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Successful treatment of canine leukocyte adhesion deficiency by foamy virus vectors

Nature Medicine volume 14, pages 93–97 (2008)Cite this article

Abstract

Recent successes in treating genetic immunodeficiencies have demonstrated the therapeutic potential of stem cell gene therapy1,2,3,4. However, the use of gammaretroviral vectors in these trials led to insertional activation of nearby oncogenes and leukemias in some study subjects, prompting studies of modified or alternative vector systems5. Here we describe the use of foamy virus vectors to treat canine leukocyte adhesion deficiency (CLAD). Four of five dogs with CLAD that received nonmyeloablative conditioning and infusion of autologous, CD34+ hematopoietic stem cells transduced by a foamy virus vector expressing canine CD18 had complete reversal of the CLAD phenotype, which was sustained more than 2 years after infusion. In vitro assays showed correction of the lymphocyte proliferation and neutrophil adhesion defects that characterize CLAD. There were no genotoxic complications, and integration site analysis showed polyclonality of transduced cells and a decreased risk of integration near oncogenes as compared to gammaretroviral vectors. These results represent the first successful use of a foamy virus vector to treat a genetic disease, to our knowledge, and suggest that foamy virus vectors will be effective in treating human hematopoietic diseases.

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Figure 1: In vitro analysis of foamy virus vector ΔΦMscvCD18.
Figure 2: Expression and function of transduced CLAD peripheral blood cells after infusion.
Figure 3: Correction of the CLAD phenotype in vivo.
Figure 4: Integration sites in FV vector–treated dogs.

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Acknowledgements

We thank W. Telford and V. Kapoor for assistance with flow cytometry, A. Sowers for assistance with nonmyeloablative irradiation, J. Taylor for advice on quantitative PCR and X. Wu for assistance with insertion site analysis programs. This work was supported by the Intramural Research Program of the US National Institutes of Health, National Cancer Institute, and Center for Cancer Research and by US National Institutes of Health grant HL53750 to D.W.R.

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Author notes

  1. David W Russell and Dennis D Hickstein: These authors contributed equally to this work.

Authors and Affiliations

  1. Experimental Transplantation and Immunology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 10 Center Drive, MSC1203, Building 10-CRC, Room 3-3330, Bethesda, 20892, Maryland, USA

    Thomas R Bauer Jr, Mehreen Hai, Laura M Tuschong, Rima L Adler, Yu-chen Gu & Dennis D Hickstein

  2. Division of Hematology, Department of Medicine, University of Washington, 1959 Northeast Pacific Street, Box 357720, Seattle, 98195, Washington, USA

    James M Allen, Iram F Khan, Erik M Olson & David W Russell

  3. Division of Veterinary Resources, Office of Research Services, National Institutes of Health, 9000 Rockville Pike, Building 28, Room 104, Bethesda, Maryland 20892, USA.,

    Tanya H Burkholder

  4. Department of Biochemistry, University of Washington, 1959 Northeast Pacific Street, Box 357720, Seattle, 98195, Washington, USA

    David W Russell

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  1. Thomas R Bauer Jr
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Contributions

T.R.B. designed experiments, performed lymphocyte and neutrophil function assays, performed transductions, analyzed all data and wrote the manuscript. J.M.A. and E.M.O. prepared and characterized foamy virus vector stocks. M.H. performed LAM-PCR studies and LM-PCR studies and participated in the CD34 stem cell collection. L.M.T. collected and analyzed flow cytometry data, participated in CD34 stem cell collection and reinfusion, performed LM-PCR cloning and analysis and participated in clinical care of dogs. I.F.K. performed quantitative PCR. R.L.A. performed LAM-PCR and LM-PCR studies. T.H.B. was responsible for clinical care of dogs. Y.-c.G. participated in CD34 stem cell collection. D.W.R. designed experiments, supervised experimentation, coordinated the foamy virus vector production and wrote the manuscript. D.D.H. designed experiments, supervised experimentation, coordinated the project and wrote the manuscript.

Corresponding authors

Correspondence to David W Russell or Dennis D Hickstein.

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Supplementary Figs. 1–3, Supplementary Tables 1–4 and Supplementary Methods (PDF 3069 kb)

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Bauer, T., Allen, J., Hai, M. et al. Successful treatment of canine leukocyte adhesion deficiency by foamy virus vectors. Nat Med 14, 93–97 (2008). https://doi.org/10.1038/nm1695

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