Vascular endothelial growth factor 2

Abstract

Disclosed are human VEGF2 polypeptides, biologically active, diagnostically or therapeutically useful fragments, analogs, or derivatives thereof, and DNA(RNA) encoding such VEGF2 polypeptides. Also provided are procedures for producing such polypeptides by recombinant techniques and antibodies and antagonists against such polypeptides. Such polypeptides may be used therapeutically for stimulating wound healing and for vascular tissue repair. Also provided are methods of using the antibodies and antagonists to inhibit tumor angiogenesis and thus tumor growth, inflammation, diabetic retinopathy, rheumatoid arthritis, and psoriasis.

1 . An isolated nucleic acid molecule comprising a polynucleotide selected from the group consisting of: 
 (A) a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) a nucleotide sequence encoding a polypeptide comprising amino acids from about −23 to about 396 in SEQ ID NO:2;  
 (b) a nucleotide sequence encoding a polypeptide comprising amino acids from about −22 to about 396 in SEQ ID NO:2;  
 (c) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 396 in SEQ ID NO:2;  
 (d) a nucleotide sequence encoding a polypeptide comprising amino acids from about −24 to about 326 in SEQ ID NO:4;  
 (e) a nucleotide sequence encoding a polypeptide comprising amino acids from about −23 to about 326 in SEQ ID NO:4;  
 (f) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 326 in SEQ ID NO:4;  
 (g) a nucleotide sequence encoding a polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit Nos. 97149 or 75698;  
 (h) a nucleotide sequence encoding the mature VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97149 or 75698;  
 (i) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), or (h); and  
 (j) a nucleotide sequence that hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), (e), (f), (g), (h), or (i), wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues; and  
   (B) a polynucleotide sequence encoding a VEGF2 polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has a sequence selected from the group consisting of: 
 (k) a nucleotide sequence encoding a polypeptide comprising amino acids from about −23 to about 396 in SEQ ID NO:2;  
 (l) a nucleotide sequence encoding a polypeptide comprising amino acids from about −22 to about 396 in SEQ ID NO:2;  
 (m) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 396 in SEQ ID NO:2;  
 (n) a nucleotide sequence encoding a polypeptide comprising amino acids from about −24 to about 326 in SEQ ID NO:4;  
 (o) a nucleotide sequence encoding a polypeptide comprising amino acids from about −23 to about 326 in SEQ ID NO:4;  
 (p) a nucleotide sequence encoding a polypeptide comprising amino acids from about 1 to about 326 in SEQ ID NO:4;  
 (q) a nucleotide sequence encoding a polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit Nos. 97149 or 75698;  
 (r) a nucleotide sequence encoding the mature VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit No. 97149 or 75698; and  
 (s) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), or (h).  
   
     
     
         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 for producing a VEGF2 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 VEGF2 polypeptide having an amino acid sequence selected from the group consisting of: 
 (A) an amino acid sequence at least 95% identical to a sequence selected from the group consisting of: 
 (a) amino acids from about −23 to about 396 in SEQ ID NO:2;  
 (b) amino acids from about −22 to about 396 in SEQ ID NO:2;  
 (c) amino acids from about 1 to about 396 in SEQ ID NO:2;  
 (d) amino acids from about −24 to about 326 in SEQ ID NO:4;  
 (e) amino acids from about −23 to about 326 in SEQ ID NO:4;  
 (f) amino acids from about 1 to about 326 in SEQ ID NO:4;  
 (g) the amino acid sequence of the VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clones contained in ATCC™ Deposit Nos. 97149 or 75698;  
 (h) the amino acid sequence of the mature VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit Nos. 97149 or 75698; and  
   (B) an isolated VEGF2 polypeptide wherein, except for at least one conservative amino acid substitution, said polypeptide has a sequence selected from the group consisting of: 
 (i) amino acids from about −23 to about 396 in SEQ ID NO:2;  
 (j) amino acids from about −22 to about 396 in SEQ ID NO:2;  
 (k) amino acids from about 1 to about 396 in SEQ ID NO:2;  
 (l) amino acids from about −24 to about 326 in SEQ ID NO:4;  
 (m) amino acids from about −23 to about 326 in SEQ ID NO:4;  
 (n) amino acids from about 1 to about 326 in SEQ ID NO:4;  
 (o) the amino acid sequence of the VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clones contained in ATCC™ Deposit Nos. 97149 or 75698; and  
 (p) the amino acid sequence of the mature VEGF2 polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC™ Deposit Nos. 97149 or 75698.  
   
     
     
         8 . An antibody against the polypeptide of  claim 7 .  
     
     
         9 . A method of making an antibody comprising introducing a polypeptide comprising at least 50 contiguous amino acids of the polypeptide of  claim 7  into an animal and recovering said antibody.  
     
     
         10 . A method of making an antibody comprising screening a single chain or Fab expression library to identify an antibody that specifically binds a polypeptide comprising at least 50 amino acids of the polypeptide of  claim 7  and recovering said antibody from said library.  
     
     
         11 . A compound effective as an agonist for the polypeptide of  claim 7 .  
     
     
         12 . A compound effective as an antagonist against the polypeptide of  claim 7 .  
     
     
         13 . A pharmaceutical composition comprising the polypeptide of  claim 7  and a pharmaceutically acceptable carrier.  
     
     
         14 . A method for the treatment of a patient comprising administering a therapeutically effective amount of the polypeptide of  claim 7 .  
     
     
         15 . A method for the treatment of a patient comprising administering a therapeutically effective amount of the agonist of  claim 11 .  
     
     
         16 . A method for the treatment of a patient comprising administering a therapeutically effective amount of the antagonist of  claim 12 .  
     
     
         17 . 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.  
     
     
         18 . A process for diagnosing a disease or a susceptibility to a disease related to expression of the polypeptide of  claim 7  comprising: 
 determining a mutation in the nucleic acid sequence encoding said polypeptide.    
     
     
         19 . A diagnostic process comprising: 
 analyzing for the presence of the polypeptide of  claim 7  in a sample derived from a host.    
     
     
         20 . A method for identifying compounds which bind to and activate or inhibit a receptor for the polypeptide of  claim 7  comprising: 
 contacting a cell expressing on the surface thereof a receptor for the polypeptide, said receptor being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said receptor, with a compound to be screened under conditions to permit binding to the receptor; and    determining whether the compound binds to and activates or inhibits the receptor by detecting the presence or absence of a signal generated from the interaction of the compound with the receptor.