Gene Ther 1993 4: 9–16

    Article  CAS  Google Scholar 

  4. Chertkov JL et al. The hematopoietic stromal microenvironment promotes retrovirus-mediated gene transfer into hematopoietic stem cells Stem Cells 1993 11: 218–227

    Article  CAS  Google Scholar 

  5. Broxmeyer HE et al. Cord blood transplantation and the potential for gene therapy. Gene transduction using a recombinant adeno-associated viral vector Ann NY Acad Sci 1995 770: 105–115

    Article  CAS  Google Scholar 

  6. Culver K et al. Lymphocytes as cellular vehicles for gene therapy in mouse and man Proc Natl Acad Sci USA 1991 88: 3155–3159

    Article  CAS  Google Scholar 

  7. Weiss LP . Functional organization of the hematopoietic tissues In: Hoffman R et al (eds) . Hematology: Basic Principles and Practice Churchill Livingston: New York 1991 pp 82–97

    Google Scholar 

  8. Quesenberry PJ et al. In vitro and in vivo studies of stromal niches Blood Cells 1994 20: 97–104

    CAS  PubMed  Google Scholar 

  9. Clark BR, Keating A . Biology of bone marrow stroma (review) Ann NY Acad Sci 1995 770: 70–78

    Article  CAS  Google Scholar 

  10. Lerat H et al. Role of stromal cells and macrophages in fibronectin biosynthesis and matrix assembly in human long-lerm marrow cultures Blood 1993 82: 1480–1492

    CAS  PubMed  Google Scholar 

  11. Matthews KE et al. Bead transfection: rapid and efficient gene transfer into marrow stromal and other adherent mammalian cells Exp Hematol 1993 21: 697–702

    CAS  PubMed  Google Scholar 

  12. Hauser SP, Waldron JA, Udupa KB, Lipschitz DA . Morphological characterization of stromal cell types in hematopoietically active long-term murine bone marrow culture J Histochem Cytochem 1995 43: 371–379

    Article  CAS  Google Scholar 

  13. Keating A et al. Donor origin of the in vitro haematopoietic microenvironment after marrow transplantation in man Nature 1982 298: 280–283

    Article  CAS  Google Scholar 

  14. Piersma AH, Ploemacher RE, Brockbank KG . Transplantation of bone marrow fibroblastoid stromal cells in mice via the intravenous route Br J Haematol 1983 54: 285–290

    Article  CAS  Google Scholar 

  15. Van Zant G, Holland BP, Eldridge PW, Chen JJ . Genotype-restricted growth and aging patterns in hematopoietic stem cell populations of allophenic mice J Exp Med 1990 171: 1547–1565

    Article  CAS  Google Scholar 

  16. Simmons PJ, Przepiorka D, Thomas ED, Torok-Storb B . Host origin of marrow stromal cells following allogeneic bone marrow transplantation Nature 1987 328: 429–432

    Article  CAS  Google Scholar 

  17. Golde DW et al. Origin of human bone marrow fibroblasts Br J Haematol 1980 44: 183–187

    Article  CAS  Google Scholar 

  18. Wilson FD et al. Cytogenetic studies on bone marrow fibroblasts from a male–female hematopoietic chimera. Evidence that stromal elements in human transplantation recipients are of host type Transplantation 1978 25: 87–88

    Article  CAS  Google Scholar 

  19. Simmons PJ, Zannettino A, Gronthos S, Leavesley D . Potential adhesion mechanisms for localization of haemopoietic progenitors to bone marrow stroma Leuk Lymphoma 1994 12: 353–363

    Article  CAS  Google Scholar 

  20. Dittel BN, LeBien TW . Reduced expression of vascular adhesion molecule-1 on bone marrow stromal cells isolated from marrow transplant recipients correlates with reduced capacity to support human B lymphopoiesis in vitro Blood 1995 86: 2833–2841

    CAS  PubMed  Google Scholar 

  21. Huss R, Smith FO, Myerson DH, Deeg HJ . Homing and immunogenicity of murine stromal cells transfected with xenogeneic MHC class II genes Cell Transplant 1995 4: 483–491

    Article  CAS  Google Scholar 

  22. Onyia JE, Clapp DW, Long H, Hock JM . Osteoprogenitor cells as targets for ex vivo gene transfer J Bone Min Res 1998 13: 20–30

    Article  CAS  Google Scholar 

  23. Cherington V et al. Retroviral vector-modified bone marrow stromal cells secrete biologically active factor IX in vitro and transiently deliver therapeutic levels of human factor IX to the plasma of dogs after reinfusion Hum Gene Ther 1998 9: 1397–1407

    Article  CAS  Google Scholar 

  24. Chuah MK et al. Bone marrow stromal cells as targets for gene therapy of hemophilia A Hum Gene Ther 1998 9: 353–365

    Article  CAS  Google Scholar 

  25. Pereira RF et al. Marrow stromal cells as a source of progenitor cells for nonhematopoietic tissues in transgenic mice with a phenotype of osteogenesis imperfecta Proc Nat Acad Sci USA 1998 95: 1142–1147

    Article  CAS  Google Scholar 

  26. Bahner I et al. Infection of human marrow stroma by human immunodeficiency virus-1 (HIV-1) is both required and sufficient for HIV-1-induced hematopoietic suppression in vitro: demonstration by gene modification of primary human stroma Blood 1997 90: 1787–1798

    CAS  PubMed  Google Scholar 

  27. Rettinger SD et al. In vivo transduction with retrovirus during hepatic in flow occlusion J Surg Res 1993 54: 418–425

    Article  CAS  Google Scholar 

  28. Dexter TM, Allen TD, Lajtha LG . Conditions controlling the proliferation of haemopoietic stem cells in vitro J Cell Physiol 1977 91: 335–344

    Article  CAS  Google Scholar 

  29. Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning Laboratory Manual, 2nd edn Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY 1989

    Google Scholar 

  30. Lubin MB et al. Precise gene dosage determination by polymerase chain reaction: theory, methodology, and statistical approach Mol Cell Probes 1991 5: 307–317

    Article  CAS  Google Scholar 

  31. Southern EM . Detection of specific sequences among DNA fragments separated by gel electrophoresis J Mol Biol 1975 98: 503–517

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by a grant from American Cancer Society (DHP 153) and USPHS National Heart Blood and Lung Institute (HL-55695) and National Cancer Institute (CA71092) and the Phi Beta Psi Sorority to NCM. We thank the office of grants and Scientific publication at the Arkansas Cancer Research Center (Paula Card-Higginson and Mary Dornhoffer) for assistance with the manuscript.

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Authors and Affiliations

  1. Central Arkansas Veterans Healthcare System, Myeloma and Transplantation Research Center, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    L Ding, S Lu, R B Batchu & N C Munshi

  2. Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, AR, USA

    R L Saylors III

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  1. L Ding
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  2. S Lu
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  3. R B Batchu
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  4. R L Saylors III
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Ding, L., Lu, S., Batchu, R. et al. Bone marrow stromal cells as a vehicle for gene transfer. Gene Ther 6, 1611–1616 (1999). https://doi.org/10.1038/sj.gt.3300973

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