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

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

Research output: Contribution to journalArticle

13 Scopus citations

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.

Original languageEnglish (US)
JournalJCI Insight
Volume3
Issue number12
DOIs
StatePublished - Jun 21 2018

Keywords

  • Diagnostics
  • Epigenetics
  • Hepatology
  • Transplantation

ASJC Scopus subject areas

  • Medicine(all)

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    Lehmann-Werman, R., Magenheim, J., Moss, J., Neiman, D., Abraham, O., Piyanzin, S., Zemmour, H., Fox, I., Dor, T., Grompe, M., Landesberg, G., Loza, B. L., Shaked, A., Olthoff, K., Glaser, B., Shemer, R., & Dor, Y. (2018). Monitoring liver damage using hepatocyte-specific methylation markers in cell-free circulating DNA. JCI Insight, 3(12). https://doi.org/10.1172/jci.insight.120687