Humanized anti-DKK2 antibody and uses thereof

Abstract

The present invention relates to the discovery that inhibition of Dickkopf2 (DKK2) increases CD8+ cytotoxic T lymphocyte (CTL) activity, attenuates tumor, and hence suppresses tumor formation. Thus, in various embodiments described herein, the methods of the invention relate to methods of treating cancer by administering to a patient an effective amount of a humanized anti-DKK2 antibody, methods for providing anti-tumor immunity in a subject, methods of stimulating a T cell mediated immune response to a cell population or a tissue and suppressing tumor in a subject. Additionally, the current invention includes methods of diagnosing a cancer or a predisposition of developing a cancer or a metastasis and methods for determining the use of immunotherapy treatment or cancer vaccine for treating cancer. Furthermore, the invention encompasses a pharmaceutical composition for treating cancer as well as a kit for carrying out the aforementioned methods.

1 . A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of a humanized anti-Dickkopf2 (anti-DKK2) antibody or fragment thereof in a pharmaceutical acceptable carrier. 
     
     
         2 . The method of  claim 1 , wherein the cancer comprises a tumor comprising cells that express an adenomatosis polyposis coli (APC) mutation. 
     
     
         3 . The method of  claim 1 , wherein the humanized anti-DKK2 antibody possesses neutralizing activity. 
     
     
         4 . The method of  claim 1 , wherein the humanized anti-DKK2 antibody targets a DDK2 neutralizing epitope comprising the amino acid sequence SEQ ID NO: 5. 
     
     
         5 . The method of  claim 1 , wherein the humanized anti-DKK2 antibody comprises at least one of the amino acid sequences selected from the group consisting of SEQ ID NOs: 1, 2 and 3. 
     
     
         6 . The method of  claim 1 , wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, intestinal cancer, pancreatic cancer, and esophageal cancer. 
     
     
         7 . The method of  claim 1 , wherein the cancer is metastatic. 
     
     
         8 . The method of  claim 1 , further comprising administering to the subject an additional agent selected from the group consisting of a chemotherapeutic agent, an anti-cell proliferation agent, an immunotherapeutic agent and any combination thereof. 
     
     
         9 . The method of  claim 8 , wherein the additional agent is a programmed cell death 1 (PD-1) antibody. 
     
     
         10 . The method of  claim 8 , wherein the humanized anti-DKK2 antibody and the additional agent are co-administered to the subject. 
     
     
         11 . The method of  claim 8 , wherein the humanized anti-DKK2 antibody and the additional agent are co-formulated and are co-administered to the subject. 
     
     
         12 . The method of  claim 1 , wherein the route of administration is selected from the group consisting of inhalation, oral, rectal, vaginal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic, intrathecal, and any combination thereof. 
     
     
         13 . A pharmaceutical composition for treating a cancer in a subject, the pharmaceutical composition comprising a humanized anti-Dickkopf2 (anti-DKK2) antibody or fragment thereof and a pharmaceutical acceptable carrier. 
     
     
         14 . The pharmaceutical composition of  claim 13 , wherein the cancer comprises a tumor comprising cells that express an adenomatosis polyposis  coli  (APC) mutation. 
     
     
         15 . The pharmaceutical composition of  claim 13 , wherein the humanized anti-DKK2 antibody possesses neutralizing activity. 
     
     
         16 . The pharmaceutical composition of  claim 13 , wherein the humanized anti-DKK2 antibody targets a DKK2 neutralizing epitope that comprises the amino acid sequence SEQ ID NO: 5. 
     
     
         17 . The pharmaceutical composition of  claim 13 , wherein the humanized anti-DKK2 antibody comprises at least one of the amino acid sequences selected from the group consisting of SEQ ID NOs: 1, 2 and 3. 
     
     
         18 . The pharmaceutical composition of  claim 13 , comprising an additional agent selected from the group consisting of a chemotherapeutic agent, an anti-cell proliferation agent, an immunotherapeutic agent and any combination thereof. 
     
     
         19 . The pharmaceutical composition of  claim 13 , wherein the additional agent is a programmed cell death 1 (PD-1) antibody. 
     
     
         20 . The pharmaceutical composition of  claim 13 , wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, intestinal cancer, pancreatic cancer, and esophageal cancer. 
     
     
         21 . The pharmaceutical composition of  claim 13 , wherein the cancer is metastatic. 
     
