Human Extracellular Matrix-1

Abstract

A human ECM-1 polypeptide and DNA (RNA) encoding such polypeptide and a procedure for producing such polypeptide by recombinant techniques is disclosed. Also disclosed are methods for utilizing such polypeptide for stimulating the differentiation in growth of osteoblasts and osteoclasts, which may be used to promote the healing of bone fractures and de novo bone formation, for osteoporosis, for and to promote angiogenesis. Antagonists to the polypeptide of the present invention are also disclosed which may be utilized to treat osteodystrophy, osteohypertrophy, osteoma, osteoblastoma and cancers. Diagnostic assays for identifying mutations in nucleic acid sequence encoding a polypeptide of the present invention.

1 . An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:
 (a) a nucleotide sequence encoding the hECM-1 polypeptide consisting of amino acid residues 1 to 540 of SEQ ID NO:2;   (b) a nucleotide sequence encoding the hECM-1 polypeptide consisting of amino acid residues 2 to 540 of SEQ ID NO:2;   (c) a nucleotide sequence encoding the HECM-1 polypeptide consisting of amino acid residues 20 to 540 of SEQ ID NO:2;   (d) a nucleotide sequence encoding the cytostatin II polypeptide consisting of the full-length polypeptide encoded by the cDNA contained in ATCC™ Deposit Number 97302;   (e) a nucleotide sequence encoding the hECM-1 polypeptide consisting of the full-length polypeptide minus the N-terminal methionine encoded by the cDNA contained in ATCC™ Deposit Number 97302;   (f) a nucleotide sequence encoding the hECM-1 polypeptide consisting of the mature form of the polypeptide encoded by the cDNA contained in ATCC™ Deposit Number 97302; and   (g) a nucleotide sequence encoding the hECM-1-SV1 polypeptide consisting of amino acid residues 1 to 415 of SEQ ID NO:9.   
     
     
         2 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector. 
     
     
         3 . A recombinant vector produced by the method of  claim 2 . 
     
     
         4 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 3  into a host cell. 
     
     
         5 . A recombinant host cell produced by the method of  claim 4 . 
     
     
         6 . A recombinant method of producing a hECM-1 polypeptide, comprising culturing the recombinant host cell of  claim 5  under conditions such that said polypeptide is expressed and recovering said polypeptide. 
     
     
         7 . An isolated hECM-1 polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of:
 (a) the amino acid sequence of the hECM-1 polypeptide consisting of amino acid residues 1 to 540 of SEQ ID NO:2;   (b) the amino acid sequence of the hECM-1 polypeptide consisting of amino acid residues 2 to 540 of SEQ ID NO:2;   (c) the amino acid sequence of the hECM-1 polypeptide consisting of amino acid residues 20 to 540 of SEQ ID NO:2;   (d) the amino acid sequence of the hECM-1 polypeptide consisting of the full-length polypeptide encoded by the cDNA contained in ATCC™ Deposit Number 97302;   (e) the amino acid sequence of the hECM-1 polypeptide consisting of the full-length polypeptide minus the N-terminal methionine encoded by the cDNA contained in ATCC™ Deposit Number 97302;   (f) the amino acid sequence of the hECM-1 polypeptide consisting of the mature form of the polypeptide encoded by the cDNA contained in ATCC™ Deposit Number 97302;   (g) the amino acid sequence of the hECM-1-SV1 polypeptide consisting of amino acid residues 1 to 415 of SEQ ID NO:9.   
     
     
         8 . The polypeptide of  claim 7 , wherein said polypeptide comprises a heterologous polypeptide. 
     
     
         9 . The polypeptide of  claim 7 , wherein said polypeptide is glycosylated. 
     
     
         10 . A composition comprising the polypeptide of  claim 7  and a pharmaceutically acceptable carrier. 
     
     
         11 . An isolated polypeptide produced by the method comprising:
 (a) expressing the polypeptide of  claim 7  in a recombinant host cell comprising a nucleic acid molecule encoding said polypeptide; and   (b) recovering said polypeptide.   
     
     
         12 . An isolated antibody or fragment thereof that specifically binds to a hECM-1 polypeptide of  claim 7 . 
     
     
         13 . An agonist to the polypeptide of  claim 7 . 
     
     
         14 . An antagonist to the polypeptide of  claim 7 . 
     
     
         15 . A method for the treatment of a patient having need of hECM-1 comprising: administering to the patient a therapeutically effective amount of the polypeptide of  claim 7 . 
     
     
         16 . A method of stimulating angiogenesis in a patient comprising administering to the patient the polypeptide of  claim 7 . 
     
     
         17 . A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:
 (a) determining a mutation in the polynucleotide of  claim 1 .   
     
     
         18 . A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:
 (a) determining the presence or amount of expression of the hECM-1 polypeptide using the antibody or fragment thereof of  claim 12 ; and   (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or amount of expression of the hECM-1 polypeptide.   
     
     
         19 . A method of inhibiting angiogenesis in a patient comprising administering to said subject the antibody of  claim 12 .