Biomarkers for disease progression in melanoma

Abstract

The present invention relates inter alia to therapeutic agents for use in the treatment of melanoma, methods of diagnosing an increased risk of metastasis in a subject suffering from melanoma, methods of treating such subjects, diagnostic assays and kits. More particularly, in certain embodiments the invention relates to identifying whether a subject suffering from melanoma has an increased risk of metastasis by determining the expression of Ambra-1 and Loricrin in a tissue sample obtained from the subject.

What is claimed is: 
     
         1 . A method comprising:
 a) obtaining tissue overlying a primary melanoma from a subject suffering from melanoma;   b) determining the expression of Ambra-1 and Loricrin in the tissue overlying a primary melanoma;   c) comparing the expression obtained in (b) with a reference tissue or levels obtained therefrom, and   d) (i) following a normal recognized care pathway if expression of Ambra-1 and Loricrin is normal or increased, or
 (ii) treating the subject with a systemic anti-cancer treatment regime if expression of Ambra-1 and Loricrin is decreased or lost. 
   
     
     
         2 . The method of  claim 1 , wherein the method comprises detecting a co-occurrence of a decrease in the expression of Ambra-1 and a decrease in Loricrin in the tissue sample compared to the reference levels, or a loss of expression of Ambra-1 and a loss of expression of Loricrin in the tissue sample. 
     
     
         3 . The method according to  claim 1 , wherein the systemic anti-cancer treatment regime is for preventing, inhibiting or delaying metastasis or decreasing the risk of metastasis in the subject. 
     
     
         4 . The method according to  claim 1 , wherein the systemic anti-cancer treatment regime comprises administering a therapeutic agent to the subject. 
     
     
         5 . A method of treating a subject suffering from melanoma, the method comprising administering a therapeutic agent to the subject, wherein the subject has been identified as having decreased or loss of expression of Ambra-1 and Loricrin in a tissue sample obtained from the subject. 
     
     
         6 . The method according to  claim 5 , wherein the method is for preventing, inhibiting or delaying metastasis or decreasing the risk of metastasis in the subject. 
     
     
         7 . The method according to  claim 5 , wherein the subject, prior to identification, was ineligible for therapeutic agent treatment. 
     
     
         8 . The method according to  claim 4 , wherein:
 (i) the therapeutic agent is administered to the subject no more than 12 weeks after the subject has been identified as having decreased or loss of expression of Ambra-1 and Loricrin in the tissue sample; or   (ii) the therapeutic agent is a chemotherapeutic agent such as Dacarbazine (DTIC), Temozolomide, Nab-paclitaxel, Paclitaxel, Carmustine (BCNU), Cisplatin, Carboplatin, Vinblastine, interleukin 2, interferon alpha, antibodies or B-Raf inhibitors.   
     
     
         9 . A method of treating melanoma in a subject, the method comprising:
 (a) determining the expression of Ambra-1 and Loricrin in tissue overlying a primary melanoma from the subject;   (b) comparing the expression obtained in (a) with a reference tissue or levels obtained therefrom, and   if there is a decrease in the expression of Ambra-1 and Loricrin in the tissue sample compared to the reference tissue or levels, or a loss of expression of Ambra-1 and Loricrin, administering a therapeutic agent to the subject.   
     
     
         10 . The method of  claim 9 , wherein the method comprises detecting a co-occurrence of a decrease in the expression of Ambra-1 and a decrease in Loricrin in the tissue sample compared to the reference levels, or a loss of expression of Ambra-1 and a loss of expression of Loricrin in the tissue sample. 
     
     
         11 . The method according to  claim 9 , wherein:
 (i) the therapeutic agent is administered to the subject no more than 12 weeks after the step of determining a decrease or loss of expression of Ambra-1 and Loricrin in the tissue sample; or   (ii) the therapeutic agent is a chemotherapeutic agent such as Dacarbazine (DTIC), Temozolomide, Nab-paclitaxel, Paclitaxel, Carmustine (BCNU), Cisplatin, Carboplatin, Vinblastine, interleukin 2, interferon alpha, antibodies and B-Raf inhibitors.   
     
     
         12 . The method according to  claim 1 , wherein the reference levels are levels of Ambra-1 and Loricrin expression that are characteristic of normal tissue. 
     
     
         13 . The method according to  claim 1 , wherein the reference tissue comprises normal tissue. 
     
     
         14 . The method according to  claim 13 , wherein the normal tissue is epidermis from a site which does not include a primary melanoma. 
     
     
         15 . The method according to  claim 13 , wherein the reference tissue is an internal reference. 
     
     
         16 . The method according to  claim 13 , wherein the normal tissue is from a site adjacent to the primary melanoma. 
     
     
         17 . The method according to  claim 1 , wherein the tissue sample comprises tissue overlying a primary melanoma and a portion of normal epidermis adjacent to the primary melanoma. 
     
     
         18 . The method according to  claim 1 , wherein the expression of Ambra-1 and Loricrin in the tissue sample is from about 25% to about 75% of the respective reference level or less than about 25% of the respective reference level. 
     
     
         19 . The method according to  claim 1 , wherein the expression of Ambra-1 and Loricrin in the tissue sample is determined by visual assessment or by an automatic slide scanner. 
     
     
         20 . The method according to  claim 1 , wherein determining the expression of Ambra-1 and Loricrin in tissue overlying a primary melanoma comprises:
 contacting tissue overlying a primary melanoma with a first ligand specific for Ambra-1, wherein the presence of Ambra-1 creates an Ambra-1-ligand complex;   contacting the tissue overlying a primary melanoma with a second ligand specific for Loricrin, wherein the presence of Loricrin creates a Loricrin-ligand complex; and   detecting and/or quantifying the Ambra-1-ligand complex and the Loricrin-ligand complex.   
     
     
         21 . The method according to  claim 20 , wherein the method comprises contacting a first section of the tissue sample with the first ligand and contacting a second section of the tissue sample with the second ligand. 
     
     
         22 . The method of  claim 20 , wherein the first ligand comprises an anti-Ambra-1 antibody or aptamer and the second ligand comprises an anti-Loricrin antibody or aptamer. 
     
     
         23 . The method according to  claim 1 , wherein the tissue sample comprises keratinocytes overlying the primary melanoma and the method comprises determining the expression of Ambra-1 and Loricrin in the keratinocytes. 
     
     
         24 . The method according to  claim 1 , wherein the subject is suffering from American Joint Commission on Cancer (AJCC) stage 1a, stage 1b, stage 2a, stage 2b or stage 2c melanoma. 
     
     
         25 . The method according to  claim 1 , wherein the subject is a human or animal.