Methods for detecting for the presence of tumor cells and for screening for anti-tumor agents

Abstract

A method and kit for detecting the presence of tumor cells in an animal by detecting expression of VEGF-B 186 , determining an increased expression of VEGF-B 186 , or by determining an increased expression of VEGF-B 186 relative to expression of VEGF-B 167 . The kit comprises a receptacle adapted to receive a sample and a means for detecting expression of VEGF-B 186 from the sample. Also, a method for screening for anti-tumor agents.

1 - 16 . (canceled)  
     
     
         17 . A kit for detecting the presence of tumor cells in a mammal, comprising: 
 a receptacle adapted to receive a sample; and    a means for detecting expression of VEGF-B 186  in said sample,    whereby detection of expression of VEGF-B 186  is indicative of the presence of tumor cells.    
     
     
         18 . A kit for detecting the presence of tumor cells in a mammal, comprising: 
 a receptacle adapted to receive a sample;    a means for determining an expression level of VEGF-B 186  in said sample; and    a means for comparing said expression level of VEGF-B 186  with a control level of VEGF-B 186  in an animal absent tumors,    wherein determination of an increased expression level of VEGF-B 186  over the control expression level is indicative of the presence of tumor cells.    
     
     
         19 . A method for screening for anti-tumor agents, comprising: 
 applying a candidate agent to a tumor cell; and    detecting the level of expression of VEGF-B 186  in the tumor cell,    wherein a decrease in the expression level indicates that the candidate agent is an anti-tumor agent.    
     
     
         20 . A kit according to  claim 17 , further comprising a means for detecting expression level of VEGF-B 167 .  
     
     
         21 . A kit according to  claim 17 , wherein the means for detecting VEGF-B 186  comprises suitable agents for detecting VEGF-B 186  at nucleic acid level.  
     
     
         22 . A kit according to  claim 21 , wherein the means for detecting VEGF-B 186  comprises suitable agents for detecting VEGF-B 186  at RNA level.  
     
     
         23 . A kit according to  claim 17 , wherein the means for detecting VEGF-B 186  comprises suitable agents for detecting VEGF-B 186  at protein level.  
     
     
         24 . A kit according to  claim 23 , wherein the means for detecting VEGF-B 186  at protein level comprises a VEGF-B 186 -specific antibody.  
     
     
         25 . A kit according to  claim 24 , wherein said VEGF-B 186 -specific antibody is a monoclonal antibody.  
     
     
         26 . A kit according to  claim 24 , wherein the antibody is labeled covalently or noncovalently with a detectable label.  
     
     
         27 . A kit according to  claim 26 , wherein the detectable label is selected from the group consisting of supermagnetic agent, a paramagnetic agent, an electron dense agent, an ecogenic agent, and a radioactive agent.  
     
     
         28 . A kit according to  claim 26 , wherein the detectable label is selected from the group consisting of  125 I,  32 P, an enzymatic label, and a fluorimetric labels.  
     
     
         29 . A kit according to  claim 28 , wherein the enzymetic label is horseradish peroxidase.  
     
     
         30 . A kit according to  claim 28 , wherein the fluorimetric label is fluorescein-5-isothiocyanate (FITC).  
     
     
         31 . A kit according to  claim 20 , wherein the means for detecting VEGF-B 167  comprises suitable agents for detecting VEGF-B 167  at nucleic acid level.  
     
     
         32 . A kit according to  claim 31 , wherein the means for detecting VEGF-B 167  comprises suitable agents for detecting VEGF-B 167  at RNA level.  
     
     
         33 . A kit according to  claim 20 , wherein the means for detecting VEGF-B 167  comprises suitable agents for detecting VEGF-B 167  at protein level.  
     
     
         34 . A kit according to  claim 33 , wherein the means for detecting VEGF-B 167  at protein level comprises a VEGF-B 167 -specific antibody.  
     
     
         35 . A kit according to  claim 34 , wherein said VEGF-B 167 -specific antibody is a monoclonal antibody.  
     
     
         36 . A kit according to  claim 34 , wherein the antibody is labeled covalently or noncovalently with a detectable label.  
     
     
         37 . A kit according to  claim 36 , wherein the detectable label is selected from the group consisting of supermagnetic agent, a paramagnetic agent, an electron dense agent, an ecogenic agent, and a radioactive agent.  
     
     
         38 . A kit according to  claim 36 , wherein the detectable label is selected from the group consisting of  125 I,  32 P, an enzymatic label, and a fluorimetric labels.  
     
     
         39 . A kit according to  claim 38 , wherein the enzymetic label is horseradish peroxidase.  
     
     
         40 . A kit according to  claim 38 , wherein the fluorimetric label is fluorescein-5-isothiocyanate (FITC).  
     
     
         41 . An isoform-specific antibody that reacts with VEGF-B 167  but not VEGF-B 186 .  
     
     
         42 . The isoform-specific antibody of  claim 41 , which is a monoclonal antibody.  
     
     
         43 . The isoform-specific antibody of  claim 41 , which is a humanized antibody.  
     
     
         44 . The isoform-specific antibody of  claim 41 , which is a chimeric antibody.  
     
     
         45 . The isoform-specific antibody of  claim 41 , which is labeled covalently or noncovalently with a detectable label.  
     
