Investigating Protein Oxidation in Hepatocyte Responses to Drugs and Xenobiotics
The goal of our research is to uncover the role of protein oxidation in hepatocyte responses to drugs, xenobiotics, and genotoxic compounds. Understanding these mechanisms is crucial for advancing knowledge of liver physiology and pathology, as well as for improving drug safety assessments.
We have developed a 3D cell culture model based on a hepatocarcinoma cell line, using multicellular spheroids propagated in rotary bioreactors. This system closely mimics the liver’s microenvironment, providing a more physiologically relevant platform for studying liver function and responses to external stimuli.
We integrate proteomics to evaluate changes in protein expression and post-translational modifications, alongside lipidomic analysis to investigate lipid metabolism and accumulation. Additionally, we have developed advanced redox-proteomics approaches that enable the detection of specific oxidative modifications, such as S-nitrosylation, S-sulfenylation, and protein carbonylation. These tools allow us to study oxidative damage at unprecedented levels of precision, providing insights into the molecular pathways impacted by oxidative stress in hepatocytes.
Related publications:
Havelund JF, Wojdyla K, Davies MJ, Jensen ON, Møller IM, Rogowska-Wrzesinska A. A biotin enrichment strategy identifies novel carbonylated amino acids in proteins from human plasma. Journal of Proteomics. 2017 Mar 6;156:40-51. DOI: 10.1016/j.jprot.2016.12.019
Wojdyla K, Wrzesinski K, Williamson J, Fey SJ, Rogowska-Wrzesinska A. Acetaminophen-induced S-nitrosylation and S-sulfenylation signalling in 3D cultured hepatocarcinoma cell spheroids. Toxicology Research. 2016;5(3):905-920. DOI: 10.1039/c5tx00469a
Wojdyla K, Williamson J, Roepstorff P, Rogowska-Wrzesinska A. The SNO/SOH TMT strategy for combinatorial analysis of reversible cysteine oxidations. Journal of Proteomics. 2015 Jan 15;113:415-434. DOI: 10.1016/j.jprot.2014.10.015
Wojdyla K, Rogowska-Wrzesinska A. Differential alkylation-based redox proteomics - Lessons learnt. Redox Biology. 2015 Aug 5;6:240-252. DOI: 10.1016/j.redox.2015.08.005
Nedić O, Rogowska-Wrzesinska A, Rattan SIS. Standardization and quality control in quantifying non-enzymatic oxidative protein modifications in relation to ageing and disease: Why is it important and why is it hard? Redox Biology. 2015 Apr 14;5:91-100. DOI: 10.1016/j.redox.2015.04.001