Method of treatment for lymphedema comprising administering a polynucleotide encoding VEGF-D
The present invention provides materials and methods for screening for and treating hereditary lymphedema in human subjects.
1. A method of treatment for lymphedema, comprising: administering to a patient with lymphedema a therapeutically effective amount of a polynucleotide that comprises a nucleotide sequence that encodes a vascular endothelial growth factor D (VEGF-D) protein that binds and stimulates wild-type VEGFR-3; and wherein said therapeutically effective amount of said polynucleotide is administered locally at a site of edema in the patient.
2. The method of claim 1 , wherein the VEGF-D protein comprises amino acids 92-205 of SEQ ID NO: 6.
3. The method of claim 1 , wherein the VEGF-D protein comprises amino acids 101-196 of SEQ ID NO: 6.
4. The method of claim 1 , wherein said VEGF-D protein comprises a member selected from the group consisting of: (a) a polypeptide comprising the amino acid sequence of SEQ ID NO: 6; and (b) a polypeptide that comprises an amino acid sequence that comprises a continuous portion of SEQ ID NO: 6 sufficient to permit the polypeptide to bind and stimulate wild-type human VEGFR-3.
5. The method of claim 1 , wherein the VEGF-D protein comprises amino acids 93-201 of SEQ ID NO: 6.
6. The method according to claim 5 , wherein the polynucleotide further includes a nucleotide sequence encoding a secretory signal peptide fused in-frame with the nucleotide sequence encoding the VEGF-D protein.
7. The method according to claim 5 , wherein the polynucleotide further includes a promoter and/or enhancer sequence operatively linked upstream of the nucleotide sequence that encodes the VEGF-D protein.
8. The method according to claim 6 , wherein the polynucleotide further includes a promoter and/or enhancer sequence operatively linked upstream of the nucleotide sequence that encodes the secretory signal peptide and the VEGF-D protein.
9. The method according to claim 7 , wherein the polynucleotide further includes a polyadenylation sequence operatively linked downstream of the nucleotide sequence that encodes the VEGF-D protein.
10. The method according to claim 8 , wherein the polynucleotide further includes a polyadenylation sequence operatively linked downstream of the nucleotide sequence that encodes the secretory signal peptide and the VEGF-D protein.
11. The method according to claim 1 , comprising administering a vector to the patient, wherein the vector comprises the polynucleotide.
12. The method according to claim 4 , comprising administering a vector to the patient, wherein the vector comprises the polynucleotide.
13. The method according to claim 12 , wherein the vector is a replication-deficient retroviral vector.
14. The method according to claim 12 , wherein the vector is selected from the group consisting of replication-deficient lentivurus vectors, adeno-associated viral vectors, adenoviral vectors, and combinations thereof.
15. The method according to claim 12 , wherein the vector further includes at least one sequence operatively linked to the polynucleotide selected from the group consisting of:
a nucleotide sequence encoding a secretory signal peptide fused in-frame with the nucleotide sequence encoding the VEGF-D protein;
a promoter and/or enhancer sequence operatively linked upstream of the nucleotide sequence that encodes the VEGF-D protein, or operatively linked upstream of the nucleotide sequence that encodes the secretory signal peptide and the VEGF-D protein; and
a polyadenylation sequence operatively linked downstream of the nucleotide sequence that encodes the VEGF-D protein.
16. The method of claim 12 , wherein said administering of said therapeutically effTective amount of said polynucleotide induces VEGFR-3 signaling in the lymphatic endothelia of the patient.
17. The method of claim 12 , wherein said administering of said therapeutically effecTive amount of said polynucleotide reduces edema in a limb of said patient.
18. The method of claim 12 , wherein said administering of said therapeutically effective amount of said polynucleotide reduces accumulation of lymph fluids in said patient.
19. The method of claim 12 . wherein said polynucleotide is administered via intravenous injection.
20. The method of claim 12 , wherein said polynucleotide is administered via intramuscular injection.