Adenoviral gene vector tethering to nanoparticle surfaces results in receptor-independent cell entry and increased transgene expression.
The present studies investigated the hypothesis that affinity immobilization of replication-defective adenoviruses (Ad) on the surfaces of biodegradable nanoparticles (NP) can improve transduction through uncoupling cellular uptake from the coxsackie-adenovirus receptor (CAR). Ad was tethered to the surfaces of polylactide-based NP that were surface-activated using a photoreactive polyallylamine-benzophenone-pyridyldithiocarboxylate polymer, which enabled (via thiol chemistry) the covalent attachment of Ad-binding proteins, either the recombinant D1 domain of CAR or an adenoviral knob-specific monoclonal antibody. Gene transfer by NP-Ad complexes was studied in relation to cellular uptake as a function of cell type and the character of NP-Ad binding. NP-Ad complexes, but not Ad applied with or without control nonimmune IgG-modified NP, significantly increased green fluorescent protein reporter expression in endothelioma and endothelial and arterial smooth muscle cells (SMC) in direct correlation to the extent of NP-Ad internalization. CAR-independent uptake of NP-Ad was confirmed by demonstrating inhibition of free Ad- but not NP-Ad complex-mediated transduction by knob protein. Complexes formulated with an Ad encoding inducible nitric oxide synthase inhibited growth of cultured SMC to a significantly greater extent than those with (GFP)Ad or (NULL)Ad or free vector. It is concluded that Ad-specific affinity tethering to biodegradable NP can significantly increase the level of gene expression via a CAR-independent uptake mechanism.
Chorny
Michael
M
Division of Cardiology, The Children's Hospital of Philadelphia, Philadelphia, PA 19104-4318, USA.
Fishbein
Ilia
I
Alferiev
Ivan S
IS
Nyanguile
Origene
O
Gaster
Richard
R
Levy
Robert J
RJ
D000256:Adenoviridae / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001158:Arteries / Q000166:cytology; D049109:Cell Proliferation; D002478:Cells, Cultured; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D004705:Endocytosis; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry* / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D007074:Immunoglobulin G / Q000737:chemistry; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D049329:Nanostructures / Q000737:chemistry*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics; D010777:Photochemistry; D011073:Polyamines / Q000737:chemistry; D011091:Polyesters / Q000737:chemistry; D051381:Rats; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes
D000906:Antibodies; C568773:Clmp protein, rat; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D007074:Immunoglobulin G; D011073:Polyamines; D011091:Polyesters; D011991:Receptors, Virus; C514706:polyallylamine-benzophenone-maleimidocarboxylate; C514705:polyallylamine-benzophenone-pyridyldithiocarboxylate polymer; D049452:Green Fluorescent Proteins; C033616:poly(lactide); D052247:Nitric Oxide Synthase Type II