Viral vectors for the treatment of retinal dystrophy

Abstract

The present invention relates to viral vectors that are capable of delivering a heterologous gene to the retina and in particular delivering RLBP1 to RPE and Müller cells of the retina. The invention also relates nucleic acids useful for producing viral vectors, compositions comprising the viral vectors and uses of the compositions and viral vectors. The invention also relates to methods of delivering and/or expressing a heterologous gene to the retina, improving the rate of dark adaption in a subject and treating RLBP1-associated retinal dystrophy.

1 . A viral vector having a vector genome comprising a retinaldehyde binding protein 1 (RLBP1) coding sequence, wherein said vector is adapted to direct expression of an RLBP1 coding sequence in retinal pigment epithelial (RPE) cells and Muller cells of the retina. 
     
     
         2 . The viral vector of  claim 1 , wherein said vector comprises an adeno-associated virus (AAV) serotype 2 or 8 capsid. 
     
     
         3 . The viral vector of  claim 2 , said vector genome comprising in the 5′ to 3′ direction:
 (i) a 5′ ITR; 
 (ii) a recombinant nucleotide sequence comprising an RLBP1 coding sequence; and 
 (iii) a 3′ ITR. 
 
     
     
         4 . The viral vector of  claim 3  where in the recombinant nucleotide sequence comprises in the 5′ to 3′ direction: (i) a promoter; (ii) an RLBP1 coding sequence; and (iii) an SV40 polyA sequence. 
     
     
         5 . The viral vector of 4, wherein the nucleotide sequence of said promoter is selected from SEQ ID NO: 3 and 10. 
     
     
         6 . The viral vector of  claim 4  wherein the vector genome comprises in the 5′ to 3′ direction nucleic acid sequences selected from the group consisting of:
 a) SEQ ID NO: 2, 10, 5, 6, 8, and 9; 
 b) SEQ ID NO: 2, 11, 5, 6, 8, 14, 9; 
 c) SEQ ID NO: 2, 22, 5, 6, 8, 23, and 9; and 
 d) SEQ ID NO: 2, 3, 4, 5, 6, 8, 23, and 9. 
 
     
     
         7 . The viral vector of  claim 2 , wherein said vector genome comprises in the 5′ to 3′ direction:
 (i) a 5′ ITR; 
 (ii) a first recombinant nucleotide sequence; 
 (iii) a non-resolvable ITR; 
 (iv) a second recombinant nucleotide sequence; and 
 (v) a 3′ ITR, 
 wherein the first and second recombinant nucleotide sequences are self-complementary. 
 
     
     
         8 . The viral vector of  claim 7  where in the second recombinant nucleotide sequence comprises in the 5′ to 3′ direction: (i) a promoter; (ii) an RLBP1 coding sequence; and (iii) an SV40 polyA sequence. 
     
     
         9 . The viral vector of 8, wherein the nucleotide sequence of said promoter comprises SEQ ID NO: 3. 
     
     
         10 . The viral vector of  claim 8  wherein the vector genome comprises nucleic acid sequences in the 5′ to 3′ direction of: SEQ ID NO: 36, 62, 63, 64, 65, 66, 1, 3, 4, 5, 6, 8, and 9. 
     
     
         11 . A viral vector comprising a vector genome capable of expressing a heterologous gene in RPE cells and Muller cells of the retina, said vector comprising a) an AAV8 or AAV2 capsid and b) said vector genome comprising an RLBP1 promoter operably linked to the heterologous gene, wherein said RLBP1 promoter has a sequence selected from SEQ ID NOs: 3 and 10. 
     
     
         12 . The viral vector of  claim 11 , wherein the vector genome is self-complementary. 
     
     
         13 . A composition comprising the viral vector of any of the preceding  claims  . 
     
     
         14 . The composition of  claim 13  further comprising a pharmaceutically acceptable excipient. 
     
     
         15 . A method of expressing a heterologous gene in retinal cells, the method comprising contacting said retinal cells with a viral vector comprising: a) an AAV2 or AAV8 capsid, and b) a vector genome comprising an RLBP1 promoter of SEQ ID NO: 3 or SEQ ID NO: 10 operably linked to the heterologous gene. 
     
     
         16 . The method of  claim 15 , wherein the retinal cells are RPE cells and Muller cells. 
     
     
         17 . The method of  claim 16 , wherein the vector genome is a self-complementary genome. 
     
     
         18 . The method of any of  claims 15-17 , wherein the heterologous gene is RLBP1. 
     
     
         19 . A nucleic acid comprising a gene cassette, said gene cassette comprises, in the 5′ to 3′ direction:
 (i) a 5′ ITR or a non-resolvable ITR; 
 (ii) a recombinant nucleotide sequence comprising an RLBP1 coding sequence; and 
 (iii) a 3′ITR. 
 
     
     
         20 . The nucleic acid of  claim 19  that is a plasmid. 
     
     
         21 . The nucleic acid of  claim 19 , wherein:
 (i) said 5′ ITR or said non-resolvable ITR have a nucleic acid sequence of SEQ ID NO: 2 or 1, respectively;   (ii) said recombinant nucleic acid sequence comprises a promoter having a nucleic acid sequence selected from: SEQ ID NO: 3, 10, 11, 12 and 22 operably linked to said RLBP1 coding sequence having a nucleic acid sequence of SEQ ID NO: 3; or 
(ii) said 3′ ITR has a nucleic acid sequence of SEQ ID NO: 9. 
     
     
         22 . The nucleic acid of  claim 19 , wherein the gene cassette comprises a nucleic acid sequence selected from: SEQ ID NO: 51, 52, 53, 54, and 55. 
     
     
         23 . The nucleic acid of  claim 19 , further comprising the nucleic acid sequence selected from SEQ ID NO: 26, 27, 28, 29, 30 and 50. 
     
     
         24 . The nucleic acid of  claim 19 , wherein the gene cassette comprises, in the 5′ to 3′ direction, the sequences selected from:
 a) SEQ ID NO: 2, 10, 5, 6, 8, and 9, 
 b) SEQ ID NO: 2, 11, 5, 6, 8, 14 and 9, 
 c) SEQ ID NO: 2, 22, 5, 6, 8, 23 and 9, 
 d) SEQ ID NO: 2, 3, 4, 5, 6, 8, 23 and 9, or 
 e) SEQ ID NO: 1, 3, 4, 5, 6, 8, and 9. 
 
     
     
         25 . A method of treating a subject having RLBP1-associated retinal dystrophy, the method comprising administering to a subject the composition of  claim 13 . 
     
     
         26 . A method of improving the rate of dark adaption in a subject having RLBP1-associated retinal dystrophy, the method comprising administering to a subject the composition of  claim 13 . 
     
     
         27 . A method of expressing an RLBP1 coding sequence in RPE cells and Muller cells in the retina of a subject having RLBP1-associated retinal dystrophy, said method comprising the step of: contacting the retina of said subject with the viral vector of  claim 6  or  10 . 
     
     
         28 . The composition of  claim 13  for use in treating a subject having RLBP1-associated retinal dystrophy. 
     
     
         29 . The composition of  claim 13  for use in improving the rate of dark adaption in a subject having RLBP1-associated retinal dystrophy. 
     
     
         30 . The viral vector of  claim 6  or  10  for use in expressing an RLBP1 coding sequence in RPE cells and Muller cells in the retina of a subject having RLBP1-associated retinal dystrophy.