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Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type

Gene Therapy volume 10, pages 1289–1296 (2003)Cite this article

Abstract

The objective of the study was to analyze and compare the abilities of various human cell types with inherently dissimilar osteogenic potentials to induce heterotopic bone formation following ex vivo transduction with two distinct adenoviral vectors encoding bone morphogenetic protein type 2 (BMP2). The cells comprised primary human bone marrow mesenchymal stem cells (BM-MSCs), primary human skin fibroblasts (SFs), and a human diploid fetal lung cell line (MRC-5). The vectors included adenovirus type 5 or a chimeric adenovirus type 5 with the fiber gene of adenovirus type 35 (Ad5F35-BMP2), both demonstrating significantly different expression of BMP2 in vitro. The experimental groups consisted of the three human cell types transduced with each of the two adenoviral vectors. Using nonobese diabetic severe combined immunodeficiency (NOD/SCID) mice, the transduced cells were injected intramuscularly following ex vivo adenoviral transduction. The nature and extent of heterotopic bone formation were analyzed radiographically and histologically. At 14 days postinjection, abundant, highly mineralized bone was formed in mice injected with Ad5F35-BMP2-transduced cells irrespective of the cell type. There was no statistically significant difference in the amount of bone formed between BM-MSCs, SFs, and MRC-5 cells transduced with Ad5F35-BMP2, as assessed from bone surface area on biplanar plain radiography. Substantially lesser amounts or no bone could be detected in mice injected with cells transduced with Ad5-BMP2. Immunohistochemical analysis confirmed the presence of human cells in muscle as early as 2 days postdelivery; however, at 6–7 days after injection, the transduced cells could not be detected in surrounding muscle, or in the heterotopic bone, indicating the host origin of the newly formed bone. The results of the study demonstrate no significant difference in osteoinductive properties between BM-MSCs, SFs, and MRC-5 cells transduced ex vivo with the same type of adenovirus encoding BMP2. The level of BMP2 expression appears to be a crucial factor determining the extent of heterotopic bone formation and was significantly affected by the type of adenovirus used. In the cell types studied, Ad5F35-BMP2 was more efficacious than Ad5-BMP2 in providing adequate levels of BMP2 for efficient osteoinduction.

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References

  1. Lieberman JR et al. Regional gene therapy with a BMP-2-producing murine stromal cell line induces heterotopic and orthotopic bone formation in rodents. J Orthop Res 1998; 16: 330–339.

    Article  CAS  Google Scholar 

  2. Riew KD et al. Induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene in a rabbit spinal fusion model. Calcif Tissue Int 1998; 63: 357–360.

    Article  CAS  Google Scholar 

  3. Alden TD et al. In vivo endochondral bone formation using a bone morphogenetic protein 2 adenoviral vector. Hum Gene Ther 1999; 10: 2245–2253.

    Article  CAS  Google Scholar 

  4. Krebsbach PH, Gu K, Franceschi RT, Rutherford RB . Gene therapy-directed osteogenesis: BMP-7-transduced human fibroblasts form bone in vivo. Hum Gene Ther 2000; 11: 1201–1210.

    Article  CAS  Google Scholar 

  5. Varady P et al. Morphologic analysis of BMP-9 gene therapy-induced osteogenesis. Hum Gene Ther 2001; 12: 697–710.

    Article  CAS  Google Scholar 

  6. Okubo Y et al. In vitro and in vivo studies of a bone morphogenetic protein-2 expressing adenoviral vector. J Bone Joint Surg 2001; 83A (Suppl 1): S99–S104.

    Google Scholar 

  7. Engstrand T et al. Transient production of bone morphogenetic protein 2 by allogeneic transplanted transduced cells induces bone formation. Human Gene Ther 2000; 11: 205–211.

    Article  CAS  Google Scholar 

  8. Lee JY et al. Enhancement of bone healing based on ex vivo gene therapy using human muscle-derived cells expressing bone morphogenetic protein 2. Hum Gene Ther 2002; 13: 1201–1211.

