Abstract: The invention pertains to a method for enhancing bone density or formation. In accordance with the method, a nucleic acid encoding a secreted alkaline phosphatase (SEAP) is administered to a cell in a region of a bone such that the nucleic acid is expressed to produce the SEAP, whereby bone density or formation is enhanced within the region. The method can be employed to produce a bone graft having a cell harboring an exogenous nucleic acid encoding a SEAP. To facilitate the inventive method, the invention provides a recombinant viral vector having a nucleic acid encoding a SEAP. Optionally, a nucleic acid encoding an angiogenic protein and/or a nucleic acid encoding an osteogenic protein is employed in conjunction with the nucleic acid encoding a SEAP.

Abstract: The invention provides a composition and a method for preserving a non-enveloped viral vector. The composition comprises (a) trehalose, (b) a divalent metal salt, a cationic polymer, or a combination thereof, (c) a multiplicity of non-enveloped viral vector particles, and (d) a liquid carrier. Non-enveloped virus particles are stable in the composition in a liquid form, at elevated temperatures, for a sustained period of time.

Abstract: The present invention provides a method of identifying a gene product. The method comprises providing a multiplicity of cells comprising a first gene product. Preferably, the first gene product is produced in the multiplicity of cells by expressing a first exogenous nucleic acid sequence encoding the first gene product. A library of second nucleic acid sequences encoding second gene products is then introduced into the multiplicity of cells. The second nucleic acid sequences are expressed in the multiplicity of cells to produce the second gene products such that the first gene product and at least one of the second gene products contact. The method further comprises causing a complex to form between the first gene product, an affinity molecule that binds the first gene product, and at least one of the second gene products, and subsequently retrieving the complex. At least one second gene product of the complex then is identified.

Abstract: The present invention provides methods for administering an adenoviral gene transfer vector comprising an exogenous gene to an animal. One method involves utilizing systemic neutralizing antibodies to neutralize the adenoviral gene transfer vector outside a targeted muscle. Another method involves the repeat administration of an adenoviral gene transfer vector to a skeletal muscle.

Abstract: A method of enriching a solution of an adenovirus comprising applying a mixed solution comprising an adenovirus and at least one undesired type of biomolecule to an anion exchange chromatography resin containing a binding moiety selected from the group consisting of dimethylaminopropyl, dimethylaminobutyl, dimethylaminoisobutyl, and dimethylaminopentyl and eluting the adenovirus from the chromatography resin. Also provided is a method of purifying an adenovirus from adenovirus-infected cells comprising lysing such cells, applying the lysate to a single chromatography resin, eluting the adenovirus from the chromatography resin, and collecting a fraction containing adenovirus that is substantially as pure as triple CsCl density gradient-purified adenovirus.

Abstract: An adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a radiation inducible promoter operably linked to the nucleic acid sequence coding for TNF. This invention also provides an adenoviral vector comprising (a) an adenoviral genome deficient in the E4 region of the adenoviral genome, (b) a nucleic acid sequence coding for TNF, and (c) a spacer element of at least 15 base pairs in the E4 region of the adenoviral genome. A method of producing an adenoviral vector and a method of treating a tumor or cancer in a host comprising administering an anti-cancer or anti-tumor effective amount of the adenoviral vector of the present invention also is provided.

Abstract: The invention provides a chimeric adenovirus fiber protein including a nonnative amino acid sequence, and a chimeric adenovirus fiber protein lacking a native amino acid receptor-binding sequence. The chimeric protein trimerizes when produced in a mammalian cell.

Abstract: The inventive method of producing a eukaryotic viral vector comprises contacting a eukaryotic cell, which comprises a unique enzyme that nicks or cleaves a DNA molecule, with a recombinant phage vector, or contacting a eukaryotic cell, which does not comprise a unique enzyme that nicks or cleaves a DNA molecule, simultaneously or sequentially, in either order, with (i) a unique enzyme that nicks or cleaves a DNA molecule, and (ii) a recombinant phage vector. The recombinant phage vector comprises the DNA molecule comprising (a) a eukaryotic viral vector genome comprising a coding sequence, (b) a phage packaging site that is not contained within the eukaryotic viral vector genome, and (c) a promoter that is operably linked to the coding sequence.

Abstract: The invention provides adenoviral coat proteins comprising various non-native ligands. Further, the present invention provides an adenoviral vector that elicits less reticulo-endothelial system (RES) clearance in a host animal than a corresponding wild-type adenovirus. Also provided by the invention is a system comprising a cell having a non-native cell-surface receptor and a virus having a non-native ligand, wherein the non-native ligand of the virus binds the non-native cell-surface receptor of the cell. Using this system, a virus can be propagated. Further provided by the invention is a method of controlled gene expression utilizing selectively replication competence, a method of assaying for gene function, a method of isolating a nucleic acid, and a method of identifying functionally related coding sequences. Additionally, the invention provides a cell-surface receptor, which facilitates internalization.

Abstract: The invention provides a method for treating cancer in a human comprising administering to the human a dose of a pharmaceutical composition comprising (i) a pharmaceutically acceptable carrier and (ii) an adenoviral vector comprising a nucleic acid sequence encoding TNF-&agr; operably linked to a promoter, wherein the dose comprises about 1×107 to about 4×1012 particle units (pu) of replication-deficient adenoviral vector, at least once in a therapeutic period comprising up to 10 weeks, whereby the cancer in human is treated. The invention further provides a method of treating a human for multiple tumors, wherein the method comprises contacting a first tumor with a dose of the pharmaceutical composition at least once in a therapeutic period comprising up to about 10 weeks, whereby the human is treated for the first tumor and one or more additional tumors.

