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 - 24 . (canceled) 
     
     
         25 . A recombinant lentiviral vector comprising a first therapeutic gene that encodes a first amino acid sequence that inhibits angiogenesis and a second therapeutic gene that encodes a second amino acid sequence that inhibits angiogenesis, wherein said first therapeutic gene is different from said second therapeutic gene. 
     
     
         26 . The recombinant lentiviral vector of  claim 25 , wherein said first amino acid sequence and said second amino acid sequence are each selected from the group consisting of 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, the monokine-induced by interferon-gamma (Mig), the interferon-alpha inducible protein 10 (IP10), soluble FLT-1 (fins-like tyrosine kinase 1 receptor), and kinase insert domain receptor (KDR). 
     
     
         27 . The recombinant lentiviral vector of  claim 25 , wherein one of said therapeutic genes encodes an endostatin. 
     
     
         28 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes endostatin XV. 
     
     
         29 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes endostatin XVIII. 
     
     
         30 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes angiostatin. 
     
     
         31 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes kringle 1-5. 
     
     
         32 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes PEX. 
     
     
         33 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes the C-terminal hemopexin domain of matrix metalloproteinase-2. 
     
     
         34 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes the kringle 5 domain of human plasminogen. 
     
     
         35 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes Mig. 
     
     
         36 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes IP10. 
     
     
         37 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes soluble FLT-1. 
     
     
         38 . The recombinant lentiviral vector of  claim 26 , wherein one of said therapeutic genes encodes KDR. 
     
     
         39 . The recombinant lentiviral vector of  claim 26 , wherein the first therapeutic gene encodes endostatin XVIII and the second therapeutic gene encodes angiostatin. 
     
     
         40 . The recombinant lentiviral vector of  claim 26 , wherein the first therapeutic gene encodes Mig and the second therapeutic gene encodes IP10. 
     
     
         41 . The recombinant lentiviral vector of  claim 25 , wherein the lentiviral vector comprises an IRES (internal ribosome entry site) element between said first therapeutic gene and said second therapeutic gene so that said first amino acid sequence and said second amino acid sequence are produced from a single transcript. 
     
     
         42 . The recombinant lentiviral vector of  claim 25 , wherein said lentiviral vector encodes a fusion protein comprising the first amino acid sequence and the second amino acid sequence. 
     
     
         43 . The recombinant lentiviral vector of  claim 42 , further comprising a linker between the first amino acid sequence and the second amino acid sequence. 
     
     
         44 . The recombinant lentiviral vector of  claim 43 , wherein the linker is an elastin peptide linker. 
     
     
         45 . The recombinant lentiviral vector of  claim 42 , wherein the lentiviral vector encodes a fusion protein of endostatin XVIII and angiostatin. 
     
     
         46 . The recombinant lentiviral vector of  claim 42 , wherein the lentiviral vector encodes a fusion protein of endostatin XVIII and the kringle 5 domain of human plasminogen. 
     
     
         47 . The recombinant lentiviral vector of  claim 42 , wherein the lentiviral vector encodes a fusion protein of Mig and IP10. 
     
     
         48 . The recombinant lentiviral vector of  claim 25 , wherein said first therapeutic gene and said second therapeutic gene are under the control of two separate promoters known to be active in human retinal cells, human corneal epithelial cells, or human retinal pigment epithelial cells.