Treatment of retinitis pigmentosa

Abstract

A polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGRORF15), wherein the RPGRORF15-encoding nucleotide sequence has been codon optimised to increase fidelity of replication of the sequence.

1 .- 21 . (canceled) 
     
     
         22 . A polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR ORF15 ), wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to increase fidelity of replication of the sequence. 
     
     
         23 . The polynucleotide of  claim 22 , wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to reduce a number of purine nucleotides in comparison to a wild type sequence of RPGR ORF15 . 
     
     
         24 . The polynucleotide of  claim 22 , wherein the sequence encoding the RPGR ORF15  comprises a nucleotide sequence encoding an amino acid sequence that has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the amino acid sequence of SEQ ID NO: 1. 
     
     
         25 . The polynucleotide of  claim 23 , wherein the RPGR ORF15 -encoding nucleotide sequence does not comprise a purine nucleotide at a position corresponding to nucleotide 33, 58, 59, 114, 123, 129, 181, 182, 213, 219, 226, 227, 237, 267, 285, 306, 309, 315, 324, 330, 339, 400, 401 , 444, 456, 478, 480, 510, 594, 606, 618, 639, 697, 726, 744, 777, 807, 852, 877, 879, 888, 891, 921, 930, 960, 1042, 1050, 1116, 1140, 1183, 1184, 1194, 1197, 1221, 1249, 1251, 1257, 1273, 1276, 1281, 1290, 1293, 1357, 1372, 1373, 1413, 1446, 1452, 1464, 1474, 1475, 1482, 1519, 1520, 1542, 1584, 1590, 1599, 1608, 1653, 1668, 1674, 1689, 1692, 1734, 1761, 1776, 1788, 1803, 1824, 1845, 1854, 1881, 1893, 1902, 1909, 1910, 1929, 1932, 1960, 1987, 1988, 1992, 1998, 2020, 2031, 2047, 2049, 2062, 2067, 2076, 2121, 2167, 2169, 2190, 2193, 2208, 2229, 2259, 2283, 2298, 2323, 2343, 2346, 2361, 2373, 2382, 2388, 2403, 2409, 2410, 2412, 2448, 2457, 2472, 2476, 2478, 2505, 2520, 2547, 2574, 2622, 2634, 2661, 2673, 2706, 2712, 2763, 2775, 2796, 2811, 2817, 2832, 2838, 2871, 2886, 2892, 2898, 2910, 2922, 2931, 2937, 2958, 2970, 2997, 3033, 3039, 3069, 3075, 3117, 3129, 3156, 3166, 3235, 3246, 3273, 3285, 3306, 3321, 3369, 3399, 3405 and/or 3438 of SEQ ID NO: 2. 
     
     
         26 . The polynucleotide of  claim 22 , wherein the RPGR ORF15 -encoding nucleotide sequence is operably linked to a polynucleotide comprising a promoter element capable of driving expression of RPGR ORF15  or a functional variant thereof. 
     
     
         27 . The polynucleotide of  claim 22 , wherein the RPGR ORF15 -encoding nucleotide sequence is operably linked to a rhodopsin kinase (GRK1) promoter. 
     
     
         28 . The polynucleotide of  claim 27 , wherein the GRK1 promoter comprises the nucleotide sequence of SEQ ID NO: 7 or a functional variant having at least 90 or 95% identity thereto. 
     
     
         29 . A viral vector comprising a polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR ORF15 ), wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to increase fidelity of replication of the sequence. 
     
     
         30 . The viral vector of  claim 29 , wherein the viral vector is an adeno-associated viral (AAV), retroviral lentiviral or adenoviral vector. 
     
     
         31 . The viral vector of  claim 30 , wherein the viral vectoris an AAV vector 
     
     
         32 . The viral vector of  claim 29 , wherein the viral vector is in the form of a viral particle. 
     
     
         33 . The viral vector of  claim 29 , wherein the viral vector is a transcapsidated AAV vector. 
     