Abstract: The present invention provides a method for reducing the size of a tumor in a human comprising (a) directly injecting into the tumor, via multiple injections to different points of the tumor, a dose of a pharmaceutical composition comprising (i) a pharmaceutically acceptable carrier and (ii) a replication-deficient adenoviral vector comprising a nucleic acid sequence encoding TNF-&agr; operably linked to a promoter, wherein the dose comprises about 1×107 to about 4×1012 3 particle units (pu) of replication-deficient adenoviral vector, two or more times over a therapeutic period comprising up to 10 weeks, and (b) administering a dose of ionizing radiation over the duration of the therapeutic period, whereby the size of the tumor is reduced.

Abstract: The invention provides a composition comprising particles of an adenoviral vector comprising deficiencies in two or more gene functions required for viral replication, wherein at least one of the deficiencies is of a gene function of the E1 region of the adenoviral genome and (b) a carrier therefor, with relatively high ratios of (i) the number of particles of the adenoviral vectors to the number of particles of E1-revertant replication-deficient adenoviral vectors not comprising one or more of the deficiencies in gene functions of the E1 region of the adenoviral and (ii) the number of particles of the adenoviral vectors to the number of particles of replication-competent adenoviral vectors, as well as a method of preparing such a composition.

Abstract: The invention provides a composition comprising a gene transfer vector which comprises a nucleic acid sequence encoding a protein and a carrier therefore. The composition is characterized by a relatively high ratio of the gene transfer vector to the protein in the composition.

Abstract: The present invention provides a method of modulating neovascularization in an animal. The method comprises administering to the animal two or more nucleic acid sequences, each nucleic acid sequence encoding at least one angiogenesis-modulation factor that acts upon a different angiogenic process, such that the nucleic acid sequences are expressed to produce the angiogenesis-modulation factors to modulate neovascularization in the animal. Modulating neovascularization includes the induction of neovascularization or, in the alternative, the inhibition or reduction of neovascularization.

Abstract: The invention provides cells, particularly NCI-H460 cells and Calu-1 cells, for the propagation of replication-deficient adenoviral vectors. The cells are lung carcinomas with either a wild-type p53 gene or a heterozygous K-ras mutation. The cells comprise at least one adenoviral nucleic acid sequence, which upon expression produces a gene product that complements for at least one essential gene function of one or more regions of an adenoviral genome so as to propagate a replication-deficient adenoviral vector comprising an adenoviral genome deficient in the at least one essential gene function of the one or more regions when present in the cell.

Abstract: The present invention is directed to a method of prophylactically or therapeutically treating an animal for at least one ocular-related disorder, e.g., ocular neovascularization or age-related macular degeneration. The method comprises contacting an ocular cell with an expression vector comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and the same or different nucleic acid sequence encoding a neurotrophic agent. The method also can comprise contacting an ocular cell with different expression vectors, each comprising a nucleic acid sequence encoding an inhibitor of angiogenesis and/or a nucleic acid sequence encoding a neurotrophic agent. In addition, the present invention provides a viral vector comprising a nucleic acid sequence encoding pigment epithelium-derived factor (PEDF) or a therapeutic fragment thereof.

Abstract: The invention provides a cell and a method of using the cell for the propagation of a replication-deficient adenoviral vector, wherein the cellular genome comprises a nucleic acid sequence whose expression produces a gene product that complements a replication-deficient adenoviral vector. The nucleic acid sequence is operatively linked to a chimeric expression control sequence comprising at least a functional portion of a CMV immediate early promoter/enhancer region and/or at least a functional portion of an adenoviral promoter, wherein the chimeric expression control sequence is upregulated by one or more viral proteins not produced by the nucleic acid sequence.

Abstract: The invention provides therapeutic fusion proteins which include a first peptide portion comprising a first non-heparin binding VEGF peptide portion and a second non-VEGF peptide portion covalently associated with the first peptide portion, which first and second peptide portions separately promote angiogenesis, bone growth, wound healing, or any combination thereof. Further provided are polynucleotides encoding such fusion proteins, vectors including such polynucleotides, methods of making such proteins, and methods of promoting angiogenesis, bone growth, and/or wound healing using such proteins, polynucleotides, and vectors.

Abstract: The present invention provides a method and a composition for preserving a virus. The virus is placed in a liquid carrier with a stabilizing agent selected from the group consisting of polysorbate 80, L-arginine, polyvinylpyrrolidone, trehalose, and combinations thereof. The liquid composition can be maintained at a temperature above 0° C. for a significant period of time while maintaining a satisfactory degree of viral activity.

Abstract: The present invention provides a recombinant adenovirus comprising coat proteins that lack native binding. In particular, the present invention provides a recombinant adenovirus comprising a penton base protein and a fiber protein, wherein the penton base protein and the fiber protein lack native binding. The present invention further provides a recombinant adenovirus comprising (a) a penton base protein that lacks native binding and (b) a nonnative amino acid sequence that binds a cell-surface binding site.