     
         22 . A method for providing anti-tumor immunity in a subject, the method comprising administering to the subject an effective amount of a humanized anti-Dickkopf2 (anti-DKK2) antibody or fragment thereof with a pharmaceutical acceptable carrier. 
     
     
         23 . The method of  claim 22 , further comprising further administering to the subject an additional agent selected from the group consisting of a chemotherapeutic agent, an anti-cell proliferation agent, an immunotherapeutic agent and any combination thereof. 
     
     
         24 . The method of  claim 23 , wherein the additional agent is a programmed cell death 1 (PD-1) antibody. 
     
     
         25 . The method of  claim 23 , wherein the humanized anti-DKK2 antibody and the additional agent are co-administered to the subject. 
     
     
         26 . A method for stimulating a T cell-mediated immune response to a cell population or tissue in a subject, the method comprising administering to the subject an effective amount of a humanized anti-Dickkopf2 (anti-DKK2) antibody or fragment thereof with a pharmaceutical acceptable carrier. 
     
     
         27 . The method of  claim 26 , wherein the humanized anti-DKK2 antibody targets a DDK2 neutralizing epitope comprising the amino acid sequence SEQ ID NO: 5. 
     
     
         28 . The method of  claim 26 , wherein the humanized anti-DKK2 antibody comprises at least one of the amino acid sequences of SEQ ID NOs: 1, 2 and 3. 
     
     
         29 . The method of  claim 26 , wherein the T cell-mediated immune response is a CD8 +  cytotoxic T lymphocyte (CTL) response. 
     
     
         30 . A method of diagnosing a cancer or a predisposition for developing a cancer in a subject, the method comprising determining the expression level of a DKK2 gene in a biological sample from the subject, wherein an increase in the expression level of DKK2 in the biological sample from the subject as compared with the level of DKK2 expression in a control biological sample from a subject not having a cancer is an indication that the subject has a cancer or a predisposition for developing a cancer, and wherein when a cancer or a predisposition for developing a cancer is detected in a subject, a humanized anti-DKK2 antibody treatment is recommended for the subject. 
     
     
         31 . The method of  claim 30 , wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, intestinal cancer, pancreatic cancer, and esophageal cancer. 
     
     
         32 . The method of  claim 30 , wherein the expression level of DKK2 in the biological sample from the subject is at least 10% greater than the normal control level. 
     
     
         33 . The method of  claim 30 , wherein the expression level of DKK2 in the biological sample from the subject or normal control is determined using a method selected from the group consisting of detecting mRNA of the gene, detecting a protein encoded by the gene, and detecting a biological activity of the protein encoded by the gene. 
     
     
         34 . A method for determining the efficacy of a humanized anti-DKK2 antibody treatment for cancer in a subject in need thereof, the method comprising determining the expression level of Dickkopf2 (DKK2) gene in a biological sample from the subject, wherein an increase in the expression level of DKK2 in the biological sample from the subject as compared with the level of DKK2 expression in a control biological sample from a subject not having a cancer is an indication that the humanized anti-DKK2 antibody treatment is effective, and wherein when the humanized anti-DKK2 antibody treatment is determined to be effective, an additional treatment is recommended for the subject. 
     
     
         35 . The method of  claim 34 , wherein the additional treatment comprises at least one selected from the group consisting of chemotherapy, radiation therapy, immunotherapy and cancer vaccine therapy. 
     
     
         36 . The method of  claim 34 , wherein the expression level of DKK2 in the biological sample from the subject is at least 10% greater than the normal control level. 
     
     
         37 . The method of  claim 34 , wherein the expression level is determined by a method selected from the group consisting of detecting mRNA of the gene, detecting a protein encoded by the gene, and detecting a biological activity of the protein encoded by the gene. 
     
     
         38 . The method of  claim 34 , wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, intestinal cancer, pancreatic cancer, and esophageal cancer. 
     
     
         39 . The method of  claim 1 , wherein the subject is a human. 
     
     
         40 . (canceled) 
     
     
         41 . A composition comprising a humanized anti-Dickkopf2 (anti-DKK2) antibody targeting a DKK2 epitope comprising the amino acid sequence SEQ ID NO: 5. 
     
     
         42 . A kit for diagnosing a cancer or a predisposition for developing a cancer or a metastasis in a subject, the kit comprising a humanized anti-DKK2 antibody targeting a DKK2 epitope comprising the amino acid sequence SEQ ID NO: 5. 
     
     
         43 . The kit of  claim 42 , wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, gastric cancer, intestinal cancer, pancreatic cancer, and esophageal cancer.