Recombinant adeno-associated viral vectors for treating Bietti Crystalline Dystrophy
The present disclosure provides a recombinant adeno-associated vector comprising a codon-optimized sequence encoding CYP4V2 linked to selected gene expression regulatory sequences and its use in treating Bietti Crystalline Dystrophy (BCD).
1. An isolated nucleic acid molecule, comprising the nucleotide sequence of SEQ ID NO: 8, 9, 15, 16 or 17, and that encodes human CYP4V2 polypeptide.
2. The isolated nucleic acid molecule of claim 1 , wherein the nucleotide sequence is SEQ ID NO: 8 or SEQ ID NO: 16.
3. The isolated nucleic acid molecule of claim 1 , further comprising a promoter operatively linked to the 5′ of the nucleotide sequence encoding CYP4V2.
4. The isolated nucleic acid molecule of claim 3 , wherein the promoter is a CAG promoter.
5. The isolated nucleic acid molecule of claim 1 , further comprising a polyadenylation sequence at the 3′ of the nucleotide sequence encoding CYP4V2.
6. The isolated nucleic acid molecule of claim 5 , wherein the polyadenylation (polyA) sequence is bovine growth hormone (bGH) polyA, synthesized polyA (SPA) or Simian Virus 40 (SV40) polyA.
7. The isolated nucleic acid molecule of claim 1 , further comprising a Woodchuck Hepatitis Virus posttranscriptional regulatory element (WPRE).
8. The isolated nucleic acid molecule of claim 1 , comprising the nucleotide sequence of SEQ ID NO:25 having SEQ ID NO:8, the nucleotide sequence of SEQ ID NO:26 having SEQ ID NO:9, the nucleotide sequence of SEQ ID NO:32 having SEQ ID NO:15, the nucleotide sequence of SEQ ID NO:33 having SEQ ID NO:16, or the nucleotide sequence of SEQ ID NO:34 having SEQ ID NO:17.
9. A recombinant AAV (rAAV) vector, comprising the nucleic acid molecule of claim 1 .
10. The recombinant AAV vector of claim 9 , comprising two inverted terminal repeats.
11. The recombinant AAV vector of claim 10 , comprising two AAV2 inverted terminal repeats.
12. A viral particle comprising the recombinant AAV vector of claim 9 packaged into an AAV capsid.
13. The viral particle of claim 12 , wherein the AAV capsid is AAV8 capsid.
14. A pharmaceutical composition comprising the viral particle of claim 12 , and a pharmaceutically acceptable excipient.
15. A method of alleviating a symptom of Bietti Crystalline Dystrophy (BCD) in a mammalian subject, comprising administering via subretinal injection of an effective amount of the rAAV vector of claim 9 to the subject.