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Long-term correction of hemophilia A mice following lentiviral mediated delivery of an optimized canine factor VIII gene

Gene Therapy volume 24, pages 742–748 (2017)Cite this article

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Abstract

Current therapies for hemophilia A include frequent prophylactic or on-demand intravenous factor treatments which are costly, inconvenient and may lead to inhibitor formation. Viral vector delivery of factor VIII (FVIII) cDNA has the potential to alleviate the debilitating clotting defects. Lentiviral-based vectors delivered to murine models of hemophilia A mediate phenotypic correction. However, a limitation of lentiviral-mediated FVIII delivery is inefficient transduction of target cells. Here, we engineer a feline immunodeficiency virus (FIV) -based lentiviral vector pseudotyped with the baculovirus GP64 envelope glycoprotein to mediate efficient gene transfer to mouse hepatocytes. In anticipation of future studies in FVIII-deficient dogs, we investigated the efficacy of FIV-delivered canine FVIII (cFVIII). Codon-optimization of the cFVIII sequence increased activity and decreased blood loss as compared to the native sequence. Further, we compared a standard B-domain deleted FVIII cDNA to a cDNA including 256 amino acids of the B-domain with 11 potential asparagine-linked oligosaccharide linkages. Restoring a partial B-domain resulted in modest reduction of endoplasmic reticulum (ER) stress markers. Importantly, our optimized vectors achieved wild-type levels of phenotypic correction with minimal inhibitor formation. These studies provide insights into optimal design of a therapeutically relevant gene therapy vector for a devastating bleeding disorder.

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Acknowledgements

We acknowledge the support of the University of Iowa Genomics Division, Viral Vector Core, and Cell Morphology Core. This work was supported by the National Institutes of Health: R44 HL081976 (WCR and PBM) and P01 HL051670 (PBM). Core facilities at the University of Iowa were partially supported by the National Institutes of Health: P01 HL51670, P01 HL091842, and the Center for Gene Therapy for Cystic Fibrosis P30 DK54759.

Author contributions

PBM, WCR, PLS and JMS designed the experiments. JMS, MJP, CGA, MB, DTR and ALC. collected the data. JMS and DTR analyzed the data. JMS and PLS wrote the manuscript. All authors approve the submitted and final versions for publication.

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Authors and Affiliations

  1. Stead Family Department of Pediatrics, Carver College of Medicine, University of Iowa, Iowa City, IA, USA

    J M Staber, M J Pollpeter, P L Sinn & P B McCray

  2. Pappajohn Biomedical Institute, University of Iowa, Iowa City, IA, USA

    J M Staber, M J Pollpeter, A L Cooney, P L Sinn & P B McCray

  3. Department of Virogenics, San Diego, CA, USA

    C-G Anderson, M Burrascano & W C Raschke

  4. Center for Gene Therapy of Cystic Fibrosis and Other Genetic Diseases, Iowa City, IA, USA

    A L Cooney, P L Sinn & P B McCray

  5. Department of Microbiology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA

    A L Cooney & P B McCray

  6. Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, USA

    D T Rutkowski

Authors
  1. J M Staber
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  2. M J Pollpeter
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Corresponding author

Correspondence to J M Staber.

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Competing interests

CGA, MB and WCR are employees of Virogenics, Inc. Janice Staber has received honorarium from Baxalta.

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Staber, J., Pollpeter, M., Anderson, CG. et al. Long-term correction of hemophilia A mice following lentiviral mediated delivery of an optimized canine factor VIII gene. Gene Ther 24, 742–748 (2017). https://doi.org/10.1038/gt.2017.67

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