    Article  CAS  Google Scholar 

  9. Gazit D et al. Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy. J Gene Med 1999; 1: 121–133.

    Article  CAS  Google Scholar 

  10. Musgrave DS et al. Ex vivo gene therapy to produce bone using different cell types. Clin Orthop 2000; 378: 290–305.

    Article  Google Scholar 

  11. Olmsted EA et al. Adenovirus-mediated BMP2 expression in human bone marrow stromal cells. J Cell Biochem 2001; 82: 11–21.

    Article  CAS  Google Scholar 

  12. Olmsted-Davis EA et al. Use of a chimeric adenovirus vector enhances BMP2 production and bone formation. Hum Gene Ther 2002; 13: 1337–1347.

    Article  CAS  Google Scholar 

  13. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP . Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 1997; 64: 295–312.

    Article  CAS  Google Scholar 

  14. Davis AR et al. Construction of adenoviral vectors. Mol Biotechnol 2001; 18: 63–70.

    Article  CAS  Google Scholar 

  15. Gutekunst CA et al. The cellular and subcellular localization of huntingtin-associated protein 1 (HAP1): comparison with huntingtin in rat and human. J Neurosci 1998; 18: 7674–7686.

    Article  CAS  Google Scholar 

  16. Farinas I, Yoshida CK, Backus C, Reichardt LF . Lack of neurotrophin-3 results in death of spinal sensory neurons and premature differentiation of their precursors. Neuron 1996; 17: 1065–1078.

    Article  CAS  Google Scholar 

  17. Reubinoff BE et al. Neural progenitors from human embryonic stem cells. Nat Biotechnol 2001; 19: 1134–1140.

    Article  CAS  Google Scholar 

  18. Turgeman G et al. Engineered human mesenchymal stem cells: a novel platform for skeletal cell mediated gene therapy. J Gene Med 2001; 3: 240–251.

    Article  CAS  Google Scholar 

  19. Pittenger MF et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143–147.

    Article  CAS  Google Scholar 

  20. Caplan AI . Mesenchymal stem cells. J Orthop Res 1991; 9: 641–650.

    Article  CAS  Google Scholar 

  21. Bruder SP, Fink DJ, Caplan AI . Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy. J Cell Biochem 1994; 56: 283–294.

    Article  CAS  Google Scholar 

  22. Lennon DP et al. Dilution of human mesenchymal stem cells with dermal fibroblasts and the effects on in vitro and in vivo osteochondrogenesis. Dev Dyn 2000; 219: 50–62.

    Article  CAS  Google Scholar 

  23. Jacobs JP, Jones CM, Baille JP . Characteristics of a human diploid cell designated MRC-5. Nature 1970; 227: 168–170.

    Article  CAS  Google Scholar 

  24. Yotnda P et al. Efficient infection of primitive hematopoietic stem cells by modified adenovirus. Gene Therapy 2001; 8: 930–937.

    Article  CAS  Google Scholar 

  25. Shayakhmetov DM, Papayannopoulou T, Stamatoyannopoulos G, Lieber A . Efficient gene transfer into human CD34(+) cells by a retargeted adenovirus vector. J Virol 2000; 74: 2567–2583.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Dr Malcolm Brenner for his insightful review of the manuscript and Dr Robert McAlhany for virus preparation. We also thank Genetics Institute for the anti-BMP2 monoclonal antibody (h3b2/17.8.1). This work was supported by National Institutes of Health Grants R03AR47463-01 (EAO-D) and R21AR484-01 (ARD).

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Authors and Affiliations

  1. Department of Orthopaedic Surgery, Baylor College of Medicine, Houston, TX, USA

    Z Gugala, E A Olmsted-Davis, R W Lindsey & A R Davis

  2. Department of Pediatrics, Baylor College of Medicine, Houston, TX, USA

    E A Olmsted-Davis & A R Davis

  3. Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA

    E A Olmsted-Davis & A R Davis

  4. Department of Bone Pathology, Armed Forces Institute of Pathology, Washington, DC, USA

    F H Gannon

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  1. Z Gugala
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  5. A R Davis
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Gugala, Z., Olmsted-Davis, E., Gannon, F. et al. Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type. Gene Ther 10, 1289–1296 (2003). https://doi.org/10.1038/sj.gt.3302006

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  • DOI: https://doi.org/10.1038/sj.gt.3302006

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