Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor promotes collateral development in a rabbit model of hind limb ischemia

Gene Therapy volume 8, pages 837–845 (2001)Cite this article

Abstract

Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor (bFGF), a new strategy for the treatment of chronic vascular occlusive disease, was examined in a rabbit model of hind limb ischemia. The left femoral artery was completely excised to induce an ischemic state in the hind limb of male rabbits. Simultaneously, a skin section was resected from the wound, and host fibroblasts were cultured. The cultured fibroblasts were infected with adenovirus vector containing modified human bFGF cDNA with the secretory signal sequence (AxCAMAssbFGF) or LacZ cDNA (AxCALacZ). At 21 days after femoral artery excision, the gene-transduced fibroblasts were administered through the left internal iliac artery. The fibroblasts significantly accumulated in the ischemic hind limb, and the AxCAMAssbFGF-treated cells secreted bFGF for less than 14 days without elevation of systemic bFGF level. At 28 days after cell administration, calf blood pressure ratio, angiographic score, capillary density of muscle tissue and blood flow of the left internal iliac artery were determined, and animals with AxCAMAssbFGF-treated cells showed significantly greater development of collateral vessels, as compared with those with AxCALacZ-treated cells. These findings suggest that adenovirus-mediated ex vivo gene transfer of bFGF was effective for improvement of chronic limb ischemia.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

References

  1. Sasayama S, Fujita M . Recent insight into coronary collateral circulation Circulation 1992 85: 1197–1204

    Article  CAS  Google Scholar 

  2. Carmeliet P . Mechanisms of angiogenesis and arteriogenesis Nat Med 2000 6: 389–395

    Article  CAS  Google Scholar 

  3. Hockel M et al. Therapeutic angiogenesis Arch Surg 1993 128: 423–429

    Article  CAS  Google Scholar 

  4. Takeshita S et al. Therapeutic angiogenesis. A single intra-arterial bolus of vascular endothelial growth factor augments revascularization in a rabbit ischemic hind limb model J Clin Invest 1994 93: 662–670

    Article  CAS  Google Scholar 

  5. Unger EF et al. Basic fibroblast growth factor enhances myocardial collateral flow in a canine model Am J Physiol 1994 266: H1588–H1595

    CAS  PubMed  Google Scholar 

  6. Feldman LJ et al. Low-efficacy of percutaneous adenovirus-mediated arterial gene transfer in the atherosclerotic rabbit J Clin Invest 1995 95: 2662–2671

    Article  CAS  Google Scholar 

  7. Tsurumi Y et al. Direct intramuscular gene transfer of naked DNA encoding vascular endothelial growth factor augments collateral development and tissue perfusion Circulation 1996 94: 3281–3290

    Article  CAS  Google Scholar 

  8. Bauters C et al. Physiological assessment of augmented vascularity induced by VEGF in ischemic rabbit hindlimb Am J Physiol 1994 267: H1263–H1271

    CAS  Google Scholar 

  9. Abraham JA et al. Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor Science 1986 233: 545–548

    Article  CAS  Google Scholar 

  10. Sasada R, Seno M, Watanabe T, Igarashi K . Expression of modified bFGF cDNAs in mammalian cells Ann NY Acad Sci 1991 638: 149–160

    Article  CAS  Google Scholar 

  11. Kass-Eisler A et al. Quantitative determination of adenovirus-mediated gene delivery to rat cardiac myocytes in vitro and in vivo Proc Natl Acad Sci USA 1993 90: 11498–11502

    Article  CAS  Google Scholar 

  12. Arras M et al. The delivery of angiogenic factors to the heart by microsphere therapy Nat Biotechnol 1998 16: 159–162

    Article  CAS  Google Scholar 

  13. Friesel RE, Maciag T . Molecular mechanisms of angiogenesis: fibroblast growth factor signal transduction FASEB J 1995 9: 919–925

    Article  CAS  Google Scholar 

  14. Losordo DW et al. Gene therapy for myocardial angiogenesis: initial clinical results with direct myocardial injection of phVFGF165 as sole therapy for myocardial ischemia Circulation 1998 98: 2800–2804

