Systemic inflammation reprograms human hepatic phenotype and suppresses absorption, distribution, metabolism, and excretion gene expression: Validation in a 3-dimensional human liver spheroid system.

Inflammation profoundly alters hepatic drug metabolism and transport, requiring careful consideration in pharmacotherapy to minimize adverse outcomes. In this study, we compared global transcriptomic profiles of macroscopically and histologically normal liver tissue from patients undergoing tumor metastasis resection (n = 37) and critically ill patients requiring intensive care (n = 50), alongside an advanced 3-dimensional primary human liver spheroid model. Transcriptomic analysis revealed clear segregation between cohorts, identifying 6250 differentially expressed genes and a pronounced inflammatory signature in livers from critically ill patients, marked by elevated acute-phase proteins, including C-reactive protein, serum amyloid A1, and serum amyloid A2. Three-dimensional primary human liver spheroids were analyzed with increasing cellular complexity: hepatocyte monocultures, hepatocyte-nonparenchymal cell cocultures, and triple cultures incorporating liver sinusoidal endothelial cells, with or without free fatty acid supplementation to mimic metabolic stress. Incorporation of nonparenchymal cells and liver sinusoidal endothelial cells markedly upregulated C-reactive protein expression and induced transcriptomic shifts closely mirroring inflamed human liver tissue. Pathway enrichment and targeted analysis of absorption, distribution, metabolism, and excretion genes demonstrated consistent inflammation-driven downregulation of cytochrome P450 enzymes, phase II enzymes, transporters, and nuclear receptors critical for drug metabolism and biliary efflux-patterns nearly identical to those observed between inflamed liver tissue and tissue obtained by tumor resection. These findings highlight substantial phenotype changes in macroscopically and histologically normal livers and validate advanced multicellular 3-dimensional human liver spheroids as physiologically relevant models for recapitulating inflammation-induced perturbations in hepatic pharmacokinetics, offering predictive tools for improved preclinical safety assessment and personalized dosing in patients with systemic inflammation. SIGNIFICANCE STATEMENT: Inflammation profoundly alters hepatic drug metabolism and transport, requiring careful consideration in pharmacotherapy to minimize adverse outcomes. This study highlights substantial phenotype changes caused by inflammation in macroscopically and histologically normal livers and validates advanced multicellular 3-dimensional human liver spheroids as physiologically relevant models for recapitulating inflammation-induced perturbations in hepatic pharmacokinetics.