Methods to produce rod-derived cone viability factor (rdcvf)

Abstract

The invention is related to methods of producing rod-derived cone viability factor (RdCVF). This invention also relates to the treatment of an ocular disease in a mammal using RdCVF. Also provided are expression vectors for high secreted expression of RdCVF of using nucleotide sequences encoding heterologous signal proteins and optionally markers for furin cleavage.

1 - 46 . (canceled) 
     
     
         47 . A method of treating Dry Age-related Macular Degeneration (Dry AMD) in a mammal, the method comprising:
 (1) administering to a diseased eye of the mammal a nucleic acid encoding a RdCVF fusion protein, where the nucleic acid encoding the RdCVF fusion protein comprises:
 a) a RdCVF-encoding nucleotide sequence; 
 b) a signal peptide-encoding nucleotide sequence positioned upstream of the RdCVF-encoding nucleotide sequence, wherein the signal peptide-encoding sequence encodes the signal peptide of human growth hormone (hGH), brain-derived neurotrophic factor (BDNF), or β-glucoronidase (GUSB), and directs secretion of expressed RdCVF; and 
 c) a nucleotide sequence encoding a furin cleavage marker located downstream of the signal peptide-encoding nucleotide sequence and upstream of the RdCVF-encoding nucleotide sequence; and 
   (2) expressing the RdCVF fusion protein at a therapeutically effective amount in the diseased eye to treat Dry AMD.   
     
     
         48 . The method of  claim 47 , wherein the nucleic acid encoding the RdCVF fusion protein is an expression vector that is a recombinant adeno-associated virus (AAV) or adenovirus (Ad). 
     
     
         49 . (canceled) 
     
     
         50 . The method of  claim 47 , wherein the signal peptide-encoding sequence and the furin cleavage marker sequence are separated by a sequence encoding an amino acid linker. 
     
     
         51 . The method of  claim 47 , wherein the RdCVF-encoding nucleotide sequence lacks the initiating methionine coding sequence. 
     
     
         52 . (canceled) 
     
     
         53 . A method of treating Dry Age-related Macular Degeneration (Dry AMD)in a mammal, the method comprising:
 (1) administering to a diseased eye of the mammal a nucleic acid encoding a RdCVF fusion protein, where the nucleic acid encoding the RdCVF fusion protein comprises:
 a) a RdCVF-encoding nucleotide sequence lacking the initiating methionine coding sequence; 
 b) an exogenous signal peptide-encoding nucleotide sequence positioned upstream of the RdCVF-encoding nucleotide sequence, wherein the exogenous signal peptide-encoding sequence is selected from the group consisting of: human growth hormone (hGH), brain-derived neurotrophic factor (BDNF), and β-glucoronidase (GUSB); and 
   (2) expressing the RdCVF fusion protein at a therapeutically effective amount in the diseased eye to treat the Dry AMD.   
     
     
         54 . The method of  claim 53 , wherein the nucleic acid encoding the RdCVF fusion protein is an expression vector that is a recombinant adeno-associated virus (AAV) or adenovirus (Ad). 
     
     
         55 . (canceled)