Biodistribution and retargeting of FX-binding ablated adenovirus serotype 5 vectors.
A major limitation for adenoviral transduction in vivo is the profound liver tropism of adenovirus type 5 (Ad5). Recently, we demonstrated that coagulation factor X (FX) binds to Ad5-hexon protein at high affinity to mediate hepatocyte transduction after intravascular delivery. We developed novel genetically FX-binding ablated Ad5 vectors with lower liver transduction. Here, we demonstrate that FX-binding ablated Ad5 predominantly localize to the liver and spleen 1 hour after injection; however, they had highly reduced liver transduction in both control and macrophage-depleted mice compared with Ad5. At high doses in macrophage-depleted mice, FX-binding ablated vectors transduced the spleen more efficiently than Ad5. Immunohistochemical studies demonstrated transgene colocalization with CD11c(+), ER-TR7(+), and MAdCAM-1(+) cells in the splenic marginal zone. Systemic inflammatory profiles were broadly similar between FX-binding ablated Ad5 and Ad5 at low and intermediate doses, although higher levels of several inflammatory proteins were observed at the highest dose of FX-binding ablated Ad5. Subsequently, we generated a FX-binding ablated virus containing a high affinity Ad35 fiber that mediated a significant improvement in lung/liver ratio in macrophage-depleted CD46(+) mice compared with controls. Therefore, this study documents the biodistribution and reports the retargeting capacity of FX binding-ablated Ad5 vectors in vitro and in vivo.
Alba
Raul
R
British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow, UK.
Bradshaw
Angela C
AC
Coughlan
Lynda
L
Denby
Laura
L
McDonald
Robert A
RA
Waddington
Simon N
SN
Buckley
Suzanne M K
SM
Greig
Jenny A
JA
Parker
Alan L
AL
Miller
Ashley M
AM
Wang
Hongjie
H
Lieber
Andre
A
van Rooijen
Nico
N
McVey
John H
JH
Nicklin
Stuart A
SA
Baker
Andrew H
AH
D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D018925:Chemokines / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005170:Factor X / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D008099:Liver / Q000378:metabolism / Q000821:virology; D008168:Lung / Q000378:metabolism / Q000821:virology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012703:Serotyping; D013154:Spleen / Q000378:metabolism / Q000821:virology; D013997:Time Factors; D014018:Tissue Distribution; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D036022:Capsid Proteins; D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators; D011994:Recombinant Proteins; C018315:hexon capsid protein, Adenovirus; D005170:Factor X; D001616:beta-Galactosidase