TGF-BETA RECEPTOR TYPE II VARIANTS AND USES THEREOF
In certain aspects, the present disclosure relates to polypeptides comprising a truncated, ligand-binding portion of the extracellular domain of TβRII polypeptide useful to selectively antagonize a TβRII ligand. The disclosure further provides compositions and methods for use in treating or preventing TGFβ associated disorders.
1 . A TβRII fusion polypeptide comprising a first amino acid sequence from the extracellular domain of TβRII and a heterologous amino acid sequence, wherein the first amino acid sequence consists of an amino acid sequence at least 80% identical to:
a) a sequence beginning at any of positions 23 to 35 of SEQ ID NO: 5 and ending at any of positions 153 to 159 of SEQ ID NO: 5 or
b) a sequence beginning at any of positions 23 to 60 of SEQ ID NO: 6 and ending at any of positions 178 to 184 of SEQ ID NO: 6.
2 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 23 of SEQ ID NO: 5 and ending at position 159 of SEQ ID NO: 5.
3 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 29 of SEQ ID NO: 5 and ending at position 159 of SEQ ID NO: 5.
4 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 35 of SEQ ID NO: 5 and ending at position 159 of SEQ ID NO: 5.
5 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 23 of SEQ ID NO: 5 and ending at position 153 of SEQ ID NO: 5.
6 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 29 of SEQ ID NO: 5 and ending at position 153 of SEQ ID NO: 5.
7 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 35 of SEQ ID NO: 5 and ending at position 153 of SEQ ID NO: 5.
8 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 23 of SEQ ID NO: 6 and ending at positions 184 of SEQ ID NO: 6.
9 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 29 of SEQ ID NO: 6 and ending at position 184 of SEQ ID NO: 6.
10 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 23 of SEQ ID NO: 6 and ending at position 178 of SEQ ID NO: 6.
11 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of the sequence beginning at position 29 of SEQ ID NO: 6 and ending at position 178 of SEQ ID NO: 6.
12 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence consists of a sequence that has a D at the position corresponding to position 36 of SEQ ID NO: 47 and/or a K at the position corresponding to position 76 of SEQ ID NO: 47.
13 . A TβRII fusion polypeptide comprising a first amino acid sequence at least 80% identical to the sequence of SEQ ID NO: 7 or SEQ ID NO: 13, or active fragment thereof, and a second heterologous portion, wherein the first amino acid sequence has a D at the position corresponding to position 36 of SEQ ID NO: 47 and/or a K at the position corresponding to position 76 of SEQ ID NO: 47.
14 . The TβRII fusion polypeptide of claim 13 wherein the first amino acid sequence comprises an N-terminal truncation of 1-12 amino acids corresponding to amino acids 1-12 of SEQ ID NO: 7 or 1-37 amino acids corresponding to amino acids 1-37 of SEQ ID NO: 13.
15 . The TβRII fusion polypeptide of claim 13 , wherein the first amino acid sequence comprises an N-terminal truncation of 6 amino acids corresponding to amino acids 1-6 of SEQ ID NO: 7 or SEQ ID NO: 13.
16 . The TβRII fusion polypeptide of claim 13 , wherein the first amino acid sequence comprises an N-terminal truncation of 12 amino acids corresponding to amino acids 1-12 of SEQ ID NO: 7 or 37 amino acids corresponding to amino acids 1-37 of SEQ ID NO: 13.
17 . The TβRII fusion polypeptide of claim 13 , wherein the first amino acid sequence comprises a C-terminal truncation of 1-6 amino acids corresponding to amino acids 137-132 of SEQ ID NO: 7 or amino acids 162-157 of SEQ ID NO: 13.
18 . The TβRII fusion polypeptide of claim 13 , wherein the first amino acid sequence comprises a C-terminal truncation of 6 amino acids corresponding to amino acids 132-137 of SEQ ID NO: 7 or amino acids 157-162 of SEQ ID NO: 13.
19 . The TβRII fusion polypeptide of claim 1 , wherein the first amino acid sequence comprises an insertion corresponding to SEQ ID NO: 18 between the residues corresponding to positions 117 and 118 of SEQ ID NO: 47.
20 . The TβRII fusion polypeptide of claim 1 , wherein the heterologous portion comprises one or more polypeptide portions that enhance one or more of: in vivo stability, in vivo half life, uptake/administration, tissue localization or distribution, formation of protein complexes, and/or purification.
21 . The TβRII fusion polypeptide of claim 1 , wherein the heterologous portion comprises a polypeptide portion selected from: an immunoglobulin Fc domain and a serum albumin.
22 . The TβRII fusion polypeptide of claim 21 , wherein the immunoglobulin Fc domain is joined to the TβRII polypeptide by a linker.
23 . The TβRII fusion polypeptide of claim 1 , wherein the polypeptide includes one or more modified amino acid residues selected from: a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, an amino acid conjugated to a lipid moiety, and an amino acid conjugated to an organic derivatizing agent.
24 . The TβRII fusion polypeptide of claim 23 , wherein the polypeptide is glycosylated.
25 . A TβRII fusion polypeptide comprising a first amino acid sequence consisting of a portion of the extracellular domain of TβRII that comprises an amino acid sequence that is at least 95% identical to an amino acid sequence selected from SEQ ID NOs: 7-17 and 47-49, and a second heterologous portion.
26 - 35 . (canceled)
36 . The polypeptide of claim 1 , wherein the polypeptide binds human GDF15 with an equilibrium dissociation constant (K D ) less than 1×10 −8 M.
37 . The polypeptide of claim 1 , wherein the polypeptide has a glycosylation pattern characteristic of expression of the polypeptide in CHO cells.
38 . A homodimer comprising two polypeptides as defined in claim 1 .
39 . An isolated polynucleotide comprising a coding sequence for the polypeptide of claim 1 .
40 . A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim 39 .
41 . A cell transformed with an isolated polynucleotide of claim 35 .
42 . (canceled)
43 . (canceled)
44 . A pharmaceutical preparation comprising the polypeptide of claim 1 and a pharmaceutically acceptable excipient.
45 . A method of modulating the response of a cell to a TGFβ superfamily member, the method comprising exposing the cell to a polypeptide of claim 1 .
46 . A method of treating a disease or condition associated with a TGFβ superfamily member in a patient in need thereof, the method comprising administering to the patient an effective amount of a polypeptide of claim 1 .
47 - 69 . (canceled)