Function and regulation of adamts-1

Abstract

The present invention relates to ADAMTS-1 and uses thereof. The present invention also relates to fragments of ADAMTS-1 and methods of inhibiting cell growth and metastasis. The present invention also provide methods of identifying inhibitors and activators relating to the function of ADAMTS-1.

What is claimed is: 
     
         1 . An isolated polypeptide fragment of ADAMTS-1 that inhibits tumor growth and/or metastasis, wherein the fragment consists of SEQ ID Nos: 5, 7, 9, and or 11. 
     
     
         2 . A pharmaceutical composition comprising a polypeptide of  claim 1 . 
     
     
         3 . A composition comprising at least two different polypeptide fragment of ADAMTS-1 that inhibit cell proliferation or metastasis, wherein said fragment comprises SEQ ID Nos: 5, 7, 9, and/or 11. 
     
     
         4 . The position of  claim 3  wherein said composition is a pharmaceutical composition. 
     
     
         5 . An isolated polynucleotide encoding a polypeptide fragment of ADAMTS-1 wherein said fragment inhibits tumor growth and/or metastasis, wherein said polynucleotide comprises SEQ ID NO: 6, 8, 10, or 12. 
     
     
         6 . A pharmaceutical composition comprising the isolated polynucleotide of  claim 6 . 
     
     
         7 . The isolated polynucleotide of  claim 6  wherein said polynucleotide is a vector or plasmid. 
     
     
         8 . A method for identifying an inhibitor or an activator of ADAMTS-1 auto-cleavage comprising performing a test assay comprising:
 a) contacting ADAMTS-1 with a test compound under conditions in which ADAMTS-1 undergoes cleavage in the absence of a test compound;   b) measuring cleavage level of ADAMTS-1; and   c) comparing the cleavage level to cleavage level of ADAMTS-1 in the absence of the test compound, wherein a decrease in auto-cleavage indicates that the test compound is a cleavage inhibitor or wherein an increase in auto-cleavage indicates that the test compound is a cleavage activator.   
     
     
         9 . The method of  claim 8  wherein said the test compound is contacted with a cell comprising ADAMTS-1. 
     
     
         10 . The method of  claim 9 , further comprising performing, a negative control assay which comprises contacting a cell that does not comprise ADAMNTS-1 or a cell that comprises a cleavage resistant mutant of ADAMTS-1. 
     
     
         11 . The method of  claim 9 , further comprising performing a positive control assay which comprises contacting a cell comprising ADAMTS-1 a positive control compound and measuring cleavage. 
     
     
         12 . The method of  claim 8 , further comprising measuring the cleavage of ADAMTS-1 in the absence of the test compound. 
     
     
         13 . A method for identifying a heparin inhibitor comprising:
 a) contacting a composition comprising heparin and ADAMTS-1 with a test compound under conditions in which ADAMTS-1 undergoes auto-cleavage and/or proteolytic cleavage in absence of heparin;   b) measuring cleavage level of ADAMTS-1; and   c) comparing cleavage level of ADAMTS-1 in the absence of the test compound;   wherein an increase in the cleavage of ADAMTS-1 indicates that the compound is a heparin inhibitor.   
     
     
         14 . A method of identifying a metalloproteinase inhibitor comprising:
 a) contacting a ADAMTS-1 polypeptide or fragment thereof comprising metalloproteinase activity with a test compound under conditions which metalloproteinase activity is detected in the absence of the test compound.   b) measuring metalloproteinase activity level of ADAMTS-1; and   c) comparing the metalloproteinase activity level of ADAMTS-1 in the presence or absence of the test compound,   wherein a decrease in metalloproteinase activity indicates the test compound is a metalloproteinase inhibitor.   
     
     
         15 . The method of  claim 14  wherein said fragment comprises SEQ ID NO: 5, 7, 9, and/or 11. 
     
     
         16 . The method of  claim 14  wherein the metalloproteinase activity of ADAMTS-1 is compared to a fragment or mutant of ADAMTS-1 that has no metalloproteinase activity. 
     
     
         17 . The method of  claim 16  wherein said fragment or mutant of ADAMTS-1 that has no metalloproteinase activity comprises SEQ ID NO 31, 33, 35, and/or 36. 
     
     
         18 . A method of treating cancer in an individual comprising administering to the individual a therapeutically effective amount of a polypeptide fragment of ADAMTS-1 and/or a nucleic acid that encodes a polypeptide fragment of ADAMTS-1 that inhibits cell proliferation and/or metastasis. 
     
     
         19 . The method of  claim 18  wherein the polypeptide fragment comprises a TSP type-I motif. 
     
     
         20 . The method  claim 18  wherein the fragment comprises SEQ ID NO: 5, 7, 9 and/or 11. 
     
     
         21 . The method of  claim 18  wherein the nucleic acid molecule encoding the polypeptide fragment comprises SEQ ID NO: 6, 8, 10, and/or 11. 
     
     
         22 . The method of  claim 18  wherein said polypeptide fragment of ADAMTS-1 comprises the spacer/Cys-rich and/or spacer domain of ADAMTS-1 or a nucleic acid molecule encoding a polypeptide fragment of ADAMTS-1 comprising the spacer/Cys-rich and/or spacer domain of ADAMTS-1. 
     
     
         23 . The method of  claim 22  wherein said fragment comprises SEQ ID NO: 21 and/or 23. 
     
     
         24 . The method of  claim 22  wherein said nucleic acid molecule comprises SEQ ID NO: 22 and/or 24. 
     
     
         25 . A method of mating cancer comprising administering an inhibitor of the metalloproteinase activity of ADAMTS-1. 
     
     
         26 . The method of  claim 25  wherein the inhibitor is a metalloproteinase defective polypeptide of ADAMTS-1 or a nucleic acid molecule encoding a metalloproteinase defective polypeptide of ADAMTS-1. 
     
     
         27 . The method of  claim 26  wherein the metalloproteinase defective polypeptide of comprises SEQ ID NO: 29, 31, 33, and/or 35. 
     
     
         28 . The method of  claim 26  wherein nucleic acid molecule comprises SEQ ID NO: 30, 32, 34, and/or 36. 
     
     
         29 . The method of  claim 25  wherein said inhibitor is an antibody that binds to ADAMTS-1.