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Gene therapy for new bone formation using adeno-associated viral bone morphogenetic protein-2 vectors

Gene Therapy volume 10, pages 1345–1353 (2003)Cite this article

Abstract

Previous reports have suggested that bone morphogenetic protein (BMP) gene therapy could be applied for in vivo bone regeneration. However, these studies were conducted either using immunodeficient animals because of immunogenicity of adenovirus vectors, or using ex vivo gene transfer technique, which is much more difficult to handle. Adeno-associated virus (AAV) is a replication-defective virus without any association with immunogenicity and human disease. This study was conducted to investigate whether orthotopic new bone formation could be induced by in vivo gene therapy using AAV-based BMP2 vectors. To test the feasibility of this approach, we constructed an AAV vector carrying human BMP2 gene. Mouse myoblast cells (C2C12) transduced with this vector could produce and secrete biologically active BMP2 protein and induce osteogenic activity, which was confirmed by ELISA and alkaline phosphatase activity assay. For in vivo study, AAV-BMP2 vectors were directly injected into the hindlimb muscle of immunocompetent Sprague–Dawley rats. Significant new bone under X-ray films could be detected as early as 3 weeks postinjection. The ossification tissue was further examined by histological and immunohistochemical analysis. This study is, to our knowledge, the first to establish the feasibility of AAV-based BMP2 gene therapy for endochondral ossification in immunocompetent animals.

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Acknowledgements

We thank the Laboratory Animal Unit, The University of Hong Kong for helping us with animal holding and maintenance. Also, we are grateful to Mr David Wilmshurst, the technical writer in The University of Hong Kong, for his kind review on this manuscript. This study was supported by a University Research Grant (2000–2001) from The University of Hong Kong.

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  1. Y Chen and K D K Luk: The first two authors contributed equally to this work

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Faculty of Medicine, The University of Hong Kong, Hong Kong

    Y Chen, K D K Luk, K M C Cheung, W W Lu & J C Y Leong

  2. Institute of Molecular Biology, The University of Hong Kong, Hong Kong

    R Xu, M C Lin & H-F Kung

Authors
  1. Y Chen
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  2. K D K Luk
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  3. K M C Cheung
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  4. R Xu
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  5. M C Lin
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  6. W W Lu
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  7. J C Y Leong
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  8. H-F Kung
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Chen, Y., Luk, K., Cheung, K. et al. Gene therapy for new bone formation using adeno-associated viral bone morphogenetic protein-2 vectors. Gene Ther 10, 1345–1353 (2003). https://doi.org/10.1038/sj.gt.3301999

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