DNA from both high-capacity and first-generation adenoviral vectors remains intact in skeletal muscle.
Previous studies of the use of adenoviral vectors in animal models of gene therapy have focused on the immune response against transduced cells as the major limiting factor to long-term transgene expression. In this study we eliminated the variable of immunity induced by expression of the transgene in order to investigate vector DNA stability of both first-generation and high-capacity adenoviral vectors after gene transfer to skeletal muscle. Transgene expression from a high-capacity adenoviral vector remained at a high level for at least 20 weeks and was accompanied by persistence of intact vector genomes. In contrast, transgene expression from a first-generation adenoviral vector markedly diminished by 6 weeks after gene transfer and was accompanied by mild and variable inflammatory cell infiltrates. Surprisingly, despite this loss of transgene expression, the first-generation adenoviral vector genomes persisted like the high-capacity adenoviral vector genomes. Therefore, in the absence of immunity to transgene proteins, loss of expression from the first-generation vector was due to inhibition of transgene expression rather than to the elimination of vector-containing cells. DNA stability and persistent expression of the high-capacity adenoviral vector supports the potential of this vector for clinical applications of muscle gene transfer.
Department of Neurology, University of Pittsburgh, PA 15213, USA.
H H
HH
Chen
L M
LM
Mack
S Y
SY
Choi
M
M
Ontell
S
S
Kochanek
P R
PR
Clemens
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D003587:Cytomegalovirus / Q000235:genetics; D016189:Dystrophin / Q000235:genetics; D004801:Eosine Yellowish-(YS) / Q000032:analysis; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006416:Hematoxylin / Q000032:analysis; D008264:Macrophages / Q000276:immunology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000378:metabolism*; D009132:Muscles / Q000737:chemistry; D020079:Terminal Repeat Sequences / Q000235:genetics; D013997:Time Factors; D001616:beta-Galactosidase / Q000737:chemistry
D016189:Dystrophin; D001616:beta-Galactosidase; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin