title: Genome-scale detection of hypermethylated CpG islands in circulating cell-free DNA of hepatocellular carcinoma patients.
abstract: Despite advances in DNA methylome analyses of cells and tissues, current techniques for genome-scale profiling of DNA methylation in circulating cell-free DNA (ccfDNA) remain limited. Here we describe a methylated CpG tandems amplification and sequencing (MCTA-Seq) method that can detect thousands of hypermethylated CpG islands simultaneously in ccfDNA. This highly sensitive technique can work with genomic DNA as little as 7.5 pg, which is equivalent to 2.5 copies of the haploid genome. We have analyzed a cohort of tissue and plasma samples (n = 151) of hepatocellular carcinoma (HCC) patients and control subjects, identifying dozens of high-performance markers in blood for detecting small HCC (≤ 3 cm). Among these markers, 4 (RGS10, ST8SIA6, RUNX2 and VIM) are mostly specific for cancer detection, while the other 15, classified as a novel set, are already hypermethylated in the normal liver tissues. Two corresponding classifiers have been established, combination of which achieves a sensitivity of 94% with a specificity of 89% for the plasma samples from HCC patients (n = 36) and control subjects including cirrhosis patients (n = 17) and normal individuals (n = 38). Notably, all 15 alpha-fetoprotein-negative HCC patients were successfully identified. Comparison between matched plasma and tissue samples indicates that both the cancer and noncancerous tissues contribute to elevation of the methylation markers in plasma. MCTA-Seq will facilitate the development of ccfDNA methylation biomarkers and contribute to the improvement of cancer detection in a clinical setting.
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Wen
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Jingyi
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Surgery, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.|Ninth School of Clinical Medicine, Peking University, Beijing 100044, China.|School of Oncology, Capital Medical University, Beijing 100069, China.
authors.forename: Huahu
authors.initials: H
authors.lastname: Guo
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China.
authors.forename: Shengmin
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China.
authors.forename: Dafang
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Zhu
authors.affiliation: Center of Therapeutic Research for Liver Cancer, Beijing 302 Hospital, Beijing 100039, China.
authors.forename: Jianhui
authors.initials: J
authors.lastname: Qu
authors.affiliation: Department of Hepatobiliary Surgery, Beijing DiTan Hospital, Capital Medical University, Beijing 100015, China.
authors.forename: Limin
authors.initials: L
authors.lastname: Guo
authors.affiliation: Department of Surgery, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.|Ninth School of Clinical Medicine, Peking University, Beijing 100044, China.|School of Oncology, Capital Medical University, Beijing 100069, China.
authors.forename: Dexiao
authors.initials: D
authors.lastname: Du
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|BIMCR, Peking University, Beijing 100871, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Jin
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|BIMCR, Peking University, Beijing 100871, China.
authors.forename: Yuhao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Shen
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|BIMCR, Peking University, Beijing 100871, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Ge
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|Ministry of Education Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking University, Beijing 100871, China.|Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
authors.forename: Fuchou
authors.initials: F
authors.lastname: Tang
authors.affiliation: Biodynamic Optical Imaging Center (BIOPIC), College of Life Sciences, Peking University, Beijing 100871, China.|Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.|College of Engineering, Peking University, Beijing 100871, China.
authors.forename: Yanyi
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of Surgery, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.|Ninth School of Clinical Medicine, Peking University, Beijing 100044, China.|School of Oncology, Capital Medical University, Beijing 100069, China.
authors.forename: Jirun
authors.initials: J
authors.lastname: Peng
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis* / Q000235:genetics*; D018899:CpG Islands; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000097:blood*; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D057089:Pathology, Molecular; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004273:DNA, Neoplasm