Adeno-associated viral vectors for the treatment and prevention of diabetes

Abstract

The subject invention concerns materials and methods for gene therapy. One aspect of the invention pertains to vectors which can be used to effect genetic therapy in animals or humans having genetic disorders where expression of high levels of a protein of interest are required to treat or correct the disorder. The subject invention also pertains to methods for treating animals or humans in need of gene therapy to treat or correct a genetic disorder. The materials and methods of the invention can be used to provide therapeutically effective levels of a protein that is non-functional, or that is absent or deficient in the animal or human to be treated. In one embodiment, the materials and methods can be used to treat alpha-1-antitrypsin deficiency.

We claim:  
     
         1 . A method for providing an animal with a therapeutically effective amount of a serum protein, said method comprising introducing into cells of said animal an effective amount of viral particles or vector, wherein said viral particles or viral vector comprises a polynucleotide encoding said protein.  
     
     
         2 . The method according to  claim 1 , wherein said animal is a mammal.  
     
     
         3 . The method according to  claim 2 , wherein said mammal is a human.  
     
     
         4 . The method according to  claim 1 , wherein said vector is an adeno-associated virus vector.  
     
     
         5 . The method according to  claim 1 , wherein said vector comprises a promoter sequence capable of driving expression of said polynucleotide encoding said protein.  
     
     
         6 . The method according to  claim 5 , wherein said promoter sequence is selected from the group consisting of CMV promoter sequences, hybrid CMV enhancer/β-actin promoter sequences, EF1 promoter sequences, U1a promoter sequences and U1b promoter sequences.  
     
     
         7 . The method according to  claim 5 , wherein said promoter sequence is an inducible promoter selected from the group consisting of Tet-inducible promoters and VP16-LexA promoter.  
     
     
         8 . The method according to  claim 5 , wherein said vector further comprises an enhancer sequence.  
     
     
         9 . The method according to  claim 8 , wherein said enhancer is a synthetic enhancer.  
     
     
         10 . The method according to  claim 1 , wherein said animal has a condition that results in a defective protein or a deficiency of said protein encoded by said polynucleotide.  
     
     
         11 . The method according to  claim 1 , wherein said animal has a condition that can be ameliorated or treated by said protein encoded by said polynucleotide.  
     
     
         12 . The method according to  claim 1 , wherein said protein encoded by said polynucleotide is selected from the group consisting of anti-proteases, enzymes, structural proteins, coagulase factors, interleukins, cytokines, growth factors, interferons, and lymphokines.  
     
     
         13 . The method according to  claim 1 , wherein said cells are myofibers, myoblasts, hepatocytes, or lung cells.  
     
     
         14 . The method according to  claim 1 , wherein said polynucleotide encodes human alpha-1-antitrypsin protein, or a biologically active fragment or variant thereof.  
     
     
         15 . The method according to  claim 4 , wherein said polynucleotide encodes human alpha-1-antitrypsin protein, or a biologically active fragment or variant thereof.  
     
     
         16 . The method according to  claim 1 , wherein said viral particles are introduced into said cells or tissue by infection or injection.  
     
     
         17 . The method according to  claim 1 , wherein said vector is introduced into said cells by transfection or injection.  
     
     
         18 . The method according to  claim 1 , wherein said viral particles or vector is introduced into said cells in vitro and said treated cells are introduced into said animal.  
     
     
         19 . The method according to  claim 1 , wherein said viral particles or vector is introduced into said cells in vivo.  
     
     
         20 . The method according to  claim 19 , wherein said viral particles or vector is injected into muscle.  
     
     
         21 . The method according to  claim 19 , wherein said viral particles or vector is injected into portal or peripheral vein.  
     
     
         22 . The method according to  claim 19 , wherein said viral particles or vector is injected intratracheally or inhaled into the lungs.  
     
     
         23 . The method according to  claim 15 , wherein said vector is selected from the group consisting of dE-AT, E-AT, C-AT, C-AT2, p43C-AT, p43CB-AT, p43C-AT-IN, p43msENC-AT, p43rmsENC-AT, p43msENCB-AT and p43rmsENCB-AT.  
     
     
         24 . A recombinant viral vector comprising a polynucleotide encoding a protein capable of providing a therapeutic effect to an animal when expressed in said animal.  
     
     
         25 . The vector according to  claim 24 , wherein said animal is a mammal.  
     
     
         26 . The vector according to  claim 25 , wherein said mammal is a human.  
     
     
         27 . The vector according to  claim 26 , wherein said vector is an adeno-associated virus vector.  
     
     
         28 . The vector according to  claim 24 , wherein said vector comprises a promoter sequence capable of driving expression of said polynucleotide encoding said protein.  
     
     
         29 . The vector according to  claim 28 , wherein said promoter sequence is selected from the group consisting of CMV promoter sequences, hybrid CMV enhancer/β-actin promoter sequences, EF1 promoter sequences, U1a promoter sequences and U1b promoter sequences.  
     
     
         30 . The vector according to  claim 24 , wherein said polynucleotide encodes human alpha-1-antitrypsin protein, of a biologically active fragment or variant thereof.  
     
     
         31 . The vector according to  claim 27 , wherein said polynucleotide encodes human alpha-1-antitrypsin protein, of a biologically active fragment or variant thereof.  
     
     
         32 . The vector according to  claim 31 , wherein said vector is selected from the group consisting of dE-AT, E-AT, C-AT, C-AT2, p43C-AT, p43CB-AT, p43C-AT-IN, p43msENC-AT, p43rmsENC-AT, p43msENCB-AT and p43rmsENCB-AT.  
     
     
         33 . The vector according to  claim 28 , wherein said promoter sequence is an inducible promoter selected from the group consisting of Tet-inducible promoters and VP16-LexA promoter.  
     
     
         34 . The vector according to  claim 28 , wherein said vector further comprises an enhancer sequence.  
     
     
         35 . The vector according to  claim 34 , wherein said enhancer is a synthetic enhancer.  
     
     
         36 . The vector according to  claim 24 , wherein said protein encoded by said polynucleotide is selected from the group consisting of anti-proteases, enzymes, structural proteins, coagulase factors, interleukins, cytokines, growth factors, interferons, and lymphokines.  
     
     
         37 . A viral particle comprising the vector of  claim 24 .  
     
     
         38 . A cell comprising the vector of  claim 24 .  
     
     
         39 . The cell according to  claim 38 , wherein said cell is a myofiber, myoblast, hepatocyte, or lung cell.  
     
     
         40 . A method for treating alpha-1-antitrypsin deficiency in an animal, said method comprising introducing into cells of said animal a vector according to  claim 24 , wherein said polynucleotide of said vector encodes alpha-1-antitrypsin protein, or a biologically active fragment or variant thereof.