Frataxin expression constructs

Abstract

The invention provides polynucleotides, vectors and viruses expressing frataxin and methods of treating Friedreich's Ataxia.

1 - 56 . (canceled) 
     
     
         57 . A pharmaceutical composition comprising
 (i) a therapeutically effective amount of a recombinant AAV vector comprising a nucleotide sequence encoding a frataxin (FXN) protein comprising the amino acid sequence of SEQ ID NO: 1, or a fragment thereof; and   (ii) empty capsids at a percentage of about 25% to about 95% cp/cp or empty capsids at a percentage of about 0% cp/cp to about 25% cp/cp,   wherein the nucleotide sequence encoding the frataxin (FXN) protein is operably linked to control elements that direct the transcription and translation thereof.   
     
     
         58 . The pharmaceutical composition of  claim 57 , wherein the pharmaceutical composition comprises empty capsids at a percentage of about 50% cp/cp to about 75% cp/cp. 
     
     
         59 . The pharmaceutical composition of  claim 57 , wherein the nucleotide sequence encoding the frataxin protein of SEQ ID NO: 1 differs from SEQ ID NO: 2 due to genetic code degeneracy. 
     
     
         60 . The pharmaceutical composition of  claim 57 , wherein the amino acid threonine at the third position of the frataxin protein of SEQ ID NO: 1 is encoded by a degenerate codon having the nucleotide sequence of ACU, ACC, ACA or ACG. 
     
     
         61 . The pharmaceutical composition of  claim 57 , wherein the amino acid threonine at the third position of the frataxin protein of SEQ ID NO: 1 is encoded by a degenerate codon having the nucleotide sequence of ACC. 
     
     
         62 . The pharmaceutical composition of  claim 60 , wherein the control element comprises a constitutive promoter or tissue-specific promoter. 
     
     
         63 . The pharmaceutical composition of  claim 62 , wherein the constitutive promoter comprises a phosphoglycerate kinase (PGK1) promoter. 
     
     
         64 . The pharmaceutical composition of  claim 62 , wherein the tissue-specific promoter comprises a muscle-specific promoter. 
     
     
         65 . The pharmaceutical composition of  claim 60 , comprising about 1×10 9  to about 2.5×10 14  vg/mL of the recombinant AAV vector. 
     
     
         66 . A method for increasing frataxin (FXN) levels in a subject in need thereof comprising
 administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising:   (i) a recombinant AAV vector effective at increasing frataxin (FXN) levels in a subject with a deficiency of frataxin protein, said recombinant AAV vector comprising the nucleotide sequence encoding a frataxin (FXN) protein comprising the amino acid sequence of SEQ ID NO: 1 or a fragment thereof, and   (ii) empty capsids at a percentage of about 25% to about 95% cp/cp or empty capsids at a percentage of about 0% cp/cp to about 25% cp/cp,   wherein the nucleotide sequence encoding the frataxin protein of SEQ ID NO: 1 is operably linked to control elements that direct the transcription and translation thereof.   
     
     
         67 . The method of  claim 66 , wherein the subject is diagnosed with Friedreich's ataxia. 
     
     
         68 . The method of  claim 66 , wherein the pharmaceutical composition is administered to the subject's brain. 
     
     
         69 . The method of  claim 68 , wherein the pharmaceutical composition is administered to the subject's brain by intraparenchymal, intrathecal, intraventricular, or epidural injection. 
     
     
         70 . The method of  claim 66 , wherein the nucleotide sequence encoding the frataxin protein of SEQ ID NO: 1 differs from SEQ ID NO: 2 due to genetic code degeneracy. 
     
     
         71 . The method of  claim 66 , wherein the amino acid threonine at the third position of the frataxin protein of SEQ ID NO: 1 is encoded by a degenerate codon having the nucleotide sequence of ACU, ACC, ACA or ACG. 
     
     
         72 . The method of  claim 66 , wherein the amino acid threonine at the third position of the frataxin protein of SEQ ID NO: 1 is encoded by a degenerate codon having the nucleotide sequence of ACC. 
     
     
         73 . The method of  claim 66 , wherein the subject comprises an adult aged about 18 years to about 60 years, or a child aged about 3 years or less, about 10 years or less, or about 17 years or less. 
     
     
         74 . The method of  claim 66 , wherein the increase in frataxin protein (FXN) levels improves at least one of the subject's symptoms caused by the deficiency in frataxin protein. 
     
     
         75 . The method of  claim 74 , wherein the at least one of the subject's symptoms comprises a loss of a motor skill. 
     
     
         76 . The method of  claim 74 , wherein the increase in frataxin protein (FXN) levels results in an increase from baseline in FARS total and FARS Neuro, 25-foot walk test, GAITRite Walkway System, Biodex Balance System SD, 9-hole peg test, or combination thereof.