Influence of coagulation factor x on in vitro and in vivo gene delivery by adenovirus (Ad) 5, Ad35, and chimeric Ad5/Ad35 vectors.
The binding of coagulation factor X (FX) to the hexon of adenovirus (Ad) 5 is pivotal for hepatocyte transduction. However, vectors based on Ad35, a subspecies B Ad, are in development for cancer gene therapy, as Ad35 utilizes CD46 (which is upregulated in many cancers) for transduction. We investigated whether interaction of Ad35 with FX influenced vector tropism using Ad5, Ad35, and Ad5/Ad35 chimeras: Ad5/fiber(f)35, Ad5/penton(p)35/f35, and Ad35/f5. Surface plasmon resonance (SPR) revealed that Ad35 and Ad35/f5 bound FX with approximately tenfold lower affinities than Ad5 hexon-containing viruses, and electron cryomicroscopy (cryo-EM) demonstrated a direct Ad35 hexon:FX interaction. The presence of physiological levels of FX significantly inhibited transduction of vectors containing Ad35 fibers (Ad5/f35, Ad5/p35/f35, and Ad35) in CD46-positive cells. Vectors were intravenously administered to CD46 transgenic mice in the presence and absence of FX-binding protein (X-bp), resulting in reduced liver accumulation for all vectors. Moreover, Ad5/f35 and Ad5/p35/f35 efficiently accumulated in the lung, whereas Ad5 demonstrated poor lung targeting. Additionally, X-bp significantly reduced lung genome accumulation for Ad5/f35 and Ad5/p35/f35, whereas Ad35 was significantly enhanced. In summary, vectors based on the full Ad35 serotype will be useful vectors for selective gene transfer via CD46 due to a weaker FX interaction compared to Ad5.
Greig
Jenny A
JA
British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow G12 8TA, UK.
Buckley
Suzanne Mk
SM
Waddington
Simon N
SN
Parker
Alan L
AL
Bhella
David
D
Pink
Rebecca
R
Rahim
Ahad A
AA
Morita
Takashi
T
Nicklin
Stuart A
SA
McVey
John H
JH
Baker
Andrew H
AH
D000256:Adenoviridae / Q000235:genetics* / Q000648:ultrastructure; D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D020285:Cryoelectron Microscopy; D005170:Factor X / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics* / Q000648:ultrastructure; D006801:Humans; D051916:Membrane Cofactor Protein / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D014161:Transduction, Genetic / Q000379:methods*
D051916:Membrane Cofactor Protein; D005170:Factor X