LINE-1 methylation in plasma DNA as a biomarker of activity of DNA methylation inhibitors in patients with solid tumors.
Multiple clinical trials are investigating the use of the DNA methylation inhibitors azacitidine and decitabine for the treatment of solid tumors. Clinical trials in hematological malignancies have shown that optimal activity does not occur at their maximum tolerated doses but selection of an optimal biological dose and schedule for use in solid tumor patients is hampered by the difficulty of obtaining tumor tissue to measure their activity. Here we investigate the feasibility of using plasma DNA to measure the demethylating activity of the DNA methylation inhibitors in patients with solid tumors. We compared four methods to measure LINE-1 and MAGE-A1 promoter methylation in T24 and HCT116 cancer cells treated with decitabine treatment and selected Pyrosequencing for its greater reproducibility and higher signal to noise ratio. We then obtained DNA from plasma, peripheral blood mononuclear cells, buccal mucosa cells and saliva from ten patients with metastatic solid tumors at two different time points, without any intervening treatment. DNA methylation measurements were not significantly different between time point 1 and time point 2 in patient samples. We conclude that measurement of LINE-1 methylation in DNA extracted from the plasma of patients with advanced solid tumors, using Pyrosequencing, is feasible and has low within patient variability. Ongoing studies will determine whether changes in LINE-1 methylation in plasma DNA occur as a result of treatment with DNA methylation inhibitors and parallel changes in tumor tissue DNA.
Aparicio
Ana
A
Department of Genitourinary Medical Oncology, University of Texas, MD Anderson Cancer Center, Houston, TX 77030, USA. aaparicio@mdanderson.org
North
Brittany
B
Barske
Lindsey
L
Wang
Xuemei
X
Bollati
Valentina
V
Weisenberger
Daniel
D
Yoo
Christine
C
Tannir
Nizar
N
Horne
Erin
E
Groshen
Susan
S
Jones
Peter
P
Yang
Allen
A
Issa
Jean-Pierre
JP
D000951:Antigens, Neoplasm / Q000235:genetics; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000235:genetics*; D045744:Cell Line, Tumor; D002610:Cheek; D019175:DNA Methylation / Q000187:drug effects*; D015254:DNA Modification Methylases / Q000037:antagonists & inhibitors*; D004273:DNA, Neoplasm / Q000097:blood / Q000378:metabolism*; D000077209:Decitabine; D006801:Humans; D020084:Long Interspersed Nucleotide Elements; D058950:Melanoma-Specific Antigens; D009061:Mouth Mucosa / Q000378:metabolism; D009363:Neoplasm Proteins / Q000235:genetics; D009369:Neoplasms / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012463:Saliva / Q000378:metabolism; D017422:Sequence Analysis, DNA
D000951:Antigens, Neoplasm; D000964:Antimetabolites, Antineoplastic; D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; C072011:MAGEA1 protein, human; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D000077209:Decitabine; D015254:DNA Modification Methylases; D001374:Azacitidine