TADG-15: an extracellular serine protease overexpossed in carcinomas

Abstract

The present invention provides DNA encoding a TADG-15 protein as well as a TADG-15 protein. Also provided is a vector capable of expressing the DNA of the present invention adapted for expression in a recombinant cell and regulatory elements necessary for expression of the DNA in the cell. The present invention further provides for methods of inhibiting TADG-15 expression and/or protease activity, methods of detecting TADG-15 mRNA and/or protein and methods of screening for TADG-15 inhibitors. Additionally, the present invention provides for cell-specific targeting via TADG-15 and methods of vaccinating an individual against TADG-15. The methods described are useful in the diagnosis, treatment and prevention of cancer, particularly breast and ovarian cancer.

1 . A vector comprising a DNA encoding a tumor antigen-derived gene (TADG-15) protein and regulatory elements necessary for expression of the DNA in a cell; wherein the DNA is positioned in reverse orientation relative to the regulatory elements such that TADG-15 antisense mRNA is produced, wherein the DNA is:
 (a) isolated DNA which encodes a TADG-15 protein;   (b) isolated DNA which hybridizes under high stringency conditions to the isolated DNA of (a) above and which encodes a TADG-15 protein; or   (c) isolated DNA differing from the isolated DNAs of (a) and (b) above in codon sequence due to the degeneracy of the genetic code, and which encodes a TADG-15 protein.   
     
     
         2 . The vector of  claim 1 , wherein the DNA has the sequence shown in SEQ ID NO: 1. 
     
     
         3 . The vector of  claim 1 , wherein the TADG-15 protein has the amino acid sequence shown in SEQ ID NO: 2. 
     
     
         4 . A host cell transfected with the vector of  claim 1 . 
     
     
         5 . The host cell of  claim 4 , wherein the cell is a bacterial cell, a mammalian cell, a plant cell, or an insect cell. 
     
     
         6 . The host cell of  claim 5 , wherein the bacterial cell is  E. coli.    
     
     
         7 . A method of inhibiting expression of TADG-15 in a cell, comprising:
 introducing the vector of  claim 1  into a cell, wherein expression of the vector produces TADG-15 antisense mRNA in the cell, wherein the TADG-15 antisense mRNA hybridizes to endogenous TADG-15 mRNA, thereby inhibiting expression of TADG-15 in the cell.   
     
     
         8 . An oligonucleotide having the nucleotide sequence complementary to the DNA sequence of  claim 1 . 
     
     
         9 . A method of treating a neoplastic state in an individual in need of such treatment, comprising:
 administering to the individual an effective dose of the oligonucleotide of  claim 8 .   
     
     
         10 . The method of  claim 9 , wherein the neoplastic state is ovarian cancer, breast cancer, lung cancer, colon cancer, prostate cancer, or other cancers in which TADG-15 is overexpressed. 
     
     
         11 . A method of inhibiting a TADG-15 protein in a cell, comprising:
 introducing an antibody into a cell, wherein the antibody is specific for a TADG-15 protein or a fragment thereof, wherein binding of the antibody to the TADG-15 protein inhibits the TADG-15 protein.   
     
     
         12 . The method of  claim 11 , wherein the antibody is linked to a therapeutic moiety, said method further comprising targeting the therapeutic moiety to cells in an individual. 
     
     
         13 . The method of  claim 12 , wherein the therapeutic moiety is a radioisotope, a toxin, a chemotherapeutic agent, an immune stimulant, or a cytotoxic agent. 
     
     
         14 . The method of  claim 12 , wherein the individual suffers from ovarian cancer, lung cancer, prostate cancer, colon cancer, or other cancers in which TADG-15 is overexpressed. 
     
     
         15 . An immunogenic composition, comprising an immunogenic fragment of a TADG-15 protein and an appropriate adjuvant. 
     
     
         16 . The immunogenic composition of  claim 15 , wherein the fragment is a 9-residue fragment up to a 20-residue fragment. 
     
     
         17 . The immunogenic composition of  claim 16 , wherein the 9-residue fragment has a sequence shown in SEQ ID Nos. 2, 19, 20, 21, 29, 39, 49, 50, 59, 79, 80, 81, 82, 83, 84, 89, or 90. 
     
     
         18 . A method of vaccinating an individual against TADG-15, comprising:
 inoculating an individual with the TADG-15 protein or immunogenic fragment thereof of  claim 15 , wherein the TADG-15 protein or immunogenic fragment thereof lacks TADG-15 protease activity, wherein the inoculation with the TADG-15 protein or immunogenic fragment thereof elicits an immune response in the individual, thereby vaccinating the individual against TADG-15.   
     
     
         19 . The method of  claim 18 , wherein the individual has cancer, is suspected of having cancer or is at risk of getting cancer. 
     
     
         20 . A method of screening for compounds that inhibit TADG-15, comprising:
 (a) contacting a sample with a compound, wherein the sample comprises TADG-15 protein; and   (b) assaying for TADG-15 protease activity, wherein a decrease in the TADG-15 protease activity in the presence of the compound relative to TADG-15 protease activity in the absence of the compound is indicative of a compound that inhibits TADG-15.