Binding of adenoviral fiber knob to the coxsackievirus-adenovirus receptor is crucial for transduction of fetal muscle.
Adenoviral (Ad) infection involves attachment mediated by the Ad fiber protein binding to the coxsackievirus-adenovirus receptor (CAR) of a target cell and internalization facilitated by the interaction of the Ad penton base protein with alpha(v) integrins. To understand the relative importance of the Ad binding and internalization steps for the transduction of fetal skeletal muscle, we used a panel of genetically modified vectors that specifically ablate the fiber-CAR interaction (AdL.F*), the penton base-alpha(v) integrin interaction (AdL.PB*), or both (AdL.PB*F*) to transduce embryonic day 16 (E-16) mouse muscle in vivo and primary E-16 muscle cells in vitro. Quantification of transgene expression and vector genome copies revealed a striking absence of E-16 muscle transduction by AdL.F* and AdL.PB*F*. In contrast, fetal muscle transduction with AdL.PB* was not significantly different than with the unmodified vector. Similar results were observed with in vitro Ad infection studies in primary E-16 muscle cells. From these data we conclude that the fiber-CAR interaction is important for the transduction of fetal muscle by Ad vectors. The high dependence on fiber-CAR binding will impact the development of strategies for Ad vector retargeting to achieve muscle-specific transduction in utero.
Bilbao
Roberto
R
Department of Neurology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
Srinivasan
Suchitra
S
Reay
Daniel
D
Goldberg
Laura
L
Hughes
Tiffany
T
Roelvink
Peter W
PW
Einfeld
David A
DA
Wickham
Thomas J
TJ
Clemens
Paula R
PR
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001665:Binding Sites; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D003673:Defective Viruses / Q000235:genetics; D005260:Female; D005333:Fetus; D005822:Genetic Vectors; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000196:embryology*; D009154:Mutation; D032448:Myoblasts, Skeletal / Q000378:metabolism*; D011247:Pregnancy; D011991:Receptors, Virus / Q000378:metabolism*; D014161:Transduction, Genetic
C568772:CLMP protein, mouse; D036022:Capsid Proteins; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011991:Receptors, Virus; C018315:hexon capsid protein, Adenovirus