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 . 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.  
     
     
         2 . The composition of  claim 1  wherein said EG-VEGF polypeptide is a native sequence EG-VEGF.  
     
     
         3 . The composition of  claim 2  wherein said native sequence EG-VEGF is human.  
     
     
         4 . The composition of  claim 1  wherein said agonist or antagonist is an anti-EG-VEGF-antibody.  
     
     
         5 . The composition of  claim 1  wherein said agonist or antagonist is an anti-EG-VEGF-antibody fragment.  
     
     
         6 . The composition of  claim 1  wherein said agonist or antagonist is a small molecule.  
     
     
         7 . The composition of  claim 1  wherein said antagonist is an antisense molecule.  
     
     
         8 . The composition of  claim 1  further comprising a vascular endothelial cell growth factor (VEGF), or an agonist or antagonist thereof.  
     
     
         9 . The composition of  claim 8  wherein said VEGF is a native sequence VEGF polypeptide.  
     
     
         10 . The composition of  claim 9  wherein said native sequence VEGF polypeptide is human.  
     
     
         11 . An article of manufacture, comprising: 
 a container;    a label on the container; and    a composition comprising an active agent contained within the container,    wherein said active agent is selected from the group consisting of (a) an EG-VEGF polypeptide, (b) an agonist of an EG-VEGF polypeptide, and (c) an antagonist of an EG-VEGF polypeptide; and the label on the container indicates that the composition is effective for treating of a condition that is associated with a hormone producing endothelial tissue.    
     
     
         12 . The article of manufacture of  claim 11  wherein said condition is associated with steroidogenic endothelial cells within an endocrine gland.  
     
     
         13 . The article of manufacture of  claim 11  wherein said tissue is ovarian, testicular, cervical, adrenal, placental or prostate tissue.  
     
     
         14 . The article of manufacture of  claim 11  wherein said condition is infertility.  
     
     
         15 . The article of manufacture of  claim 11  wherein said condition is polycystic ovary syndrome.  
     
     
         16 . The article of manufacture of  claim 11  wherein said condition is cancer.  
     
     
         17 . A method for identifying a compound that binds to EG-VEGF comprising: 
 a) contacting a candidate compound with EG-VEGF; and    b) determining whether said candidate compound binds to said EG-VEGF.    
     
     
         18 . The method of  claim 17  wherein said EG-VEGF is a native sequence EG-VEGF.  
     
     
         19 . The method of  claim 17  wherein said EG-VEGF is a fragment of a native sequence EG-VEGF.  
     
     
         20 . The method of  claim 17  wherein said EG-VEGF is an amino acid sequence variant of a native sequence EG-VEGF.  
     
     
         21 . The method of  claim 20  wherein said amino acid sequence variant has at least about 85% sequence identity to the sequence of amino acid residues from about 1 or about 20 to about 105, inclusive, of FIG. 2 (SEQ ID NO: 2).  
     
     
         22 . The method of  claim 21  wherein said amino acid sequence variant is a conservative substitution variant.  
     
     
         23 . The method of  claim 17  wherein said EG-VEGF is present as a fusion protein.  
     
     
         24 . The method of  claim 17  wherein the ability of said candidate compound to compete with a molecule known to bind EG-VEGF is measured.  
     
     
         25 . The method of  claim 24  wherein said candidate compound is contacted with a whole cell or a cell membrane fraction expressing the coding sequence of EG-VEGF.  
     
     
         26 . 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.    
     
     
         27 . The method of  claim 26  wherein said compound inhibits a biological activity of said EG-VEGF.  
     
     
         28 . The method of  claim 26  wherein said compound enhances a biological activity of said EG-VEGF.  
     
     
         29 . The method of  claim 26  wherein said biological activity is the ability to induce phosphorylation of a kinase involved in cell proliferation or survival.  
     
     
         30 . The method of  claim 29  wherein said kinase is a MAP kinase.  
     
     
         31 . The method of  claim 30  wherein said MAP kinase is ERKI or ERK2.  
     
     
         32 . The method of  claim 26  wherein said biological activity is cell proliferation activity.  
     
     
         33 . The method of  claim 26  wherein said biological activity is the induction of chemotaxis.  
     
     
         34 . The method of  claim 26  wherein said biological activity is angiogenesis.  
     
     
         35 . The method of  claim 26  wherein said biological activity is the induction of cell differentiation.  
     
     
         36 . The method of  claim 26  wherein said biological activity is the induction of endothelial cell fenestration.  
     
