Targeting adenovirus gene delivery to activated tumour-associated vasculature via endothelial selectins.
Clinical experience with adenovirus vectors has highlighted the need for improved delivery and targeting. Tumour-associated endothelium offers an additional mechanism for enhanced viral uptake into tumours which is accessible for systemic gene delivery. Building on expertise in using polymer 'stealthed' viruses for targeting in vivo, adenovirus expressing luciferase (Adluc) was coated with an amino-reactive polymer based on poly [N-(2-hydroxypropyl) methacrylamide] to ablate normal infection pathways. Direct linkage of a monoclonal antibody against E-selectin (MHES) demonstrated E-selectin-specific transduction of tumour necrosis factor-α (TNF-α)-activated endothelial cells. A two-component targeting system using protein G was developed, to provide optimal antibody orientation. We report an enhancement in transduction of TNF-α-activated endothelium in vitro and ex vivo in a human umbilical vein cord model using the MHES antibody. Similarly a virus retargeted using a chimeric P-selectin Glycoprotein Ligand-1-Fc fusion (PSGL-1) protein showed better circulation kinetics and significant uptake into HepG2 xenografts following systemic administration in mice, with 36-fold higher genome copies, compared with non-modified virus. Immunohistochemistry staining of tumour sections from mice treated with PSGL-1-retargeted virus showed a co-localisation of firefly luciferase with CD31 suggesting selective endothelial targeting. Employment of optimal viral modification using protein G will enable exploration and comparison of alternative targeting ligands targeting tumour-associated endothelium.
Bachtarzi
Houria
H
Department of Clinical Pharmacology, University of Oxford, Old Road Campus, Headington, Oxford OX3 7DQ, UK. houria.bachtarzi@clinpharm.ox.ac.uk
Stevenson
Mark
M
Šubr
Vladimir
V
Ulbrich
Karel
K
Seymour
Leonard W
LW
Fisher
Kerry D
KD
D000178:Acrylamides / Q000378:metabolism; D000256:Adenoviridae / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D019040:E-Selectin / Q000276:immunology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005260:Female; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D049409:Luciferases, Firefly / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology / Q000628:therapy* / Q000821:virology; D019007:P-Selectin / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D019087:Selectins / Q000276:immunology* / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014470:Umbilical Cord / Q000187:drug effects / Q000378:metabolism / Q000821:virology; D019562:Viral Load; D023041:Xenograft Model Antitumor Assays
D000178:Acrylamides; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D019040:E-Selectin; C034284:IgG Fc-binding protein, Streptococcus; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D019087:Selectins; D014409:Tumor Necrosis Factor-alpha; D049409:Luciferases, Firefly; C032976:N-(2-hydroxypropyl)methacrylamide