Bmp-4 peptides and method of use

Abstract

The invention relates to truncated BMP-4 growth factors and variants thereof. The invention also relates to methods of making and using the truncated BMP-4 growth factors.

What is claimed is: 
     
         1 . A method of promoting differentiation of cells into osteoblasts or chondrocytes, the method comprising treating the cells with a peptide selected from the group consisting of
 a) a peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2,   b) a peptide of 115 residues or less, said peptide comprising an amino acid sequence of SEQ ID NO: 2, and   c) a peptide with an amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 2,   wherein each of peptides (a)-(c) has osteoinductive or chondroinductive activity.   
     
     
         2 . The method of  claim 1 , wherein the cells are progenitor cells or adult stem cells. 
     
     
         3 . The method of  claim 2 , wherein the progenitor cells or the adult stem cells are derived from placenta, bone marrow, adipose tissue, blood vessel, amniotic fluid, synovial fluid, synovial membrane, pericardium, periosteum, dura, peripheral blood, umbilical blood, menstrual blood, baby teeth, nucleus pulposus, brain, skin, hair follicle, intestinal crypt, neural tissue, or muscle. 
     
     
         4 . The method of  claim 1 , wherein the cells are induced pluripotent stem cells. 
     
     
         5 . A method of promoting osteogenesis of cells in a tissue, the method comprising treating the tissue with a peptide selected from the group consisting of
 a) a peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2,   b) a peptide of 115 residues or less, said peptide comprising an amino acid sequence of SEQ ID NO: 2, and   c) a peptide with an amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 2,   wherein each of peptides (a)-(c) has osteogenic activity.   
     
     
         6 . The method of  claim 5 , wherein the osteogenic activity of cells in the treated tissue is greater than the osteogenic activity of cells in untreated tissue. 
     
     
         7 . The method of  claim 5  or  6 , wherein the tissue is bone tissue or connective tissue. 
     
     
         8 . An isolated peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2, wherein the peptide has osteoinductive activity or chondroinductive activity. 
     
     
         9 . The isolated peptide of  claim 8 , wherein the amino acid sequence comprises an amino acid sequence 100% identical to the amino acid sequence of SEQ ID NO: 2. 
     
     
         10 . The isolated peptide of  claim 9 , wherein the peptide consists of the amino acid sequence of SEQ ID NO: 2. 
     
     
         11 . A fusion protein comprising (a) the peptide of any one of  claims 8 - 10 , and (b) a second peptide, wherein the second peptide comprises an amino acid sequence that is less than 70% identical to the amino acid sequence of SEQ ID NO: 3. 
     
     
         12 . A composition comprising the peptide of  claim 8  and at least one protease. 
     
     
         13 . The composition according to  claim 12 , wherein the at least one protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, BMP-1 (bone morphogenetic protein-1), MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         14 . A method of making the peptide of any one of  claims 8 - 10  comprising contacting mature BMP-4 with a protease under conditions that promote protein cleavage and harvesting the peptide. 
     
     
         15 . The method according to  claim 14 , wherein the at least one protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, BMP-1 (bone morphogenetic protein-1), MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         16 . A vector encoding the peptide of any one of  claims 8 - 11 . 
     
     
         17 . A host cell comprising the vector of  claim 16 . 
     
     
         18 . A method of making the peptide of any one of  claims 8 - 11  comprising culturing a host cell under conditions suitable for protein expression and harvesting the protein, wherein the host cell comprises a vector that encodes said peptide. 
     
     
         19 . A pharmaceutical composition comprising the peptide of any one of  claims 8 - 11 . 
     
     
         20 . A method of increasing a cellular growth factor activity, the method comprising treating a cell with at least one protease and the growth factor, wherein the growth factor is a peptide comprising an amino acid sequence of SEQ ID NO: 2, wherein the peptide has osteoinductive or chondroinductive activity. 
     
     
         21 . The method according to  claim 20 , wherein the peptide is selected from the group consisting of
 a) a peptide of 115 residues or less, said peptide comprising an amino acid sequence at least 95% identical to the amino acid sequence of SEQ ID NO: 2,   b) a peptide of 115 residues or less, said peptide comprising an amino acid sequence of SEQ ID NO: 2, and   c) a peptide with an amino acid sequence consisting of the amino acid sequence of SEQ ID NO: 2.   
     
     
         22 . The method according to  claim 20 , wherein the peptide is a fusion protein comprising (a) the peptide comprising the amino acid sequence of SEQ ID NO: 2, and (b) a second peptide, wherein the second peptide comprises an amino acid sequence that is less than 70% identical to the amino acid sequence of SEQ ID NO: 3. 
     
     
         23 . The method according to  claim 20 , wherein the composition comprises two or more proteases. 
     
     
         24 . The method according to  claim 20 , wherein the at least one protease is selected from the group consisting of collagenase, clostripain, dispase, trypsin, BMP-1 (bone morphogenetic protein-1), MMP-13 (matrix metalloproteinase-13), and a mixture thereof. 
     
     
         25 . The method according to  claim 23 , wherein the at least one protease is collagenase. 
     
     
         26 . The method according to  claim 20 , wherein the cells are progenitor cells or adult stem cells. 
     
     
         27 . The method according to  claim 25 , wherein the progenitor cells or the adult stem cells are derived from placenta, bone marrow, adipose tissue, blood vessel, amniotic fluid, synovial fluid, synovial membrane, pericardium, periosteum, dura, peripheral blood, umbilical blood, menstrual blood, baby teeth, nucleus pulposus, brain, skin, hair follicle, intestinal crypt, neural tissue, or muscle. 
     
     
         28 . The method according to  claim 20 , wherein the cells are induced pluripotent stem cells.