Method of Treating or Retarding the Development of Blindness
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.