     
         34 . The viral vector of  claim 30 , wherein the viral vector comprises an AAV2 genome and AAV5 (AAV2/5) or AAV8 (AAV2/8) capsid proteins. 
     
     
         35 . The viral vector of  claim 30 , wherein the viral vector is an AAV vector comprising an AAV2 genome and/or an AAV8 capsid protein. 
     
     
         36 . A viral vector production system comprising a set of polynucleotides encoding components required for production of a viral vector, wherein the viral vector genome comprises a polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR ORF15 ), wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to increase fidelity of replication of the sequence. 
     
     
         37 . A viral vector production cell comprising the polynucleotide of claim  1 . 
     
     
         38 . A viral vector production cell comprising the viral vector production system of  claim 36 . 
     
     
         39 . A process for producing a viral vector comprising introducing the polynucleotide of claim  1 , into a cell and culturing the cell under conditions suitable for the production of the viral vector. 
     
     
         40 . The process of  claim 40 , wherein the cell is a HEK293 or HEK293T cell. 
     
     
         41 . A cell transfected with the polynucleotide of claim  1 . 
     
     
         42 . A cell transduced by the viral vector of  claim 29 . 
     
     
         43 . A pharmaceutical composition comprising the polynucleotide of claim  1 , in combination with pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         44 . A pharmaceutical composition comprising the viral vector of  claim 29 , in combination with pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         45 . A method of treating o ting retinitis pigmentosa comprising, administering to a subject:
 a polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR ORF15 ); or a viral vector comprising the polynucleotide comprising the nucleotide sequence encoding RPGR ORF15 ,   wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to increase fidelity of replication of the sequence.   
     
     
         46 . A method of reducing photoreceptor cell death in a subject suffering from or at risk of developing retinitis pigmentosa comprising, administering to the subject:
 a polynucleotide comprising a nucleotide sequence encoding the retinitis pigmentosa GTPase regulator ORF15 isoform (RPGR ORF15 ); or a viral vector comprising the polynucleotide comprising the nucleotide sequence encoding RPGR ORF15 ,   wherein the RPGR ORF15 -encoding nucleotide sequence has been codon optimized to increase fidelity of replication of the sequence.   
     
     
         47 . The method of  claim 45 , wherein the retinitis pigmentosa is X-linked retinitis pigmentosa. 
     
     
         48 . The method of  claim 45 , wherein the polynucleotide or viral vector is administered to the eye of the subject by subretinal, direct retinal or intravitreal injection. 
     
     
         49 . The method of  claim 48 , wherein the subretinal injection comprises the steps:
 (a) administering a solution to the subject by subretinal injection in an amount effective to at least partially detach the retina to form a subretinal bleb, wherein the solution does not comprise the polynucleotide or viral vector; and   (b) administering a composition by subretinal injection into the bleb formed by step (a), wherein the comprises the polynucleotide or viral vector.   
     
     
         50 . The method of  claim 45 , wherein the polynucleotide or viral vector is administered to the subject in a single dose. 
     
     
         51 . The polynucleotide of claim  1 , which:
 (a) reduces or avoids the generation of alternatively spliced variants and/or truncated RPGR ORF15  proteins relative to a polynucleotide comprising the wild type RPGR ORF 5  gene,   (b) increases the stability and/or fidelity of replication of a nucleotide sequence comprising the RPGR ORF15  gene relative to a nucleotide sequence comprising the wild type RPGR ORF15  gene;   (c) effects higher rates of translation and/or protein expression of RPGR ORF15  protein relative a polynucleotide comprising the wild type RPGR ORF15  gene, and/or   (d) increases the yield of an RPGR ORF 5 -encoding nucleotide sequence relative to a nucleotide sequence comprising the wild type RPGR ORF 5  gene.   
     
     
         52 . A composition comprising a nucleic acid comprising
 (a) a sequence encoding a rhodopsin kinase promoter, and   (b) a sequence encoding a retinitis pigmentosa GTPase regulator ORF15 isoform (RPGRORF15) and comprising a nucleotide sequence that has at least 80% identity to the nucleic acid sequence of SEQ ID NO: 3.