Gene therapy for macular degeneration
The invention provides compositions and methods for treatment of age-related macular degeneration, including gene therapy employing vectors and transgenes expressing protective CFH polypeptide and CFHT polypeptide sequences.
1 . A viral vector comprising:
(i) a recombinant polynucleotide transgene comprising a polynucleotide sequence that has at least 98% identity to SEQ ID NO:3, wherein the polynucleotide sequence encodes a truncated complement factor H (CFHT) polypeptide, wherein the CFHT polypeptide comprises isoleucine (I) at position 62 and tyrosine (Y) at position 402 a;
(ii) a CBA promoter operably linked to the polynucleotide transgene, wherein the CBA promoter comprises a polynucleotide sequence having at least 98% identity to any of SEQ ID NO: 12-14;
(iii) a Bovine Growth Factor (bGH) polyadenylation sequence; and
(iv) left and right inverted terminal repeat (ITR) sequences,
wherein the viral vector is an adeno-associated virus 2 (AAV2) vector, and
wherein introduction of the polynucleotide transgene into a mammalian cell results in expression of the CFHT polypeptide.
2 . The viral vector of claim 1 wherein the CFHT polypeptide comprises
(a) residues 1-431 of SEQ ID NO:21; or
(b) a variant CFHT with at least 98% identity to residues 1-431 of SEQ ID NO:21.
3 . The viral vector of claim 1 wherein the promoter is a CBA promoter comprising SEQ ID NO:13.
4 . The viral vector of claim 1 wherein the polyadenylation sequence is a Bovine Growth Factor (bGH) polyadenylation sequence comprising SEQ ID NO: 29.
5 . The viral vector of claim 1 wherein the CFHT polypeptide is encoded by a polynucleotide sequence having at least 99% identity to SEQ ID NO:3.
6 . The viral vector of claim 1 , wherein the CBA promoter comprises the polynucleotide sequence set forth as any one of SEQ ID NO:12-14.
7 . The viral vector of claim 1 wherein the CFHT polypeptide comprises (a) residues 1-449 of SEQ ID NO:4; or (b) a variant CFHT with at least 98% identity to residues 1-449 of SEQ ID NO: 4.
8 . The viral vector of claim 1 wherein one of the left and right ITRs comprises SEQ ID NO: 18 and the other ITR comprises the reverse complement of SEQ ID NO: 18.
9 . The viral vector of claim 8 wherein the CFHT polypeptide is encoded by a polynucleotide sequence comprising SEQ ID NO:3.
10 . The viral vector of claim 1 wherein the CFHT polypeptide comprises SEQ ID NO:21, the promoter comprises SEQ ID NO:13; and the bGH polyadenylation sequence comprises SEQ ID NO:29.
11 . The viral vector of claim 10 wherein the polynucleotide sequence encoding the CFHT polypeptide comprises SEQ ID NO:3.
12 . A pharmaceutical composition comprising a therapeutic amount of the viral vector of claim 1 , and a pharmaceutically acceptable carrier or excipient.
13 . A method comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 12 to a subject in need of treatment for AMD, wherein the pharmaceutical composition is administered subretinally, intravitreally, intravascularly, extraocularly, or to the choroid.
14 . The method of claim 13 wherein the truncated CFHT polypeptide is expressed by retinal pigment epithelial (RPE) cells.
15 . A method comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 12 to reduce a subject's risk of developing AMD, wherein the subject is selected to be homozygous or heterozygous for a Chromosome 1 CFH risk allele, wherein the subject comprises the following CFH proteins encoded by the CFH risk alleles present in the subject: VV62/HH402/EE936, VV62/YH402/ED936, IV62/YH402/EE936, VV62/YH402/EE936, VV62/YY402/DD936/IV62/YY402/ED936, VV62/YY402/ED936, 1162/YY402/EE936, IV62/YY402/EE936, or VV62/YY402/EE936.
16 . The method of claim 15 wherein the CFH proteins encoded by the CFH risk alleles present in the subject comprise VV62/HH402/EE936, VV62/YH402/ED936, VV62/YH402/EE936, or IV62/YH402/EE936, and the subject is heterozygous or homozygous for a CFHR3/1 no deletion risk allele.