Composition and method for treating complement-mediated disease
Provided herein is a recombinant viral vector having packaged therein an expression cassette comprising an engineered complement regulator factor H (fH), wherein the gene encodes a hfH protein variant comprising short consensus repeats (SCRs) 1-4, 6-8, and 17-20. Pharmaceutical compositions containing this vector and plasmids comprising nucleic acid sequences that encode the fH protein variant are also provided.
1. A recombinant viral vector having packaged therein an expression cassette comprising a nucleic acid sequence that encodes SEQ ID NO: 48 operably linked to expression control sequences which direct expression thereof.
2. The recombinant viral vector according to claim 1 , wherein the vector is an adeno-associated virus (AAV) vector, an adenovirus vector, an RNA virus vector, a lentivirus vector, or a vaccinia virus vector.
3. A recombinant AAV vector comprising an AAV capsid and having packaged therein an expression cassette comprising an engineered complement regulator factor H (fH) gene operably linked to expression control sequences which direct expression thereof, wherein said fH gene encodes the amino acid sequence of SEQ ID NO: 48.
4. The recombinant AAV vector according to claim 3 , wherein the capsid is an AAV1, AAV2, AAV5, AAV8, rh64R1, AAV9, or rh10 capsid.
5. The recombinant AAV vector according to claim 3 , wherein the expression control sequences comprise a liver-specific promoter.
6. The recombinant AAV vector according to claim 3 , wherein the expression control sequences comprise a promoter that is tissue specific for the eye.
7. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the recombinant viral vector according to claim 1 .
8. The recombinant AAV vector according to claim 3 , wherein the expression control sequences comprise a Kozak sequence.
9. The recombinant AAV vector according to claim 3 , wherein the expression control sequences comprise a chicken beta-actin promoter.
10. The recombinant AAV vector according to claim 3 , wherein the expression control sequences comprise a polyadenylation (polyA) signal sequence.
11. The recombinant AAV vector according to claim 10 , wherein the polyA signal sequence is a rabbit beta globin polyA signal sequence.
12. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the recombinant AAV vector according to claim 3 .
13. A plasmid comprising an expression cassette comprising a nucleic acid sequence encoding a fH protein variant comprising SEQ ID NO: 48 operably linked to expression control sequences which direct expression thereof.
14. The plasmid according to claim 13 , wherein the expression cassette further comprises a 5′ ITR and a 3′ ITR.
15. The plasmid according to claim 14 , wherein the 5′ ITR and 3′ ITR are from AAV2.
16. The plasmid according to claim 13 , wherein the expression control sequences comprise a Kozak sequence.
17. The plasmid according to claim 13 , wherein the expression control sequences comprise a chicken beta-actin promoter.
18. The plasmid according to claim 13 , wherein the expression control sequences comprise a polyA signal sequence.
19. The plasmid according to claim 18 , wherein the polyA signal sequence is a rabbit beta-globin polyA signal sequence.
20. An isolated host cell comprising the plasmid according to claim 13 .
21. A recombinant AAV vector comprising an AAV capsid having packaged therein an expression cassette comprising:
(a) an AAV 5′ ITR;
(b) a chicken beta-actin promoter with CMV enhancer;
(c) a nucleic acid sequence that encodes SEQ ID NO: 48;
(d) a polyA signal sequence; and
(e) an AAV 3′ ITR.
22. The recombinant AAV vector according to claim 21 , wherein the capsid is an AAV1, AAV2, AAV5, AAV8, rh64R1, AAV9, or rh10 capsid.
23. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the recombinant AAV vector according to claim 21 .