DIFFERENTIALLY-METHYLATED REGIONS OF THE GENOME USEFUL AS MARKERS OF EMBRYO-ADULT TRANSITIONS
The present invention relates to compositions and methods for the assay, diagnosis, prognosis or monitoring of the embryonic, fetal, and adult epigenetic states of a human genome. The disclosed methods are useful in monitoring the progress of in vitro and in vivo cellular reprogramming and the diagnosis, prognosis or monitoring of cancer in an individual. Specifically, the invention provides methods for the detection and interpretation of observed differential DNA methylation patterns and associated epigenetic modifications to core histones in determining the developmental status of human cells for the detection and characterization of cancer cells and determining optimum therapeutic modalities.
1 . A method to determine the developmental staging of cells that were the source of a sample of human DNA comprised of the steps: 1) identifying DMRs differentially-methylated in embryonic (pre-fetal) cells compared to their fetal (prenatal) or adult counterparts, 2) determining whether a sample of human DNA contains methylated or unmethylated CpG epigenetic marks within said DMRs, 2) use of said markers for the diagnosis, prognosis and/or treatment of cancer.
2 . The method of claim 1 , wherein the use of the information relating to the methylated or unmethylated CpG epigenetic marks is directed to determining the completeness of the in vitro transcriptional reprogramming of cells to pluripotency (iPS cell reprogramming) or the in vivo reprogramming of cells and tissues to reverse aging or to induce tissue regeneration (iTR) in diverse tissues in the body.
3 . The method of claim 1 , wherein said embryonic (pre-fetal), fetal (prenatal), or postnatal (adult) cells are human.
4 . The method of claim 1 , wherein said methylated or unmethylated CpG epigenetic marks are identified from blood-derived cfDNA.
5 . The method of claim 1 , wherein said methylated or unmethylated CpG epigenetic marks are identified from blood-derived cfDNA through the use of methylation-specific restriction endonuclease digestion.
6 . The method of claim 1 , wherein said methylated or unmethylated CpG epigenetic marks are identified from blood-derived cfDNA through the use of methylation-specific PCR primers.
7 . The method of determining the optimum therapeutic strategy for a given cancer in humans comprised of the steps: 1) obtaining DNA from the tumor, 2) determining the relative methylation of one or more of the DMRs of the present invention in said DNA sample, 3) determining whether said DMRs are statistically correlated with pre- or post-EFT, 4) treating pre-EFT cells with chemotherapy, iCM, or radiation therapy, 5) treating post-EFT cells as CSCs and treating with iTR.