Methods and compositions for treatment of cartilage and disc tissue pathologies

Abstract

An isolated polypeptide comprising a peptide selected from: i) DHX1SDNYT, wherein X1 is L or H (SEQ ID NO:3); ii) a conservative variant of i) iii) a fragment of i) or ii); wherein the conservative variant and/or fragment retains biological activity and the peptide is 15 or less amino acids as well as recombinant cells, and uses thereof.

1 . An isolated polypeptide comprising a peptide selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 30) 
                 
                     
                   i) DHX 1 X 2 X 3 X 4 X 5 X 6 ; 
                 
             
                
                
               
            
           
         
         X 1  is any amino acid, optionally L, H, R Q; 
         X 2  is S or L; 
         X 3  is D, S or N; 
         X 4  is N or D; 
         X 5  is Y or S; and/or 
         X 6  is T or Y; 
         ii) a conservative variant of i), and 
         iii) a fragment of i) or ii); 
         wherein the conservative variant and/or fragment retains biological activity and the peptide is 15 or less amino acids. 
       
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the peptide consists of DHX 1 SDNYT (SEQ ID NO:1), wherein X 1  is L or H or a conservative variant thereof that retains biological activity. 
     
     
         3 . The isolated polypeptide of  claim 1 , comprising i) a peptide sequence consisting of DHLSDNYT (SEQ ID NO:2) or a conservative variant thereof that retains biological activity, or ii) a peptide sequence consisting of DHHSDNYT (SEQ ID NO:3) or a conservative variant thereof that retains biological activity. 
     
     
         4 . The isolated polypeptide of  claim 1 , wherein the peptide is 4, 5, 6, 7, 9, 10, 11, 12, 13, 14 or 15 amino acids. 
     
     
         5 . The isolated polypeptide of  claim 1 , comprising a peptide selected from: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 6) 
                 
                     
                   i) DHX 1 X 2 X 3 X 4 X 5 X 6 X 7 X 8 X 9 DX 10 X 11 X 12 X 13   
                 
                     
                   or 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 4) 
                 
                     
                   DHX 1 SDNYTX 2 DHDRX 3 I; 
                 
             
                
                
                
                
                
                
               
            
           
         
         X 1  is any amino acid, optionally L, H, R Q; 
         X 2  is S or L; 
         X 3  is D, S, or N; 
         X 4  is N or D: 
         X 5  is Y or S; 
         X 6  is T or Y; 
         X 7  is L, V, or T; 
         X 8  is any amino acid, optionally D, G, N, or P; 
         X 9  is H, Y, or P; 
         X 10  is R or Q; 
         X 11  is A, V, or D; 
         X 12  is I or R; and/or 
         X 13  is H or V; 
         ii) a conservative variant of i); and 
         iii) a fragment of i) or ii) 
         wherein the conservative variant and/or fragment retains biological activity. 
       
     
     
         6 . The isolated polypeptide of  claim 5 , wherein the sequence is:
 i) DHLSDNYTLDHDRAI (SEQ ID NO: 9) or a conservative variant and/or fragment thereof that retains biological activity or has at least 80% sequence identity to SEQ ID NO: 9, or   ii) DHHSDNYTVDHDRVI (SEQ ID NO: 10) or a conservative variant and/or fragment thereof that retains biological activity or has at least 80% sequence identity to SEQ ID NO: 10.   
     
     
         7 . The isolated polypeptide of  claim 1 , wherein the peptide is conjugated to a stabilizing moiety and/or carrier. 
     
     
         8 . An isolated nucleic acid that encodes the polypeptide of  claim 1 . 
     
     
         9 . A vector comprising the isolated nucleic acid of  claim 8 . 
     
     
         10 . A recombinant cell expressing the polypeptide of  claim 1 . 
     
     
         11 . The recombinant cell of  claim 10 , wherein the cell is a chondrocyte lineage cell, a stem cell or a disc cell. 
     
     
         12 . A pharmaceutical composition comprising the isolated polypeptide of  claim 1  and a pharmaceutically acceptable carrier, stabilizing agent, or diluent. 
     
     
         13 . A pharmaceutical composition comprising a scaffold formed of a biocompatible material comprising the isolated polypeptide of  claim 1  and a pharmaceutically acceptable carrier, stabilizing agent, or diluent. 
     
     
         14 . A method of inducing matrix synthesis in a cartilage, chondrocyte cell, and/or disc cell or in a tissue comprising a cartilage, chondrocyte cell, and/or disc cell, the method comprising incubating/culturing the cartilage, chondrocyte cell, and/or disc cell with an effective amount of the isolated polypeptide of  claim 1  under conditions to induce proteoglycan and/or collagen synthesis, thus producing an induced cartilage, chondrocyte cell, and/or disc cell with increased matrix synthesis. 
     
     
         15 . A method of producing cartilage, a chondrocyte cell, and/or disc tissue for implanting into a subject, the method comprising incubating/culturing the cartilage, chondrocyte cell, and/or disc cell with an effective amount of the isolated polypeptide of  claim 1  under conditions to induce proteoglycan and collagen synthesis, thus producing an induced cartilage, chondrocyte cell, and/or disc cell with increased matrix synthesis, wherein a substantially pure population of the induced cartilage, chondrocyte cell, and/or disc cell can be isolated. 
     
     
         16 . A method of alleviating a symptom of and/or treating a cartilage and/or disc disorder, comprising administering to a subject in need thereof the isolated polypeptide of  claim 1 . 
     
     
         17 . The method of  claim 16 , wherein the cartilage and/or disc disorder is intervertebral disc degeneration or an inflammatory or degenerative joint disease, wherein the inflammatory or degenerative joint disease comprises arthritis, undesirable osteogenesis, and/or calcification. 
     
     
         18 . A method of alleviating a symptom of and/or treating a cartilage and/or disc disorder, comprising administering to a subject in need thereof the recombinant cell of  claim 10 . 
     
     
         19 . A method of alleviating a symptom of and/or treating a cartilage and/or disc disorder, comprising administering to a subject in need thereof the pharmaceutical composition of  claim 12 .