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Persistent expression of canine factor IX in hemophilia B canines

Gene Therapy volume 6, pages 1695–1704 (1999)Cite this article

Abstract

We previously demonstrated that direct intramuscular injection of recombinant adeno-associated virus (rAAV) carrying the human FIX (hFIX) cDNA can safely be administered to hemophilic B canines and express human factor IX protein; however, the functional activity of the hFIX protein could not be assessed due to anti-human FIX antibody (inhibitor) formation. To test the therapeutic efficacy of rAAV in hemophilic dogs, rAAV type 2 (rAAV2) carrying canine FIX (cFIX) cDNA was injected into the skeletal muscle of two dogs at doses of 1012–13particles. Circulating cFIX protein levels were maintained for 1 year at levels of 1–2% of normal. Hemostatic correction (WBCT and APTT) paralleled plasma FIX antigen levels. Both dogs still required plasma infusion for spontaneous and traumatic bleeding events. Inhibitors to cFIX protein were not detected in either animal by Bethesda assay. Neutralizing antibodies directed against AAV-2 capsid were pronounced and persistent. Vector DNA and mRNA transcripts were detected only at the injected skeletal muscle tissue. Analysis of both high and low molecular weight DNA identified both replicative episomal and integrated AAV species. These results demonstrate that persistent secretion of the FIX transgene protein, necessary for successful gene therapy of hemophilia B, can be achieved using the parvovirus-based rAAV vector

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Acknowledgements

We would like to thank Dr Robert Bagnell for his technical assistance, Ms Pam McElveen, Dan Ewell and the staff at the Francis Owen Blood Research Laboratory for their excellent care and handling of the animals, and the UNC Vector Core for helpful discussions. PEM is a Judith Graham Pool Fellow of the National Hemophilia Foundation. CEW is a recipient of the Lucille B Markey Charitable Trust.

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Authors and Affiliations

  1. UNC Gene Therapy Center, University of North Carolina, Chapel Hill, NC, USA

    H Chao, R J Samulski, P E Monahan & C E Walsh

  2. Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC, USA

    D A Bellinger & T C Nichols

  3. Department of Pediatrics, University of North Carolina, Chapel Hill, NC, USA

    P E Monahan

  4. Department of Medicine, University of North Carolina, Chapel Hill, NC, USA

    T C Nichols & C E Walsh

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  1. H Chao
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Chao, H., Samulski, R., Bellinger, D. et al. Persistent expression of canine factor IX in hemophilia B canines. Gene Ther 6, 1695–1704 (1999). https://doi.org/10.1038/sj.gt.3301024

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