Transmembrane serine protease overexpressed in ovarian carcinoma and uses thereof

Abstract

The present invention provides a transmembrane serine protease TADG-12 protein, splice variants of the TADG-12 protein and DNA fragments encoding such proteins. Also provided are vectors and host cells capable of expressing the DNAs. The present invention further provides various methods of early detection and therapies of associated ovarian and other malignancies by utilizing the DNAs and/or proteins disclosed herein.

1 . A DNA fragment encoding a Tumor Associated Differentially-Expressed Gene-12 (TADG-12) protein or splicing variant of TADG-12, said DNA comprising:
 (a) an isolated DNA fragment encoding a TADG-12 protein that has the amino acid sequence of SEQ ID NO: 2;   (b) an isolated DNA fragment encoding a TADG-12 splicing variant that has an amino acid sequence comprising part of SEQ ID NO. 2; and   (c) an isolated DNA fragment differing from the isolated DNA fragments of (a) and (b) above in codon sequence due to the degeneracy of the genetic code, and which encodes a TADG-12 protein or a splicing variant of TADG-12.   
     
     
         2 . The DNA fragment of  claim 1 , wherein the DNA fragment has a sequence shown in SEQ ID NO: 1, 3 or 155. 
     
     
         3 . The DNA fragment of  claim 1 , wherein the variant TADG-12 protein has an amino acid sequence shown in SEQ ID NO: 4 or 154. 
     
     
         4 . A vector comprising the DNA fragment of  claim 1  and regulatory elements necessary for expressing said DNA in a cell. 
     
     
         5 . The vector of  claim 4 , wherein said DNA fragment encodes a TADG-12 protein having the amino acid sequence shown in SEQ ID NO: 2, 4 or 154. 
     
     
         6 . A host cell transfected with the vector of  claim 4 , said vector expressing a TADG-12 protein. 
     
     
         7 . The host cell of  claim 6 , wherein the cell is a bacterial cell, a mammalian cell, a plant cell, or an insect cell. 
     
     
         8 . An antisense oligonucleotide complementary to the DNA fragment encoding a splicing variant of TADG-12 protein of  claim 1 . 
     
     
         9 . An isolated and purified TADG-12 splicing variant protein encoded for by a DNA comprising:
 (a) isolated DNA encoding a TADG-12 splicing variant, said variant protein having an amino acid sequence comprising part of SEQ ID NO. 2; and   (b) isolated DNA differing from the isolated DNA of (a) above in codon sequence due to the degeneracy of the genetic code, and which encodes a splicing variant of TADG-12.   
     
     
         10 . The isolated and purified TADG-12 splicing variant protein of  claim 9 , wherein the TADG-12 protein has an amino acid sequence shown in SEQ ID NO: 154. 
     
     
         11 . An antibody directed against the TADG-12 splicing variant protein of  claim 9 . 
     
     
         12 . A method for detecting malignant hyperplasia in a biological sample, comprising:
 (a) isolating mRNA from the sample; and   (b) detecting in the sample mRNA encoding a TADG-12 splicing variant protein, wherein the presence of the mRNA in said sample indicates the presence of malignant hyperplasia, wherein the absence of the mRNA in the sample indicates the absence of malignant hyperplasia.   
     
     
         13 . The method of  claim 12 , wherein the detection method is PCR amplification, Northern blot, dot blot or a DNA array chip. 
     
     
         14 . The method of  claim 13 , wherein the PCR amplification uses primers having a nucleotide sequence shown in SEQ ID NOS: 28-31. 
     
     
         15 . The method of  claim 12 , wherein the TADG-12 splicing variant protein has an amino acid sequence shown in SEQ ID NO: 154. 
     
     
         16 . The method of  claim 12 , wherein said biological sample is blood, urine, saliva, tears, interstitial fluid, ascites fluid, tumor tissue biopsy, or circulating tumor cells. 
     
     
         17 . A method for detecting malignant hyperplasia in a biological sample, comprising:
 (a) isolating protein from the sample; and   (b) detecting in said sample a TADG-12 splicing variant protein, wherein the presence of the TADG-12 splicing variant protein in said sample indicates the presence of malignant hyperplasia, wherein the absence of the TADG-12 protein variant in said sample indicates the absence of malignant hyperplasia.   
     
     
         18 . The method of  claim 17 , wherein the detection method is Western blot, dot blot, ELISA sandwich assay, radioimmunoassay or flow cytometry analysis. 
     
     
         19 . The method of  claim 17 , wherein the TADG-12 protein has an amino acid sequence shown in SEQ ID NO: 154. 
     
     
         20 . The method of  claim 17 , wherein the biological sample is blood, urine, saliva, tears, interstitial fluid, ascites fluid, tumor tissue biopsy, or circulating tumor cells. 
     
     
         21 . A method of targeted therapy to an individual, comprising:
 administering to an individual a compound having a targeting moiety and a therapeutic moiety, wherein the targeting moiety is specific for a TADG-12 splicing variant protein.   
     
     
         22 . The method of  claim 21 , wherein the targeting moiety is an antibody directed against a TADG-12 splicing variant protein or a ligand or ligand binding domain that binds a TADG-12 splicing variant protein. 
     
     
         23 . The method of  claim 21 , wherein the TADG-12 protein has an amino acid sequence shown in SEQ ID NO: 154. 
     
     
         24 . The method of  claim 23 , wherein the therapeutic moiety is a radioisotope, a toxin, a chemotherapeutic agent, an immune stimulant, or a cytotoxic agent. 
     
     
         25 . The method of  claim 23 , wherein the individual suffers from ovarian cancer, lung cancer, prostate cancer, or colon cancer. 
     
     
         26 . A method of vaccinating an individual against TADG-12, comprising:
 inoculating the individual with a TADG-12 splicing variant protein or fragment thereof, wherein the inoculation elicits an immune response in the individual, thereby vaccinating said individual against TADG-12.   
     
     
         27 . The method of  claim 26 , wherein the individual has a cancer, is suspected of having a cancer or is at risk of getting a cancer. 
     
     
         28 . The method of  claim 26 , wherein the TADG-12 splicing variant protein has an amino acid sequence shown in SEQ ID NO: 154.