     
         37 . The method of  claim 26  wherein said candidate compound is contacted with a whole cell or a cell membrane fraction expressing the coding sequence of EG-VEGF.  
     
     
         38 . The method of  claim 37  wherein said cell is a recombinant host cell engineered to express said EG-VEGF.  
     
     
         39 . The method of  claim 26  wherein said candidate compound is contacted with an isolated EG-VEGF.  
     
     
         40 . The method of  claim 39  wherein said EG-VEGF is immobilized on a solid support.  
     
     
         41 . A compound identified by the method of  claim 17  or  claim 26 .  
     
     
         42 . A method of identifying a receptor for EG-VEGF, said method comprising combining EG-VEGF with a composition comprising cell membrane material wherein said EG-VEGF complexes with a receptor on said cell membrane material, and identifying said receptor as a EG-VEGF receptor.  
     
     
         43 . The method of  claim 42  wherein EG-VEGF binds to said receptor, and said method further includes a step of crosslinking said EG-VEGF and said receptor.  
     
     
         44 . A method of inducing cell proliferation, comprising contacting said cells with EG-VEGF in an amount effective to induce proliferation of said cells.  
     
     
         45 . The method of  claim 44  wherein said cells are endothelial cells.  
     
     
         46 . The method of  claim 45  wherein said endothelial cells are steroidogenic endothelial cells.  
     
     
         47 . The method of  claim 46  wherein said endothelial cells are cells of a steroidogenic gland.  
     
     
         48 . The method of  claim 45  further comprising contacting said cells with VEGF.  
     
     
         49 . A method of inducing cell proliferation, said method comprising introducing a nucleic acid encoding EG-VEGF into said cells in an amount effective to induce cell proliferation.  
     
     
         50 . The method of  claim 49  wherein said cells are endothelial cells.  
     
     
         51 . The method of  claim 50  wherein said endothelial cells are steroidogenic endothelial cells.  
     
     
         52 . The method of  claim 51  wherein said endothelial cells are cells of a steroidogenic gland.  
     
     
         53 . The method of  claim 50  further comprising introducing a nucleic acid encoding VEGF to said cells.  
     
     
         54 . A method of inducing chemotaxis in cells, comprising contacting said cells with EG-VEGF in an amount effective to induce chemotaxis.  
     
     
         55 . The method of  claim 54  wherein said cells are endothelial cells.  
     
     
         56 . The method of  claim 55  wherein said cells are steroidogenic endothelial cells.  
     
     
         57 . The method of  claim 56  wherein said cells are endothelial cells of a steroidogenic gland.  
     
     
         58 . The method of  claim 55  further comprising contacting said cells with VEGF.  
     
     
         59 . A method of inducing chemotaxis in cells, said method comprising introducing a nucleic acid encoding EG-VEGF into said cells in an amount effective to induce chemotaxis.  
     
     
         60 . The method of  claim 59  wherein said cells are endothelial cells.  
     
     
         61 . The method of  claim 60  wherein said cells are steroidogenic endothelial cells.  
     
     
         62 . The method of  claim 60  further comprising introducing a nucleic acid encoding VEGF to said cells.  
     
     
         63 . A method of treating an individual for a condition associated with hormone producing tissue, said method comprising administering to said individual a composition comprising EG-VEGF or an agonist or antagonist thereof in an amount effective to treat said condition.  
     
     
         64 . The method of  claim 63  further comprising administering VEGF or an agonist or antagonist thereof to said individual.  
     
     
         65 . A method of treating an individual for a condition associated with hormone producing tissue, said method comprising administering to said individual a composition comprising a nucleic acid encoding EG-VEGF or an agonist or antagonist thereof in an amount effective to treat said condition.  
     
     
         66 . The method of  claim 65  further comprising administering a composition comprising a nucleic acid encoding VEGF or an agonist or antagonist thereof to said individual.  
     
     
         67 . A method of inducing angiogenesis in hormone producing tissue, comprising contacting said tissue with EG-VEGF or an agonist thereof in an amount effective to induce angiogenesis.  
     
     
         68 . The method of  claim 67  further comprising contacting said tissue with VEGF or an agonist thereof.  
     
     
         69 . A method of inducing angiogenesis in hormone producing tissue, said method comprising introducing a nucleic acid molecule encoding EG-VEGF or an agonist thereof into said hormone producing tissue in an amount effective to induce angiogenesis.  
     
