Method for molecular diagnosis of tumor angiogenesis and metastasis

Abstract

The invention is directed to methods of assessing the molecular stage of an disease characterized by abnormal angiogenesis. More particularly, the invention is directed to a method of determining the metastatic potential of a tumor by determining the presence of specific molecular markers. The invention is based in part on the discovery that the key marker for determining the eventual metastasis of a tumor is vascular endothelia growth factor. Knowledge of the particular markers present in a tumor condition allows the physician to tailor cancer treatment and manage the disease condition.

What is claimed is:  
     
         1 . A method of diagnosing metastatic disease in a human comprising: 
 a) removing a sample of cells from the human;    b) assaying the sample for VEGF transcriptional activity; and    c) determining the existence of metastatic disease characterized by the induction of VEGF transcriptional activity.    
     
     
         2 . The method of    claim 1    wherein the cells are tumor cells.  
     
     
         3 . The method of    claim 2    wherein the tumor cells are obtained through a surgical debulking of the tumor.  
     
     
         4 . The method of    claim 1    wherein the transcriptional activity is measured by hybridization of RNA from the cells to a labeled complementary nucleic acid.  
     
     
         5 . The method of    claim 1    wherein the transcriptional activity is measured by polymerase chain reaction.  
     
     
         6 . The method of    claim 1    further comprising assaying the sample for the presence of VEGF protein.  
     
     
         7 . The method of    claim 1    further comprising assaying the sample for the expression of tyrosine kinase receptors involved in angiogenesis.  
     
     
         8 . The method of    claim 7    wherein the tyrosine kinase receptor is chosen from the group consisting of the KDR/flk-1 receptor, the flt-1 receptor, and/or the tek/tie-2 receptor.  
     
     
         9 . The method of    claim 1    further comprising assaying the sample for the expression of hypoxia induced factor 1.  
     
     
         10 . The method of    claim 1    further comprising assaying for the abnormal expression of an oncogene.  
     
     
         11 . A method of diagnosing metastatic disease in a human consisting of: 
 a) removing a sample of tissue or fluid from the human;    b) assaying for the presence of VEGF protein in the sample; and    c) determining the existence of metastatic disease by the presence of VEGF protein, wherein the abnormal presence of VEGF protein indicates the presence of metastatic disease.    
     
     
         12 . The method of    claim 11    wherein the sample is obtained through a surgical debulking of the tumor.  
     
     
         13 . The method of    claim 11    wherein the fluid is blood.  
     
     
         14 . The method of    claim 11    wherein the presence of VEGF is determined using an anti-VEGF antibody.  
     
     
         15 . The method of    claim 14    wherein the anti-VEGF antibody is used in an ELISA assay.  
     
     
         16 . The method of    claim 11    further comprising assaying the sample for the expression of tyrosine kinase receptors involved in angiogenesis.  
     
     
         17 . The method of    claim 16    wherein the tyrosine kinase receptors are chosen from the group consisting of the KDR/flk-1 receptor, the flt-1 receptor, and/or the tek/tie-2 receptor.  
     
     
         18 . The method of    claim 11    further comprising assaying the sample for the expression of hypoxia induced factor.  
     
     
         19 . The method of    claim 11    further comprising assaying for the expression of an oncogene.  
     
     
         20 . A method of diagnosing metastasis in a human comprising: 
 a) detectably labeling a ligand which specifically recognizes VEGF;    b) administering the labeled ligand to the human; and    c) detecting the localization of the labeled antibody or fusion protein in the human, wherein the abnormal localization of VEGF is indicative of a metastatic disease.