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Sustained expression of human factor VIII in mice using a parvovirus-based vector.
1UNC Gene Therapy Center, and Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Blood
|February 26, 2000
Summary
This study demonstrates sustained expression of human factor VIII (hFVIII) in mice using recombinant adeno-associated virus (rAAV) vectors. This gene therapy approach shows promise for treating hemophilia A by enabling liver cells to produce functional hFVIII.
Area of Science:
- Gene Therapy
- Hematology
- Molecular Biology
Background:
- Hemophilia A treatment requires persistent therapeutic levels of human factor VIII (hFVIII).
- Recombinant adeno-associated virus (rAAV) vectors offer potential for stable gene transduction in target tissues like the liver.
Purpose of the Study:
- To investigate sustained hFVIII expression in immunocompetent mice using rAAV vectors.
- To evaluate the efficacy of liver-specific transgene expression linked to B-domain-deleted hFVIII (BDD-hFVIII) cDNA.
Main Methods:
- Generated rAAV/BDD-hFVIII virion particles using a transfection scheme that eliminates adenovirus.
- Transduced cell lines (293 and HepG2) and performed in vivo experiments in C57BL/6 and NOD/scid mice via portal vein injection.
- Assessed hFVIII production using Coatest and APTT assays, and detected transgene mRNA in the liver.
Main Results:
- Functional BDD-hFVIII protein was produced in transduced cells.
- NOD/scid mice achieved up to 27% of normal human plasma hFVIII levels, while C57BL/6 mice developed anti-hFVIII antibodies.
- Transgene mRNA was primarily detected in the liver with no observed pathologic abnormalities.
Conclusions:
- Sustained hFVIII expression was achieved in mice using rAAV vectors.
- This approach demonstrates potential for hemophilia A treatment by enabling liver-based hFVIII production.
- Further research is warranted to optimize delivery and immune response mitigation for clinical application.
