Toxicoproteomics of HepG2 Reveals Translational Biomarkers of Paracetamol-Induced Liver Injury and Recovery

ABSTRACT: Drug-induced liver injury (DILI) is a leading cause of acute liver failure (ALF) and a major obstacle in drug development, reflecting the limited predictive value of animal models and lack of robust,mechanism-based biomarkers. Human in vitro systems, coupled with unbiased omics approaches, have emerged as a potential alternative strategy. Here, quantitative toxicoproteomics was applied to paracetamol (acetaminophen; APAP)-exposed HepG2 cells to identify translational biomarkers of APAP-induced DILI. Cells were exposed to the IC₅₀ concentration of APAP for 8 h (injury phase), followed by drug withdrawal and a 24 h recovery phase. LC–MS/MS analysis quantified 1,478 proteins and revealed increased protein secretion upon APAP exposure. Notably, comparison of the proteomic dataset against available enriched proteomic profiles of ALF patients’ serum confirmed the clinical relevance of the molecular signature obtained. Injury-phase upregulated proteins, including KRT18 and HMGB1, recapitulated known clinical DILI biomarkers. Conversely, recovery was characterized by enrichment of the liver regeneration markers AFP, APOA1 and HP, enabling discrimination between injury and regeneration. HPD, S100A6 and TXNRD1 emerged as candidate diagnostic biomarkers, while SERPINA1 and complement components (C3, C5) showed prognostic potential. Collectively, this study demonstrates that toxicoproteomics applied to human in vitro hepatic models, validated against patient-derived data, enables the discovery of novel translational biomarkers of liver injury and recovery.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Cardiovascular System Disease

SUBMITTER: Rui Vitorino  

LAB HEAD: Joana Miranda

PROVIDER: PXD072667 | Pride | 2026-07-27

REPOSITORIES: Pride