Patent EA-002544-B1 (EA)
Title: Ret ligand (retl) for stimulating neutral and renal growth
Inventor: SANOCOLA-NADEL MICHELE
IPC: A61K47/48
Filed: 1997-05-07
Abstract: l. Nucleic acid encoding a c-Ret ligand selected from rat RetL1 (SEQ ID No:2), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No: 13), murine RetL3 (SEQ ID No: 17), and human RetL3 (SEQ ID No:21). 2. Nucleic acid of claim 1 encoding an amino acid substitution variant of a c-Ret ligand selected from rat RetL1 (SEQ ID No:2), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No: 13), murine RetL3 (SEQ ID No: 17), and human RetL3 (SEQ ID No:21). 3. Nucleic acid of claim 2 wherein said substitution variant, when aligned with a c-Ret ligand selected from rat RetL1 (SEQ ID No:2), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No: 13), murine RetL3 (SEQ ID No: 17), and human RetL3 (SEQ ID No:21), shares at least 40% sequence similarity therewith, further wherein said substitution variant shares at least 80% of aligned cystein e residues with said c-Ret ligand. 4. Nucleic acid of claim 2 wherein said substitution variant, when aligned with a c-Ret ligand selected from rat RetL1 (SEQ ID No:2), human RetL1 (SEQ ID No:11), human RetL2 (SEQ ID No:13), murine RetL3 (SEQ ID No:17), and human RetL3 (SEQ ID No 21), shares at least 80% sequence similarity therewith. 5. Nucleic acid encoding soluable variant of a rat RetL1 (SEQ ID No:2), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No:13), murine RetL3 (SEQ ID No:17), and human RetL3 (SEQ ID No:21). 6. Nucleic acid having a nucleotide sequence selected from the group comprising rat retL cDNA (SEQ ID No:1), ), partial human retL1 cDNA (SEQ ID No:8), human retL1 cDNA (SEQ ID No: 10), human retL2 (SEQ ID No: 12), murine retL3 (SEQ ID No: 16), partial human retL3 (SEQ ID No:18), and human retL3 (SEQ ID No:20). 7. A vector having the nucleic acid of claim 1, 2, 5 or 6 present as a n insert therein, said vector optionally comprising an expression control sequence operably linked to said insert. 8. A prokaryotic host cell, comprising the vector of claim 7. 9. A eukaryotic host cell, comprising the vector of claim 7. 10. A polypeptide production method, comprising the steps of culturing host cells of claim 8 or 9 in a cell culture medium and, recovering a c-Ret ligand polypeptide expressed from the vector insert within said host cells. 11. A c-Ret ligand polypeptide, said polypeptide having been processed by removal of a hydrophobic N-terminal signal sequence ret ligand polypeptide selected from the group comprising rat RetL1 (SEQ ID No:2), partial human RetL1 (SEQ ID No:9), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No: 13), partial murine RetL3 (SEQ ID No: 15), murine RetL3 (SEQ ID No: 17), partial human RetL3 (SEQ ID No:19), and human RetL3 (SEQ ID No:21). 12. A c-Ret ligand polypeptide of claim 11, which when bound to c-Ret, triggers dimerization or autophosphorylation thereof. 13. c-Ret ligand selected from: (a) rat RetL1 (SEQ ID No:2), partial human RetL1 (SEQ ID No:9), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No: 13), partial murine RetL3 (SEQ ID No: 15), murine RetL3 (SEQ ID No: 17), partial human RetL3 (SEQ ID No:19), and human RetL3 (SEQ ID No:21); or  (b) amino acid substitution variants of rat RetL1 (SEQ ID No:2), partial human RetL1 (SEQ ID No:9), human RetL1 (SEQ ID No: 11), human RetL2 (SEQ ID No:13), partial murine RetL3 (SEQ ID No:15), murine RetL3 (SEQ ID No:17), partial human RetL3 (SEQ ID No:19), and human RetL3 (SEQ ID No:21). 14. A soluble, truncated c-Ret ligand polypeptide comprising at least about 400 amino acids, having a hydrophobic C-terminal sequence. 15. A fusion protein comprising a c-Ret ligand polypeptide of claim 12, 13, 14, 15 or 16, fused to an immunoglobulin polyeptide or a toxin polypeptide, imageable compound or radionuclide. 16. A polyclonal antibody that binds specifically to a c-Ret ligand polypeptide of claim 11, 12, 13 or 14. 17. A monoclonal antibody that binds specifically to a c-Ret ligand polypeptide of claim 11, 12, 13 or 14. 18. An antibody that blocks binding of said c-Ret ligand polypeptide to said c-R et polypeptide. 19. Use of a c-Ret ligand polypeptide of claim 11, 12, 13 or 14 for treating or therapy of renal or neurodegenerative deseases. 