Designer ligands of tgf-beta superfamily

Abstract

The present disclosure relates to chimeric polypeptide having TGF-beta activity, nucleic acids encoding the polypeptides, and host cells for producing the polypeptides.

What is claimed is: 
     
         1 . A recombinant TGF-β ligand polypeptide having at least 95% sequence identity to an amino acid sequence comprising
 a first segment of amino acid residues comprising amino acid residues 1 to 32 of SEQ ID NO:2 (“1b”); 
 a second segment of amino acid residues comprising amino acid residues 33 to x 16  of SEQ ID NO:2 (“1b”), 
 wherein: x 16  is residue 45, 46, 47, of 48 of SEQ ID NO: 2; 
 a third segment of amino acid residues comprising amino acid residues x 17  to x 1  of SEQ ID NO:2 (“3b”), or amino acid residues x 2  to x 3  of SEQ ID NO:5 (“3a”), 
 wherein: x 17  is x 16 +1 of SEQ ID NO: 2,
 x 1  is residue 65, 66, 67, or 68 of SEQ ID NO:2, 
 x 2  is residue 42, 43, 44, or 45 of SEQ ID NO:5, and 
 x 3  is residue 61, 62, 63, or 64 of SEQ ID NO:5; and 
 
 a fourth segment of amino acid residues comprising amino acid residues x 4  to x 5  of SEQ ID NO:5 (“4a”) or amino acid residues x 6  to x 7  of SEQ ID NO:2 (“4b”), 
 wherein: x 4  is x 1  of SEQ ID NO: 5 if the third segment comprises 3b, or x 4  is x 3 +1 of SEQ ID NO: 5 if the third segment comprises 3a,
 x 5  is residue 78, 79, 80, 81, 82, 83 or 84 of SEQ ID NO:5, 
 x 6  is x 3  of SEQ ID NO: 2 if the third segment comprises 3a, or x 6  is x 1 +1 of SEQ ID NO: 2 if the third segment comprises 3b, 
 x 7  is residue 76, 77, 78, 79, 80, 81 or 82 of SEQ ID NO:2, and; 
 
 a fifth segment of amino acid residues comprising amino acid residues x 8  to x 9  of SEQ ID NO:5 (“5a”) or amino acid residues x 10  to x 11  of SEQ ID NO:2 (“5b”), 
 wherein: x 8  is x 5  of SEQ ID NO: 5 if the fourth segment comprises 4b, or x 8  is x 5 +1 of SEQ ID NO: 5 if the fourth segment comprises 4a,
 x 9  is residue 90, 91, 92, 93, 94, 95 or 96 of SEQ ID NO:5, 
 x 10  is x 7  of SEQ ID NO: 2 if the fourth segment comprises 4a, or x 10  is 
  x 7 +1 of SEQ ID NO: 2 if the fourth segment comprises 4b, 
 x 11  is residue 88, 89, 90, 91, 92, 93, or 94 of SEQ ID NO:2, and 
 
 a sixth segment comprising amino acid residues x 12  to x 13  of SEQ ID NO:5 (“6a”) or amino acid residues x 14  to x 15  of SEQ ID NO:2 (“6b”), 
 wherein: x 12  is x 9  of SEQ ID NO: 5 if the fifth segment comprises 5b, or x 12  is x 9 +1 of SEQ ID NO: 5 if the fifth segment comprises 5a,
 x 13  is residue 114, 115, or 116 of SEQ ID NO:5, 
 x 14  is x 11  of SEQ ID NO: 2 if the fifth segment comprises 5a, or x 14  is 
  x 11 +1 of SEQ ID NO: 2 if the fifth segment comprises 5b, 
 x 15  is residue 112, 113, or 114 of SEQ ID NO:2; 
 
 wherein the recombinant TGF-β ligand polypeptide activates cell proliferation or activity associated with SMAD pathway. 
 
     
     
         2 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 17  to x 1  of SEQ ID NO:2 (“3b”), x 17  is x 16 +1 of SEQ ID NO: 2.   
     
     
         3 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 2  to x 3  of SEQ ID NO:5 (“3a”).   
     
     
         4 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 2  to x 3  of SEQ ID NO:5 (“3a”);   the fourth segment comprises amino acid residues x 4  to x 5  of SEQ ID NO:5 (“4a”), wherein x 6  is x 3 +1 of SEQ ID NO: 5;   the fifth segment comprises amino acid residues x 8  to x 9  of SEQ ID NO:5 (“5a”), wherein x 8  is x 5 +1 of SEQ ID NO: 5; and   the sixth segment comprises amino acid residues x 12  to x 13  of SEQ ID NO:5 (“6a”), wherein x 12  is x 9 +1 of SEQ ID NO: 5.   
     
