Fusion proteins comprising pdgf and vegf binding portions and methods of using thereof

Abstract

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 - 50 . (canceled) 
     
     
         51 . A method of delivering a viral vector to a subject, comprising administering an rAAV particle comprising a nucleic acid encoding a fusion protein;
 wherein the fusion protein comprises (a) an extracellular portion of a PDGF receptor comprising at least Ig-like domains D1-D3 of a PDGF receptor beta (PDGFRβ), (b) an extracellular portion of a VEGF receptor 1 (Flt-1) comprising at least an Ig-like domain D2, and (c) a multimerization domain, wherein the fusion proteins bind to a PDGF and a VEGF, and the fusion protein is arranged from N-terminal to C-terminus in the following order: (a), (b), and (c);   wherein the fusion protein does not comprise an Ig-like domain D3 of a VEGFR2.   
     
     
         52 . The method of  claim 51 , wherein the subject has macular degeneration or proliferative diabetic retinopathy. 
     
     
         53 . The method of  claim 52 , wherein the subject has macular degeneration that is wet age-related macular degeneration or dry age-related macular degeneration. 
     
     
         54 . The method of  claim 51 , wherein the rAAV particle is administered by intravitreal injection to the subject. 
     
     
         55 . The method of  claim 51 , wherein the rAAV particle comprises capsid proteins of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, or AAVrh10. 
     
     
         56 . The method of  claim 55 , wherein the nucleic acid comprises an ITR from a serotype different from the serotype of the capsid. 
     
     
         57 . The method of  claim 56 , wherein the ITR is an ITR of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, or AAVrh10. 
     
     
         58 . The method of  claim 51 , wherein the subject has cancer. 
     
     
         59 . The method of  claim 51 , wherein the subject has rheumatoid arthritis, osteoarthritis, or asthma. 
     
     
         60 . The method of  claim 51 , wherein the subject has uveitis or corneal neovascularization. 
     
     
         61 . The method of  claim 51 , wherein (a) comprises:
 (i) Ig-like domains D1-D4 of a PDGF receptor beta;   (ii) Ig-like domains D1-D5 of a PDGF receptor beta; or   (iii) amino acid sequence SEQ ID NO:1, 2, or 3.   
     
     
         62 . The method of  claim 51 , wherein (b) comprises:
 (i) Ig-like domains D1-D3 of a VEGFR1 (Flt-1); or   (ii) amino acid sequence SEQ ID NO:4 or 5.   
     
     
         63 . The method of  claim 51 , wherein the fusion protein further comprises at least one of a linker peptide between the extracellular portion of the PDGF receptor and the extracellular portion of the VEGF receptor and a peptide linker between the extracellular portion of the VEGF receptor and the multimerization domain. 
     
     
         64 . The method of  claim 63 , wherein the peptide linker comprises the amino acid sequence selected from the group consisting of SEQ ID NOs: 42-50. 
     
     
         65 . The method of  claim 51 , wherein the multimerization domain is:
 (i) a Fc region of an antibody;   (ii) 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, IgG3, or IgG4; or   (iii) a Fc region of an antibody comprising the amino acid sequence of SEQ ID NO:6.   
     
     
         66 . The method of  claim 51 , wherein the fusion protein is in a dimeric or a multimeric form. 
     
     
         67 . The method of  claim 51 , wherein the rAAV particle is administered as a pharmaceutical composition comprising the rAAV particle and a pharmaceutically acceptable carrier. 
     
     
         68 . A method of delivering a viral vector to a subject, comprising administering by intravitreal injection an rAAV particle comprising a nucleic acid encoding a fusion protein;
 wherein the fusion protein comprises (a) an extracellular portion of a PDGF receptor comprising at least Ig-like domains D1-D3 of a PDGF receptor beta (PDGFRβ), (b) an extracellular portion of a VEGF receptor 1 (Flt-1) comprising at least an Ig-like domain D2, and (c) a multimerization domain, wherein the fusion proteins bind to a PDGF and a VEGF, and the fusion protein is arranged from N-terminal to C-terminus in the following order: (a), (b), and (c);   wherein the fusion protein further comprises at least one of a linker peptide between the extracellular portion of the PDGF receptor and the extracellular portion of the VEGF receptor and a peptide linker between the extracellular portion of the VEGF receptor and the multimerization domain;   wherein the fusion protein does not comprise an Ig-like domain D3 of a VEGFR2.   
     
     
         69 . The method of  claim 68 , wherein the wherein the rAAV particle comprises capsid proteins of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, or AAVrh10. 
     
     
         70 . The method of  claim 69 , wherein the nucleic acid comprises an ITR from a serotype different from the serotype of the capsid, and wherein the ITR is an ITR of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, or AAVrh10. 
     
     
         71 . The method of  claim 68 , wherein the subject has macular degeneration or proliferative diabetic retinopathy.