Viral vectors with modified tropism

Abstract

The present invention relates to gene therapy. In particular, therapeutic agents, therapeutic gene products, and compositions are disclosed. Various systems and methods useful in targeting and delivering non-native nucleotide sequences to specific cells are disclosed, wherein virus-antibody-ligand conjugates are used to facilitate targeting and delivery.

We claim:  
     
         1 . A tropism-modified adenoviral vector system that specifically targets cells expressing a preselected receptor, comprising: 
 an antibody or fragment thereof that binds an adenoviral capsid protein;    a targeting ligand that binds the preselected receptor; and    an adenovirus containing a nucleic acid molecule that encodes a therapeutic gene product under the control of a promoter;    wherein the ligand is conjugated to the antibody or fragment thereof and wherein the antibody or fragment thereof is bound to the adenovirus.    
     
     
         2 . The vector of  claim 1 , wherein said promoter is a tissue-specific promoter.  
     
     
         3 . The vector of  claim 1 , wherein said targeting ligand is a polypeptide reactive with an FGF receptor.  
     
     
         4 . The vector of  claim 3 , wherein said polypeptide reactive with an FGF receptor is an antibody or fragment thereof.  
     
     
         5 . The vector of  claim 4 , wherein the antibody is 11A8.  
     
     
         6 . The vector of  claim 3 , wherein said polypeptide reactive with an FGF receptor is selected from the group consisting of FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8, FGF-9, FGF-11, FGF-13, FGF-14, and FGF-15.  
     
     
         7 . The vector of  claim 3 , wherein said polypeptide reactive with an FGF receptor is FGF-2.  
     
     
         8 . The vector of  claim 3 , wherein said polypeptide reactive with an FGF receptor is KGF.  
     
     
         9 . The vector of  claim 1 , wherein said targeting ligand is selected from the group consisting of a polypeptide reactive with a VEGF receptor, a polypeptide reactive with a PDGF receptor, and a polypeptide reactive with an EGF receptor.  
     
     
         10 . The vector of  claim 1 , wherein the native tropism of said vector is ablated.  
     
     
         11 . The vector of  claim 1 , wherein the therapeutic gene product is a cytocide or a prodrug.  
     
     
         12 . The vector of  claim 1 , wherein the therapeutic gene product enhances cellular proliferation.  
     
     
         13 . The vector of  claim 1 , wherein the therapeutic gene product is a biologically active protein or polypeptide that augments or complements an endogenous protein.  
     
     
         14 . The vector of  claim 1 , wherein the therapeutic gene product enhances cellular differentiation.  
     
     
         15 . The vector of  claim 1 , wherein the therapeutic gene product is a molecule which enhances tissue repair or regeneration.  
     
     
         16 . The vector of  claim 1 , wherein the therapeutic gene product is a molecule which stimulates a protective immune response.  
     
     
         17 . The vector of  claim 10 , wherein the prodrug is thymidine kinase, nitroreductase, or cytosine deaminase.  
     
     
         18 . A pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, wherein the vector includes a nucleic acid molecule encoding a therapeutic gene product under the control of a promoter.  
     
     
         19 . The composition according to  claim 18 , wherein the ligand is a polypeptide reactive with an FGF receptor.  
     
     
         20 . The composition according to  claim 19 , wherein the polypeptide reactive with an FGF receptor is selected from the group consisting of FGF-1, FGF-2, FGF-3, FGF-4, FGF-5, FGF-6, FGF-7, FGF-8, FGF-9, FGF-11, FGF-13, FGF-14, and FGF-15.  
     
     
         21 . The composition according to  claim 19 , wherein the polypeptide reactive with an FGF receptor is FGF-2.  
     
     
         22 . The composition according to  claim 19 , wherein the polypeptide reactive with an FGF receptor is KGF.  
     
     
         23 . The composition according to  claim 19 , wherein the polypeptide reactive with an FGF receptor is an antibody.  
     
     
         24 . The composition according to  claim 23 , wherein the antibody is a single-chain antibody.  
     
     
         25 . The composition according to  claim 18 , wherein the ligand is genetically fused with an adenoviral capsid protein.  
     
