Vesicular monoamine transporter gene therapy in Parkinson's disease

Abstract

The present invention provides methods and compositions for the therapeutic intervention of Parkinson's disease. More particularly, methods of making and sequestering dopamine are disclosed. Additionally, methods of genetically modifying donor cells by gene transfer for grafting into the central nervous system to treat defective, diseased or damaged cells are disclosed. Methods and compositions for carrying out such gene transfer and grafting are described.

1 - 54 . (canceled) 
     
     
         55 . A viral vector comprising nucleotide sequences encoding tyrosine hydroxylase (TH), GTP-cyclohydrolase I (GTPCH) and aromatic amino acid dopa decarboxylase (AADC), wherein the nucleotide sequences are operably linked by one or more Internal Ribosome Entry Site(s) (IRES). 
     
     
         56 . The vector of  claim 55  is lentiviral. 
     
     
         57 . The vector of  claim 55 , wherein the nucleotide sequences form a multicistronic construct. 
     
     
         58 . The viral vector of  claim 55  further comprising a nucleotide sequence encoding a vesicular monoamine transporter (VMAT). 
     
     
         59 . A method for producing dopamine in cells of a mammal, the method comprising:
 (a) obtaining a multicistronic viral vector comprising nucleotide sequences encoding tyrosine hydroxylase (TH), GTP-cyclohydrolase I (GTPCH) and aromatic amino acid dopa decarboxylase (AADC), wherein the nucleotide sequences are operably linked by one or more Internal Ribosome Entry Site(s) (IRES) and wherein the nucleotide sequences are under the transcriptional control of one or more promoters active in eukaryotic cells; and   (b) transforming a plurality of cells of the mammal with the viral vector in vivo by injecting the viral vector into the mammal at a site in need thereof.   
     
     
         60 . The method of  claim 59  wherein the viral vector is lentiviral. 
     
     
         61 . The method of  claim 59  further comprising providing a nucleotide sequence encoding a vesicular monoamine transporter (VMAT) to the viral vector. 
     
     
         62 . The method of  claim 59  further comprising administering L-DOPA to the mammal. 
     
     
         63 . A method of treating Parkinson's disease in a human subject comprising:
 (a) obtaining a multicistronic viral vector comprising nucleotide sequences encoding tyrosine hydroxylase (TH), GTP-cyclohydrolase I (GTPCH) and aromatic amino acid dopa decarboxylase (AADC), wherein the nucleotide sequences are operably linked by one or more Internal Ribosome Entry Site(s) (IRES) and wherein the nucleotide sequences are under the transcriptional control of one or more promoters active in eukaryotic cells; and   (b) transforming a plurality of cells of the mammal with the viral vector in vivo by injecting the viral vector into the mammal at a site in need thereof to treat Parkinson's disease.   
     
     
         64 . The method of  claim 63  further comprising providing a nucleotide sequence encoding a vesicular monoamine transporter (VMAT) and transforming the VMAT into a plurality of cells of the mammal. 
     
     
         65 . The method of  claim 63  further comprising administering L-DOPA to the mammal. 
     
     
         66 . The method of  claim 63 , wherein the viral vector is lentiviral.