Reduction of Natural Adenovirus Tropism to Mouse Liverby Fiber-Shaft Exchange in Combination with both CAR- andαv Integrin-BindingAblation

ABSTRACT The primary receptor, the coxsackievirus and adenovirus receptor (CAR), and the secondary receptor, αv integrins, are the tropism determinants of adenovirus (Ad) type 5. Inhibition of the interaction of both the fiber with CAR and the penton base with the αv integrin appears to be crucial to the development of targeted Ad vectors, which specifically transduce a given cell population. In this study, we developed Ad vectors with ablation of both CAR and αv integrin binding by mutating the fiber knob and the RGD motif of the penton base. We also replaced the fiber shaft domain with that derived from Ad type 35. High transduction efficiency in the mouse liver was suppressed approximately 130- to 270-fold by intravenous administration of the double-mutant Ad vectors, which mutated two domains each of the fiber knob and shaft and the RGD motif of the penton base compared with those of conventional Ad vectors (type 5). Most significantly, the triple-mutant Ad vector containing the fiber knob with ablation of CAR binding ability, the fiber shaft of Ad type 35, and the penton base with a deletion of the RGD motif mediated a >30,000-fold lower level of mouse liver transduction than the conventional Ad vectors. This triple-mutant Ad vector also mediated reduced transduction in other organs (the spleen, kidney, heart, and lung). Viral DNA analysis showed that systemically delivered triple-mutant Ad vector was primarily taken up by liver nonparenchymal cells and that most viral DNAs were easily degraded, resulting in little gene expression in the liver. These results suggest that the fiber knob, fiber shaft, and RGD motif of the penton base each plays an important role in Ad vector-mediated transduction to the mouse liver and that the triple-mutant Ad vector exhibits little tropism to any organs and appears to be a fundamental vector for targeted Ad vectors.

[1]  G. Nemerow,et al.  Adenovirus serotype 5 fiber shaft influences in vivo gene transfer in mice. , 2003, Human Gene Therapy.

[2]  M. Magnusson,et al.  Adenovirus stripping: a versatile method to generate adenovirus vectors with new cell target specificity. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[3]  H. Mizuguchi,et al.  Generation of fiber‐modified adenovirus vectors containing heterologous peptides in both the HI loop and C terminus of the fiber knob , 2003, The journal of gene medicine.

[4]  H. Hamada,et al.  Reduction of Natural Adenovirus Tropism to the Liver by both Ablation of Fiber-Coxsackievirus and Adenovirus Receptor Interaction and Use of Replaceable Short Fiber , 2003, Journal of Virology.

[5]  M. Magnusson,et al.  Genetic retargeting of adenovirus vectors: functionality of targeting ligands and their influence on virus viability , 2002, The journal of gene medicine.

[6]  H. Mizuguchi,et al.  CAR- or αv integrin-binding ablated adenovirus vectors, but not fiber-modified vectors containing RGD peptide, do not change the systemic gene transfer properties in mice , 2002, Gene Therapy.

[7]  D. Curiel,et al.  Gene transfer to ovarian cancer versus normal tissues with fiber-modified adenoviruses. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[8]  M. Rollence,et al.  In vivo hepatic adenoviral gene delivery occurs independently of the coxsackievirus-adenovirus receptor. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[9]  H. Mizuguchi,et al.  Enhanced antitumor effect and reduced vector dissemination with fiber-modified adenovirus vectors expressing herpes simplex virus thymidine kinase , 2002, Cancer Gene Therapy.

[10]  H. Mizuguchi,et al.  Adenovirus vectors containing chimeric type 5 and type 35 fiber proteins exhibit altered and expanded tropism and increase the size limit of foreign genes. , 2002, Gene.

[11]  I. Kovesdi,et al.  Reducing the Native Tropism of Adenovirus Vectors Requires Removal of both CAR and Integrin Interactions , 2001, Journal of Virology.

[12]  H. Mizuguchi,et al.  Efficient gene transfer by fiber-mutant adenoviral vectors containing RGD peptide. , 2001, Biochimica et biophysica acta.

[13]  F. Cosset,et al.  Organ distribution of gene expression after intravenous infusion of targeted and untargeted lentiviral vectors , 2001, Gene Therapy.

[14]  R. Alemany,et al.  CAR-binding ablation does not change biodistribution and toxicity of adenoviral vectors , 2001, Gene Therapy.

[15]  T. Mayumi,et al.  Optimization of transcriptional regulatory elements for constructing plasmid vectors. , 2001, Gene.

[16]  M. Kay,et al.  A simplified system for constructing recombinant adenoviral vectors containing heterologous peptides in the HI loop of their fiber knob , 2001, Gene Therapy.

[17]  D. Curiel,et al.  Genetic Targeting of an Adenovirus Vector via Replacement of the Fiber Protein with the Phage T4 Fibritin , 2001, Journal of Virology.

[18]  Teruhiko Yoshida,et al.  Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity , 2001, Gene Therapy.

