TY - JOUR
T1 - Preclinical in vivo evaluation of pseudotyped adeno-associated virus vectors for liver gene therapy
AU - Grimm, Dirk
AU - Zhou, Shangzhon
AU - Nakai, Hiroyuki
AU - Thomas, Clare E.
AU - Storm, Theresa A.
AU - Fuess, Sally
AU - Matsushita, Takashi
AU - Allen, James
AU - Surosky, Richard
AU - Lochrie, Michael
AU - Meuse, Leonard
AU - McClelland, Alan
AU - Colosi, Peter
AU - Kay, Mark A.
PY - 2003/10/1
Y1 - 2003/10/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=0141482003&partnerID=8YFLogxK
U2 - 10.1182/blood-2003-02-0495
DO - 10.1182/blood-2003-02-0495
M3 - Article
C2 - 12791653
AN - SCOPUS:0141482003
SN - 0006-4971
VL - 102
SP - 2412
EP - 2419
JO - Blood
JF - Blood
IS - 7
ER -