High copper levels induce oxidative stress and inflammatory processes in a cell culture model of Wilson's disease.

Wilson’s disease (WD) is a genetic disorder resulting from mutations in the ATP7B gene that lead to copper overload in hepatocytes. To gain a deeper understanding of the cellular mechanisms underlying WD, a reanalysis of the GSE1073236 dataset was conducted and validated by comparative analyses in a cell culture model of HepG2 and HepG2 ATP7B knockout (ATP7B-KO) cells, as well as in primary hepatocytes from WD patients and controls. Increased expression levels of genes associated with autophagy, oxidative stress and inflammation were observed in copper-treated HepG2 ATP7B-KO cells and in primary hepatocytes from patients with WD. Furthermore, copper increased the secretion of IL1B, TNF, GM-CSF, and IL8 in HepG2 ATP7B-KO cells. Accordingly, copper exposure enhanced NFKB promoter activity 6 h after treatment. However, the transcriptional activity of AP1 and NFKB was reduced in these cells 24 h after treatment with 0.6mM CuCl2. In addition, both HepG2 and HepG2 ATP7B-KO cells showed increased oxidative stress and H2O2 levels after copper treatment, indicating that reactive oxygen species could play a role in WD. In the WD cell culture model, critical mechanisms behind copper-related cell death highlight the importance of this model in developing molecular targets for future therapeutic strategies.