Methods for treating cancer using anti-Wnt2 monoclonal antibodies and siRNA

Abstract

This invention relates to methods of inhibiting the growth of cells, in particular cancer cells, that overexpress Wnt2. The methods comprise contacting the cell with an agent that binds to Wnt2 mRNA or Wnt2 protein, interferes with Wnt2 signaling or inhibits binding of the Wnt2 protein to another protein, such as a Frizzled receptor.

1 . A method of inhibiting proliferation of a cell that overexpresses a Wnt2, comprising contacting the cell with an amount of agent that inhibits Wnt2 signaling effective to inhibit proliferation of the cell.  
     
     
         2 . The method of  claim 1  in which the agent is a siRNA.  
     
     
         3 . The method of  claim 1  in which the agent is an anti-Wnt2 antibody.  
     
     
         4 . The method of  claim 3  in which the anti-Wnt2 antibody is a monoclonal antibody.  
     
     
         5 . The method of  claim 3  in which the anti-Wnt2 antibody comprises: 
 (i) aV L CDR1 amino acid sequence as shown in SEQ ID NO:56, SEQ ID NO:104 or SEQ ID NO:125;    (ii) a V L CDR2 amino acid sequence as shown in SEQ ID NO:58;    (iii) a V L CDR3 amino acid sequence as shown in SEQ ID NO:60; SEQ ID NO:84, SEQ ID NO:107, SEQ ID NO:126 or SEQ ID NO:138;    (iv) a V H CDR1 amino acid sequence as shown in SEQ ID NO:63 or SEQ ID NO:87;    (v) a V H CDR2 amino acid sequence as shown in SEQ ID NO:65 or SEQ ID NO:89; and    (vi) a V H CDR3 amino acid sequence as shown in SEQ ID NO:67, SEQ ID NO:91 or SEQ ID NO:110.    
     
     
         6 . The method of  claim 3  in which the anti-Wnt2 antibody specifically binds a polypeptide comprising an amino acid sequence corresponding to amino acid residues 49-63 of human Wnt2 (SEQ ID NO:2).  
     
     
         7 . The method of  claim 6  in which the anti-Wnt2 antibody specifically binds a polypeptide comprising SSQRQLCHRPDVMR (SEQ ID NO:2).  
     
     
         8 . The method of  claim 3  in which the anti-Wnt2 antibody competes for binding a Wnt2 with a second anti-Wnt2 antibody that specifically binds a polypeptide comprising an amino acid sequence corresponding to amino acid residues 49-63 of human Wnt2 (SEQ ID NO:2).  
     
     
         9 . The method of  claim 8  in which the anti-Wnt2 antibody competes for binding a Wnt2 with a second anti-Wnt2 antibody that specifically binds a polypeptide comprising SSQRQLCHRHPDVMR (SEQ ID NO:2).  
     
     
         10 . The method of  claim 1  which is practiced in vitro.  
     
     
         11 . The method of  claim 1  which is practiced in vivo.  
     
     
         12 . The method of  claim 1  in which the cell is a cancer cell.  
     
     
         13 . The method of  claim 12  in which the cancer cell is selected from the group consisting of breast, ovarian, colorectal, gastric, lung, kidney, bladder, prostate, uterine, thyroid, pancreatic, cervical, esophageal, mesothelioma, head and neck, hepatocellular, melanoma, brain, vulval, testicular, sarcoma, intestine, skin, leukemia, and lymphoma cancer cells.  
     
     
         14 . A method of inducing apoptosis of a cell that overexpresses a Wnt2, comprising contacting the cell with an amount of an agent that inhibits Wnt2 signaling effective to induce apoptosis of the cell.  
     
     
         15 . A method of inhibiting Wnt2 signaling in a cell, comprising contacting a cell that overexpresses a Wnt2 with an amount of an anti-Wnt2 antibody effective to inhibit Wnt2 signaling.  
     
     
         16 . A method of treating a disease associated with Wnt2 signaling, comprising administering to a subject in need of such treatment an amount of an agent that inhibits Wnt2 signaling effective to treat the disease.  
     
     
         17 . The method of  claim 16  in which the agent is an anti-Wnt2 antibody.  
     
     
         18 . The method of  claim 16  in which the disease is cancer.  
     
     
         19 . The method of  claim 18  in which the cancer is selected from the group consisting of breast, ovarian, colorectal, gastric, lung, kidney, bladder, prostate, uterine, thyroid, pancreatic, cervical, esophageal, mesothelioma, head and neck, hepatocellular, melanoma, brain vulval, testicular, sarcoma, intestine, skin, leukemia, and lymphoma cancer cells.  
     
     
         20 . A method of treating cancer that overexpresses Wnt2, comprising administering to a subject an amount of an anti-Wnt2 antibody effective to treat the cancer.  
     
     
         21 . The method of  claim 20  in which the anti-Wnt2 antibody is a monoclonal antibody.  
     
