Vector for expressing alpha-N-acetyl-galactosaminidase and method of treating MPS I by stereotactic injection into the brain of a mammal

Abstract

A purified nucleic acid molecule which is capable of expressing a lysosomal enzyme wherein said nucleic acid molecule comprises at least a sequence coding for said lysosomal enzyme and a promoter highly active in the brain inserted upstream from said sequence.

1 - 22 . (canceled)  
     
     
         23 . An AAV vector for the expression of α-N-acetyl-glucosaminidase, wherein the vector comprises (a) the nucleic acid coding sequence for α-N-acetyl-glucosaminidase, as found in C.N.C.M I-2891; (b) a phosphoglycerate kinase gene promoter; and (c) a woodchuck hepatitis virus posttranscriptional regulatory element, and wherein the coding sequence is operably linked to (b) and (c).  
     
     
         24 . A method of transforming a brain cell of a mammal in vivo comprising: (a) providing a vector comprising: (i) a nucleic acid sequence encoding α-N-acetyl-glucosaminidase, as found in C.N.C.M. I-2891; (ii) a phosphoglycerate kinase gene promoter; and (iii) a woodchuck hepatitis virus posttranscriptional regulatory element; and (b) delivering the vector to the brain of the mammal by stereotactic injection; wherein the nucleic acid, promoter, and regulatory element are in operable linkage and the cell is local to the stereotactic injection site.  
     
     
         25 . A cell transformed in vitro with the vector of claim  37 , wherein the cell expresses α-N-acetyl-glucosaminidase from the vector sequence.  
     
     
         26 . A method of treating or preventing the accumulation of plaques associated with MPS I, wherein the method comprises administering by stereotactic injection, into the brain of the mammal, an AAV vector comprising (a) a phosphoglycerate kinase gene promoter sequence; (b) a woodchuck hepatitis virus posttranscriptional regulatory element; and (c) an α-L-iduronidase-encoding sequence operably linked to (a) and (b), wherein brain cells local to the injection site express α-N-acetyl-glucosaminidase from the vector sequences.  
     
     
         27 . A method of treating or preventing MPS III disease in a mammal, wherein the method comprises administering by stereotactic injection into the brain of the mammal, an AAV vector comprising (a) a phosphoglycerate kinase gene promoter sequence; (b) a woodchuck hepatitis virus posttranscriptional regulatory element; and (c) an α-N-acetyl-glucosaminidase-encoding sequence operably linked to (a) and (b), wherein brain cells local to the injection site express α-N-acetyl-glucosaminidase from the vector sequences.  
     
     
         28 . A method of providing α-N-acetyl-glucosaminidase to the brain of a mammal comprising administering, by stereotactic injection into the brain of the mammal, an AAV vector comprising (a) a sequence encoding α-N-acetyl-glucosaminidase; (b) an operably linked phosphoglycerate kinase gene promoter sequence located upstream from (a); (c) an operably linked woodchuck hepatitis virus posttranscriptional regulatory element; and (d) two AAV terminal repeat sequences flanking (a), (b), and (c), wherein the α-N-acetyl-glucosaminidase is delivered to areas of the brain of the mammal distal to the injection site.