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Coxsackievirus and adenovirus receptor (CAR)-positive immature osteoblasts as targets of adenovirus-mediated gene transfer for fracture healing

Gene Therapy volume 10, pages 1623–1628 (2003)Cite this article

Abstract

Adenovirus vectors are expected to be a powerful tool for gene therapy to treat severe fractures. Adenovirus invades cells through binding to the coxsackievirus and adenovirus receptor (CAR) on the cell membrane. CAR expression is low in normal adult animals, but it is induced on regenerating cells in some experimental models. We made a rib fracture model in mice and evaluated the histological changes and CAR mRNA expression by RT-PCR 1, 5, 10, 14, and 21 days after the fracture. CAR mRNA was expressed exclusively in the fractured ribs at each time point, but not in the normal ribs. We detected the CAR protein immunohistochemically in fibroblast-like cells in the fracture callus on days 10 and 14 after fracture. In situ hybridization showed that these fibroblast-like cells expressed mRNA of type I collagen and osteopontin, but not osteocalcin, defining the cells as immature osteoblasts. We then transferred small doses (104–108 PFU) of lacZ-expressing adenovirus vector into immature osteoblasts on day 14. β-galactosidase was detected only on the immature osteoblasts at every dose. Immature osteoblasts play an important role in the matrix replacement step in fracture healing. CAR-mediated gene transfer into immature osteoblasts can be reasonable for adenovirus-mediated treatment of fracture healing.

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Authors and Affiliations

  1. Division of Orthopedic Surgery, Department of Regenerative and Transplant Medicine, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

    T Ito, K Tokunaga, H Kawashima, H Kitahara, T Horikoshi, A Ogose & N Endo

  2. Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan

    H Maruyama

  3. Research Laboratory for Molecular Genetics, Niigata University, Niigata, Japan

    Y Hotta & R Kuwano

  4. Division of Molecular Metabolism and Diabetes, Department of Internal Medicine, Tohoku University Graduate School of Medicine, Tohoku, Japan

    H Katagiri

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  1. T Ito
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  2. K Tokunaga
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  3. H Maruyama
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  4. H Kawashima
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  5. H Kitahara
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  6. T Horikoshi
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  7. A Ogose
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  8. Y Hotta
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  9. R Kuwano
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  10. H Katagiri
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  11. N Endo
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Ito, T., Tokunaga, K., Maruyama, H. et al. Coxsackievirus and adenovirus receptor (CAR)-positive immature osteoblasts as targets of adenovirus-mediated gene transfer for fracture healing. Gene Ther 10, 1623–1628 (2003). https://doi.org/10.1038/sj.gt.3302060

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  • DOI: https://doi.org/10.1038/sj.gt.3302060

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