Method for the diagnosis, prognosis and treatment of cancer metastasis

Abstract

This study describes a method to determine the likelihood of the development of metastasis in a subject suffering from cancer, in addition to a method to design a customized therapy in a subject suffering from cancer, in particular breast, colon, lung, kidney and thyroid cancer, based on the determination of the expression level of one or more genes whose expression is modulated by an increase in c-MAF expression. It also describes a method for the identification of marker genes with a propensity for metastatic cancer based on inducing the modulation of the c-MAF expression Finally, the use of PTHLH and PODXL inhibitors and RERG activators in the treatment and/or prevention of the cancer, in particular breast, colon, lung, kidney and thyroid cancer.

1 . In vitro method to predict the metastasis of breast cancer in a subject which involves determining the level of expression, in a sample of tumoral tissue taken from said subject, of one or more genes whose expression is modulated in response to an increase in the levels of the expression of c-MAF in said tumor in which the altered levels of expression of said one or more genes with respect to a reference value are indicative of a high risk of the development of metastasis. 
     
     
         2 . Method according to  claim 1  in which said one or more genes whose expression is modulated in response to an increase in the levels of expression of c-MAF is selected from the group formed of the genes taken up in Table 1 and the genes indicated in Table 2 and in which the modulation of the levels of expression of said one or more genes is an increase to the expression of one or more genes taken up in Table 1 and/or a decrease in the expression of one or more genes indicated in Table 2. 
     
     
         3 . Method according to  claim 2  in which said gene whose expression is modulated in response to an increase in the levels of expression of c-MAF is selected from the group of PTHLH, PODXL and RERG. 
     
     
         4 . In vitro method for the design of a personalized therapy for a subject affected with breast cancer, involving the determination of the level of expression of a tumoral tissue sample taken from said subject, of one or more genes whose expression is modulated in response to an increase in the levels of expression of c-MAF in which altered levels of expression of said one or more genes with respect to a reference value are indicative that said subject is appropriate to receive a treatment for the prevention of metastasis. 
     
     
         5 . Method according to  claim 4  in which said one or more genes whose expression is modulated in response to an increase in the levels of expression of c-MAF are selected from the group formed by one or more of the genes included in Table 1 and/or one or more of the genes indicated in Table 2 and in which the modulation of the levels of expression of one or more of the genes from Table 1 represents an increase in the expression and/or where the modulation of the levels of expression of one or more of the genes from Table 2 represents a decrease in expression. 
     
     
         6 . Method according to  claim 5  in which said gene whose expression is modulated in response to an increase in the levels of expression of c-MAF is selected from the group of PTHLH, PODXL and RERG. 
     
     
         7 . Method according to  claims 1  through  6 , in which the breast cancer is selected from the group consisting of an ER+ cancer and a triple-negative cancer. 
     
     
         8 . Method according to any of the  claims 1  to  7  in which the metastasis is a bone metastasis. 
     
     
         9 . Method according to  claim 9 , in which the bone metastasis is an osteolytic metastasis. 
     
     
         10 . Method according to any of the  claims 1  through  9  in which the determination of the levels of expression of said one or more genes whose expression is modulated in response to an increase in the levels of expression of c-MAF is performed/occurs during the determination of the levels of expression of the RNAm of said gene. 
     
     
         11 . Method according to any of the  claims 1  through  9  in which the determination of the levels of expression of said one or more genes is modulated in response to an increase in the levels of expression of c-MAF is performed during the determination of the levels of expression of the polypeptide encoded by said gene. 
     
     
         12 . The use of an agent to inhibit the expression of a gene or the activity of the product of the expression of said gene for the preparation of a medication for the treatment and/or prevention of the metastasis of breast cancer in which said gene is characterized because its expression in breast cancer tumor cells increases in response to an increase in the levels of expression of c-MAF in said cells or decreases in response to a decrease in the levels of expression of c-MAF in said cells. 
     
     
         13 . Use according to  claim 12 , in which the agent that inhibits the expression of a gene is selected from an RNAip specific to said gene, an antisense oligonucleotide, specific to said gene, a ribozyme specific for said gene or on which the agent that inhibits the activity of the product of the expression of said gene is selected from the group formed by an inhibitor antibody specific for said product of the expression, a dominant-negative variant of said product of the expression and an inhibitory peptide from said product of the expression. 
     
