Hemojuvelin fusion proteins and uses thereof
The present invention provides a hemojuvelin (HJV) fusion protein (e.g., a human HJV.Fc) protein, polynucleotides and vectors encoding such proteins, and methods for making such proteins. Also provided are methods for treating iron-related disorders which include administration of a HJV fusion protein to a patient in need thereof.
1. A fusion protein comprising:
(a) a hemojuvelin (HJV) polypeptide or fragment thereof, wherein said polypeptide or fragment thereof has at least 95% amino acid sequence identity to a portion of the HJV protein of SEQ ID NOs: 2, 3 or 4 and is at least 50 amino acids in length; and
(b) a first fusion partner, wherein said first fusion partner is conjugated to said HJV polypeptide or fragment thereof, wherein said first fusion partner is an IgG1 Fc; and
(c) a second fusion partner.
2. The fusion protein of claim 1 , wherein said first fusion partner is fused to the N-terminus or to the C-terminus of the HJV protein fragment.
3. The fusion protein of claim 1 , wherein said IgG1 Fc is human IgG1 Fc.
4. The fusion protein of claim 1 , wherein said IgG1 Fc is at least 95% identical to SEQ ID NO: 6.
5. The fusion protein of claim 1 , wherein said HJV fragment is a soluble fragment.
6. The fusion protein of claim 1 , wherein said HJV fragment lacks the C-terminal GPI anchoring domain.
7. The fusion protein of claim 1 , wherein said HJV fragment lacks the N-terminal signal sequence.
8. The fusion protein of claim 1 , wherein said HJV fragment lacks both the C-terminal GPI anchoring domain and the N-terminal signal sequence.
9. The fusion protein of claim 1 , further comprising purification or detection tag.
10. The fusion protein of claim 9 , wherein the purification or detection tag is selected from the group consisting of detectable proteins, DNA binding domains, gene activation domains, purification tags and secretion signal peptides.
11. The fusion protein of claim 1 , wherein the first fusion partner is conjugated to said HJV polypeptide or fragment via a covalent bond.
12. The fusion protein of claim 11 , wherein said HJV fragment lacks the N terminal signal sequence.
13. The fusion protein of claim 1 , wherein the HJV polypeptide or fragment thereof is not SEQ ID NO: 62, 63 or 64.
14. The fusion protein of claim 1 , wherein said fusion protein has enhanced proteolytic stability.
15. The fusion protein of claim 14 , wherein said enhanced proteolytic stability is conferred by a sequence alteration at the amino acid corresponding to amino acid 172 of isoform A of human HJV.
16. The fusion protein of claim 1 , wherein said fusion protein has an amino acid sequence with at least 95% identity to the sequence of SEQ ID NO: 10.
17. The fusion protein of claim 16 , wherein said fusion protein has an amino acid sequence comprising the sequence of SEQ ID NO: 10.
18. The fusion protein of claim 16 , wherein said fusion protein has an amino acid sequence consisting of the sequence of SEQ ID NO: 10.
19. The fusion protein of claim 1 , wherein said fusion protein has an amino acid sequence with at least 95% identity to the sequence of SEQ ID NO: 7.
20. The fusion protein of claim 19 , wherein said fusion protein has an amino acid sequence comprising the sequence of SEQ ID NO: 7.
21. The fusion protein of claim 1 , wherein said fusion protein has an amino acid sequence with at least 95% identity to the sequence of SEQ ID NO: 1.
22. The fusion protein of claim 21 , wherein said fusion protein has an amino acid sequence comprising the sequence of SEQ ID NO: 1.
23. A pharmaceutical composition comprising the fusion protein of claim 1 and a pharmaceutically acceptable carrier.
24. The fusion protein of claim 1 , wherein the second fusion partner comprises a HJV polypeptide or fragment thereof, wherein wherein said polypeptide or fragment thereof has at least 95% amino acid sequence identity to a portion of the HJV protein of SEQ ID NOs. 2, 3 or 4 and is at least 50 amino acids in length fused to a polypeptide comprising an amino acid sequence 95% identical to IgG1 Fc.
25. The fusion protein of claim 24 , wherein the second polypeptide or fragment is conjugated to said first fusion partner via a crosslinker.
26. The fusion protein of claim 25 , wherein the crosslinker is a DSS crosslinker.
27. The fusion protein of claim 25 , wherein wherein the second polypeptide or fragment is conjugated to said first fusion partner via one or more disulfide bonds.
28. The fusion protein of claim 24 , wherein the IgG1 Fc is at least 95% identical to SEQ ID NO: 6.
29. The fusion protein of claim 1 , further comprising a BMP-2.
30. A method for producing the fusion protein of claim 1 comprising:
(a) introducing into a cell a vector comprising a polynucleotide encoding the fusion protein operably linked to a promoter; and
(b) culturing said cell under conditions where said protein is expressed.
31. The method of claim 30 , further comprising purifying said protein of step (b).
32. An isolated polynucleotide encoding the fusion protein of claim 1 .
33. A vector comprising the polynucleotide of claim 32 .
34. The vector of claim 33 , wherein the vector is a viral vector.
35. The vector of claim 34 , wherein the viral vector is selected from the group consisting of an adenoviral vector, a poxvirus vector and a lentiviral vector.
36. The vector of claim 33 , wherein the vector comprises a tissue- or cell-type specific promoter is a muscle or liver specific promoter.
37. A pharmaceutical composition comprising the vector of claim 33 and a pharmaceutically acceptable carrier.
38. An isolated host cell comprising the vector of claim 33 .