PCA3, PCA3 Genes, and Methods of Use

Abstract

The present invention relates, in general, to a prostate-specific antigen, PCA3. In particular, the present invention relates to nucleic acid molecules coding for the PCA3 protein; purified PCA3 proteins and polypeptides; recombinant nucleic acid molecules; cells containing the recombinant nucleic acid molecules; antibodies having binding affinity specifically to PCA3 proteins and polypeptides; hybridomas containing the antibodies; nucleic acid probes for the detection of nucleic acids encoding PCA3 proteins; a method of detecting nucleic acids encoding PCA3 proteins or polypeptides in a sample; kits containing nucleic acid probes or antibodies; bioassays using the nucleic acid sequence, protein or antibodies of this invention to diagnose, assess, or prognose a mammal afflicted with prostate cancer; therapeutic uses; and methods of preventing prostate cancer in an animal.

1 . An isolated DNA molecule which is:
 (a) a DNA molecule comprising the PCA3 nucleotide sequence of the polynucleotide clone contained in the deposit at the Centraal voor Schimmelcultures as accession number CBS 682.97;   (b) a DNA molecule comprising the PCA3 nucleotide sequence of the polynucleotide clone contained in the deposit at the Centraal voor Schimmelcultures as accession number CBS 100521;   (c) a DNA molecule comprising a PCA3 nucleotide sequence at least 90% identical to the PCA3 nucleotide sequence of SEQ ID NO: 1, 3, 4 or 6;   (d) a DNA molecule that hybridizes under high stringency conditions to the PCA3 nucleotide sequence of SEQ ID NO: 1, 3, 4 or 6 at a site spanning at least one PCA3 exon-exon junction, wherein said PCA3 exon-exon junction corresponds to nucleotide positions:
 (i) 98-99, 263-264, 446-447, or 985-986 of SEQ ID NO: 1; 
 (ii) 98-99, 281-282, or 820-821 of SEQ ID NO: 3; 
 (iii) 98-99 or 281-282 of SEQ ID NO: 4; or 
 (iv) 120-121, 285-286, 468-469, 1007-1008, 2066-2067, or 2622-2623 of SEQ ID NO: 6; 
   (e) a DNA molecule comprising at least 10 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6 and spanning at least one of the PCA3 exon-exon junctions defined in (d); or   (f) a DNA molecule fully complementary to any one of (a) to (e).   
     
     
         2 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) hybridizes to at least 10 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         3 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) hybridizes to at least 12 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         4 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) hybridizes to at least 15 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         5 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) hybridizes to at least 18 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         6 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) hybridizes under high stringency conditions to the PCA3 nucleotide sequence of SEQ ID NO: 1, 3, 4 or 6 at a site spanning at least the PCA3 exon-exon junction corresponding to nucleotide positions 446-447 of SEQ ID NO: 1, or positions 468-469 of SEQ ID NO: 6. 
     
     
         7 . The isolated DNA molecule of  claim 1 , wherein said high stringency conditions in (d) comprise hybridization in 0.2×SSC and 0.1% SDS at 68° C. 
     
     
         8 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (e) comprises at least 12 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         9 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (e) comprises at least 15 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         10 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (e) comprises at least 18 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         11 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (e) spans at least the PCA3 exon-exon junction corresponding to nucleotide positions 446-447 of SEQ ID NO: 1, or positions 468-469 of SEQ ID NO: 6. 
     
     
         12 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) or (e) has a length of between 15 to 500 nucleotides. 
     
     
         13 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule in (d) or (e) is a probe or a primer. 
     
     
         14 . The isolated DNA molecule of  claim 1  comprising:
 (a) the nucleotide sequence of PCA3 exon 1 corresponding to positions 1-98 of SEQ ID NO: 1, or positions 1-120 of SEQ ID NO: 6; 
 (b) the nucleotide sequence of PCA3 exon 3 corresponding to positions 264-446 of SEQ ID NO: 1, or positions 286-468 of SEQ ID NO: 6; 
 (c) the nucleotide sequence of PCA3 exon 4a corresponding to positions 447-985 of SEQ ID NO: 1, or positions 469-1007 of SEQ ID NO: 6; 
 (d) the nucleotide sequence of PCA3 exon 4b corresponding to positions 986-2037 of SEQ ID NO: 1, or positions 1008-2066 of SEQ ID NO: 6; 
 (e) the nucleotide sequence of PCA3 exon 4c corresponding to positions 2067-2622 of SEQ ID NO: 6; and/or 
 (f) the nucleotide sequence of PCA3 exon 4d corresponding to positions 2623-3582 of SEQ ID NO: 6. 
 
     
     
         15 . The isolated DNA molecule of  claim 1 , wherein said DNA molecule comprises the nucleotide sequence of SEQ ID NO: 1, 3, 4 or 6. 
     
     
         16 . The isolated DNA molecule of  claim 1  comprising a detectable label. 
     
     
         17 . The isolated DNA molecule of  claim 1  which is cDNA or a fragment thereof. 
     
     
         18 . A kit for detecting the presence of a PCA3 nucleic acid in a sample, said kit comprising at least one container having disposed therein the nucleic acid molecule of  claim 13 . 
     
     
         19 . An isolated DNA molecule that hybridizes under high stringency conditions to the PCA3 nucleotide sequence of SEQ ID NO: 1, 3, 4 or 6 at a site spanning at least one PCA3 exon-exon junction, wherein said PCA3 exon-exon junction corresponds to nucleotide positions:
 (i) 98-99, 263-264, 446-447, or 985-986 of SEQ ID NO: 1;   (ii) 98-99, 281-282, or 820-821 of SEQ ID NO: 3;   (iii) 98-99 or 281-282 of SEQ ID NO: 4; or   (iv) 120-121, 285-286, 468-469, 1007-1008, 2066-2067, or 2622-2623 of SEQ ID NO: 6;   
       or an isolated DNA molecule that is fully complementary thereto. 
     
     
         20 . An isolated DNA molecule comprising at least 10 contiguous nucleotides of SEQ ID NO: 1, 3, 4 or 6 and at least one PCA3 exon-exon junction, wherein said PCA3 exon-exon junction corresponds to nucleotide positions:
 (i) 98-99, 263-264, 446-447, or 985-986 of SEQ ID NO: 1;   (ii) 98-99, 281-282, or 820-821 of SEQ ID NO: 3;   (iii) 98-99 or 281-282 of SEQ ID NO: 4; or   (iv) 120-121, 285-286, 468-469, 1007-1008, 2066-2067, or 2622-2623 of SEQ ID NO: 6;   
       or an isolated DNA molecule that is fully complementary thereto.