Method of Treating or Retarding the Development of Blindness

Abstract

A method for treating an ocular disorder characterized by the defect or absence of a normal gene in the ocular cells of a human or animal subject involves administering to the subject by subretinal injection an effective amount of a recombinant adeno-associated virus carrying a nucleic acid sequence encoding the normal gene under the control of a promoter sequence which expresses the product of the gene in the ocular cells. The ocular cells are preferably retinal pigment epithelial (RPE) cells, and the gene is preferably an RPE-specific gene, e.g., RPE65. The promoter is one that can express the gene product in the RPE cells. Compositions for subretinal administration are useful in this method.

1 . A method for treating an ocular disorder characterized by the defect or absence of a normal retinal pigment specific epithelial 65 (RPE65) gene in ocular cells of a subject, said method comprising:
 administering to said subject by subretinal injection an effective amount of a recombinant adeno-associated virus serotype 1 (rAAV1) or 2 (rAAV2) carrying a nucleic acid sequence encoding said normal RPE65 gene under the control of a promoter sequence which expresses the product of said RPE65 gene in said ocular cells.   
   
   
       2 . The method according to  claim 1 , wherein said normal RPE65 gene is obtained from the same subject species as the subject being treated. 
   
   
       3 . The method according to  claim 1 , wherein said promoter is a cell-specific promoter. 
   
   
       4 . The method according to  claim 1 , wherein said promoter is the chicken beta actin promoter/CMV enhancer. 
   
   
       5 . The method according to  claim 1 , wherein said ocular cells are retinal pigment epithelial cells. 
   
   
       6 . A composition for treating an ocular disorder characterized by the defect or absence of a normal retinal specific pigment epithelial 65 (RPE65) gene in the retinal pigment epithelial (RPE) cells of a subject, said composition comprising:
 (i) an effective amount of a recombinant adeno-associated virus (rAAV) serotype 1 (rAAV1) or 2 (rAAV2) carrying a nucleic acid sequence encoding said normal RPE65 gene under the control of a promoter sequence which expresses the product of said RPE65 gene in said RPE cells;   (ii) at least one carrier; and   (iii) additional components suitable for subretinal injection.   
   
   
       7 . The method according to  claim 6 , wherein said effective amount is 1×10 9  to 2×10 12  rAAV infectious units in a volume of about 150 to about 800 μl. 
   
   
       8 . The method according to  claim 7 , wherein said effective amount is about 1×10 10  to 2×10 11  rAAV infectious units in a volume of about 250 to about 500 μl. 
   
   
       9 . A method for treating Leber congenital amaurosis in a human, said method comprising:
 administering to said human by subretinal injection an effective amount of a recombinant adeno-associated virus (rAAV) serotype 1 or 2 carrying a nucleic acid sequence encoding a normal retinal specific pigment epithelial 65 (RPE65) gene under the control of a promoter sequence which expresses the product of said RPE65 gene in retinal pigment epithelial (RPE) cells which contain a mutated version of said RPE65 gene, wherein expression of said normal RPE65 gene provides to said RPE cells the product necessary to restore or maintain vision in said human.   
   
   
       10 . The method according to  claim 9 , wherein said promoter is cell-specific. 
   
   
       11 . The method according to  claim 9 , wherein said rAAV carries the normal RPE65 gene. 
   
   
       12 . The method according to  claim 9 , wherein said effective amount is 1×10 9  to 2×10 12  rAAV infectious units in a volume of about 150 to about 800 μl. 
   
   
       13 . The method according to  claim 12 , wherein said effective amount is about 1×10 10  to 2×10 11  rAAV infectious units in a volume of about 250 to about 500 μl. 
   
   
       14 . A method for treating an ocular disorder characterized by the defect or absence of a normal retinal specific pigment epithelial 65 (RPE65) gene in the retinal pigment epithelial (RPE) cells of a subject selected from the group consisting of a primate, canine, and human, said method comprising:
 administering to said subject by subretinal injection an effective amount of a recombinant adeno-associated virus serotype 2 (rAAV2) carrying a nucleic acid sequence encoding said normal RPE65 gene under the control of a promoter sequence which expresses the product of said RPE65 gene in said RPE cells.   
   
   
       15 . The method according to  claim 14 , wherein said subject is a canine or human. 
   
   
       16 . The method according to  claim 14 , wherein said effective amount is 1×10 9  to 2×10 12  rAAV2 infectious units in a volume of about 150 to about 800 μl. 
   
   
       17 . The method according to  claim 16 , wherein said effective amount is 1×10 10  to 2×10 11  rAAV2 infectious units in a volume of about 250 to about 500 μl. 
   
   
       18 . A method for treating an ocular disorder characterized by the defect or absence of a normal retinal specific pigment epithelial 65 (RPE65) gene in the retinal pigment epithelial cells (RPE) of a subject, said method comprising:
 administering to said subject by subretinal injection of a recombinant adeno-associated virus serotype 1 (rAAV1) carrying a nucleic acid sequence encoding said normal RPE65 gene under the control of a promoter sequence which expresses the product of said RPE65 gene in said RPE cells,   wherein said rAAV1 is present in an amount of 1×10 9  to 2×10 12  rAAV1 infectious units in a volume of about 150 to about 800 μl.   
   
   
       19 . The method according to  claim 18 , wherein said rAAV1 amount is about 1×10 10  to 2×10 11  rAAV1 infectious units in a volume of about 250 to about 500 μl.