Modelling Ferroptosis in a Human Microglial Line by Sequential Exposure to Iron and GPX4 Inhibition

Area of Science:

• Neuroscience
• Cell Biology
• Biochemistry

Background:

• Excessive iron accumulation is linked to neurodegenerative diseases (NDDs).
• Ferroptosis, an iron-dependent cell death, is implicated in NDD pathogenesis.
• Microglia, brain immune cells, buffer iron but are vulnerable to ferroptosis, worsening neuroinflammation.

Purpose of the Study:

• To investigate the molecular mechanisms driving ferroptosis in human microglia.
• To establish and utilize a human microglial ferroptosis model for multiomic analysis.

Main Methods:

• Established a human microglial (HMC3 cell line) ferroptosis model.
• Performed integrated multiomic profiling (lipidomics, transcriptomics, proteomics).
• Utilized ferrostatin-1 (Fer-1) as a rescue agent to identify key pathways.

Main Results:

• The model recapitulated ferroptosis hallmarks: increased reactive oxygen species (ROS) and lipid peroxidation, reversed by Fer-1.
• Identified dysregulated lipid species, including sterol accumulation (e.g., 7-oxo-cholesterol) and polyunsaturated fatty acid (PUFA) oxidation.
• Revealed upregulation of the mevalonate pathway and cholesterol metabolism, with some genes reversed by Fer-1, indicating a preferroptotic signature.

Conclusions:

• Developed a novel human microglial ferroptosis model for studying early molecular events.
• Discovered specific lipid and metabolic alterations associated with microglial ferroptosis.
• Findings suggest potential therapeutic targets for neuroprotection by modulating microglial ferroptosis and maintaining cellular homeostasis.