Rage Fusion Proteins And Methods Of Use
Disclosed are RAGE fusion proteins comprising RAGE polypeptide sequences linked to a second, non-RAGE polypeptide. The RAGE fusion protein may utilize a RAGE polypeptide domain comprising a RAGE ligand binding site and an interdomain linker directly linked to an immunoglobulin C H 2 domain. Such fusion proteins may provide specific, high affinity binding to RAGE ligands. Also disclosed is the use of the RAGE fusion proteins as therapeutics for RAGE-mediated pathologies.
1 - 52 . (canceled)
53 . A method of treating in a subject inflammation and/or rejection associated with transplantation of at least one of a tissue or a plurality of cells from a first site to a second site comprising administering to the subject a fusion protein comprising a RAGE polypeptide directly linked to a polypeptide comprising a C H 2 domain of an immunoglobulin or a portion of a C H 2 domain of an immunoglobulin, and wherein the RAGE fusion protein comprises a ligand binding site.
54 . The method of claim 53 , wherein the RAGE polypeptide comprises a RAGE interdomain linker linked to a RAGE immunoglobulin domain such that the C-terminal amino acid of the RAGE immunoglobulin domain is linked to the N-terminal amino acid of the RAGE interdomain linker, and the C-terminal amino acid of the RAGE interdomain linker is directly linked to the N-terminal amino acid of a polypeptide comprising the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin.
55 . The method of claim 53 , wherein the RAGE ligand binding site comprises the amino acid sequence as set forth in SEQ ID NO: 9 or a sequence at least 90% identical thereto, or the amino acid sequence as set forth in SEQ ID NO: 10 or a sequence at least 90% identical thereto.
56 . The method of claim 54 , wherein the RAGE polypeptide comprising a RAGE interdomain linker linked to a RAGE immunoglobulin domain comprises a fragment of a full-length RAGE protein.
57 . The method of claim 53 , further comprising a first RAGE immunoglobulin domain and a first RAGE interdomain linker linked to a second RAGE immunoglobulin domain and a second RAGE interdomain linker, such that the N-terminal amino acid of the first RAGE interdomain linker is linked to the C-terminal amino acid of the first RAGE immunoglobulin domain, the N-terminal amino acid of the second RAGE immunoglobulin domain is linked to the C-terminal amino acid of the first RAGE interdomain linker, the N-terminal amino acid of the second RAGE interdomain linker is linked to the C-terminal amino acid of the second RAGE immunoglobulin domain, and the C-terminal amino acid of the second RAGE interdomain linker is directly linked to the N-terminal amino acid of the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin.
58 . The method of claim 53 , wherein the RAGE fusion protein comprises the amino acid sequence as set forth in any one of SEQ ID NO: 33, SEQ ID NO: 34, or SEQ ID NO: 56.
59 . The method of claim 53 , wherein the RAGE fusion protein comprises the amino acid sequence as set forth in SEQ ID NO: 33 without the C-terminal lysine amino acid residue, the amino acid sequence as set forth in SEQ ID NO: 34 without the C-terminal lysine amino acid residue, or the amino acid sequence as set forth in SEQ ID NO: 56 without the C-terminal lysine amino acid residue.
60 . The method of claim 53 , wherein the RAGE interdomain linker directly linked to the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin comprises the amino acid sequence as set forth in SEQ ID NO: 22 or a sequence at least 90% identical thereto, or the amino acid sequence as set forth in SEQ ID NO: 24 or a sequence at least 90% identical thereto.
61 . The method of claim 53 , comprising a single RAGE immunoglobulin domain linked via a RAGE interdomain linker to the N-terminal amino acid of the polypeptide comprising the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin.
62 . The method of claim 61 , wherein the RAGE fusion protein comprises the amino acid sequence as set forth in any one of SEQ ID NO: 36, SEQ ID NO: 37, or SEQ ID NO: 57.
63 . The method of claim 61 , wherein the RAGE fusion protein comprises the amino acid sequence as set forth in SEQ ID NO: 36 without the C-terminal lysine amino acid residue, the amino acid sequence as set forth in SEQ ID NO: 37 without the C-terminal lysine amino acid residue, or the amino acid sequence as set forth in SEQ ID NO: 57 without the C-terminal lysine amino acid residue.
64 . The method of claim 53 , wherein the RAGE interdomain linker directly linked to the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin, comprises the amino acid sequence as set forth in SEQ ID NO: 21 or a sequence at least 90% identical thereto, or the amino acid sequence as set forth in SEQ ID NO: 23 or a sequence at least 90% identical thereto.
65 . The method of claim 53 , wherein the method of administration comprises at least one of intravenous administration, intraperitoneal administration or subcutaneous administration of the RAGE fusion protein to the subject.
66 - 73 . (canceled)
74 . The method of claim 53 , wherein the fusion protein does not include an immunoglobulin Fc hinge region.
75 . The method of claim 53 , wherein the first and second sites are in different subjects.
76 . The method of claim 53 , wherein the first and second sites are in the same subject.
77 . The method of claim 74 , wherein the transplanted cells or tissue comprise a cell or tissue of a pancreas, skin, liver, kidney, heart, bone marrow, blood, bone, muscle, artery, vein, cartilage, thyroid, nervous system, or stem cells.
78 . A method of preventing transplant rejection of a cell, a plurality of cells, or a tissue, the method comprising administering a therapeutically effective amount of a fusion protein comprising a RAGE polypeptide directly linked to a polypeptide comprising a C H 2 domain of an immunoglobulin or a portion of a C H 2 domain of an immunoglobulin and wherein the RAGE fusion protein comprises a ligand binding site.
79 . A method of treating osteoporosis in a subject comprising administering to the subject a fusion protein comprising a RAGE polypeptide directly linked to a polypeptide comprising a C H 2 domain of an immunoglobulin or a portion of a C H 2 domain of an immunoglobulin, and wherein the RAGE fusion protein comprises a ligand binding site.
80 . The method of claim 79 , wherein the RAGE fusion protein does not include an immunoglobulin Fc hinge region.
81 . The method of claim 79 , wherein the RAGE fusion protein comprises a first RAGE immunoglobulin domain and a first RAGE interdomain linker linked to a second RAGE immunoglobulin domain and a second RAGE interdomain linker, such that the N-terminal amino acid of the first RAGE interdomain linker is linked to the C-terminal amino acid of the first RAGE immunoglobulin domain, the N-terminal amino acid of the second RAGE immunoglobulin domain is linked to the C-terminal amino acid of the first RAGE interdomain linker, the N-terminal amino acid of the second RAGE interdomain linker is linked to the C-terminal amino acid of the second RAGE immunoglobulin domain, and the C-terminal amino acid of the second RAGE interdomain linker is directly linked to the N-terminal amino acid of the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin.
82 . The method of claim 79 , wherein the RAGE fusion protein comprises a single RAGE immunoglobulin domain linked via a RAGE interdomain linker to the N-terminal amino acid of the polypeptide comprising the C H 2 domain of the immunoglobulin or the portion of the C H 2 domain of the immunoglobulin.