     
         46 . The isoform-specific antibody of  claim 45 , wherein the detectable label is selected from the group consisting of supermagnetic agent, a paramagnetic agent, an electron dense agent, an ecogenic agent, and a radioactive agent.  
     
     
         47 . The isoform-specific antibody of  claim 45 , wherein the detectable label is selected from the group consisting of  125 I,  32 P, an enzymatic label, and a fluorimetric labels.  
     
     
         48 . The isoform-specific antibody of  claim 47 , wherein the enzymetic label is horseradish peroxidase.  
     
     
         49 . The isoform-specific antibody of  claim 47 , wherein the fluorimetric label is fluorescein-5-isothiocyanate (FITC).  
     
     
         50 . A pharmaceutical composition comprising an antibody of  claim 41 , and a pharmaceutically acceptable excipient.  
     
     
         51 . An isoform-specific antibody that reacts with VEGF-B 186  but not VEGF-B 167 .  
     
     
         52 . The isoform-specific antibody of  claim 51 , which is a monoclonal antibody.  
     
     
         53 . The isoform-specific antibody of  claim 41 , which is a humanized antibody.  
     
     
         54 . The isoform-specific antibody of  claim 41 , which is a chimeric antibody.  
     
     
         55 . The isoform-specific antibody of  claim 52 , which is labeled covalently or noncovalently with a detectable label.  
     
     
         56 . The isoform-specific antibody of  claim 55 , wherein the detectable label is selected from the group consisting of supermagnetic agent, a paramagnetic agent, an electron dense agent, an ecogenic agent, and a radioactive agent.  
     
     
         57 . The isoform-specific antibody of  claim 55 , wherein the detectable label is selected from the group consisting of  125 I,  32 P, an enzymatic label, and a fluorimetric labels.  
     
     
         58 . The isoform-specific antibody of  claim 57 , wherein the enzymetic label is horseradish peroxidase.  
     
     
         59 . The isoform-specific antibody of  claim 57 , wherein the fluorimetric label is fluorescein-5-isothiocyanate (FITC).  
     
     
         60 . A pharmaceutical composition comprising an antibody of  claim 51 , and a pharmaceutically acceptable excipient.  
     
     
         61 . A method for inhibiting VEGF-B 167  mediated angiogenesis in a mammal in need thereof, comprising administering to said mammal an effective amount of a VEGF-B 167  antagonist.  
     
     
         62 . A method according to  claim 61 , wherein the VEGF-B 167  antagonist is a VEGF-B 167 -specific antibody.  
     
     
         63 . A method according to  claim 61 , wherein the VEGF-B 167  antagonist comprises a small molecule antagonist.  
     
     
         64 . A method according to  claim 61 , wherein the VEGF-B 167  antagonist comprises a nucleic acid molecule that is anti-sense to a polynucleotide sequence encoding VEGF-B 167 .  
     
     
         65 . A method according to  claim 61 , wherein the method is for inhibiting the proliferation of angiogenesis-dependent tumor, or for inhibiting new blood vessel formation in diabetic retinopathy, psoriasis, arthopathies or a vascular tumor.  
     
     
         66 . A method according to  claim 61 , wherein the vascular tumor is haemangioma.  
     
     
         67 . A method for inhibiting VEGF-B 186  mediated angiogenesis in a mammal in need thereof, comprising administering to said mammal an effective amount of a VEGF-B 186  antagonist.  
     
     
         68 . A method according to  claim 67 , wherein the VEGF-B 167  antagonist comprises a VEGF-B 186 -specific antibody.  
     
     
         69 . A method according to  claim 67 , wherein the VEGF-B 167  antagonist comprises a small molecule antagonist.  
     
     
         70 . A method according to  claim 67 , wherein the VEGF-B 167  antagonist comprises a nucleic acid that is anti-sense molecule to a polynucleotide sequence encoding VEGF-B 186 .  
     
     
         71 . A method according to  claim 67 , wherein the method is for inhibiting the proliferation of angiogenesis-dependent tumor, or for inhibiting new blood vessel formation in diabetic retinopathy, psoriasis, arthopathies or a vascular tumor.  
     
     
         72 . A method according to  claim 67 , wherein the vascular tumor is haemangioma.  
     
     
         73 . A method for inhibiting tumor cell formation or tumor cell growth, the method comprising administering to a patient in need thereof an antagonist to VEGF-B 186 .  
     
     
         74 . A method according to  claim 73 , wherein the antagonist comprises an isoform-specific antibody to VEGF-B 186 .  
     
     
         75 . A method according to  claim 73 , wherein the VEGF-B 186  antagonist comprises a small molecule antagonist.  
     
     
         76 . A method according to  claim 73 , wherein the VEGF-B 186  antagonist comprises a nucleic acid molecule that is anti-sense to a polynucleotide sequence encoding VEGF-B 186 .  
     
     
         77 . A method according to  claim 73 , wherein the tumor cell is selected from the group consisting of fibrosarcoma, melanoma, Lewis lung carcinoma, glioma, and pheochromocytoma.  
     
     
         78 . A method according to  claim 73 , wherein the tumor cell is benign or malignant.  
     
     
         79 . A method according to  claim 73 , wherein the patient is human.