     
         14 . Use according to  claim 12  or  13  in which the gene whose expression increases in response to an increase in the levels of expression of c-MAF in a breast tumor or whose expression decreases in response to a decrease in the levels of expression of c-MAF in a breast tumor is selected from the genes described in Table 1. 
     
     
         15 . Use according to  claim 14  in which the gene whose expression increases in response to an increase in the levels of expression of c-MAF in a breast tumor is the PHTLH gene or the PODXL gene. 
     
     
         16 . Use of an agent to stimulate the expression of a gene or the activity of the product of the expression of said gene for the preparation of a medication for the treatment and/or prevention of the metastasis of breast cancer, in which said gene is characteristic because its expression in breast cancer tumor cells decreases in response to an increase in the levels of expression of c-MAF in said cells or because its expression increases in response to a decrease in the levels of expression of c-MAF in said cells. 
     
     
         17 . Use according to  claim 16 , in which the agent that stimulates the expression of said gene is a polynucleotide that contains the encoded sequence of said gene or in which the agent that stimulates the activity of the product of said gene is a polypeptide encoded by said gene. 
     
     
         18 . Use according to  claim 17  in which the gene whose expression decreases in response to an increase in c-MAF expression levels in a breast tumor or whose expression increases in response to a decrease in c-MAF expression levels in a breast tumor is selected from the genes described in Table 2. 
     
     
         19 . Use according to  claim 18  in which the gene whose expression decreases in response to an increase in c-MAF expression levels in a breast tumor is the RERG gene. 
     
     
         20 . Use according to any of the  claims 12  to  19 , in which the breast cancer is selected from a group formed by ER+ cancer and triple negative cancer. 
     
     
         21 . Use according to  claims 12  to  20 , in which metastasis is bone metastasis. 
     
     
         22 . Use according to  claim 21 , in which the bone metastasis is oseolytic metastasis. 
     
     
         23 . In vitro method for the identification of a gene marker for propensity to metastasis in a subject suffering from breast cancer that involves
 (i) determining the expression levels of a candidate gene and of c-MAF in a breast cancer primary tumor sample and   (ii) determining the change in the expression level of said gene in a population of breast cancer cells in response to a modulation to the expression of the c-MAF gene   
       in which if the expression levels of said gene are statistically significantly correlated to the expression of c-MAF in the breast cancer primary tumor sample and the change in the expression levels in response to the modulation of c-MAF gene expression are statistically significantly correlated to the change in the levels of said gene, this is indicative that said gene is a marker for propensity to metastasis in a subject. 
     
     
         24 . Method according to  claim 23  in which the expression of the given gene determined in step (i) is directly correlated with the levels of c-MAF in the primary tumor sample and if the change in the expression levels in response to modulation of c-MAF gene expression is directly correlated to said modulation this is indicative that the high levels of said gene are indicative of propensity to metastasis. 
     
     
         25 . Method according to  claim 23  in which the expression of the given gene determined in step (i) is inversely correlated with the levels of c-MAF in the primary tumor sample and if the change in the expression levels in response to modulation of c-MAF gene expression is inversely correlated to said modulation this is indicative that the reduced levels of said gene are indicative of propensity to metastasis. 
     
     
         26 . Method according to any of the  claims 23  to  25  in which the modulation of c-MAF expression is an increase in the expression. 
     
     
         27 . Method according to  claim 26  in which the increase in expression is carried out by expression in the cell population of the c-MAF short isoform and/or the long isoform. 
     
     
         28 . Method according to any of the  claims 23  to  25  in which the modulation of c-MAF expression levels is a decrease in the expression. 
     
     
         29 . Method according to  claim 28  in which c-MAF short isoform and/or the c-MAF long isoform expression is decreased. 
     
     
         30 . Method according to any of the  claims 23  to  29  in which the breast cancer cells and/or the primary tumor are ER+ or triple negative. 
     
     
         31 . Method according to any of the  claims 23  to  30  in which the metastasis is bone metastasis. 
     
     
         32 . Method according to any of the  claims 23  to  31  in which the breast cancer cells are from an established cell line. 
     
     
         33 . Method according to  claim 32 , the established cell line is line MCF7.