Preclinical in vivo evaluation of pseudotyped adeno-associated virus vectors for liver gene therapy

Blood. 2003 Oct 1;102(7):2412-9. doi: 10.1182/blood-2003-02-0495. Epub 2003 Jun 5.


We report the generation and use of pseudotyped adeno-associated viral (AAV) vectors for the liver-specific expression of human blood coagulation factor IX (hFIX). Therefore, an AAV-2 genome encoding the hfIX gene was cross-packaged into capsids of AAV types 1 to 6 using efficient, large-scale technology for particle production and purification. In immunocompetent mice, the resultant vector particles expressed high hFIX levels ranging from 36% (AAV-4) to more than 2000% of normal (AAV-1, -2, and -6), which would exceed curative levels in patients with hemophilia. Expression was dose- and time-dependent, with AAV-6 directing the fastest and strongest onset of hFIX expression at all doses. Interestingly, systemic administration of 2 x 1012 vector particles of AAV-1, -4, or -6 resulted in hFIX levels similar to those achieved by portal vein delivery. For all other serotypes and particle doses, hepatic vector administration yielded up to 84-fold more hFIX protein than tail vein delivery, corroborated by similarly increased vector DNA copy numbers in the liver, and elicited a reduced immune response against the viral capsids. Finally, neutralization assays showed variable immunologic cross-reactions between most of the AAV serotypes. Our technology and findings should facilitate the development of AAV pseudotype-based gene therapies for hemophilia B and other liver-related diseases.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Antibodies / blood
  • Capsid
  • Carcinoma, Hepatocellular
  • DNA, Viral / pharmacokinetics
  • Factor IX / genetics*
  • Gene Dosage
  • Gene Expression Regulation, Viral
  • Genetic Therapy / methods*
  • Genetic Vectors / administration & dosage*
  • Genetic Vectors / immunology
  • Genetic Vectors / pharmacokinetics
  • Hemophilia B / therapy*
  • Humans
  • Kidney / cytology
  • Liver Diseases / therapy*
  • Liver Neoplasms
  • Mice
  • Mice, Inbred C57BL
  • Promoter Regions, Genetic
  • Tissue Distribution
  • Tumor Cells, Cultured


  • Antibodies
  • DNA, Viral
  • Factor IX