     
         22 . The method of  claim 20  in which the anti-Wnt2 antibody comprises: 
 (i) aV L CDR1 amino acid sequence as shown in SEQ ID NO:56, SEQ ID NO:104 or SEQ ID NO:125;    (ii) a V L CDR2 amino acid sequence as shown in SEQ ID NO:58;    (iii) a V L CDR3 amino acid sequence as shown in SEQ ID NO:60; SEQ ID NO:84, SEQ ID NO:107, SEQ ID NO:126 or SEQ ID NO:138;    (iv) a V H CDR1 amino acid sequence as shown in SEQ ID NO:63 or SEQ ID NO:87;    (v) a V H CDR2 amino acid sequence as shown in SEQ ID NO:65 or SEQ ID NO:89; and    (vi) a V H CDR3 amino acid sequence as shown in SEQ ID NO:67, SEQ ID NO:91 or SEQ ID NO:110.    
     
     
         23 . The method of  claim 20  in which the anti-Wnt2 antibody specifically binds a polypeptide comprising an amino acid sequence corresponding to amino acid residues 49-63 of human Wnt2 (SEQ ID NO:2).  
     
     
         24 . The method of  claim 23  in which the anti-Wnt2 antibody specifically binds a polypeptide comprising SSQRQLCHRPDVMR (SEQ ID NO:2).  
     
     
         25 . The method of  claim 20  in which the anti-Wnt2 antibody competes for binding a Wnt2 with a second anti-Wnt2 antibody that specifically binds a polypeptide comprising an amino acid sequence corresponding to amino acid residues 49-63 of human Wnt2 (SEQ ID NO:2).  
     
     
         26 . The method of  claim 25  in which the anti-Wnt2 antibody competes for binding a Wnt2 with a second anti-Wnt2 antibody that specifically binds a polypeptide comprising SSQRQLCHRHPDVMR (SEQ ID NO:2).  
     
     
         27 . A pharmaceutical composition comprising an anti-Wnt2 antibody and a pharmaceutically acceptable excipient, carrier, and/or diluent.  
     
     
         28 . The pharmaceutical composition of  claim 27  in which the anti-Wnt2 antibody is a polyclonal antibody.  
     
     
         29 . The pharmaceutical composition of  claim 27  in which the anti-Wnt2 antibody is a monoclonal antibody.  
     
     
         30 . A monoclonal anti-Wnt2 antibody that specifically binds a polypeptide comprising an amino acid sequence that corresponds to amino acid residues 49-63 of human Wnt2.  
     
     
         31 . The monoclonal anti-Wnt2 antibody of  claim 30  that competes for binding to a Wnt2 with a second anti-Wnt2 antibody that specifically binds a polypeptide comprising an amino acid sequence that corresponds to amino acid residues 49-63 of human Wnt2.  
     
     
         32 . The monoclonal antibody of  claim 30  comprising 
 (i) aV L CDR1 amino acid sequence as shown in SEQ ID NO:56, SEQ ID NO:104 or SEQ ID NO:125;    (ii) a V L CDR2 amino acid sequence as shown in SEQ ID NO:58;    (iii) a V L CDR3 amino acid sequence as shown in SEQ ID NO:60; SEQ ID NO:84, SEQ ID NO:107, SEQ ID NO:126 or SEQ ID NO:138;    (iv) a V H CDR1 amino acid sequence as shown in SEQ ID NO:63 or SEQ ID NO:87;    (v) a V H CDR2 amino acid sequence as shown in SEQ ID NO:65 or SEQ ID NO:89; and    (vi) a V H CDR3 amino acid sequence as shown in SEQ ID NO:67, SEQ ID NO:91 or SEQ ID NO:110.    
     
     
         33 . A nucleic acid encoding the monoclonal antibody of  claim 32 .  
     
     
         34 . The nucleic acid of  claim 33  comprising: 
 (i) a nucleic acid encoding aV L CDR1 selected from the group consisting of nucleic acid sequences shown in SEQ ID NO:70, SEQ ID NO:1 13 and SEQ ID NO:131;    (ii) a nucleic acid encoding aV L CDR2 selected from the group consisting of nucleic acid sequences shown in SEQ ID NO:72 and SEQ ID NO:115;    (iii) a nucleic acid encoding aV L CDR3 selected from the group of nucleic acid sequences shown in SEQ ID NO:94, SEQ ID NO:117, SEQ ID NO:134 and SEQ ID NO:143;    (iv) a nucleic acid encoding aV H CDR1 selected from the group of nucleic acid sequences shown in SEQ ID NO:77 and SEQ ID NO:97;    (v) a nucleic acid encoding aV H CDR2 selected from the group of nucleic acid sequences shown in SEQ ID NO:79 and SEQ ID NO:99; and    (vi) a nucleic acid encoding aV H CDR3 selected from the group of nucleic acid sequences shown in SEQ ID NO:101 and SEQ ID NO:120.