Mapping proteome and lipidome changes in early-onset non-alcoholic fatty liver disease using hepatic 3D spheroids

Helle Sedighi Frandsen, Joel Mario Vej-Nielsen, Lauren Elizabeth Smith, Lang Sun, Karoline Lindgaard Mikkelsen, Annemette Præstegaard Thulesen, Christina Erika Hagensen, Fuquan Yang, Adelina Rogowska-Wrzesinska*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Non-alcoholic fatty liver disease affects one-fourth of the world’s population. Central to the disease progression is lipid accumulation in the liver, followed by inflammation, fibrosis and cirrhosis. The underlying mechanism behind the early stages of the disease is poorly understood. We have exposed human hepatic HepG2/C3A cells-based spheroids to 65 μM oleic acid and 45 μM palmitic acid and employed proteomics and lipidomics analysis to investigate their effect on hepatocytes. The treatment successfully induced in vivo hallmarks of NAFLD, as evidenced by intracellular lipid accumulation and increased ATP levels. Quantitative lipidome analysis revealed an increase in ceramides, LPC and saturated triglycerides and a decrease in the ratio of PC/PE, similar to the changes observed in patients’ liver biopsies. The proteomics analysis combined with qPCR showed increased epithelial to mesenchymal transition (EMT) signalling. Activation of EMT was further validated by transcriptomics in TGF-β treated spheroids, where an increase in mesenchymal cell markers (N-cadherin and collagen expression) was found. Our study demonstrates that this model system thus closely echoes several of the clinical features of non-alcoholic fatty liver disease and can be used to investigate the underlying molecular changes occurring in the condition.

Original languageEnglish
Article number3216
JournalCells
Volume11
Issue number20
ISSN2073-4409
DOIs
Publication statusPublished - Oct 2022

Keywords

  • 3-dimensional cell culture
  • fatty liver disease
  • human hepatocytes
  • lipidomics
  • NAFLD
  • proteomics
  • spheroids
  • steatosis

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