Frataxin expression constructs

1. A method of treating at least one symptom of Friedreich's Ataxia in a subject in need thereof, said method comprising administering to said subject a therapeutically effective amount of a composition comprising:

(a) an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to:

a 5′UTR selected from 5U2, FTH1, GAPDH, and RPL6-5′Splice;

a 3′UTR selected from a SV40 early adenylation sequence, a SV40 late adenylation sequence, a human growth hormone polyadenylation sequence (hGHpA), and a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; and

a control element that directs transcription and/or translation; and

(b) a pharmaceutically acceptable excipient,

wherein the at least one symptom of Friedreich's Ataxia comprises loss of coordination in the arms and/or legs; and

wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof.

2. The method of claim 1 , wherein said frataxin is expressed in the mitochondria, cerebellum, hippocampus, anterior cortex, dorsal root ganglion, or a combination thereof of the subject.

3. The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 25% of normal levels of frataxin.

4. The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 30% of normal levels of frataxin.

5. The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 40% of normal levels of frataxin.

6. The method of claim 1 wherein said frataxin is expressed in said subject at a level of greater than 50% of normal levels of frataxin.

7. The method of claim 1 , wherein the AAV vector is an AAV5 vector.

8. The method of claim 1 , wherein:

the 5′UTR is FTH1 and the 3′UTR is a SV40 early polyadenylation sequence or a SV40 late polyadenylation sequence;

the 5′UTR is GAPDH and the 3′UTR is a SV40 late polyadenylation sequence or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7;

the 5′UTR is 5U2 and the 3′UTR is a human growth hormone polyadenylation sequence (hGHpA) or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; or

the 5′UTR is RPL6-5′Splice and the 3′UTR is a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7.

9. The method of claim 1 , wherein the control element comprises a promoter.

10. The method of claim 9 , wherein the promoter is a CMV promoter, a UBC promoter, an EF1α promoter, a PGK1 promoter, or a minimal frataxin promoter.

11. The method of claim 9 , wherein the promoter is an inducible promoter.

12. The method of claim 11 , wherein the AAV vector further comprises a gene switch that regulates the inducible promoter.

13. The method of claim 12 , wherein the gene switch is an ecdysone receptor (EcR) gene switch.

14. The method of claim 1 , wherein the pharmaceutically acceptable excipient comprises Na 2 HPO 4 , NaH 2 PO 4 , NaCl, KCl, CaCl 2 , MgCl 2 , or a combination thereof.

15. The method of claim 1 , wherein the composition has a pH of about 6.5 to about 7.5.

16. The method of claim 1 , wherein the composition comprises about 1×10 9 to about 2×10 13 g/mL of the AAV vector.

17. The method of claim 1 , wherein about 1×10 10 to about 1×10 15 vg of the AAV vector is administered to the subject.

18. The method of claim 1 , wherein about 3.7×10 10 , about 1.11×10 11 , or about 3.7×10 11 vg/g of the AAV vector is administered to the subject on a brain weight basis.

19. A method of increasing frataxin levels in a subject having a frataxin deficiency, the method comprising administering to the subject:

an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to:

a 5′UTR selected from 5U2, FTH1, GAPDH, and RPL6-5′Splice;

a 3′UTR selected from a SV40 early adenylation sequence, a SV40 late adenylation sequence, a human growth hormone polyadenylation sequence (hGHpA), and a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; and

a control element that directs transcription and/or translation,

wherein the frataxin encoded by the nucleic acid molecule is produced by the subject at a level that is at least 25% of frataxin levels produced by a subject not having a frataxin deficiency,

wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof.

20. A method of treating at least one symptom of Friedreich's Ataxia in a subject in need thereof, the method comprising administering to the subject a pharmaceutical formulation comprising:

(a) an adeno-associated viral (AAV) vector, wherein said AAV vector comprises a nucleic acid molecule encoding human frataxin operably linked to a 5′UTR, a 3′UTR, wherein:

the 5′UTR is FTH1 and the 3′UTR is a SV40 early polyadenylation sequence or a SV40 late polyadenylation sequence;

the 5′UTR is GAPDH and the 3′UTR is a SV40 late polyadenylation sequence or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7;

the 5′UTR is 5U2 and the 3′UTR is a human growth hormone polyadenylation sequence (hGHpA) or a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7; or

the 5′UTR is RPL6-5′Splice and the 3′UTR is a synthetic 3′ regulatory element having a nucleic acid sequence of SEQ ID NO:7, and

(b) phosphate buffer solution (PBS) or Dulbecco's phosphate buffer solution (DPBS),

wherein the frataxin encoded by the at least one nucleic acid molecule is produced by the subject at a level sufficient to treat at least one symptom of Friedreich's Ataxia in the subject; and

wherein the at least one symptom of Friedreich's Ataxia comprises loss of coordination in the arms and/or legs; and

wherein the administering comprises intrathecal (IT) administration, intracerebroventricular (ICV) administration, intraparenchymal administration, intracerebellar administration, or a combination thereof.

21. The method of claim 20 , wherein about 1×10 10 to about 1×10 15 vg of the AAV vector is administered to the subject.