Polypeptides Homologous to VEGF and BMP1

Abstract

The present invention involves the identification and preparation of vascular endothelial growth factor-E (VEGF-E). VEGF-E is a novel polypeptide related to vascular endothelial growth factor (VEGF) and bone morphogenetic protein 1. VEGF-E has homology to VEGF including conservation of the amino acids required for activity of VEGF. VEGF-E can be useful in wound repair, as well as in the generation and regeneration of tissue.

1 . An isolated nucleic acid comprising a nucleotide sequence encoding a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising amino acid residues 1 through 345 of  FIG. 2  (SEQ ID NO:2). 
     
     
         2 . The nucleic acid of  claim 1  comprising the sequence of nucleotides 259 through 1293 of  FIG. 1  (SEQ ID NO:1), or its complement. 
     
     
         3 . An isolated nucleic acid comprising a nucleotide sequence which hybridizes under stringent conditions to the nucleotide sequence of  claim 1 . 
     
     
         4 . A vector comprising the nucleic acid of  claim 1 . 
     
     
         5 . A host cell comprising the nucleic acid of  claim 1 . 
     
     
         6 . The host cell of  claim 5 , wherein said cell is a Chinese Hamster Ovary cell, an insect cell, an  E. coli  cell, or a yeast cell. 
     
     
         7 . The host cell of  claim 6  that is a baculovirus-infected insect cell. 
     
     
         8 . A process for producing a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising culturing the host cell of  claim 5  under conditions suitable for expression of the VEGF-E polypeptide and recovering the VEGF-E polypeptide from the cell culture. 
     
     
         9 . A polypeptide produced by the process of  claim 8 . 
     
     
         10 . A VEGF-E polypeptide comprising amino acid residues 1 through 345 of  FIG. 2  (SEQ ID NO:2). 
     
     
         11 . A VEGF-E polypeptide selected from the group consisting of:
 (a) a polypeptide comprising amino acid residues 1 through 345 of  FIG. 2  (SEQ ID NO:2); and   (b) a polypeptide fragment of (a), wherein said fragment is biologically active.   
     
     
         12 . A VEGF-E polypeptide encoded by the nucleotide sequence insert of ATCC deposit No. 209653. 
     
     
         13 . A chimeric polypeptide comprising the polypeptide of  claim 10  fused to a heterologous amino acid sequence. 
     
     
         14 . The chimeric polypeptide of  claim 13 , wherein said heterologous amino acid sequence is an epitope tag sequence or a Fc region of an immunoglobulin. 
     
     
         15 . A composition comprising the polypeptide of  claim 10  in admixture with a carrier. 
     
     
         16 . The composition of  claim 15  comprising a therapeutically effective amount of the polypeptide, wherein the carrier is a pharmaceutically acceptable carrier. 
     
     
         17 . A pharmaceutical product comprising:
 (a) the composition of  claim 15 ;   (b) a container containing said composition; and   (c) a label affixed to said container, or a package insert included in said pharmaceutical product, referring to the use of said VEGF-E polypeptide in the treatment of a cardiovascular or endothelial disorder.   
     
     
         18 . A method for treating a cardiovascular or endothelial disorder in a mammal comprising administering to the mammal an effective amount of the composition of  claim 15 . 
     
     
         19 . The method of  claim 18  wherein the disorder is cardiac hypertrophy, trauma, or a bone-related disorder. 
     
     
         20 . The method of  claim 19  wherein said cardiac hypertrophy is characterized by the presence of an elevated level of PGF 2 . 
     
     
         21 . The method of  claim 19  wherein said cardiac hypertrophy has been induced by myocardial infarction. 
     
     
         22 . The method of  claim 21  wherein said VEGF-E polypeptide administration is initiated within 48 hours following myocardial infarction. 
     
     
         23 . A method for diagnosing a disease or a susceptibility to a disease related to a mutation in a nucleic acid sequence encoding vascular endothelial cell growth factor-E (VEGF-E) comprising:
 (a) isolating a nucleic acid sequence encoding VEGF-E from a sample derived from a host; and   (b) determining a mutation in the nucleic acid sequence encoding VEGF-E.   
     
     
         24 . A method of diagnosing cardiovascular and endothelial disorders in a mammal comprising detecting the level of expression of a gene encoding a vascular endothelial cell growth factor-E (VEGF-E) polypeptide (a) in a test sample of tissue cells obtained from the mammal, and (b) in a control sample of known normal tissue cells of the same cell type, wherein a higher or lower expression level in the test sample indicates the presence of a cardiovascular or endothelial dysfunction in the mammal from which the test tissue cells were obtained. 
     
     
         25 . A method for identifying an agonist to a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising:
 (a) contacting cells and a candidate compound under conditions that allow the polypeptide to stimulate proliferation of the cells; and   (b) measuring the extent to which cell proliferation is inhibited by the compound.   
     
     
         26 . An agonist to a VEGF-E polypeptide identified by the method of  claim 25 . 
     
     
         27 . A method for identifying a compound that inhibits the expression or activity of a vascular endothelial cell growth factor-E (VEGF-E) polypeptide, comprising:
 (a) contacting a candidate compound with the polypeptide under conditions and for a time sufficient to allow the compound and polypeptide to interact; and   (b) measuring the extent to which the compound interacts with the polypeptide.   
     
     
         28 . A compound identified by the method of  claim 27 . 
     
     
         29 . An isolated antibody that binds a vascular endothelial cell growth factor-E (VEGF-E) polypeptide. 
     
     
         30 . The antibody of  claim 29  that is a monoclonal antibody. 
     
     
         31 . A method for determining the presence of a vascular endothelial cell growth factor-E (VEGF-E) polypeptide comprising exposing a cell suspected of containing the VEGF-E polypeptide to the antibody of  claim 29  and determining binding of said antibody to said cell. 
     
     
         32 . A method of diagnosing cardiovascular, endothelial, or angiogenic disorders in a mammal comprising (a) contacting the antibody of  claim 29  with a test sample of tissue cells obtained from the mammal, and (b) detecting the formation of a complex between the anti-VEGF-E antibody and the VEGF-E polypeptide in the test sample. 
     
     
         33 . An article of manufacture, comprising:
 a container;   a label on the container; and   
       a composition comprising the antibody of  claim 29  contained within the container; wherein the label on the container indicates instructions for using the antibody in a therapeutic method or diagnostic method.