Compositions and methods for diagnosing, evaluating and treating cancer
The present invention relates to methods of determining a cancer treatment prognosis for a subject in need thereof by evaluating epigenetic and genetic changes within a tumor sample from the subject. The present invention further provides methods of treating cancer in a subject by evaluating epigenetic and genetic changes within a tumor sample from the subject. In addition, the present invention provides methods of screening test agents to identify agents that decrease tumor cell plasticity.
1. A method of treating cancer in a subject comprising:
(a) obtaining a first tumor sample and a second tumor sample from the subject,
wherein the first tumor sample and the second tumor sample each comprise a plurality of cancer cells from the subject,
wherein the first tumor sample is obtained before the subject has been treated with an antitumor agent, and
wherein the second tumor sample is obtained after the subject has been treated with the antitumor agent;
(b) assaying the first tumor sample for a first level of inconsistent DNA methylation status and the second tumor sample for a second level of inconsistent DNA methylation status,
wherein the DNA methylation status is assayed at one or more regions of neighboring CpG sites having locally disordered methylation,
wherein the neighboring CpG sites comprise a collection of CpG sites:
(1) along one or more sequences in DNA, and/or
(2) at genomic loci of one or more genes,
wherein the DNA methylation status is assayed by: massive parallel sequencing with bisulfite conversion; whole-genome bisulfite sequencing; reduced representation bisulfite sequencing; microarray; or a genome-wide microarray;
and
(c) treating the subject by either:
(1) continuing the treatment of the subject with the antitumor agent when the first level of inconsistent DNA methylation status is the same as the second level of inconsistent DNA methylation status, or
(2) altering treatment of the subject with an antitumor agent when the first level of inconsistent DNA methylation status is not the same as the second level of inconsistent DNA methylation status.
2. The method of claim 1 , wherein the one or more regions of neighboring CpG sites having locally disordered methylation:
comprise CpG sites within a CpG island; or
are within a promoter.
3. The method according to claim 1 , wherein the method further comprises assaying a subclonal genetic mutation.
4. The method of claim 3 , further comprising:
a) altering the treatment with the antitumor agent or initiating a non-chemotherapeutic treatment when the subject has a subclonal genetic mutation; or
b) continuing treatment with the antitumor agent when the subject has no subclonal genetic mutation.
5. The method of claim 1 , wherein the tumor samples are from a solid tumor.
6. The method of claim 5 , wherein the solid tumor sample is from adrenocortical carcinoma, bone tumors, brain cancer, breast cancer, cervical cancer, colorectal carcinoma, desmoid tumors, desmoplastic small round cell tumors, endocrine tumors, esophageal cancer, Ewing sarcoma family tumors, gastric cancer, germ cell tumors, head or neck cancer, hepatoblastoma, hepatocellular carcinoma, lung cancer, melanoma, mesothelioma, nasopharyngeal carcinoma, neuroblastoma, non-rhabdomyosarcoma soft tissue sarcoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, skin carcinoma, testicular cancer, thyroid carcinoma, uterine cancer and Wilms tumors.
7. The method of claim 1 , wherein the cancer is CLL.
8. The method of claim 1 , wherein altering the treatment of the subject with the antitumor agent comprises:
(1) decreasing the amount of the antitumor agent being administered to the subject when the first level of inconsistent DNA methylation status is higher than the second level of inconsistent DNA methylation status; or
(2) administering a non-chemotherapeutic treatment to the subject when the first level of inconsistent DNA methylation status is lower than the second level of inconsistent DNA methylation status.
9. The method of claim 1 , wherein the tumor samples are from a hematological cancer, leukemia, or chronic lymphocytic leukemia.
10. The method according to claim 1 , wherein the antitumor agent is an angiogenesis inhibitor selected from the group consisting of angiostatin K1-3, DL-α-difluoromethyl-ornithine, endostatin, fumagillin, genistein, minocycline, staurosporine, and (±)-thalidomide.
