Gene therapy for ocular disorders

Abstract

Compositions and methods are provided for treating Leber congenital amaurosis (LCA) in a subject. In one aspect, a recombinant adeno-associated viral vector is provided which includes a nucleic acid molecule comprising a sequence encoding Lebercilin. In another aspect, Lebercilin has an amino acid sequence of SEQ ID NO: 1. In yet another aspect, the nucleic acid molecule has a sequence of SEQ ID NO: 3 or a variant thereof. In desired embodiments, the subject is human, cat, dog, sheep, or non-human primate.

1 . A nucleic acid sequence encoding human lebercilin, wherein the nucleic acid sequence is at least 90% identical to SEQ ID NO: 3. 
     
     
         2 . A recombinant adeno-associated virus (rAAV) comprising an AAV capsid protein and the nucleic acid of  claim 1 . 
     
     
         3 . The rAAV of  claim 2 , wherein the nucleotide sequence is at least 95% identical to SEQ ID NO: 3. 
     
     
         4 . The rAAV of  claim 2 , wherein the nucleotide sequence is operatively associated with expression control sequences that directs expression of the nucleotide sequence in a host cell. 
     
     
         5 . An rAAV expression cassette comprising the rAAV of  claim 4 , a 5′ AAV inverted terminal repeat (ITR), and a 3′ AAV ITR. 
     
     
         6 . A plasmid comprising the expression cassette of  claim 5 . 
     
     
         7 . The rAAV of  claim 4 , wherein the host cell is a human cell. 
     
     
         8 . The rAAV of  claim 4 , wherein the host cell is a photoreceptor cell. 
     
     
         9 . The rAAV of  claim 4 , wherein the expression control sequences comprise a rhodopsin kinase promoter sequence. 
     
     
         10 . The rAAV of  claim 4 , wherein the expression control sequences comprise a cytomegalovirus (CMV) promoter sequence or a hybrid promoter sequence comprising a CMV promoter sequence and a chicken beta actin (CBA) promoter sequence. 
     
     
         11 . The rAAV of  claim 1 , wherein the AAV capsid protein is an AAV8 capsid, or variant thereof, an AAV7 capsid, or variant thereof, an AAV5 capsid, or variant thereof, or an AAV2 capsid or variant thereof. 
     
     
         12 . The rAAV of  claim 1 , wherein the AAV capsid protein is an AAV8 capsid. 
     
     
         13 . A host cell comprising the rAAV of  claim 1 . 
     
     
         14 . A composition comprising the rAAV of  claim 1  and a carrier or excipient suitable for delivery to a plurality of ocular cells of a subject. 
     
     
         15 . The composition of  claim 14 , comprised in a lipid delivery vehicle. 
     
     
         16 . The composition of  claim 15 , wherein the lipid delivery vehicle is a liposome. 
     
     
         17 . A recombinant adeno-associated virus (rAAV) comprising an AAV8 capsid and an expression cassette comprising:
 a) a 5′ AAV ITR;   b) a rhodopsin kinase promoter;   c) a nucleotide sequence at least 90% identical to SEQ ID NO: 3; and   d) a 3′AAV ITR.   
     
     
         18 . The rAAV of  claim 17 , wherein the nucleotide sequence is at least 95% identical to SEQ ID NO: 3. 
     
     
         19 . A composition comprising the rAAV of  claim 17 , and a pharmaceutically acceptable excipient. 
     
     
         20 . The composition of  claim 19 , wherein the pharmaceutically acceptable excipient is a carrier or excipient suitable for delivery to a plurality of ocular cells of a subject. 
     
     
         21 . The composition of  claim 19 , comprised in a lipid delivery vehicle. 
     
     
         22 . The composition of  claim 21 , wherein the lipid delivery vehicle is a liposome. 
     
     
         23 . The composition of  claim 20 , wherein the subject is a human. 
     
     
         24 . A method of treating a subject with an eye disease or disorder, the method comprising administering to the subject a recombinant adeno-associated virus (rAAV) comprising an AAV capsid protein and a nucleic acid comprising a nucleotide sequence at least 90% identical to SEQ ID NO: 3.