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.

[1]  S Cusack,et al.  Kinetic Analysis of Adenovirus Fiber Binding to Its Receptor Reveals an Avidity Mechanism for Trimeric Receptor-Ligand Interactions* , 2001, The Journal of Biological Chemistry.

[2]  Robert Gurny,et al.  Poly(lactic acid) nanoparticles labeled with biologically active Neutravidin for active targeting. , 2004, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[3]  H. Cavanagh,et al.  Cell contact dependent extended release of adenovirus by microparticles in vitro. , 2001, Journal of virological methods.

[4]  J. Deisenhofer,et al.  Characterization of the knob domain of the adenovirus type 5 fiber protein expressed in Escherichia coli , 1994, Journal of virology.

[5]  M. Pandori,et al.  Adenovirus-microbead conjugates possess enhanced infectivity: a new strategy for localized gene delivery. , 2002, Virology.

[6]  A. Panet,et al.  Synchronized Infection of Cell Cultures by Magnetically Controlled Virus , 2005, Journal of Virology.

[7]  P. Lowenstein,et al.  Acute direct adenoviral vector cytotoxicity and chronic, but not acute, inflammatory responses correlate with decreased vector-mediated transgene expression in the brain. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[8]  C. chou,et al.  Is green fluorescent protein toxic to the living cells? , 1999, Biochemical and biophysical research communications.

[9]  Joel A Swanson,et al.  Drug delivery strategy utilizing conjugation via reversible disulfide linkages: role and site of cellular reducing activities. , 2003, Advanced drug delivery reviews.

[10]  M. Rots,et al.  A novel strategy to modify adenovirus tropism and enhance transgene delivery to activated vascular endothelial cells in vitro and in vivo. , 2004, Human gene therapy.

[11]  Robert Gurny,et al.  Surface modification of poly(lactic acid) nanoparticles by covalent attachment of thiol groups by means of three methods. , 2003, International journal of pharmaceutics.

[12]  Hatem Fessi,et al.  In vitro degradation of nanospheres from poly(D,L-lactides) of different molecular weights and polydispersities , 1996 .

[13]  Jun Ren,et al.  Nitric oxide synthase gene therapy for cardiovascular disease. , 2002, Japanese journal of pharmacology.

[14]  J. English,et al.  Polyglycolide and polylactide , 1998 .

[15]  T. Sano,et al.  Chemically inactivated adenoviral vectors that can efficiently transduce target cells when delivered in the form of virus-microbead conjugates , 2005, Gene Therapy.

[16]  W. Mark Saltzman,et al.  Enhancement of transfection by physical concentration of DNA at the cell surface , 2000, Nature Biotechnology.

[17]  A. Domb.  Polymeric site-specific pharmacotherapy , 1994 .

[18]  Indu Bala,et al.  PLGA nanoparticles in drug delivery: the state of the art. , 2004, Critical reviews in therapeutic drug carrier systems.

[19]  K. Leong,et al.  Coacervate microspheres as carriers of recombinant adenoviruses , 1999, Cancer Gene Therapy.

[20]  M. Bewley,et al.  Coxsackievirus and Adenovirus Receptor Amino-Terminal Immunoglobulin V-Related Domain Binds Adenovirus Type 2 and Fiber Knob from Adenovirus Type 12 , 1999, Journal of Virology.

[21]  D. Quintanar-Guerrero,et al.  Preparation techniques and mechanisms of formation of biodegradable nanoparticles from preformed polymers. , 1998, Drug development and industrial pharmacy.

[22]  F. Farzaneh,et al.  Streptavidin paramagnetic particles provide a choice of three affinity-based capture and magnetic concentration strategies for retroviral vectors. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[23]  L. Barbu-Tudoran,et al.  Effects of different application parameters on penetration characteristics and arterial vessel wall integrity after local nanoparticle delivery using a porous balloon catheter. , 2004, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[24]  D. Brough,et al.  Increased in vitro and in vivo gene transfer by adenovirus vectors containing chimeric fiber proteins , 1997, Journal of virology.

[25]  V. Kähäri,et al.  Efficient infection of tumor endothelial cells by a capsid-modified adenovirus , 2006, Gene Therapy.

[26]  Motohiro Uo,et al.  Microparticle formation and its mechanism in single and double emulsion solvent evaporation. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[27]  J. Whitsett,et al.  In vivo evaluation of the safety of adenovirus-mediated transfer of the human cystic fibrosis transmembrane conductance regulator cDNA to the lung. , 1994, Human gene therapy.

[28]  H. Mizuguchi,et al.  Approaches to improving the kinetics of adenovirus-delivered genes and gene products. , 2005, Advanced drug delivery reviews.

[29]  Jon Dobson,et al.  Improved method of recombinant AAV2 delivery for systemic targeted gene therapy. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[30]  Andrew H Baker,et al.  Designing gene delivery vectors for cardiovascular gene therapy. , 2004, Progress in biophysics and molecular biology.

[31]  A. Beaudet,et al.  Acute toxicity after high-dose systemic injection of helper-dependent adenoviral vectors into nonhuman primates. , 2004, Human gene therapy.

[32]  I. Alferiev,et al.  Cholesterol-derivatized polyurethane: characterization and endothelial cell adhesion. , 2005, Journal of biomedical materials research. Part A.

[33]  M. Kaleko,et al.  Synthesis of adenoviral targeting molecules by intein-mediated protein ligation , 2003, Gene Therapy.

[34]  Jayanth Panyam,et al.  Biodegradable nanoparticles for drug and gene delivery to cells and tissue. , 2003, Advanced drug delivery reviews.

[35]  A. Palmer,et al.  Photochemical control of the infectivity of adenoviral vectors using a novel photocleavable biotinylation reagent. , 2002, Chemistry & biology.

[36]  D. Janero,et al.  Nitric oxide and postangioplasty restenosis: pathological correlates and therapeutic potential. , 2000, Free radical biology & medicine.