Lentiviral vector-mediated gene transfer and uses thereof

Abstract

The present invention provides lentiviral vectors that are useful in human gene therapy for inherited or acquired proliferative ocular disease. It furnishes methods to exploit the ability of lentiviral vectors to transduce both mitotically active and inactive cells so that eye diseases may be treated.

1 - 15 . (canceled)  
     
     
         16 . A recombinant lentiviral vector comprising a therapeutic gene that regulates angiogenesis.  
     
     
         17 - 23 . (canceled)  
     
     
         24 . The recombinant lentiviral vector of  claim 25 , wherein said lentiviral vector is selected from the group consisting of pHR-CMV-Endo/Ang-ires-eGFP, pHR-CMV-Endo/Kringle-ires-eGFP, pHR-EF1/HTLV-EndoAng-ires-eGFP, pHR-EF1/HTLV-EndoKringle-ires-eGFP and pHR-EF1/HTLV-MigIP10-ires-eGFP.  
     
     
         25 . The recombinant lentiviral vector of  claim 16 , wherein said gene that regulates angiogenesis encodes proteins or polypeptides selected from the group consisting of a tissue inhibitor of metalloproteinase, endostatin, angiostatin, endostatin XVIII, endostatin XV, kringle 1-5, PEX, the C-terminal hemopexin domain of matrix metalloproteinase-2, the kringle 5 domain of human plasminogen, a fusion protein of endostatin and angiostatin, a fusion protein of endostatin and the kringle 5 domain of human plasminogen, the monokine-induced by interferon-gamma (Mig), the interferon-alpha inducible protein 10 (IP10), a fusion protein of Mig and IP10, soluble FLT-1 (fins-like tyrosine kinase 1 receptor), and kinase insert domain receptor (KDR).  
     
     
         26 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes kinase insert domain receptor (KDR).  
     
     
         27 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes tissue inhibitor of metalloproteinase (TIMP)-1.  
     
     
         28 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes tissue inhibitor or metalloproteinase (TIMP)-2  
     
     
         29 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes tissue inhibitor of metalloproteinase (TIMP)-3.  
     
     
         30 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes tissue inhibitor of metalloproteinase (TIMP)-4.  
     
     
         31 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes endostatin.  
     
     
         32 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes angiostatin.  
     
     
         33 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes endostatin XVIII.  
     
     
         34 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes endostatin XV.  
     
     
         35 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes the C-terminal hemopexin domain of matrix metalloproteinase-2.  
     
     
         36 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes the kringle 5 domain of human plasminogen  
     
     
         37 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes a fusion protein of endostatin and angiostatin.  
     
     
         38 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes a fusion protein of endostatin and the kringle 5 domain of human plasminogen.  
     
     
         39 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes the monokine-induced by interferon-gamma (Mig).  
     
     
         40 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes the interferon-alpha inducible protein 10 (IP10).  
     
     
         41 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes a fusion protein of Mig and IP10.  
     
     
         42 . The recombinant lentiviral vector of  claim 25 , wherein said gene that regulates angiogenesis encodes soluble FLT-1 (fins-like tyrosine kinase 1 receptor).  
     
     
         43 . The recombinant lentiviral vector of  claim 25 , wherein the gene that regulates angiogenesis encodes kringle 1-5.  
     
     
         44 . The recombinant lentiviral vector of  claim 25 , wherein the gene that regulates angiogenesis encodes PEX.  
     
     
         45 . The recombinant lentiviral vector of  claim 16 , wherein the lentiviral vector further expresses a marker gene.  
     
     
         46 . The recombinant lentiviral vector of  claim 45 , wherein said marker gene is enhanced green fluorescent protein gene.  
     
     
         47 . The recombinant lentiviral vector of  claim 16 , wherein the lentiviral vector comprises an IRES (internal ribosome entry site) element between two cloning sites so that two different proteins are produced from a single transcript.  
     
     
         48 . The recombinant lentiviral vector of  claim 25 , wherein the lentiviral vector is selected from the group consisting of pHR-CMV-KDR-ires-eGFP, pHR-CMV-Timp1-ires-eGFP, pHR-EF1/HTLV-Ang-ires-eGFP, pHR-EF1/HTLV-Endo XV-ires-eGFP, pHR-EF 1/HTLV-Kringle 1-5-ires-eGFP, pHR-EF1/HTLV-Timp1-ires-eGFP, pHR-EF1/HTLV-Timp4-ires-eGFP and pHR-EF1/HTLV-Endo XVIII-ires-eGFP.