rAAV-delivered alpha-1-antitrypsin compositions and method 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.

1 . A method for providing a diabetic mammal with a therapeutically effective amount of an α-1-antitrypsin polypeptide, said method comprising introducing into suitable cells of said diabetic mammal an effective amount of an adeno-associated viral vector or a plurality of adeno-associated viral particles comprising said vector, wherein said vector comprises a polynucleotide that encodes a mammalian α-1-antitrypsin polypeptide, and wherein said polypeptide is expressed in said cells. 
     
     
         2 .- 75 . (canceled) 
     
     
         76 . The method of  claim 1 , wherein said vector comprises a promoter operably linked to said polynucleotide. 
     
     
         77 . The method of claim  41 , wherein said promoter is a hybrid CMV enhancer/β-actin promoter. 
     
     
         78 . The method of  claim 77 , wherein said promoter is a chicken β-actin promoter. 
     
     
         79 . The method of  claim 1 , wherein said vector comprises a polynucleotide that encodes a human α-1-antitrypsin polypeptide. 
     
     
         80 . The method of  claim 79 , wherein said vector comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11. 
     
     
         81 . The method of  claim 80 , wherein said vector comprises a sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6. 
     
     
         82 . The method of  claim 81 , wherein said vector comprises the sequence of SEQ ID NO:6. 
     
     
         83 . The method of  claim 1 , wherein said diabetic mammal has a defect in, or an absence of, a biologically-active α-1-antitrypsin protein. 
     
     
         84 . The method of  claim 1 , wherein said diabetic mammal has a deficiency in the level of biologically-active α-1-antitrypsin protein. 
     
     
         85 . The method of  claim 1 , wherein said diabetic mammal is human. 
     
     
         86 . The method of  claim 1 , wherein said plurality of adeno-associated viral particles is introduced into said cells by infection or transfection. 
     
     
         87 . The method of  claim 1 , wherein said vector or said plurality of adeno-associated viral particles is introduced into cells of said diabetic mammal in vitro and the transduced cells are then introduced into said diabetic mammal. 
     
     
         88 . The method of  claim 1 , wherein said vector or said plurality of adeno-associated viral particles is introduced into said cells in vivo. 
     
     
         89 . The method of  claim 1 , wherein said vector or said plurality of adeno-associated viral particles is injected into a muscle of said diabetic mammal. 
     
     
         90 . The method of  claim 1 , wherein said vector or said plurality of adeno-associated viral particles is injected into a portal or peripheral vein of said diabetic mammal. 
     
     
         91 . A method for providing a diabetic mammal with a therapeutically effective amount of an α-1-antitrypsin polypeptide, comprising introducing into suitable cells of said diabetic mammal an effective amount of an adeno-associated viral vector or a plurality of adeno-associated viral particles comprising said vector; wherein said vector comprises a polynucleotide encoding an α-1-antitrypsin polypeptide operably linked to a chicken beta-actin promoter, and wherein said polypeptide is expressed in said cell. 
     
     
         92 . The method of  claim 91 , wherein said mammal is a human. 
     
     
         93 . The method of  claim 91 , wherein said vector comprises a polynucleotide encoding a human α-1-antitrypsin protein. 
     
     
         94 . A method for preventing Type I diabetes in a mammal, comprising introducing into suitable cells of said mammal, an adeno-associated viral vector or an adeno-associated viral particle that comprises said vector; wherein said vector comprises a polynucleotide that encodes a mammalian α-1-antitrypsin protein, and wherein said protein is expressed in said cells in an amount effective to prevent Type I diabetes in said mammal. 
     
     
         95 . The method of claim  68 , wherein said mammal is a human. 
     
     
         96 . A method for treating Type I diabetes in a mammal, comprising introducing into suitable cells of said mammal a therapeutically-effective amount of an adeno-associated viral vector or an adeno-associated viral particle that comprises said vector; wherein said vector comprises a polynucleotide encoding human α-1-antitrypsin protein, and wherein said protein is expressed in said cells. 
     
     
         97 . A method for providing a mammal diagnosed with, or at risk for developing, Type I diabetes with a therapeutically effective amount of an α-1-antitrypsin polypeptide, said method comprising providing to said mammal an effective amount of an adeno-associated viral vector or a plurality of adeno-associated viral particles comprising said vector, wherein said vector comprises a polynucleotide encoding an α-1-antitrypsin polypeptide, and wherein said polypeptide is expressed in cells of said mammal. 
     
     
         98 . The method of  claim 97 , wherein said vector comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11.