[19]  R. Eisensmith,et al.  An immunomodulatory procedure that stabilizes transgene expression and permits readministration of E1-deleted adenovirus vectors. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[20]  A. Lieber,et al.  Dependence of Adenovirus Infectivity on Length of the Fiber Shaft Domain , 2000, Journal of Virology.

[21]  R. Alemany,et al.  Blood clearance rates of adenovirus type 5 in mice. , 2000, The Journal of general virology.

[22]  H. Abe,et al.  Dependence of efficient adenoviral gene delivery in malignant glioma cells on the expression levels of the Coxsackievirus and adenovirus receptor. , 2000, Journal of neurosurgery.

[23]  V. Krasnykh,et al.  Genetic targeting of adenoviral vectors. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[24]  G. Gerken,et al.  Liver sinusoidal endothelial cells are not permissive for adenovirus type 5. , 2000, Human gene therapy.

[25]  M. Bewley,et al.  Structural analysis of the mechanism of adenovirus binding to its human cellular receptor, CAR. , 1999, Science.

[26]  I. Kovesdi,et al.  Identification of a conserved receptor-binding site on the fiber proteins of CAR-recognizing adenoviridae. , 1999, Science.

[27]  A. Beavil,et al.  Mutations in the DG Loop of Adenovirus Type 5 Fiber Knob Protein Abolish High-Affinity Binding to Its Cellular Receptor CAR , 1999, Journal of Virology.

[28]  A. Houtsmuller,et al.  Expression of Coxsackie adenovirus receptor and alphav-integrin does not correlate with adenovector targeting in vivo indicating anatomical vector barriers , 1999, Gene Therapy.

[29]  M. Kay,et al.  A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. , 1999, Human gene therapy.

[30]  C. Miller,et al.  A system for the propagation of adenoviral vectors with genetically modified receptor specificities , 1999, Nature Biotechnology.

[31]  D. Spehner,et al.  Fiberless Recombinant Adenoviruses: Virus Maturation and Infectivity in the Absence of Fiber , 1999, Journal of Virology.

[32]  M. Kay,et al.  Efficient construction of a recombinant adenovirus vector by an improved in vitro ligation method. , 1998, Human gene therapy.

[33]  M. Hashida,et al.  Targeted delivery of plasmid DNA to hepatocytes in vivo: optimization of the pharmacokinetics of plasmid DNA/galactosylated poly(L-lysine) complexes by controlling their physicochemical properties. , 1998, The Journal of pharmacology and experimental therapeutics.

[34]  G. Nemerow,et al.  Complementation of a fibre mutant adenovirus by packaging cell lines stably expressing the adenovirus type 5 fibre protein. , 1998, The Journal of general virology.

[35]  M. Kay,et al.  The role of Kupffer cell activation and viral gene expression in early liver toxicity after infusion of recombinant adenovirus vectors , 1997, Journal of virology.

[36]  M. Finegold,et al.  Macrophage depletion increases the safety, efficacy and persistence of adenovirus-mediated gene transfer in vivo , 1997, Gene Therapy.

[37]  L. Philipson,et al.  HCAR and MCAR: the human and mouse cellular receptors for subgroup C adenoviruses and group B coxsackieviruses. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[38]  J. Bergelson,et al.  Isolation of a Common Receptor for Coxsackie B Viruses and Adenoviruses 2 and 5 , 1997, Science.

[39]  Hanns Lochmüller,et al.  The route of administration is a major determinant of the transduction efficiency of rat tissues by adenoviral recombinants. , 1995, Gene therapy.

[40]  G. Nemerow,et al.  Integrin alpha v beta 5 selectively promotes adenovirus mediated cell membrane permeabilization , 1994, The Journal of cell biology.

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

[42]  G. Nemerow,et al.  Integrins α v β 3 and α v β 5 promote adenovirus internalization but not virus attachment , 1993, Cell.

[43]  J. Maizel,et al.  The polypeptides of adenovirus. I. Evidence for multiple protein components in the virion and a comparison of types 2, 7A, and 12. , 1974, Virology.

[44]  M. Perricaudet,et al.  Genetic manipulations of adenovirus type 5 fiber resulting in liver tropism attenuation , 2003, Gene Therapy.

[45]  S. Kochanek,et al.  Selective depletion or blockade of Kupffer cells leads to enhanced and prolonged hepatic transgene expression using high-capacity adenoviral vectors. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[46]  S. Cusack,et al.  Influence of adenoviral fiber mutations on viral encapsidation, infectivity and in vivo tropism , 2001, Gene Therapy.

[47]  T. Wickham,et al.  Targeting adenovirus , 2000, Gene Therapy.

[48]  C. Dinney,et al.  Biodistribution of an adenoviral vector carrying the luciferase reporter gene following intravesical or intravenous administration to a mouse , 1999, Cancer Gene Therapy.

[49]  R. Crystal,et al.  Innate immune mechanisms dominate elimination of adenoviral vectors following in vivo administration. , 1997, Human gene therapy.