Factor H variants for treatment of disease
Disclosed herein are factor H variants, vectors comprising said factor H variants, and pharmaceutical compositions comprising the same for treatment of disorders.
1. A recombinant vector having packaged therein an expression cassette comprising an engineered human complement factor H (hfH) gene operably linked to expression control sequences which direct expression thereof, wherein the hfH gene encodes a soluble hfH protein variant that retains complement regulatory function, wherein the hfH protein variant comprises:
amino acids 1 to 62 of SEQ ID NO: 3 (SCR1)—L1—amino acids 1 to 57 of SEQ ID NO: 5 (SCR2)—L2—amino acids 1 to 60 of SEQ ID NO: 7 (SCR3)—L3—amino acids 1 to 53 of SEQ ID NO: 9 (SCR4)—L4—amino acids 1 to 61 of SEQ ID NO: 13 (SCR6)—L4′—amino acids 1 to 54 of SEQ ID NO: 14 (SCR7)—L5—amino acids 1 to 58 of SEQ ID NO: 16 (SCR8)—L5″—amino acids 1 to 56 of SEQ ID NO: 33 (SCR17)—L5″′—amino acids 1 to 55 of SEQ ID NO: 35 (SCR18)—L5″″—amino acids 1 to 55 of SEQ ID NO: 37 (SCR19)—L6—amino acids 1 to 65 of SEQ ID NO: 38 (SCR20), wherein “L” refers to a linker, and each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 18 amino acids, and wherein SCR5, SCR9, SCR10, SCR11, SCR12, SCR13, SCR14, SCR15, and SCR16 are absent.
2. The recombinant vector according to claim 1 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.
3. The recombinant 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.
4. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the recombinant vector according to claim 1 .
5. The recombinant vector according to claim 1 , wherein each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 12 amino acids.
6. The recombinant vector according to claim 5 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.
7. A recombinant AAV vector comprising an AAV capsid having packaged therein an expression cassette comprising an engineered human complement regulator factor H (hfH) gene operably linked to expression control sequences which direct expression thereof, wherein the hfH gene encodes a soluble hfH protein variant that retains complement regulatory function, wherein the hfH protein variant comprises:
amino acids 1 to 62 of SEQ ID NO: 3 (SCR1)—L1—amino acids 1 to 57 of SEQ ID NO: 5 (SCR2)—L2—amino acids 1 to 60 of SEQ ID NO: 7 (SCR3)—L3—amino acids 1 to 53 of SEQ ID NO: 9 (SCR4)—L4—amino acids 1 to 61 of SEQ ID NO: 13 (SCR6)—L4′—amino acids 1 to 54 of SEQ ID NO: 14 (SCR7)—L5—amino acids 1 to 58 of SEQ ID NO: 16 (SCR8)—L5″—amino acids 1 to 56 of SEQ ID NO: 33 (SCR17)—L5″′—amino acids 1 to 55 of SEQ ID NO: 35 (SCR18)—L5″″—amino acids 1 to 55 of SEQ ID NO: 37 (SCR19)—L6—amino acids 1 to 65 of SEQ ID NO: 38 (SCR20), wherein “L” refers to a linker, and each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 18 amino acids, and wherein SCR5, SCR9, SCR10, SCR11, SCR12, SCR13, SCR14, SCR15, and SCR16 are absent.
8. The recombinant AAV vector according to claim 7 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.
9. The recombinant AAV vector according to claim 7 , wherein the AAV capsid is an AAV1, an AAV2, an AAV5, an AAV8, an rh64R1, an AAV9, or an rh10 capsid.
10. The recombinant AAV vector according to claim 7 , wherein the expression control sequences comprise a liver-specific promoter sequence.
11. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and the recombinant AAV vector according to claim 7 .
12. The recombinant AAV vector according to claim 9 , wherein the AAV capsid is an AAV2 capsid.
13. The recombinant AAV vector according to claim 7 , wherein the expression control sequences comprise a constitutive promoter sequence.
14. The recombinant AAV vector according to claim 7 , wherein the expression control sequences comprise a chicken beta-actin promoter sequence.
15. The recombinant AAV vector according to claim 7 , wherein the hfH gene is codon optimized.
16. The recombinant AAV vector according to claim 7 , wherein each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 12 amino acids.
17. The recombinant AAV according to claim 16 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.
18. A plasmid comprising a nucleotide sequence that encodes an engineered human complement regulator factor H (hfH) gene operably linked to expression control sequences which direct expression thereof, wherein the hfH gene encodes a soluble hfH protein variant that retains complement regulatory function, wherein the hfH protein variant comprises:
amino acids 1 to 62 of SEQ ID NO: 3 (SCR1)—L1—amino acids 1 to 57 of SEQ ID NO: 5 (SCR2)—L2—amino acids 1 to 60 of SEQ ID NO: 7 (SCR3)—L3—amino acids 1 to 53 of SEQ ID NO: 9 (SCR4)—L4—amino acids 1 to 61 of SEQ ID NO: 13 (SCR6)—L4′—amino acids 1 to 54 of SEQ ID NO: 14 (SCR7)—L5—amino acids 1 to 58 of SEQ ID NO: 16 (SCR8)—L5″—amino acids 1 to 56 of SEQ ID NO: 33 (SCR17)—L5″′—amino acids 1 to 55 of SEQ ID NO: 35 (SCR18)—L5″″—amino acids 1 to 55 of SEQ ID NO: 37 (SCR19)—L6—amino acids 1 to 65 of SEQ ID NO: 38 (SCR20), wherein “L” refers to a linker, and each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 18 amino acids, and wherein SCR5, SCR9, SCR10, SCR11, SCR12, SCR13, SCR14, SCR15, and SCR16 are absent.
19. The plasmid according to claim 18 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.
20. The plasmid according to claim 18 , wherein the expression control sequences comprise a constitutive promoter sequence.
21. The plasmid according to claim 18 , wherein the expression control sequences comprise a chicken beta-actin promoter sequence and/or a Kozak sequence.
22. A packaging host cell comprising the plasmid according to claim 18 .
23. The plasmid according to claim 18 , wherein each of L1, L2, L3, L4, L4′, L5, L5″, L5″′, L5″″, and L6 is independently selected from an amino acid sequence of 1 to 12 amino acids.
24. The plasmid according to claim 23 , wherein the hfH protein variant comprises an amino acid sequence at least 95% identical to amino acids 1 to 701 of SEQ ID NO: 48.