A novel strategy to modify adenovirus tropism and enhance transgene delivery to activated vascular endothelial cells in vitro and in vivo.
To assess the possibilities of retargeting adenovirus to activated endothelial cells, we conjugated bifunctional polyethylene glycol (PEG) onto the adenoviral capsid to inhibit the interaction between viral knob and coxsackie-adenovirus receptor (CAR). Subsequently, we introduced an alphav integrin-specific RGD peptide or E-selectin-specific antibody to the other functional group of the PEG molecule for the retargeting of the adenovirus to activated endothelial cells. In vitro studies showed that this approach resulted in the elimination of transgene transfer into CAR-positive cells, while at the same time specific transgene transfer to activated endothelial cells was achieved. PEGylated, retargeted adenovirus showed longer persistence in the blood circulation with area under plasma concentration-time curve (AUC) values increasing 12-fold compared to unmodified virus. Anti-E-selectin antibody-PEG-adenovirus selectively homed to inflamed skin in mice with a delayed-type hypersensitivity (DTH) inflammation, resulting in local expression of the reporter transgene luciferase. This is the first study showing the benefits of PEGylation on adenovirus behavior upon systemic administration. The approach described here can form the basis for further development of adenoviral gene therapy vectors with improved pharmacokinetics and increased efficiency and specificity of therapeutic gene transfer into endothelial cells in disease.
Ogawara
Ken-ichi
K
Department of Pharmacokinetics, Groningen University Institute for Drug Exploration, Groningen, The Netherlands.
Rots
Marianne G
MG
Kok
Robbert J
RJ
Moorlag
Henk E
HE
Van Loenen
Anne-Miek
AM
Meijer
Dirk K F
DK
Haisma
Hidde J
HJ
Molema
Grietje
G
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D019540:Area Under Curve; D002478:Cells, Cultured; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004730:Endothelium, Vascular; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009842:Oligopeptides; D011092:Polyethylene Glycols / Q000737:chemistry; D011991:Receptors, Virus; D019076:Transgenes; D018523:Tropism; D014471:Umbilical Veins / Q000166:cytology
C568771:CLMP protein, human; C568772:CLMP protein, mouse; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D009842:Oligopeptides; D011991:Receptors, Virus; D011092:Polyethylene Glycols; C047981:arginyl-glycyl-aspartic acid; D008156:Luciferases