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

Dirk Grimm, Shangzhon Zhou, Hiroyuki Nakai, Clare E. Thomas, Theresa A. Storm, Sally Fuess, Takashi Matsushita, James Allen, Richard Surosky, Michael Lochrie, Leonard Meuse, Alan McClelland, Peter Colosi, Mark A. Kay

Research output: Contribution to journalArticlepeer-review

189 Scopus citations


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 × 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.

Original languageEnglish (US)
Pages (from-to)2412-2419
Number of pages8
Issue number7
StatePublished - Oct 1 2003
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology


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