     
         70 . The method of  claim 69  further comprising introducing nucleic acid encoding VEGF or an agonist thereof into said tissue.  
     
     
         71 . A method of inducing cell differentiation in cells, comprising contacting said cells with EG-VEGF or an agonist thereof in an amount effective to induce cell differentiation.  
     
     
         72 . The method of  claim 71  further comprising contacting said cells with VEGF or an agonist thereof.  
     
     
         73 . A method of inducing cell differentiation in cells, comprising introducing a nucleic acid molecule encoding EG-VEGF into said cells in an amount effective to induce cell differentiation.  
     
     
         74 . The method of  claim 73  further comprising introducing a nucleic acid molecule encoding VEGF or an agonist thereof into said cells.  
     
     
         75 . A method of inducing fenestration in cells, contacting said cells with EG-VEGF or an agonist thereof in an amount effective to induce fenestration.  
     
     
         76 . The method of  claim 75  wherein said cells are endothelial cells.  
     
     
         77 . The method of  claim 76  wherein said cells are steroidogenic endothelial cells.  
     
     
         78 . The method of  claim 77  wherein said cells are endothelial cells of a steroidogenic gland.  
     
     
         79 . The method of  claim 76  further comprising contacting said cells with VEGF or an agonist thereof.  
     
     
         80 . A method of inducing fenestration in cells, comprising introducing a nucleic acid molecule encoding EG-VEGF or an agonist thereof into said cells in an amount effective to induce fenestration.  
     
     
         81 . The method of  claim 80  wherein said cells are endothelial cells.  
     
     
         82 . The method of  claim 81  wherein said cells are steroidogenic endothelial cells.  
     
     
         83 . The method of  claim 82  wherein said cells are endothelial cells of a steroidogenic gland.  
     
     
         84 . The method of  claim 81  further comprising introducing a nucleic acid molecule encoding VEGF into said cells.  
     
     
         85 . A method of inhibiting endothelial cell proliferation, comprising contacting said cells with an EG-VEGF antagonist in an amount effective to inhibit cell proliferation.  
     
     
         86 . A method of inhibiting chemotaxis in endothelial cells, comprising contacting said cells with an EG-VEGF antagonist in an amount effective to inhibit chemotaxis.  
     
     
         87 . A method of inhibiting angiogenesis in hormone producing tissue, comprising contacting said tissue with an EG-VEGF antagonist in an amount effective to inhibit angiogenesis.  
     
     
         88 . A method of inhibiting differentiation of cells, comprising contacting said cells with an EG-VEGF antagonist in an amount effective to inhibit cell differentiation.  
     
     
         89 . A method of regulating fertility in an individual, comprising administering an EG-VEGF antagonist to said individual in an amount effective to regulate fertility.  
     
     
         90 . The method of  claim 89  wherein said fertility is regulated by inhibiting follicle maturation.  
     
     
         91 . The method of  claim 89  wherein said fertility is regulated by inhibiting ovulation.  
     
     
         92 . The method of  claim 89  wherein said individual has or is at risk of having polycystic ovary syndrome and fertility is regulated so as to maintain fertility.  
     
     
         93 . A method of treating cancer in cells responsive to EG-VEGF in an individual, said method comprising administering an EG-VEGF antagonist to said individual in an amount effective to treat cancer.  
     
     
         94 . A method of treating cancer of the reproductive organs in an individual, comprising administering an EG-VEGF antagonist to said individual in an amount effective to inhibit cancer.  
     
     
         95 . The method of  claim 94  wherein said cancer is selected from the group consisting of ovarian cancer, testicular cancer, prostate cancer, and uterine cancer.  
     
     
         96 . The method of  claim 94  wherein said cancer is hormone-dependent cancer.  
     
     
         97 . The method of  claim 96  wherein said cancer is andogen-dependent cancer.  
     
     
         98 . The method of  claim 97  wherein said cancer is androgen-dependent prostate cancer.  
     
     
         99 . A method of treating an ovarian cyst in an individual, comprising administering an EG-VEGF antagonist to said individual in an amount effective to inhibit growth of a cyst.  
     
     
         100 . A method of regulating permeability of an endothelial cell comprising contacting said endothelial cell with an effective amount of EG-VEGF.  
     
     
         101 . The method of claim  100  further comprising contacting said endothelial cell with VEGF.  
     
     
         102 . The method of claim  100  wherein said cell is an endothelial cell of the adrenal cortex.  
     
     
         103 . The method of claim  100  wherein said cell is an endothelial cell of the ovary.