Serum microRNA signatures as "liquid biopsies" for interrogating hepatotoxic mechanisms and liver pathogenesis in human

PLoS One. 2017 May 17;12(5):e0177928. doi: 10.1371/journal.pone.0177928. eCollection 2017.


MicroRNAs (miRNAs) released into the peripheral circulation upon cellular injury have shown a promise as a new class of tissue-specific biomarkers. We were first to demonstrate that next-generation sequencing analysis of serum from human subjects with acetaminophen-induced liver injury revealed a specific signature of circulating miRNAs. We consequently hypothesized that different types of hepatic liver impairments might feature distinct signatures of circulating miRNAs and that this approach might be useful as minimally invasive diagnostic "liquid biopsies" enabling the interrogation of underlying molecular mechanisms of injury in distant tissues. Therefore we examined serum circulating miRNAs in a total of 72 serum samples from a group of 53 subjects that included patients with accidental acetaminophen overdose, hepatitis B infection, liver cirrhosis and type 2 diabetes as well as gender- and age-matched healthy subjects with no evidence of liver disease. The miRNA signatures were identified using next-generation sequencing that provided analysis for the whole miRNome, including miRNA isoforms. Compared to the healthy subjects, a total of 179 miRNAs showed altered serum levels across the diseased subjects. Although many subjects have elevated alanine aminotransferase suggesting liver impairments, we identified distinct miRNA signatures for different impairments with minimum overlap. Furthermore, the bioinformatics analysis of miRNA signatures revealed relevant molecular pathways associated with the mechanisms of toxicity and or pathogenesis of disease. Interestingly, the high proportion of miRNA isoforms present in the respective signatures indicated a new level of complexity in cellular response to stress or disease. Our study demonstrates for the first time that signatures of circulating miRNAs show specificity for liver injury phenotypes and, once validated, might become useful for diagnosis of organ pathologies as "liquid biopsies".

MeSH terms

  • Adult
  • Biomarkers / blood
  • Chemical and Drug Induced Liver Injury / genetics*
  • Diabetes Mellitus, Type 2 / genetics*
  • Female
  • Gene Expression Profiling / methods
  • Gene Expression Regulation
  • Gene Regulatory Networks
  • Genetic Predisposition to Disease
  • Hepatitis B / genetics*
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Liver Cirrhosis / genetics*
  • Male
  • MicroRNAs / blood*


  • Biomarkers
  • MicroRNAs

Grants and funding

The project was funded by Pfizer Inc. in forms of research grants to J.C.K. (University of Maastricht) and K.J.J. (University of Michigan). Pfizer Inc. provided support in the form of salaries for authors [S.J.S., M.G., D.B., P.C. and J.A.] and reviewed the final manuscript but the funding organization did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.