11. The method according to claim 1 , wherein the antitumor agent is a DNA intercalator/cross-linker selected from the group consisting of bleomycin, carboplatin, carmustine, chlorambucil, cyclophosphamide, cis-diammineplatinum(II) dichloride, melphalan, mitoxantrone, and oxaliplatin.
12. The method according to claim 1 , wherein the antitumor agent is a DNA synthesis inhibitor selected from the group consisting of (±)-amethopterin, 3-amino-1,2,4-benzotriazine 1,4-dioxide, aminopterin, cytosine β-D-arabinofuranoside, 5-fluoro-5′-deoxyuridine, 5-fluorouracil, ganciclovir, hydroxyurea, and mitomycin C.
13. The method according to claim 1 , wherein the antitumor agent is a DNA-RNA transcription regulator selected from the group consisting of actinomycin D, daunorubicin, doxorubicin, homoharringtonine, and idarubicin.
14. The method according to claim 1 , wherein the antitumor agent is an enzyme inhibitor selected from the group consisting of S(+)-camptothecin, curcumin, (−)-deguelin, 5,6-dichlorobenzimidazole 1-β-D-ribofuranoside, etoposide, formestane, fostriecin, hispidin, 2-imino-1-imidazoli-dineacetic acid, mevinolin, trichostatin A, tyrphostin AG 34, and Tyrphostin AG 879.
15. The method according to claim 1 , wherein the antitumor agent is a gene regulator selected from the group consisting of 5-aza-2′-deoxycytidine, 5-azacytidine, cholecalciferol, 4-hydroxytamoxifen, melatonin, mifepristone, raloxifene, all trans-retinal, retinoic acid, 9-cis-retinoic acid, 13-cis-retinoic acid, retinol, tamoxifen, and troglitazone.
16. The method according to claim 1 , wherein the antitumor agent is a microtubule inhibitor selected from the group consisting of colchicine, dolastatin 15, nocodazole, paclitaxel, podophyllotoxin, rhizoxin, vinblastine, vincristine, vindesine, and vinorelbine.
17. The method according to claim 1 , wherein the antitumor agent is a neoantigen.
18. The method according to claim 1 , wherein the antitumor agent is an unclassified antitumor agent selected from the group consisting of 17-(allylamino)-17-demethoxygeldanamycin, 4-amino-1,8-naphthalimide, apigenin, brefeldin A, cimetidine, dichloromethylene-diphosphonic acid, leuprolide, luteinizing hormone-releasing hormone, pifithrin-a, rapamycin, sex hormone-binding globulin, thapsigargin, and urinary trypsin inhibitor fragment.
19. The method according to claim 1 , wherein the antitumor agent is a monoclonal antibody selected from the group consisting of rituximab, alemtuzumab, ipilimumab, bevacizumab, cetuximab, panitumumab, trastuzumab, vemurafenib, imatinib mesylate, erlotinib, gefitinib, vismodegib, 90Y-ibritumomab tiuxetan, 131I-tositumomab, ado-trastuzumab emtansine, lapatinib, pertuzumab, ado-trastuzumab emtansine, regorafenib, sunitinib, denosumab, sorafenib, pazopanib, axitinib, dasatinib, nilotinib, bosutinib, ofatumumab, obinutuzumab, ibrutinib, idelalisib, crizotinib, afatinib dimaleate, ceritinib, tositumomab, ibritumomab tiuxetan, brentuximab vedotin, bortezomib, siltuximab, trametinib, dabrafenib, pembrolizumab, carfilzomib, ramucirumab, cabozantinib, and vandetanib.
20. The method according to claim 1 , wherein the antitumor agent is a cytokine selected from the group consisting of interferons, interleukins, and hematopoietic growth factors.
21. The method according to claim 20 , wherein the antitumor agent is selected from INF-α, IL-2, aldesleukin, erythropoietin, granulocyte-macrophage colony-stimulating factor, and granulocyte colony-stimulating factor.
