Chromosomal integration of adenoviral vector DNA in vivo.
So far there has been no report of any clinical or preclinical evidence for chromosomal vector integration following adenovirus (Ad) vector-mediated gene transfer in vivo. We used liver gene transfer with high-capacity Ad vectors in the FAH(Deltaexon5) mouse model to analyze homologous and heterologous recombination events between vector and chromosomal DNA. Intravenous injection of Ad vectors either expressing a fumarylacetoacetate hydrolase (FAH) cDNA or carrying part of the FAH genomic locus resulted in liver nodules of FAH-expressing hepatocytes, demonstrating chromosomal vector integration. Analysis of junctions between vector and chromosomal DNA following heterologous recombination indicated integration of the vector genome through its termini. Heterologous recombination occurred with a median frequency of 6.72 x 10(-5) per transduced hepatocyte, while homologous recombination occurred more rarely with a median frequency of 3.88 x 10(-7). This study has established quantitative and qualitative data on recombination of adenoviral vector DNA with genomic DNA in vivo, contributing to a risk-benefit assessment of the biosafety of Ad vector-mediated gene transfer.
Stephen
Sam Laurel
SL
Center for Molecular Medicine, University of Cologne, 50931 Cologne, Germany.
Montini
Eugenio
E
Sivanandam
Vijayshankar Ganesh
VG
Al-Dhalimy
Muhseen
M
Kestler
Hans A
HA
Finegold
Milton
M
Grompe
Markus
M
Kochanek
Stefan
S
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002875:Chromosomes / Q000235:genetics* / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000821:virology; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D016662:Virus Integration / Q000235:genetics*
D017931:DNA Primers; D004279:DNA, Viral; D011994:Recombinant Proteins; D006867:Hydrolases; C020156:fumarylacetoacetase