Fusion proteins comprising PDGF and VEGF binding portions and methods of using thereof
The present provides fusion proteins comprising PDGF and VEGF binding portions, and recombinant viral particles encoding the fusion proteins. Compositions comprising the fusion proteins and viral particles as well as methods of using the same are also provided.
1. A fusion protein comprising (a) Ig-like domains D1-D3 of a PDGF receptor beta (PDGFRβ), (b) an extracellular portion of an Ig-like domain D2 of a VEGF receptor 1 (FLT1), and (c) a multimerization domain, wherein the fusion proteins binds to a PDGF and a VEGF, and the fusion protein is arranged from N-terminus to C-terminus in the following order: (a), (b) and (c) and wherein the fusion protein does not comprise an Ig-like domain D2 of a VEGFR2.
2. The fusion protein of claim 1 , wherein the extracellular portion of the PDGFR comprises:
(a) the Ig-like domains D1-D4 of the PDGFR;
(b) the Ig-like domains D1-D5 of the PDGFR; or
(c) the amino acid sequence SEQ ID NO:1, 2, or 3.
3. The fusion protein of claim 1 , wherein the extracellular portion of the VEGF receptor comprises:
(a) Ig-like domains D1-D3 of a VEGFR1 (FLT-1); or
(b) the amino acid sequence of SEQ ID NO:4 or 5.
4. The fusion protein of claim 1 , wherein the fusion protein further comprises a linker peptide between the extracellular portion of the PDGF receptor and the extracellular portion of the VEGF receptor, and/or a peptide linker between the extracellular portion of the VEGF receptor and the multimerization domain.
5. The fusion protein of claim 4 , wherein the peptide linker comprises the amino acid sequence selected from the group consisting of Gly 9 (SEQ ID NO:42), Glu 9 (SEQ ID NO:43), Ser 9 (SEQ ID NO:44), Gly 5 Cys-Pro t -Cys (SEQ ID NO:45), (Gly 4 -Ser) 3 (SEQ ID NO:46), Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO:47), Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO:48), Gly-Asp-Leu-Ile-Tyr-Arg-Asn-Gln-Lys (SEQ ID NO:49), and Gly 9 -Pro-Ser-Cys-Val-Pro-Leu-Met-Arg-Cys-Gly-Gly-Cys-Cys-Asn (SEQ ID NO:50).
6. The fusion protein of claim 1 , wherein the multimerization domain is:
(a) a Fc region of an antibody;
(b) a Fc region of an antibody comprising a CH3 region of IgG1, IgG2, IgG3, or IgG4, or a CH2 and a CH3 region of IgG1, IgG2, IgG3, or IgG4; or
(c) a Fc region of an antibody comprising the amino acid sequence of SEQ ID NO:6.
7. The fusion protein of claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:13 or 15.
8. The fusion protein of claim 1 , wherein the fusion protein is in a dimeric or a multimeric form.
9. A composition comprising the fusion protein claim 1 and a pharmaceutically acceptable carrier.
10. The fusion protein of claim 1 , wherein the fusion protein comprises an amino acid sequence having at least 90% identity to SEQ ID NO:13 or 15.