title: Adenovirus serotype 5 fiber shaft influences in vivo gene transfer in mice.
abstract: Adenoviral vectors used in gene therapy are predominantly derived from adenovirus serotype 5 (Ad5), which infects a broad range of cells. Ad5 cell entry involves interactions with the coxsackie-adenovirus receptor (CAR) and integrins. To assess these receptors in vivo, we mutated amino acid residues in fiber and penton that are involved in receptor interaction and showed that CAR and integrins play a minor role in hepatic transduction but that integrins can influence gene delivery to other tissues. These data suggest that an alternative entry pathway exists for hepatocyte transduction in vivo that is more important than CAR or integrins. In vitro data suggest a role for heparan sulfate glycosaminoglycans (HSG) in adenovirus transduction. The role of the fiber shaft in liver uptake was examined by introducing specific amino acid changes into a putative HSG-binding motif contained within the shaft or by preparing fiber shaft chimeras between Ad5 and Ad35 fibers. Results were obtained that demonstrate that the Ad5 fiber shaft can influence gene transfer both in vitro and to the liver in vivo. These observations indicate that the currently accepted two-step entry pathway, which involves CAR and integrins, described for adenoviral infection in vitro, is not used for hepatic gene transfer in vivo. In contrast, alpha(v) integrins influence gene delivery to the lung, spleen, heart, and kidney. The detargeted vector constructs described here may provide a foundation for the development of targeted adenoviral vectors.
authors.affiliation: Genetic Therapy, Inc., a Novartis Company, Gaithersburg, MD 20878, USA.
authors.forename: Theodore A G
authors.initials: TA
authors.lastname: Smith
authors.forename: Neeraja
authors.initials: N
authors.lastname: Idamakanti
authors.forename: Michele L
authors.initials: ML
authors.lastname: Rollence
authors.forename: Jennifer
authors.initials: J
authors.lastname: Marshall-Neff
authors.forename: Jin
authors.initials: J
authors.lastname: Kim
authors.forename: Kathy
authors.initials: K
authors.lastname: Mulgrew
authors.forename: Glen R
authors.initials: GR
authors.lastname: Nemerow
authors.forename: Michael
authors.initials: M
authors.lastname: Kaleko
authors.forename: Susan C
authors.initials: SC
authors.lastname: Stevenson
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D020816:Amino Acid Motifs; D000818:Animals; D015153:Blotting, Western; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005822:Genetic Vectors / Q000008:administration & dosage / Q000097:blood / Q000493:pharmacokinetics*; D006025:Glycosaminoglycans / Q000502:physiology; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D011991:Receptors, Virus / Q000378:metabolism; D014161:Transduction, Genetic; D014771:Virion / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: C568771:CLMP protein, human; C568772:CLMP protein, mouse; D036022:Capsid Proteins; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D006025:Glycosaminoglycans; D016023:Integrins; D009842:Oligopeptides; D011991:Receptors, Virus; C018315:hexon capsid protein, Adenovirus; C043322:penton protein, adenovirus; C047981:arginyl-glycyl-aspartic acid; D001616:beta-Galactosidase