Eg-vegf nucleic acids and polypeptides and methods of use

Abstract

The present invention is directed to novel polypeptides designated herein as EG-VEGF and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention. Also provided herein are methods of screening for modulators of EG-VEGF. Furthermore, methods and related methods of treatment are described herein which pertain to regulating cellular proliferation and chemotaxis.

What is claimed is: 
     
         1 . An antibody selected from the group consisting of anti-EG-VEGF monoclonal antibodies 1C6, 2A3, 2A8 and 4H9. 
     
     
         2 . An antibody that binds essentially the same epitope of EG-VEGF bound by an antibody selected from the group consisting of anti-EG-VEGF monoclonal antibodies 1-C6, 2A3, 2A8 and 4H9. 
     
     
         3 . The antibody of  claim 2  which is an antibody fragment. 
     
     
         4 . The antibody of  claim 3  selected from the group consisting of Fab, Fab′, F(ab) 2 , and Fv fragments. 
     
     
         5 . The antibody of  claim 2  which is a chimeric antibody. 
     
     
         6 . The antibody of  claim 2  which is humanized. 
     
     
         7 . The antibody of  claim 2  which is human. 
     
     
         8 . The antibody of  claim 2  which is a bispecific antibody. 
     
     
         9 . The antibody of  claim 8  wherein said bispecific antibody has binding specificity for VEGF. 
     
     
         10 . The antibody of  claim 2  which is detectably labeled. 
     
     
         11 . A composition of matter comprising (a) an EG-VEGF polypeptide, (b) an agonist of an EG-VEGF polypeptide, or (c) an antagonist of an EG-VEGF polypeptide, in admixture with a pharmaceutically acceptable carrier. 
     
     
         12 . The composition of  claim 11  wherein said EG-VEGF polypeptide is a native sequence EG-VEGF. 
     
     
         13 . The composition of  claim 12  wherein said native sequence EG-VEGF is human. 
     
     
         14 . The composition of  claim 11  wherein said agonist or antagonist is an anti-EG-VEGF-antibody. 
     
     
         15 . The composition of  claim 14  wherein said antagonist is an anti-EG-VEGF antibody selected from the group consisting of monoclonal antibodies 1C6, 2A3, 2A8 and 4H9. 
     
     
         16 . The composition of  claim 14  wherein said antagonist is an anti-EG-VEGF antibody that binds essentially the same epitope of EG-VEGF bound by an antibody selected from the group consisting of anti-EG-VEGF monoclonal antibodies 1C6, 2A3, 2A8 and 4H9. 
     
     
         17 . The composition of  claim 11  wherein said agonist or antagonist is an anti-EG-VEGF antibody fragment. 
     
     
         18 . The composition of  claim 17  wherein said antibody fragment is selected from the group consisting of Fab, Fab′, F(ab) 2 , and Fv fragments. 
     
     
         19 . The composition of  claim 11  wherein said antagonist is an antisense molecule. 
     
     
         20 . The composition of  claim 11  further comprising a vascular endothelial growth factor (VEGF), or an agonist or antagonist thereof. 
     
     
         21 . The composition of  claim 20  wherein said VEGF is a native sequence VEGF polypeptide. 
     
     
         22 . The composition of  claim 21  wherein said native sequence VEGF is human. 
     
     
         23 . An article of manufacture, comprising:
 a container;   a label on the container; and   a composition comprising an anti-EG-VEGF antibody binding essentially the same epitope as an antibody selected from the group consisting of monoclonal antibodies 1C6, 2A3, 2A8 and 4H9.   
     
     
         24 . The article of manufacture of  claim 23  comprising an anti-EG-VEGF antibody selected from the group consisting of monoclonal antibodies 1C6, 2A3, 2A8 and 4H9. 
     
     
         25 . A method for identifying a compound that modulates a biological activity of EG-VEGF, comprising the steps of:
 a) contacting a candidate compound with EG-VEGF; and   b) determining an alteration in said biological activity of EG-VEGF.   
     
     
         26 . The method of  claim 25  wherein said compound inhibits a biological activity of said EG-VEGF. 
     
