Human tissue inhibitor of metalloproteinase-4

Abstract

A human tissue inhibitor of metalloproteinases-4 polypeptide and DNA (RNA) encoding such polypeptide and a procedure for producing such polypeptide by recombinant techniques. Also disclosed are methods for utilizing such polypeptide for the treatment of diseases, including arthritis and cancer. Antagonists against such polypeptides and their use as a therapeutic to resorb scar tissue are also disclosed. Diagnostic assays for detecting levels of human TIMP-4 protein and mutations in human TIMP-4 nucleic acid sequence are also disclosed.

What is claimed is:  
     
         1 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide encoding the polypeptide as set forth in SEQ ID NO:2;    (b) a polynucleotide capable of hybridizing to and which is at least 70% identical to the polynucleotide of (a); and    (c) a polynucleotide fragment of the polynucleotide of (a) or (b).    
     
     
         2 . The polynucleotide of  claim 1  wherein the polynucleotide is DNA.  
     
     
         3 . The polynucleotide of  claim 2  which encodes the polypeptide as set forth in SEQ ID NO:2.  
     
     
         4 . The polynucleotide of  claim 2  which encodes the polypeptide comprising amino acid 1 to 195 of of SEQ ID NO:2.  
     
     
         5 . The polynucleotide of  claim 2  comprising the nucleotide sequence of SEQ ID NO:1 from nucleotide 1 to 675.  
     
     
         6 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide which encodes a mature polypeptide encoded by the DNA contained in ATCC Deposit No. 75946;    (b) a polynucleotide which encodes a polypeptide expressed by the DNA contained in ATCC Deposit No. 75946;    (c) a polynucleotide capable of hybridizing to and which is at least 70% identical to the polynucleotide of (a) or (b); and    (d) a polynucleotide fragment of the polynucleotide of (a), (b) or (c).    
     
     
         7 . A vector containing the DNA of  claim 2 .  
     
     
         8 . A host cell genetically engineered with the vector of  claim 7 .  
     
     
         9 . A process for producing a polypeptide comprising: expressing from the host cell of  claim 8  the polypeptide encoded by said DNA.  
     
     
         10 . A process for producing cells capable of expressing a polypeptide comprising transforming or transfecting the cells with the vector of  claim 7 .  
     
     
         11 . A polypeptide comprising a member selected from the group consisting of (i) a polypeptide having the deduced amino acid sequence of SEQ ID NO:2 and fragments, analogs and derivatives thereof; (ii) a polypeptide comprising amino acid 1 to amino acid 195 of SEQ ID NO:2; and (iii) a polypeptide encoded by the cDNA of ATCC Deposit No. 75946 and fragments, analogs and derivatives of said polypeptide.  
     
     
         12 . A compound effective as an agonist for the polypeptide of  claim 11 .  
     
     
         13 . A compound effective as an antagonist against the polypeptide of  claim 11 .  
     
     
         14 . A method for the treatment of a patient having need of TIMP-4 comprising: 
 administering to the patient a therapeutically effective amount of the polypeptide of  claim 11 .    
     
     
         15 . The method of  claim 14  wherein said therapeutically effective amount of the polypeptide is administered by providing to the patient DNA encoding said polypeptide and expressing said polypeptide in vivo.  
     
     
         16 . A method for the treatment of a patient having need of TIMP-4 comprising: 
 administering to the patient a therapeutically effective amount of the compound of  claim 12 .    
     
     
         17 . A method for the treatment of a patient having need to inhibit TIMP-4 comprising: 
 administering to the patient a therapeutically effective amount of the antagonist of  claim 13 .    
     
     
         18 . A process for identifying compounds active as agonists or antagonists to the polypeptide of  claim 11  comprising: 
 combining an MMP, human TIMP-4, a compound to be screened and a reaction mixture containing substrate capable of degradation by the MMP, wherein said substrate is labeled; and  
 determining the ability of the compound to enhance or block the degradation of the substrate by said MMP by measuring the label released from the substrate.  
 
     
     
         19 . A process for diagnosing a disease or a susceptibility to a disease related to a mutation in the polypeptide of  claim 11  comprising: 
 determining a mutation in the human TIMP-4 nucleic acid sequence.  
 
     
     
         20 . A diagnostic process comprising: 
 analyzing for the presence of the polypeptide of  claim 11  in a sample derived from a host.    
     
     
         21 . A method of treating restenosis in a patient, comprising administering to the patient an isolated nucleic acid molecule encoding a TIMP-4 polypeptide selected from the group consisting of: 
 (a) the amino acid sequence shown as residues −29 to 195 in SEQ ID NO:2;    (b) the amino acid sequence shown as residues −28 to 195 in SEQ ID NO:2;    (c) the amino acid sequence shown as residues 1 to 195 in SEQ ID NO:2; and    (d) a fragment of the sequence described in (a) whereing a polypeptide consisting of the fragment retains protease inhibiting activity;    wherein said nucleic acid molecule is operatively linked to a transcription control sequence; and wherein the expression of said nucleic acid molecule results in an increased amount of the TIMP-4 polypeptide in an amount effective to inhibit metalloproteinase activity.    
     
     
         22 . The method of  claim 21 , wherein said isolated nucleic acidmolecule is administered to said patient in a viral vector delivery vehicle.  
     
     
         23 . The method of  claim 22 , wherein said viral vector delivery vehicle is from adenovirus.