Compositions and methods for diagnosing, evaluating and treating cancer by means of the DNA methylation status
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 calculating a proportion of discordant reads (PDR) in a tumor sample from the subject comprising:
detecting DNA methylation status by massive parallel sequencing with bisulfite conversion; whole-genome bisulfite sequencing; reduced representation bisulfite sequencing; microarray, or a genome-wide microarray at one or more regions of neighboring CpG sites in a plurality of cells in the tumor sample;
comparing the DNA methylation status in multiple regions of neighboring CpG sites along a sequence of CpG sites in DNA of the plurality of cells; and
determining a relative number of cells in the tumor sample having inconsistent DNA methylation status across the sequence of CpG sites as compared to the total number of cells in the tumor sample or a number of cells in the tumor sample having consistent DNA methylation status across the sequence of CpG sites thereby calculating the PDR in the tumor sample,
determining the PDR in the tumor sample to be less than or equal to 0.15, and
administering to the subject an antitumor agent.
2. The method according to claim 1 , wherein DNA methylation is detected by microarray or
wherein a PDR greater than 0.15 indicates that the subject is more likely to (1) develop resistance to an antitumor agent; (2) relapse after treatment with an antitumor agent; (3) develop a metastatic tumor; or (4) any combination of the above.
3. The method according to claim 1 , wherein the method further comprises performing a second PDR calculation on a second tumor sample from the subject comprising:
detecting DNA methylation status by massive parallel sequencing with bisulfite conversion; whole-genome bisulfite sequencing; reduced representation bisulfite sequencing; microarray, or a genome-wide microarray at one or more regions of neighboring CpG sites in a plurality of cells in the second tumor sample;
comparing the DNA methylation status in multiple regions of neighboring CpG sites along a sequence of CpG sites in DNA of the plurality of cells; and
determining a relative number of cells in the second tumor sample having inconsistent DNA methylation status across the sequence of CpG sites as compared to the total number of cells in the second tumor sample or a number of cells in the second tumor sample having consistent DNA methylation status across the sequence of CpG sites thereby calculating the PDR in the second tumor sample.
4. The method according to claim 3 , wherein the method further comprises the step of comparing the PDR from the first tumor sample to the PDR from the second tumor sample,
wherein the first and second tumor samples are from different tumors.
5. The method according to claim 3 , wherein the first and second tumor samples are obtained from the same tumor at different times.
6. The method according to claim 5 , wherein the subject is treated with an antitumor agent between obtaining the first tumor sample and obtaining the second tumor sample.
7. The method of claim 6 , wherein the method further comprises stopping or changing the treatment with the antitumor agent if the PDR from the second tumor sample is greater than the PDR from the first tumor sample, or continuing the treatment with the antitumor agent if the PDR from the second tumor sample is the same as or less than the PDR from the first tumor sample.
8. The method of claim 1 , wherein the method further comprises,
calculating a second PDR from a tumor sample obtained from the same tumor in the subject; and
comparing the second PDR to the first PDR;
wherein the antitumor agent is administered after obtaining the first tumor sample and before obtaining the second tumor sample, and
wherein treatment with the antitumor agent is stopped or changed if the second PDR, is different than the first PDR.
9. The method according to claim 8 , wherein the subject is treated with the antitumor agent for 3 months, 6 months, 9 months or 12 months before obtaining the second tumor sample.
10. The method according to claim 1 , 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.