Methods of treating cells containing fusion genes by genomic targeting
The present invention relates to methods for treating patients having cancer or a premalignant or neoplastic condition. It is 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 other than a prostate cancer cell, e.g., a hepatocellular cancer cell, having the fusion gene. The present invention provides methods for treating cancer patients that include performing a genome editing technique targeting a fusion gene present within one or more cells of a subject to produce an anti-cancer effect.
1. A method of treating a subject, comprising (i) determining that at least one fusion gene is present in a sample obtained from a subject; wherein the at least one fusion gene is 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 (ii) then performing a CRISPR/Cas genome editing technique targeting the one or more fusion genes within one or more cells of the subject to achieve an anti-neoplastic effect or an anti-cancer effect, wherein the CRISPR/Cas system uses a Cas9 endonuclease, wherein the subject does not have prostate cancer.
2. The method of claim 1 , wherein the subject has a pre-malignant or a neoplastic condition.
3. The method of claim 1 , wherein the subject has cancer.
4. The method of claim 3 , wherein the cancer is breast cancer, liver cancer, lung cancer, cervical cancer, endometrial cancer, pancreatic cancer, ovarian cancer, gastric cancer, thyroid cancer, glioblastoma multiforme, colorectal cancer, diffuse large B cell-lymphoma, sarcoma, acute and chronic lymphocytic leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, or esophageal adenocarcinoma.
5. The method of claim 1 , wherein the at least one fusion gene is detected by FISH analysis or by reverse transcription polymerase chain reaction.
6. The method of claim 1 , wherein the CRISPR/Cas system cleaves a sequence within the fusion gene genomic sequence to insert a nucleic acid within the at least one fusion gene to induce cell death.
7. The method of claim 1 , wherein the genome editing technique comprises: transducing the one or more cells with (i) a vector comprising a nucleic acid encoding a Cas9 protein and two guide RNAs (gRNA) and (ii) a vector comprising a donor nucleic acid and one or more targeting sequences.
8. The method of claim 7 , wherein one of the gRNAs is complementary to a region within a first gene of the at least one fusion gene and the other gRNA is complementary to a region within a second gene of the at least one fusion gene.
9. The method of claim 7 , wherein the donor nucleic acid encodes HSV-1 thymidine kinase.
10. The method of claim 7 , wherein the method further comprises administering to the subject a therapeutically effective amount of ganciclovir or valganciclovir.
11. The method of claim 7 , wherein one of the gRNAs is complementary to a region within a MAN2A1 gene of a MAN2A1-FER fusion gene and the other gRNA can be complementary to a region within a FER gene.
12. The method of claim 7 , wherein one of the gRNAs is complementary to a region within a TMEM135 gene of a TMEM135-CCDC67 fusion gene and the other gRNA can be complementary to a region within a CCDC67 gene.
13. The method of claim 12 , wherein the one or more cells of the subject are cancer cells.
14. The method of claim 13 , wherein the cancer cells are breast cancer, liver cancer, lung cancer, cervical cancer, endometrial cancer, pancreatic cancer, ovarian cancer, gastric cancer, thyroid cancer, glioblastoma multiforme, colorectal cancer, diffuse large B cell lymphoma, sarcoma, acute and chronic lymphocytic leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma or esophageal adenocarcinoma cells.
15. The method of claim 3 , wherein the cancer is not lung adenocarcinoma, glioblastoma multiforme or hepatocellular carcinoma.