22. The method according to claim 1 , wherein the antitumor agent is a targeted therapy selected from the group consisting of toremifene, fulvestrant, anastrozole, exemestane, letrozole, ziv-aflibercept, alitretinoin, temsirolimus, tretinoin, denileukin diftitox, vorinostat, romidepsin, bexarotene, pralatrexate, lenaliomide, belinostat, pomalidomide, cabazitaxel, enzalutamide, abiraterone acetate, radium 223 chloride, and everolimus.
23. The method according to claim 1 , wherein the antitumor agent is a checkpoint inhibitor selected from the group consisting of an inhibitor of the programmed death-1 pathway, an anti-PD 1 antibody, an anti-cytotoxic T-lymphocyte-associated antigen antibody, and an antibody targeting the CD28 CTLA4 Ig superfamily.
24. The method according to claim 1 , wherein the antitumor agent is selected from BTLA, LAG3, ICOS, PDL1 and KIR.
25. The method according to claim 1 , wherein the antitumor agent is an inhibitor targeting a member of the TNFR superfamily selected from the group consisting of CD40, OX40, CD137, GITR, CD27 and TIM-3.
26. The method according to claim 1 , wherein the antitumor agent is an epigenetic targeted drug selected from the group consisting of HDAC inhibitors, kinase inhibitors, DNA methyltransferase inhibitors, histone demethylase inhibitors, and histone methylation inhibitors.
27. The method according to claim 1 , wherein the antitumor agent is an epigenetic drug selected from the group consisting of azacitidine, decitabine, vorinostat, romidepsin, and ruxolitinib.
28. A method of treating cancer in a subject comprising:
(a) obtaining a first tumor sample and a second tumor sample from the subject,
wherein the first tumor sample and the second tumor sample each comprise a plurality of cancer cells from the subject,
wherein the first tumor sample is obtained before the subject has been treated with an antitumor agent, and
wherein the second tumor sample is obtained after the subject has been treated with the antitumor agent;
(b) assaying the first tumor sample for a first level of inconsistent DNA methylation status and the second tumor sample for a second level of inconsistent DNA methylation status,
wherein the DNA methylation status is assayed at one or more regions of neighboring CpG sites having locally disordered methylation,
wherein the neighboring CpG sites comprise a collection of CpG sites:
(1) along one or more sequences in DNA, and/or
(2) at genomic loci of one or more genes, and
wherein the DNA methylation status is assayed by: massive parallel sequencing with bisulfite conversion; whole-genome bisulfite sequencing; reduced representation bisulfite sequencing; microarray; or a genome-wide microarray;
and
(c) treating the subject by continuing to treat the subject with the antitumor agent when the level of the first level of inconsistent DNA methylation status is the same as the second level of inconsistent DNA methylation status or when the level of the first level of inconsistent DNA methylation status is higher than the second level of inconsistent DNA methylation status.
29. A method of treating cancer in a subject comprising:
(a) obtaining a first tumor sample and a second tumor sample from the subject,
wherein the first tumor sample and the second tumor sample each comprise a plurality of cancer cells from the subject,
wherein the first tumor sample is obtained before the subject has been treated with an antitumor agent, and
wherein the second tumor sample is obtained after the subject has been treated with the antitumor agent;
(b) assaying the first tumor sample for a first level of inconsistent DNA methylation status and the second tumor sample for a second level of inconsistent DNA methylation status,
wherein the DNA methylation status is assayed at one or more regions of neighboring CpG sites having locally disordered methylation,
wherein the neighboring CpG sites comprise a collection of CpG sites:
(1) along one or more sequences in DNA, and/or
(2) at genomic loci of one or more genes, and
wherein the DNA methylation status is assayed by: massive parallel sequencing with bisulfite conversion; whole-genome bisulfite sequencing; reduced representation bisulfite sequencing; microarray; or a genome-wide microarray;
and
(c) altering the treatment of the subject with the antitumor agent or initiating a non-chemotherapeutic treatment when the first level of inconsistent DNA methylation status is lower than the second level of inconsistent DNA methylation status.