Frizzled 2 as a target for therapeutic antibodies in the treatment of cancer

Abstract

Disclosed herein are methods of treating cancer in a subject, and methods for inhibiting growth, migration and/or invasion of a cancer cell in the subject, comprising administering to the subject a therapeutically effective amount of an antibody or antigen binding fragment thereof that downmodulales Fzd2. The antibody may specifically bind Fzd2, and may promote internalization of the Fzd2 receptor by the cancer cells and/or prevent ligand binding to Fzd2. Specific antibodies, and also specific portions of the Fzd2 molecule for antibody binding are disclosed. In one embodiment the antibody specifically binds to the epitope HGAEQICVGQNHSEDGAPAL (SEQ ID NO: 1). Specific cancers (e.g. late stage hepatocellular carcinoma), intended for treatment are provided, and include cancers that exhibit overexpression of Fzd2, and/or Wnt5a.

What is claimed: 
     
         1 . A method of inhibiting growth, migration and/or invasion of a cancer cell in a subject comprising administering to the subject a therapeutically effective amount of an agent that downmodulates Fzd2, such that the agent is delivered to the cancer cells, to thereby treat the cancer. 
     
     
         2 . The method of  claim 1 , wherein the agent is nucleic acid sequences or an antibody or antigen binding fragment thereof that specifically binds Fzd2 to downmodulate Fzd2. 
     
     
         3 . The method of  claim 2 , wherein the nucleic acid sequences are RNAi. 
     
     
         4 . The method of  claim 3 , wherein the RNAi is siRNA, shRNA, or microRNA. 
     
     
         5 . The method of  claim 1 , wherein administration is by a route selected from the group consisting of parenteral, infusion, injection, transmucosal, oral, topical, instillation, and inhalation. 
     
     
         6 . The method of  claim 2 , wherein the antibody binds to Fzd2 and promotes internalization of the Fzd2 receptor by the cancer cells and/or prevents ligand binding to Fzd2 and/or specifically binds an extracellular portion of the Fzd-2 protein, and/or the antibody specifically binds to Fzd2 within a region of Fzd2 corresponding to amino acids 24-247 of Fzd2, amino acids 125-163 of Fzd2, amino acids 134-163 of Fzd2, and/or amino acids 144-163 of Fzd2. 
     
     
         7 . The method of  claim 6 , wherein the antibody specifically binds to the epitope HGAEQICVGQNHSEDGAPAL (SEQ ID NO: 1). 
     
     
         8 . The method of  claim 1 , wherein the cancer is selected from the group consisting of gastrointestinal cancer, prostate cancer, ovarian cancer, breast cancer, head and neck cancer, lung cancer, non-small cell lung cancer, cancer of the nervous system, kidney cancer, retina cancer, skin cancer, liver cancer, pancreatic cancer, genital-urinary cancer and bladder cancer. 
     
     
         9 . The method of  claim 1 , wherein the cancer is liver cancer. 
     
     
         10 . The method of  claim 1 , wherein the cancer is late stage hepatocellular carcinoma. 
     
     
         11 . The method of  claim 1 , wherein the cancer displays overexpression of Fzd2 and/or Wnt5a. 
     
     
         12 . A method of monitoring Fzd2 downmodulation therapy for cancer that exhibits overexpression of Fzd2 or overexpression of Wnt5a in a subject, comprising:
 a) administering to the subject of an agent that downmodulates Fzd2, such that the agent is delivered to cancer cells of the subject; and   b) measuring one or more of:
 i) the expression of one or more of MMP2, MMP3, MMP9, and serpin E1 in the cancer cells before administration and after administration; and 
 ii) the phosphorylation level of one or more of STAT3, MEK1/2, ERK1/2 and srk family kinases, in the cancer cells before administration and after administration; 
   wherein reduced expression after administration indicates effective therapy and/or reduced phosphorylation after administration indicates effective therapy   
     
     
         13 . The method of  claim 12 , wherein the agent is nucleic acid sequences or an antibody or antigen binding fragment thereof that specifically binds Fzd2 to downmodulate Fzd2. 
     
     
         14 . The method of  claim 13 , wherein the nucleic acid sequences are RNAi. 
     
     
         15 . The method of  claim 12 , further comprising measuring the phosphorylation level of one or more of STAT3, MEK1/2, ERK1/2 and srk family kinases, in the cancer cells before administration and after administration, wherein reduced phosphorylation after administration indicates effective therapy. 
     
     
         16 . The method of  claim 12 , wherein the reduced expression is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. 
     
     
         17 . The method of  claim 12 , wherein measuring expression is by detection of mRNA and/or immuno-detection of protein. 
     
     
         18 . The method of  claim 12 , wherein detection of mRNA is by qPCR of mRNA of the tumor using gene specific primers. 
     
     
         19 . The method of  claim 15 , wherein measuring the phosphorylation level is by quantitative immunodetection of the phosphorylated proteins. 
     
     
         20 . A method of treating cancer in a subject comprising,
 a) measuring one or more of:
 i) the expression levels of one or more of MMP2, MMP3, MMP9, sICAM1, and SerpinE1 in the cancer, as compared to an appropriate control; and 
 ii) the phosphorylation level of one or more of STAT3, MEK1/2, ERK1/2 and srk family kinases in the cancer, as compared to an appropriate control; 
   b) administering to the subject of an agent that downmodulates Fzd2, such that the agent is delivered to the cancer cells, when the cancer has increased expression levels or phosphorylation levels, to thereby treat the cancer.   
     
     
         21 . A pharmaceutical composition comprising a therapeutic humanized antibody or antigen binding fragment thereof that specifically binds Fzd2 to downmodulate Fzd2. 
     
     
         22 . The pharmaceutical composition of  claim 21 , wherein the humanized antibody or antigen binding fragment thereof binds to Fzd2 and promotes internalization of the Fzd2 receptor by the cancer cells and/or prevents ligand binding to Fzd2 and/or specifically binds an extracellular portion of the Fzd-2 protein, and/or the antibody specifically binds to Fzd2 within a region of Fzd2 corresponding to amino acids 24-247 of Fzd2, amino acids 125-163 of Fzd2, amino acids 134-163 of Fzd2, and/or amino acids 144-163 of Fzd2. 
     
     
         23 . The pharmaceutical composition of  claim 21 , wherein the antibody or antigen binding fragment thereof specifically binds to the epitope HGAEQICVGQNHSEDGAPAL (SEQ ID NO: 1).