     
         26 . The composition according to  claim 18 , wherein the ligand is chemically conjugated to an adenoviral capsid protein.  
     
     
         27 . The composition according to  claim 18 , wherein the ligand is conjugated to an antibody or fragment thereof that binds a viral capsid protein.  
     
     
         28 . The composition according to  claim 18 , wherein the therapeutic gene product is selected from the group consisting of protein, ribozyme and antisense.  
     
     
         29 . The composition according to  claim 18 , wherein the therapeutic gene product is a cytocide.  
     
     
         30 . The composition according to  claim 18 , wherein the therapeutic gene product is a prodrug.  
     
     
         31 . A method of treating tumors, comprising administering a pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, wherein said vector includes a nucleotide sequence encoding a therapeutic gene product under the control of a promoter, wherein said therapeutic gene product is selected from the group consisting of E-cadherin, BGP, Rb, p53, CDKN2/P16/MTS1, PTEN/MMAC1, APC, p331NG1, Smad4, maspin, VHL, WT1, Men1, NF2, MXI1, and FHIT.  
     
     
         32 . A method of treating ischemia, comprising administering a pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, wherein said vector includes a nucleotide sequence encoding a therapeutic gene product under the control of a promoter, wherein said therapeutic gene product is selected from the group consisting of IGF, TGFβ1, TGF,β2, TGFβ3, HGF, VEGF 121, VEGF 165, FGF1, FGF2, FGF 4, FGF5, PDGF-A, and PDGF-B.  
     
     
         33 . A method of treating connective tissue injury, comprising administering a pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, wherein said vector includes a nucleotide sequence encoding a therapeutic gene product under the control of a promoter, wherein said therapeutic gene product is selected from the group consisting of PTH, BMP1, BMP2, BMP3, BMP4, BMP5, BMP6, BMP7, BMP8, BMP10, BMP11, mammalian BMP, and Xenopus BMP.  
     
     
         34 . A method of treating tissue injury, comprising administering a pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, wherein said vector includes a nucleotide sequence encoding a therapeutic gene product under the control of a promoter, wherein said therapeutic gene product is selected from the group consisting of Bovine VEGF, VEGF, VEGF-B, VEGF-C, Angiopoietin-1, Angiogenin, IGF-1, IGF-II, HGF, PDGF A, PDGF B, TGFB1, TGFB2, and TGFB3.  
     
     
         35 . A method of treating cancer, comprising contacting the cancer cells with a pharmaceutical composition comprising a physiologically acceptable buffer and a tropism-modified adenoviral vector presenting a ligand on its surface, 
 wherein said vector includes a nucleotide sequence encoding a therapeutic gene product under the control of a promoter,    wherein said therapeutic gene product is selected from the group consisting of HSVTK, VZVTK, nitroreductase, and cytosine deaminase; and    contacting the cancer cells with a substrate.    
     
     
         36 . The method according to any one of claims  31 - 35 , wherein the ligand is a polypeptide reactive with an FGF receptor.  
     
     
         37 . The method according to  claim 36 , wherein the polypeptide reactive with an FGF receptor is FGF-2.  
     
     
         38 . The method according to any one of claims  31 - 35 , wherein the ligand is an antibody or a fragment thereof.  
     
     
         39 . The method according to  claim 38 , wherein the antibody is a single-chain antibody.  
     
     
         40 . The method according to any one of claims  31 - 35 , wherein the ligand is conjugated to an antibody or fragment thereof that binds a viral capsid protein.  
     
     
         41 . The method according to  claim 40 , wherein the viral capsid protein is adenovirus fiber protein.  
     
     
         42 . The method according to  claim 40 , wherein the viral capsid protein is adenovirus knob protein.  
     
     
         43 . The method according to any one of claims  31 - 35 , wherein the ligand is chemically conjugated to a protein on the surface of a viral vector.  
     
     
         44 . The method according to any one of claims  31 - 35 , wherein the therapeutic gene product is selected from the group consisting of protein, ribozyme and antisense.  
     
     
         45 . The method according to any one of claims  31 - 35 , wherein the therapeutic gene product is a prodrug.