20. Use of a fusion protein of claim 15 for treating or therapy of renal or neurodegenerative deseases. 21. Use of an antibody of claims 16, 17 or 18 for treating or therapy of renal or neurodegenerative deseases. 22. A method of stimulating growth of, or limiting damage to, c-Ret expressing t issue in a subject, comprising the step of administering a therapeutically effective amount of a c-Ret ligand polypeptide of claim 11, 12, 13 or 14. 23. The method of claim 22 wherein said c-Ret ligand polypeptide is administered concurrently with a GDNF-related polypeptide. 24. The method of claim 22 wherein said c-Ret expressing tissue is renal tissue or neural tissue. 25. A method of suppressing growth of a tumor cell that expresses c-Ret, comprising the step of contacting said cell with an effective amount of a c-Ret ligand polypeptide of claim 14. 26. A method of suppressing growth of a tumor cell that expresses a c-Ret ligand polypeptide comprising the step of contacting said cell with an effective amount of a c-Ret ligand polypeptide of claim 14. 27. A method of modulating c-Ret signal transduction involving a cell expressing either a c-Ret polypeptide or a c-Ret ligand polypeptide comprising the step of contacting said cell with a c-Ret ligand polypeptide of claim 14. 28. A method of targetting a toxin imageable compound or radionuclide to a cell comprising the step of contacting said cell with a fusion protein of claim 15. 29. A method of suppressing growth of a tumor cell comprising the step of using a fusion protein of claim 15. 30. An isolated nucleic acid having a nucleotide sequence comprising a degenerative sequence selected from the group comprising rat retL cDNA (SEQ ID No:1), ), partial human retL1 cDNA (SEQ ID No:8), human retL1 cDNA (SEQ ID No: 10), human retL2 (SEQ ID No: 12), murine retL3 (SEQ ID No: 16), partial human retL3 (SEQ ID No:18), and human retL3 (SEQ ID No:20). 31. An isolated nucleic acid having a nucleotide sequence comprising a polymorphic sequence selected from the group comprising rat retL cDNA (SEQ ID No:1), ), partial human retL1 cDNA (SEQ ID No:8), human retL1 cDNA (SEQ ID No: 10), human retL2 (SEQ ID No: 12), murine retL3 (SEQ ID No: 16), partial human retL3 (SEQ ID No:18), and human retL3 (SEQ ID No:20). 32. A method of stimulating growth of, or limiting damage to, c-Ret expressing t issue in a subject, comprising the step of administering a therapeutically effective amount of an immunoglobulin fusion protein of claim 15. 33. A method of stimulating growth of, or limiting damage to, c-Ret expressing t issue in a subject, comprising the step of administering a therapeutically effective amount of an antibody of claim 16, 17 or 18. 34. A method of suppressing growth of a tumor cell that expresses c-Ret, comprising the step of contacting said cell with an effective amount of an antibody of claim 16, 17 or 18. 35. A method of modulating c-Ret signal transduction involving a cell expressing either a c-Ret polypeptide or a c-Ret ligand polypeptide comprising the step of using a fusion protein of claim 15. 36. A method of modulating c-Ret signal transduction involving a cell expressing either a c-Ret polypeptide or a c-Ret ligand polypeptide comprising the step of contacting of said cell with the antibody of claim 16, 17 or 18. 37. A method of targetting a toxin imageable compound or radionuclide to a cell comprising the step of using the antibody of claim 16, 17 or 18 conjugated to said toxin imageable compound or radionuclide. 38. A method of suppressing growth of a tumor cell expressing either a c-Ret polypeptide or a c-Ret ligand polypeptide comprising the step of contacting said cell with the antibody of claim 16, 17 or 18 conjugated to said toxin imageable compound or radionuclide. 39. A vector comprising a nucleic acid of claim 30 or 31 present as a n insert therein, said vector optionally comprising an expression control sequence operably linked to said insert. 40. A prokaryotic host cell, comprising the vector of claim 39. 41. A eukaryotic host cell, comprising the vector of claim 39. 42. A process for the production of a polypeptide product, said process comprising the step of culturing host cells of claim 40 or 41 in a cell culture medium and, recovering a c-Ret ligand polypeptide expressed from the vector insert within said host cells.