Identification of metabolic prognostic biomarkers in carbon tetrachloride-induced liver fibrosis using MAS-H1-NMR

Currently, the golden standard of liver disease diagnosis is based on histological assessments of biopsy. The use of liver biopsies, however, is not without cost or risk i.e. bleeding, perforation, death. Furthermore, liver biopsies can be subjected to sampling and inter- and intra-observer variabilities. We aim to identify a minimal invasive biomarker that can be used to differentiate the stages of liver disease. Therefore, metabolomics profiling of healthy and CCl₄-induced fibrotic mice using an updated version of Nuclear Magnetic Resonance (MAS-H¹-NMR) was performed in blood plasma.

We have investigated the fingerprints of fibrosis progression by biochemical, histopathological and metabolites as well as gene expression analysis in a time-resolved experiment. Biochemical parameters e.g. ALT and AST were increased during disease progression upon CCl₄ injection. This alteration was correlated with accumulation of extracellular matrix (ECM) and ECM-producing cells as indicated by picro-sirius red and alpha-SMA staining, respectively. Further, blood plasma-based metabolic profiling revealed that several metabolites namely, acetate, ethanol, glucose and lactate significantly reflected the disease stage as indicated by AUC (1.00) compared to heathy mice. Some metabolites e.g. betaine, phenylalanine, citrate, glutamine, alanine, pyruvate, creatinine and O-phosphocholine were also predict the disease stages, however, AUC was ranged between 0.74 and 0.963.

However, formate, valine and tyrosine were poorly predictors where, AUC ranged between 0.52 and 0.67. Further validation of those very good predictors is ongoing in clinical samples.