Engineered VEGF variants for retinal neuroprotection, promotion of axon growth and axon regeneration
The invention provides methods of promoting retinal ganglion axon formation, survival, and synapse formation.
1. An engineered vascular endothelial growth factor (VEGF) chimeric polypeptide comprising: a first VEGF polypeptide covalently linked to a second VEGF polypeptide via a first polypeptide linker comprising the amino acid sequence of SEQ ID NO: 14, and a second polypeptide linker comprising the amino acid sequence of SEQ ID NO: 15 linking the second VEGF polypeptide to a flexible transmembrane anchor.
2. The engineered VEGF chimeric polypeptide of claim 1 , wherein the flexible transmembrane anchor is a glycosylphosphatidylinisotol (GPI) anchor.
3. The engineered VEGF chimeric polypeptide of claim 1 , further comprising a signal peptide.
4. The engineered VEGF chimeric polypeptide of claim 1 , wherein a signal peptide is linked to the N-terminal end of the first VEGF polypeptide.
5. The engineered VEGF chimeric polypeptide of claim 1 , wherein each of the first and second VEGF polypeptides comprises about 100 amino acids to about 200 amino acids.
6. The engineered VEGF chimeric polypeptide of claim 5 , wherein each of the first and second VEGF polypeptides comprises about 121 amino acids.
7. The engineered VEGF chimeric polypeptide of claim 1 , wherein the first and second VEGF polypeptide comprise: isoforms, mutants, variants, derivatives, active fragments of VEGF or combinations thereof.
8. The engineered VEGF chimeric polypeptide of claim 1 , wherein each of the first and second VEGF polypeptides is a VEGF isoform comprising 121 amino acids.
9. The engineered VEGF chimeric polypeptide of claim 8 , wherein the VEGF isoform comprises the amino acid sequence set forth as SEQ ID NO: 13.
10. The engineered VEGF chimeric polypeptide of claim 1 , wherein the flexible transmembrane anchor comprises SEQ ID NO: 6.
11. An engineered vascular endothelial growth factor (VEGF) chimeric polypeptide comprising: a sequence identity that is at least 80% identical to SEQ ID NO: 1 over the entire length of SEQ ID NO: 1.
12. The engineered VEGF chimeric polypeptide of claim 11 , wherein the VEGF chimeric polypeptide comprises SEQ ID NO: 1.
13. The engineered vascular VEGF chimeric polypeptide of claim 11 , further comprising a flexible peptide linker comprising about 7 amino acids.
14. The engineered vascular VEGF chimeric polypeptide of claim 11 , wherein the VEGF polypeptide comprises about 189 amino acids.
15. A pharmaceutical composition comprising
the engineered vascular endothelial growth factor (VEGF) chimeric polypeptide of claim 1 .
16. A retinal ganglion cell (RGC) comprising a membrane anchored vascular endothelial growth factor (VEGF) chimeric polypeptide, wherein said chimeric VEGF polypeptide comprises SEQ ID NO: 1.
17. A stem cell comprising the engineered vascular endothelial growth factor (VEGF) chimeric polypeptide of claim 1 .