    Article  CAS  Google Scholar 

  15. Wilson JM . Adenoviruses as gene-delivery vehicles New Engl J Med 1996 334: 1185–1187

    Article  CAS  Google Scholar 

  16. Yang Y et al. Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy Proc Natl Acad Sci USA 1994 91: 4407–4411

    Article  CAS  Google Scholar 

  17. Baumgartner I et al. Constitutive expression of phVEGF165 after intramuscular gene transfer promotes collateral vessel development in patients with critical limb ischemia Circulation 1998 97: 1114–1123

    Article  CAS  Google Scholar 

  18. Risau W . What, if anything, is an angiogenic factor? Cancer Metast Rev 1996 15: 149–151

    Article  CAS  Google Scholar 

  19. Ozaki H et al. Basic fibroblast growth factor is neither necessary nor sufficient for the development of retinal neovascularization Am J Pathol 1998 153: 757–765

    Article  CAS  Google Scholar 

  20. Gowdak LHW et al. Adenovirus-mediated VEGF121 gene transfer stimulates angiogenesis in normoperfused skeletal muscle and preserves tissue perfusion after induction of ischemia Circulation 2000 102: 565–571

    Article  CAS  Google Scholar 

  21. Wu HM, Yuan Y, McCarthy M, Granger HJ . Acidic and basic FGFs dilate arterioles of skeletal muscle through a NO-dependent mechanism Am J Physiol 1996 271: H1087–H1094

    Article  CAS  Google Scholar 

  22. Miyake S et al. Effective generation of recombinant adenoviruses using adenovirus DNA–terminal protein complex and a cosmid bearing the full-length virus genome Proc Natl Acad Sci USA 1996 93: 1320–1324

    Article  CAS  Google Scholar 

  23. Akimoto M et al. Adenovirally expressed basic fibroblast growth factor rescues photoreceptor cells in RCS rats Invest Ophthalmol Vis Sci 1999 40: 273–279

    CAS  PubMed  Google Scholar 

  24. Kurokawa T, Sasada R, Iwane M, Igarashi K . Cloning and expression of cDNA encoding human basic fibroblast growth factor FEBS 1987 213: 189–194

    Article  CAS  Google Scholar 

  25. Takahashi JC et al. Adenovirus-mediated gene transfer of basic fibroblast growth factor induces in vitro angiogenesis Atherosclerosis 1997 132: 199–205

    Article  CAS  Google Scholar 

  26. Koyama H, Reidy MA . Reinjury of arterial lesions induces intimal smooth muscle cell replication that is not controlled by fibroblast growth factor 2 Circ Res 1997 80: 408–417

    Article  CAS  Google Scholar 

  27. Pu LQ et al. A persistent hind limb ischemia model in the rabbit J Invest Surg 1994 7: 49–60

    Article  CAS  Google Scholar 

  28. Kardami E, Fandrich RR . Basic fibroblast growth factor in atria and ventricles of the vertebrate heart J Cell Biol 1989 109: 1865–1875

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by a Grant-in Aid for Scientific Research (C) from the Ministry of Education, Science, and Culture of Japan (10671103). The studies using 111In and 51Cr were carried out at Radioisotope Center, The University of Tokyo.

Author information

Author notes

  1. N Ohara and H Koyama: The first two authors contributed equally to this paper

Authors and Affiliations

  1. Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    N Ohara, H Koyama, T Miyata & H Shigematsu

  2. Department of Molecular Medicine, Sapporo Medical University, Hokkaido, Japan

    H Hamada

  3. Department of Neurosurgery, Osaka Medical College, Osaka, Japan

    S-I Miyatake

  4. Department of Ophthalmology, Shinshu University, Nagano, Japan

    M Akimoto

Authors
  1. N Ohara
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H Koyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T Miyata
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H Hamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S-I Miyatake
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M Akimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H Shigematsu
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ohara, N., Koyama, H., Miyata, T. et al. Adenovirus-mediated ex vivo gene transfer of basic fibroblast growth factor promotes collateral development in a rabbit model of hind limb ischemia. Gene Ther 8, 837–845 (2001). https://doi.org/10.1038/sj.gt.3301475

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301475

Keywords

This article is cited by

Search

Advanced search

Quick links