Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
Monitoring liver damage using hepatocyte-specific methylation markers in cell-free circulating DNA
Roni Lehmann-Werman, … , Ruth Shemer, Yuval Dor
Roni Lehmann-Werman, … , Ruth Shemer, Yuval Dor
Published June 21, 2018
Citation Information: JCI Insight. 2018;3(12):e120687. https://doi.org/10.1172/jci.insight.120687.
View: Text | PDF
Resource and Technical Advance Hepatology Transplantation

Monitoring liver damage using hepatocyte-specific methylation markers in cell-free circulating DNA

  • Text
  • PDF
Abstract

Liver damage is typically inferred from serum measurements of cytoplasmic liver enzymes. DNA molecules released from dying hepatocytes are an alternative biomarker, unexplored so far, potentially allowing for quantitative assessment of liver cell death. Here we describe a method for detecting acute hepatocyte death, based on quantification of circulating, cell-free DNA (cfDNA) fragments carrying hepatocyte-specific methylation patterns. We identified 3 genomic loci that are unmethylated specifically in hepatocytes, and used bisulfite conversion, PCR, and massively parallel sequencing to quantify the concentration of hepatocyte-derived DNA in mixed samples. Healthy donors had, on average, 30 hepatocyte genomes/ml plasma, reflective of basal cell turnover in the liver. We identified elevations of hepatocyte cfDNA in patients shortly after liver transplantation, during acute rejection of an established liver transplant, and also in healthy individuals after partial hepatectomy. Furthermore, patients with sepsis had high levels of hepatocyte cfDNA, which correlated with levels of liver enzymes aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Duchenne muscular dystrophy patients, in which elevated AST and ALT derive from damaged muscle rather than liver, did not have elevated hepatocyte cfDNA. We conclude that measurements of hepatocyte-derived cfDNA can provide specific and sensitive information on hepatocyte death, for monitoring human liver dynamics, disease, and toxicity.

Authors

Roni Lehmann-Werman, Judith Magenheim, Joshua Moss, Daniel Neiman, Ofri Abraham, Sheina Piyanzin, Hai Zemmour, Ilana Fox, Talya Dor, Markus Grompe, Giora Landesberg, Bao-Li Loza, Abraham Shaked, Kim Olthoff, Benjamin Glaser, Ruth Shemer, Yuval Dor

×

Figure 1

Liver-specific markers.

Options: View larger image (or click on image) Download as PowerPoint
Liver-specific markers.
(A) Structure of the 3 loci (adjacent to the ITI...
(A) Structure of the 3 loci (adjacent to the ITIH4, IGF2R, and VTN genes) used as hepatocyte biomarkers. Lollipops represent CpG sites. Red indicates CpG sites represented in the Infinium HumanMethylation450K BeadChip. Arrows mark positions of PCR primers. Markers are defined by the methylation status of CpG sites between primers. (B) Methylation status of ITIH4, IGF2R, and VTN in DNA from multiple tissues. Shown is the percentage of molecules in which all CpG sites were unmethylated. (C) Spike-in experiments. Human liver DNA was mixed with human leukocyte DNA in the indicated proportions (0 to 20%), and the percentage of fully unmethylated hepatocyte markers was determined.

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts