Methods and compositions for diagnosing and treating disorders involving angiogenesis

Abstract

The present invention relates to polynucleotides associated with angiogenesis-related disorders. The present invention also relates to canine endostatin genes, novel genes associated with angiogenesis-related disorders, such as cancer. The invention encompasses endostatin nucleic acids, recombinant DNA molecules, cloned genes or degenerate variants thereof, endostatin gene products and antibodies directed against such gene products, cloning vectors containing mammalian endostatin gene molecules, and hosts that have been genetically engineered to express such molecules. The invention further relates to methods for the identification of compounds that modulate the expression of endostatin genes and gene products and to using such compounds as therapeutic agents in the treatment of angiogenesis-related disorders, e.g., cancer. The invention also relates to methods for the diagnostic evaluation, genetic testing and prognosis of angiogenesis-related disorders, e.g., cancer, and to methods and compositions for the treatment these disorders.

1 - 11 . (Cancelled)  
     
     
         12 . A transgenic, non-human animal which has been genetically engineered to contain a transgene comprising a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, an isolated nucleic acid molecule that encodes an endostatin consisting of SEQ ID NO: 2, and an isolated nucleic acid molecule that encodes and endostatin consisting of SEQ ID NO: 4.  
     
     
         13 . The transgenic, non-human animal of  claim 12 , wherein the transgene is expressed.  
     
     
         14 . An isolated polypeptide comprising an amino acid sequence of: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         15 . The antibody which binds to the isolated polypeptide of  claim 14 .  
     
     
         16 . An isolated polypeptide comprising an amino acid sequence encoded by an isolated nucleic acid molecule, which hybridizes under highly stringent conditions to the complement of a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, an isolated nucleic acid molecule that encodes an endostatin consisting of SEQ ID NO: 2, and an isolated nucleic acid molecule that encodes and endostatin consisting of SEQ ID NO: 4.  
     
     
         17 . An isolated polypeptide comprising an amino acid sequence encoded by an isolated nucleic acid molecule, which hybridizes under moderately stringent conditions to the complement of a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, an isolated nucleic acid molecule that encodes an endostatin consisting of SEQ ID NO: 2, and an isolated nucleic acid molecule that encodes and endostatin consisting of SEQ ID NO: 4.  
     
     
         18 . An isolated fusion polypeptide comprising a fusion peptide and an amino acid sequence of: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         19 . An isolated fusion polypeptide comprising a fusion peptide and an amino acid sequence encoded by an isolated nucleic acid molecule, which hybridizes under highly stringent conditions to the complement of a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, an isolated nucleic acid molecule that encodes an endostatin consisting of SEQ ID NO: 2, and an isolated nucleic acid molecule that encodes and endostatin consisting of SEQ ID NO: 4.  
     
     
         20 . An isolated fusion polypeptide comprising a fusion peptide and an amino acid sequence encoded by an isolated nucleic acid molecule, which hybridizes under moderately stringent conditions to the complement of a nucleic acid selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, an isolated nucleic acid molecule that encodes an endostatin consisting of SEQ ID NO: 2, and an isolated nucleic acid molecule that encodes and endostatin consisting of SEQ ID NO: 4.  
     
     
         21 . A method for treating an angiogenesis-related disorder in a subject comprising administering to the subject a compound which modulates the function, activity and/or expression of an endostatin sequence in the subject.  
     
     
         22 . The method of  claim 21 , wherein the compound enhances or increases the function, activity and/or expression of the endostatin sequence.  
     
     
         23 . The methods of any one of claims  21 - 22 , wherein the compound is selected from the group consisting of a small organic molecule, an antibody, a ribozyme or an antisense molecule.  
     
     
         24 . The method of any one of claims  21 - 22 , wherein the angiogenesis-related disorder is selected from the group consisting of cancer; angiogenesis-dependent cancer, comprising solid tumors, blood born tumors such as leukemias, and tumor metastases; benign tumors, comprising hemangiomas, acoustic neuromas, neurofibromas, trachomas, and pyogenic granulomas; rheumatoid arthritis; psoriasis; ocular angiogenic diseases, comprising diabetic retinopathy, retinopathy of prematurity, macular degeneration, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia, rubeosis; Osler-Webber Syndrome; myocardial angiogenesis; plaque neovascularization; telangiectasia; hemophiliac joints; angiofibroma; wound granulation; corornary collaterals; cerebral collaterals; arteriovenous malformations; ischemic limb angiogenesis; diabetic neovascularization; macular degeneration; fractures; vasculogenesis; hematopoiesis; ovulation; menstruation; and placentation.  
     
     
         25 . The method of  claim 21 , wherein the endostatin sequence encodes an amino acid sequence comprising: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         26 . The method of  claim 21 , wherein the subject is a dog.  
     
     
         27 . A method for identifying a compound which modulates expression of an endostatin sequence comprising: 
 a) contacting a test compound to a cell that expresses an endostatin sequence;    b) measuring a level of endostatin sequence expression in the cell;    c) comparing the level of endostatin sequence expression in the cell in the presence of the test compound to a level of endostatin sequence expression in the cell in the absence of the test compound,    wherein if the level of endostatin sequence expression in the cell in the presence of the test compound differs from the level of expression of the endostatin sequence in the cell in the absence of the test compound, a compound that modulates expression of an endostatin sequence is identified.    
     
     
         28 . The method of  claim 27 , wherein the endostatin sequence is endogenously expressed within the cell.  
     
     
         29 . The method of  claim 27 , wherein the endostatin sequence encodes an amino acid sequence comprising: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         30 . The method of  claim 27 , wherein the endostatin sequence comprises: 
 a) a nucleic acid as shown in SEQ ID NO: 1; or    b) a nucleic acid as shown in SEQ ID NO: 3.    
     
     
         31 . A method for identifying a compound which modulates activity of an endostatin sequence product comprising: 
 a) contacting a test compound to a cell that expresses an endostatin sequence product;    b) measuring a level of endostatin sequence product in the cell;    c) comparing the level of endostatin sequence product activity in the cell in the presence of the test compound to a level of endostatin sequence product activity in the cell in the absence of the test compound,    wherein if the level of endostatin sequence product activity in the cell in the presence of the test compound differs from the level of endostatin sequence product activity in the cell in the absence of the test compound, a compound that modulates activity of an endostatin sequence product is identified.    
     
     
         32 . The method of  claim 31 , wherein the endostatin sequence product comprises: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         33 . A method for modulating the activity and/or expression of an endostatin sequence in a cell comprising administering to the cell a compound which modulates the activity and/or expression of an endostatin sequence in the cell.  
     
     
         34 . The method of  claim 33 , wherein the compound is selected from the group consisting of a small organic molecule, an antibody, a ribozyme or an antisense molecule.  
     
     
         35 . The method of  claim 33 , wherein the endostatin sequence encodes an amino acid sequence comprising: 
 a) SEQ ID NO: 2; or    b) SEQ ID NO: 4.    
     
     
         36 . The method of  claim 33 , wherein the endostatin sequence comprises: 
 a) a nucleic acid as shown in SEQ ID NO: 1; or    b) a nucleic acid as shown in SEQ ID NO: 3.