Distinguishing PCA3 messenger RNA species in benign and malignant prostate tissues
View Patent ↗This invention concerns the discovery of two distinct PCA3 mRNA sequences. One of these sequences corresponds to a short PCA3 mRNA molecule whereas the other PCA3 RNA molecule is longer as it comprises an additional sequence between exon 3 and exon 4a. The short RNA is associated with prostate cancer whereas the long RNA sequence is associated with a non-malignant state of the prostate. Based on the differential expression levels of these two PCA3 RNA sequences, protocols for the diagnosis of prostate disease are provided. The invention also relates to therapeutic approaches to prostate cancer.
1. A prostate cancer-classifying composition comprising:
(a) a PCA3 nucleic acid molecule comprising an additional sequence between PCA3 exon 3 and PCA3 exon 4a, said additional sequence corresponding essentially to nucleotides 27 to 254 of SEQ ID NO: 1, wherein said PCA3 nucleic acid molecule is from a prostate cell of a human subject having or suspected of having prostate cancer; and
(b) at least one oligonucleotide enabling the determination of whether said PCA3 nucleic acid molecule RNA comprises or lacks said additional sequence between PCA3 exon 3 and PCA3 exon 4a,
wherein the presence or level of a PCA3 nucleic acid molecule lacking said additional sequence indicates that said prostate cells comprise prostate cancer cells.
2. The prostate cancer-classifying composition of claim 1 , wherein said determination of whether PCA3 RNA from said prostate cells or said nucleic acid preparation comprises or lacks an additional sequence between PCA3 exon 3 and PCA3 exon 4a comprises performing a hybridization and/or amplification reaction.
3. The prostate cancer-classifying composition of claim 2 , wherein said amplification reaction comprises a reaction which is: a nucleic acid sequence-based amplification reaction (NASBA), a polymerase chain reaction (PCR), a transcription-based amplification reaction, a strand displacement amplification (SDA), a ligase chain reaction (LCR), or a Qβ replicase reaction.
4. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes specifically to:
(a) PCA3 exon 3;
(b) PCA3 exon 4a;
(c) the PCA3 exon 3-exon 4a junction;
(d) the PCA3 exon 3-intron 3 junction; or
(e) the PCA3 intron 3-exon 4a junction.
5. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes specifically to a PCA3 exon 3-exon 4a junction as defined by nucleotide positions 26 and 27 of SEQ ID NO: 2.
6. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes to at least 10 consecutive nucleotides of a PCA3 nucleic acid molecule set forth in SEQ ID NO: 1 or 2.
7. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes to at least 12 consecutive nucleotides of a PCA3 nucleic acid molecule set forth in SEQ ID NO: 1 or 2.
8. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes to at least 15 consecutive nucleotides of a PCA3 nucleic acid molecule set forth in SEQ ID NO: 1 or 2.
9. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes to at least 20 consecutive nucleotides of a PCA3 nucleic acid molecule set forth in SEQ ID NO: 1 or 2.
10. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes under high stringency conditions to at least 10 consecutive nucleotides across nucleotides 26 and 27 of SEQ ID NO: 2.
11. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes under high stringency conditions to at least 15 consecutive nucleotides across nucleotides 26 and 27 of SEQ ID NO: 2.
12. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes under high stringency conditions to at least 18 consecutive nucleotides across nucleotides 26 and 27 of SEQ ID NO: 2.
13. The prostate cancer-classifying composition of claim 12 , wherein said high stringency conditions comprise a hybridization at 65° C. in 5×SSC, 5×Denhardt's solution, 1% SDS, and 100 μg/mL denatured carrier DNA.
14. The prostate cancer-classifying composition of claim 1 further comprising a DNA polymerase.
15. The prostate cancer-classifying composition of claim 1 , wherein said composition enables the determination of the malignancy status of said subject having or suspected of having prostate cancer.
16. The prostate cancer-classifying composition of claim 5 , wherein said composition enables the determination of the malignancy status of said subject having or suspected of having prostate cancer.
17. The prostate cancer-classifying composition of claim 1 , wherein said oligonucleotide hybridizes under high stringency conditions to a PCA3 nucleic acid molecule set forth in SEQ ID NO: 1 or 2, wherein said high stringency conditions comprise a hybridization at 65° C. In 5×SSC, 5×Denhardt's solution, 1% SDS, and 100 μg/mL denatured carrier DNA.
18. The prostate cancer-classifying composition of claim 17 , wherein said oligonucleotide is:
(a) at least 15 nucleotides in length;
(b) at least 18 nucleotides in length; or
(c) from at least 18 to about 50 nucleotides in length.
19. The prostate cancer-classifying composition of claim 18 , wherein said oligonucleotide hybridizes under high stringency conditions across nucleotides 26 and 27 of SEQ ID NO: 2.
20. The prostate cancer-classifying composition of claim 17 , wherein said oligonucleotide is at least 20 to about 35 nucleotides in length.
21. The prostate cancer-classifying composition of claim 17 , wherein said oligonucleotide is detectably labeled.
22. The prostate cancer-classifying composition of claim 17 , wherein said oligonucleotide hybridizes under high stringency conditions across nucleotides 26 and 27 of SEQ ID NO: 2.
23. The prostate cancer-classifying composition of claim 17 , wherein said PCA3 nucleic acid molecule is RNA or cDNA.
24. The prostate cancer-classifying composition of claim 1 , wherein said PCA3 nucleic acid molecule is RNA or cDNA.