Methods for treating cells containing fusion genes
The present invention relates to methods for treating prostate cancer patients. It is based, at least in part, on the discovery that approximately 90% of men carrying at least one of the following fusion genes: TRMT11-GRIK2, SLC45A2-AMACR, MTOR-TP53BP1, LRRC59-FLJ60017, TMEM135-CCDC67 and CCNH-05orf30 experienced prostate cancer recurrence, metastases and/or prostate cancer-specific death after radical prostatectomy (each examples of “progressive prostate cancer”), while these outcomes occurred in only 36% of men not carrying any of these fusion genes. It is also based, at least in part, on the discovery that a genome editing technique that specifically targets a fusion gene can induce cell death in a cancer cell that carries the fusion gene.
1. A method of treating a subject having prostate cancer, comprising determining whether a sample of the subject contains one or more fusion genes selected from the group consisting of TRMT11-GRIK2, SLC45A2-AMACR, MTOR-TP53BP1, LRRC59-FLJ60017, TMEM135-CCDC67, KDM4B-AC011523.2, MAN2A1-FER, PTEN-NOLC1, CCNH-C5orf30, ZMPSTE24-ZMYM4, CLTC-ETV1, ACPP-SEC13, DOCK7-OLR1, PCMTD1-SNTG1 and a combination thereof; and where if one or more fusion genes are detected in the sample then performing a CRISPR/Cas genome editing technique targeting the one or more fusion genes present within one or more prostate cancer cells of the subject,
wherein the genome editing technique comprises introducing into the one or more prostate cancer cells: (i) a vector comprising a nucleic acid encoding a Cas protein or a mutant thereof and two guide RNAs (gRNA), wherein one gRNA is complementary to a region within one gene of the fusion gene and a second gRNA is complementary to a region within a second gene of the fusion gene, and (ii) a vector comprising a donor nucleic acid and one or more targeting sequences,
wherein the two gRNAs direct binding of the Cas 9 protein or mutant thereof to a genomic sequence of the fusion gene to promote cleavage of the fusion gene and insertion of the donor nucleic acid into the fusion gene, and wherein expression of the donor nucleic acid induces cell death of the one or more prostate cancer cells.
2. The method of claim 1 , wherein the fusion gene is TMEM135-CCDC67 or CCNH-C5orf30.
3. The method of claim 1 , wherein the fusion gene is detected by FISH analysis.
4. The method of claim 1 , wherein the fusion gene is detected by reverse transcription polymerase chain reaction.
5. The method of claim 1 , wherein the donor nucleic acid encodes HSV-1 thymidine kinase.
6. The method of claim 1 further comprising administering to the subject a therapeutically effective amount of ganciclovir or valganciclovir.
7. The method of claim 1 , wherein the one or more targeting sequences are complementary to one or more sequences of the fusion gene to promote homologous recombination of and the insertion of the donor nucleic acid into the fusion gene.
8. The method of claim 1 , wherein the Cas 9 protein is Cas 9 D10A .