Methods and compositions for treatment of cartilage and disc tissue pathologies

Abstract

An isolated polypeptide comprising a peptide selected from: i) DHX 1 SDNYT, wherein X 1 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: 
       
         
           
                 
                 
               
                     
                   i) 
                 
                     
                   (SEQ ID NO: 30) 
                 
                     
                   DHX 1 X 2 X 3 X 4 X 5 X 6 ; 
                 
             
                
                
                
               
            
           
         
         X 1  is any amino acid, optionally L, H, R, or 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 
         X 6  is T or Y; 
         ii) a conservative variant of i), and 
         iii) a fragment of i) or ii); 
         wherein the conservative variant or fragment retains biological activity and the peptide is 15 or fewer 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: 
       
         
           
                 
                 
               
                     
                   i) 
                 
                     
                   (SEQ ID NO: 6) 
                 
                     
                   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, or 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 
         X 13  is H or V; 
         ii) a conservative variant of i); and 
         iii) a fragment of i) or ii) 
         wherein the conservative variant 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 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 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 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 producing an induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis, the method comprising culturing the cartilage, chondrocyte cell, or disc cell with an effective amount of the isolated polypeptide of  claim 1  under conditions to induce proteoglycan or collagen synthesis, thereby producing an induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis. 
     
     
         15 . A method of producing an induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis for implanting into a subject, the method comprising culturing the cartilage, chondrocyte cell, or disc cell with an effective amount of the isolated polypeptide of  claim 1  under conditions to induce proteoglycan and collagen synthesis, thereby producing an induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis. 
     
     
         16 . A method of alleviating a symptom of and/or treating a cartilage or disc disorder, comprising administering to a subject in need thereof the isolated polypeptide of  claim 1 . 
     
     
         17 . A method of alleviating a symptom of and/or treating a cartilage or disc disorder, comprising administering to a subject in need thereof the recombinant cell of  claim 10 . 
     
     
         18 . A method of alleviating a symptom of and/or treating a cartilage or disc disorder, comprising administering to a subject in need thereof the pharmaceutical composition of  claim 12 . 
     
     
         19 . A liposome comprising an isolated polypeptide having the sequence DHX 1 X 2 X 3 X 4 X 5 X 6  (SEQ ID NO: 30); wherein X 1  is L, H, R, or Q; X 2  is 5; X 3  is D, S or N; X 4  is N; X 5  is Y; and X 6  is T. 
     
     
         20 . An induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis for implanting into a subject, wherein the cartilage, chondrocyte, or disc cell is made by a method comprising culturing a cartilage, chondrocyte, or disc cell with an effective amount of an isolated polypeptide comprising a peptide selected from: 
       
         
           
                 
                 
               
                     
                   i) 
                 
                     
                   (SEQ ID NO: 30) 
                 
                     
                   DHX 1 X 2 X 3 X 4 X 5 X 6 ; 
                 
             
                
                
                
               
            
           
         
         X 1  is any amino acid, optionally L, H, R, or 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 
         X 6  is T or Y; 
         ii) a conservative variant of i), and 
         iii) a fragment of i) or ii); 
       
       wherein the conservative variant or fragment retains biological activity and the peptide is 15 or fewer amino acids,
 thereby producing an induced cartilage, chondrocyte cell, or disc cell with increased matrix synthesis.