Adeno-associated viral vector-mediated gene transfer of human blood coagulation factor IX into mouse liver.
Publication/Presentation Date
6-15-1998
Abstract
Recombinant adeno-associated virus vectors (AAV) were prepared in high titer (10(12) to 10(13) particles/mL) for the expression of human factor IX after in vivo transduction of murine hepatocytes. Injection of AAV-CMV-F.IX (expression from the human cytomegalovirus IE enhancer/promoter) into the portal vein of adult mice resulted in no detectable human factor IX in plasma, but in mice injected intravenously as newborns with the same vector, expression was initially 55 to 110 ng/mL. The expression in the liver was mostly transient, and plasma levels decreased to undetectable levels within 5 weeks. However, long-term expression of human F.IX was detected by immunofluorescence staining in 0.25% of hepatocytes 8 to 10 months postinjection. The loss of expression was likely caused by suppression of the CMV promoter, because polymerase chain reaction data showed no substantial loss of vector DNA in mouse liver. A second vector in which F.IX expression was controlled by the human EF1alpha promoter was constructed and injected into the portal vein of adult C57BL/6 mice at a dose of 6.3 x 10(10) particles. This resulted in therapeutic plasma levels (200 to 320 ng/mL) for a period of at least 6 months, whereas no human F.IX was detected in plasma of mice injected with AAV-CMV-F.IX. Doses of AAV-EF1alpha-F. IX of 2.7 x 10(11) particles resulted in plasma levels of 700 to 3, 200 ng/mL. Liver-derived expression of human F.IX from the AAV-EF1alpha-F.IX vector was confirmed by immunofluorescence staining. We conclude that recombinant AAV can efficiently transduce hepatocytes and direct stable expression of an F.IX transgene in mouse liver, but sustained expression is critically dependent on the choice of promoter.
Volume
91
Issue
12
First Page
4600
Last Page
4607
ISSN
0006-4971
Published In/Presented At
Nakai, H., Herzog, R. W., Hagstrom, J. N., Walter, J., Kung, S. H., Yang, E. Y., Tai, S. J., Iwaki, Y., Kurtzman, G. J., Fisher, K. J., Colosi, P., Couto, L. B., & High, K. A. (1998). Adeno-associated viral vector-mediated gene transfer of human blood coagulation factor IX into mouse liver. Blood, 91(12), 4600–4607.
Disciplines
Medicine and Health Sciences | Pediatrics
PubMedID
9616156
Department(s)
Department of Pediatrics
Document Type
Article