     
         27 . The method of  claim 25  wherein said compound enhances a biological activity of said EG-VEGF. 
     
     
         28 . The method of  claim 25  wherein said biological activity is the ability to induce phosphorylation of a kinase involved in cell proliferation or survival. 
     
     
         29 . The method of  claim 28  wherein said kinase is a MAP kinase. 
     
     
         30 . The method of  claim 29  wherein said MAP kinase is ERK1 or ERK2. 
     
     
         31 . The method of  claim 25  wherein said biological activity is the ability to induce phosphorylation of Akt or eNOS. 
     
     
         32 . The method of  claim 25  wherein said biological activity is the ability to stimulate cell proliferation. 
     
     
         33 . The method of  claim 25  wherein said biological activity is the induction of chemotaxis. 
     
     
         34 . The method of  claim 25  wherein said biological activity is the induction of angiogenesis. 
     
     
         35 . The method of  claim 25  wherein said biological activity is the induction of cell differentiation. 
     
     
         36 . The method of  claim 25  wherein said biological activity is the induction of endothelial cell fenestration. 
     
     
         37 . The method of  claim 25  wherein said biological activity is the enhancement of endothelial cell survival. 
     
     
         38 . The method of  claim 25  wherein said candidate compound is contacted with a whole cell or a cell membrane fraction expressing the coding sequence of EG-VEGF. 
     
     
         39 . The method of  claim 38  wherein said cell is a recombinant host cell engineered to express said EG-VEGF. 
     
     
         40 . The method of  claim 25  wherein said candidate compound is contacted with an isolated EG-VEGF. 
     
     
         41 . The method of  claim 40  wherein said EG-VEGF is immobilized on a solid support. 
     
     
         42 . A compound identified by the method of  claim 25 . 
     
     
         43 . A method of inducing cell proliferation, comprising contacting said cells with EG-VEGF in an amount effective to induce proliferation of said cells. 
     
     
         44 . The method of  claim 43  wherein said cells are endothelial cells. 
     
     
         45 . The method of  claim 44  wherein said endothelial cells are steroidogenic endothelial cells. 
     
     
         46 . The method of  claim 45  wherein said endothelial cells are cells of a steroidogenic gland. 
     
     
         47 . The method of  claim 43  further comprising contacting said cells with VEGF. 
     
     
         48 . A method of inducing chemotaxis in cells, comprising contacting said cells with EG-VEGF in an amount effective to induce chemotaxis. 
     
     
         49 . The method of  claim 48  wherein said cells are endothelial cells. 
     
     
         50 . The method of  claim 49  wherein said cells are steroidogenic endothelial cells. 
     
     
         51 . The method of  claim 50  wherein said cells are endothelial cells of a steroidogenic gland. 
     
     
         52 . The method of  claim 48  further comprising contacting said cells with VEGF. 
     
     
         53 . A method of enhancing cell survival comprising contacting cells with EG-VEGF in an amount effective to enhance cell survival. 
     
     
         54 . The method of  claim 53  wherein said cells are endothelial cells. 
     
     
         55 . The method of  claim 54  wherein said cells are steroidogenic endothelial cells. 
     
     
         56 . The method of  claim 55  wherein said cells are endothelial cells of a steroidogenic gland. 
     
     
         57 . The method of  claim 53  further comprising contacting said cells with VEGF. 
     
     
         58 . A method of inhibiting endothelial cell proliferation, comprising contacting said cells with an EG-VEGF antagonist in an amount effective to inhibit cell proliferation. 
     
     
         59 . The method of  claim 58  wherein said EG-VEGF antagonist is an anti-EG-VEGF antibody selected from the group consisting of monoclonal antibodies 1C6, 2A3, 2A8 and 4H9. 
     
     
         60 . A method of inhibiting chemotaxis in endothelial cells, comprising contacting said cells with an EG-VEGF antagonist in an amount effective to inhibit chemotaxis. 
     
     
         61 . The method of  claim 60  wherein said EG-VEGF antagonist is an anti-EG-VEGF antibody selected from the group consisting of monoclonal antibodies 1C6, 2A3, 2A8 and 4H9.