Chromosomal Integration of Adenoviral Vector DNA In Vivo

Abstract

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 ^Δexon5 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 × 10 ⁻⁵ per transduced hepatocyte, while homologous recombination occurred more rarely with a median frequency of 3.88 × 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.