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Redox Mechanisms Driving Skin Fibroblast-to-Myofibroblast Differentiation.

Marzieh Aminzadehanboohi1, Manousos Makridakis2, Delphine Rasti1

  • 1Department of Pathology and Immunology, Medical School, University of Geneva, 1211 Geneva, Switzerland.

Antioxidants (Basel, Switzerland)
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Transforming Growth Factor-Beta 1 (TGF-β1) drives fibroblast to myofibroblast differentiation via hydrogen peroxide (H₂O₂). This study identifies key oxidative mechanisms and proteins involved in this crucial wound healing and fibrosis process.

Keywords:
epidermal growth factorfibroblastsmyofibroblastsproteomicsredox signallingthiol oxidationtransforming growth factor β1

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Physiology

Background:

  • Transforming Growth Factor-Beta 1 (TGF-β1) is crucial for fibroblast differentiation into myofibroblasts, a key process in tissue repair and fibrosis.
  • Hydrogen peroxide (H₂O₂) acts as a signaling molecule downstream of TGF-β1, regulating cellular processes via redox modifications.
  • Understanding the oxidative mechanisms of TGF-β1 signaling is vital for comprehending fibrosis and developing therapeutic strategies.

Purpose of the Study:

  • To investigate the specific oxidative mechanisms triggered by TGF-β1 in human skin fibroblasts.
  • To identify the sources of hydrogen peroxide (H₂O₂) involved in TGF-β1-induced myofibroblast differentiation.
  • To uncover novel redox-sensitive proteins and pathways regulating fibroblast differentiation.

Main Methods:

  • Primary human skin fibroblast cultures were used to study TGF-β1 effects.
  • Gene expression analysis of redox-related genes was performed.
  • Inhibition of H₂O₂-producing enzymes (genetic and pharmacological) was evaluated.
  • Redox proteomics (OxICAT) was employed to identify oxidized proteins.

Main Results:

  • TGF-β1 upregulated oxidant-generating genes and downregulated antioxidant genes.
  • Pharmacological inhibition of a flavoenzyme with diphenyleneiodonium reduced myofibroblast differentiation and oxygen consumption.
  • Key proteins, including EGFR, filamin A, fibulin-2, and endosialin, showed increased cysteine oxidation.

Conclusions:

  • This study elucidates the sources of H₂O₂ in TGF-β1-mediated fibroblast differentiation.
  • Novel redox mechanisms involving specific protein cysteine oxidation are highlighted.
  • Findings provide insights into the molecular basis of fibrosis and tissue repair processes.