     
         5 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 2  to x 3  of SEQ ID NO:5 (“3a”);   the fourth segment comprises amino acid residues x 4  to x 5  of SEQ ID NO:5 (“4a”), wherein x 6  is x 3 +1 of SEQ ID NO: 5;   the fifth segment comprises amino acid residues x 10  to x 11  of SEQ ID NO:5 (“5b”), wherein x 10  is x 7  of SEQ ID NO: 2; and   the sixth segment comprises amino acid residues x 12  to x 13  of SEQ ID NO:5 (“6a”), wherein x 12  is x 9  of SEQ ID NO: 5.   
     
     
         6 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 17  to x 1  of SEQ ID NO:2 (“3b”), x 17  is x 16 +1 of SEQ ID NO: 2;   the fourth segment comprises amino acid residues x 6  to x 7  of SEQ ID NO:2 (“4b”), wherein x 6  is x 1 +1 of SEQ ID NO: 2;   the fifth segment comprises amino acid residues x 8  to x 9  of SEQ ID NO:5 (“5a”), wherein x 8  is x 5  of SEQ ID NO: 5; and   the sixth segment comprises amino acid residues x 12  to x 13  of SEQ ID NO:5 (“6a”), wherein x 12  is x 9 +1 of SEQ ID NO: 5.   
     
     
         7 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 17  to x 1  of SEQ ID NO:2 (“3b”), x 17  is x 16 +1 of SEQ ID NO: 2;   the fourth segment comprises amino acid residues x 6  to x 7  of SEQ ID NO:2 (“4b”), wherein x 6  is x 1 +1 of SEQ ID NO: 2;   the fifth segment comprises amino acid residues x 10  to x 11  of SEQ ID NO:2 (“5b”), wherein x 10  is x 7 +1 of SEQ ID NO: 2; and   the sixth segment comprises amino acid residues x 12  to x 13  of SEQ ID NO:5 (“6a”), wherein x 12  is x 9  of SEQ ID NO: 5.   
     
     
         8 . The recombinant polypeptide of  claim 1 , wherein
 the third segment comprises amino acid residues x 17  to x 1  of SEQ ID NO:2 (“3b”), x 17  is x 16 +1 of SEQ ID NO: 2;   the fourth segment comprises amino acid residues x 6  to x 7  of SEQ ID NO:2 (“4b”), wherein x 6  is x 1 +1 of SEQ ID NO: 2;   the fifth segment comprises amino acid residues x 8  to x 9  of SEQ ID NO:5 (“5a”), wherein x 8  is x 5  of SEQ ID NO: 5; and   the sixth segment comprises amino acid residues x 14  to x 15  of SEQ ID NO:5 (“6b”), wherein x 14  is x 11  of SEQ ID NO: 2.   
     
     
         9 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein the recombinant TGF-β ligand polypeptide has at least 97% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments. 
     
     
         10 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein the recombinant TGF-β ligand polypeptide has at least 98% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments. 
     
     
         11 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein the recombinant TGF-β ligand polypeptide has at least 99% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments. 
     
     
         12 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein the recombinant TGF-β ligand polypeptide has 100% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments. 
     
     
         13 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein the polypeptide comprises a sequence selected from the group consisting of SEQ ID NOs: 13, 15, 9, 7, 11, 114, 116, 118, 120, 122, 124, 126, 128, 130, and 132. 
     
     
         14 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein said polypeptide comprises the sequence set forth in SEQ ID NO: 9. 
     
     
         15 . The recombinant TGF-β ligand polypeptide of  claim 1 , wherein said polypeptide has at least 95% sequence identity to the sequence set forth in SEQ ID NO: 9. 
     
     
         16 . A polynucleotide comprising a sequence selected from the group consisting of SEQ ID NOs: 12, 14, 8, 6, 10, 113, 115, 117, 119, 121, 123, 125, 127, 129, and 131. 
     
     
         17 . The polynucleotide of  claim 11 , wherein said polynucleotide has at least 95% sequence identify to the sequence set forth in SEQ ID NO: 8. 
     
     
         18 . A method of activating cell proliferation or activity associated with SMAD pathway, comprising contact a cell or tissue with a recombinant TGF-β ligand polypeptide of  claim 1 . 
     
     
         19 . A method of treating a disease or disorder associated with bone, cartilage, neurological tissue, cardiac tissue, skeletal muscle or endocrine tissue in a subject in need of the treatment, comprising contacting the patient with a recombinant TGF-β ligand polypeptide of  claim 1 . 
     
     
         20 . A method of treating a cell proliferative disease or disorder in a subject in need of the treatment, comprising contacting the patient with a recombinant TGF-β ligand polypeptide of  claim 1 .