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Persulfidation alters gene regulatory programs and promotes endothelial specification.

Janina Wittig1, Ran Xu2, Fredy Delgado Lagos2

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Summary

Endogenous sulfides regulate stem cell differentiation. Lowering sulfide levels impacts cell fate, while adding sulfide preserves pluripotency or promotes vascular development in mesodermal cells.

Keywords:
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Area of Science:

  • Biochemistry
  • Cell Biology
  • Developmental Biology

Background:

  • Endogenously generated sulfides are vital signaling molecules with diverse biological effects.
  • The role of sulfides in pluripotent stem cell commitment and differentiation remains largely unexplored.
  • Protein persulfidation is a key post-translational modification influenced by sulfide levels.

Purpose of the Study:

  • To investigate the role of endogenous sulfides in induced pluripotent stem cell (iPSC) differentiation.
  • To determine how sulfide levels affect cell fate commitment in different germ layers.
  • To explore the potential of sulfide modulation for engineering stem cell fate and vascularization.

Main Methods:

  • Directed differentiation of iPSCs into three germ layers.
  • Measurement of endogenous sulfide levels during differentiation.
  • Supplementation with a sulfide donor to iPSCs and mesodermal cells.
  • Analysis of protein persulfidation, transcription factor activity, and gene expression.
  • Assessment of vascular organoid development.

Main Results:

  • Endogenous sulfide levels decrease during iPSC differentiation, with the mesodermal lineage showing the lowest capacity for sulfide generation.
  • Sulfide supplementation in iPSCs preserved pluripotency by maintaining OCT4 activity.
  • Sulfide addition to mesodermal cells enhanced WNT signaling, ETS transcription factor activity, and expression of angiogenic genes.
  • Sulfide supplementation promoted blood vessel morphogenesis in vascular organoids.

Conclusions:

  • Protein persulfidation acts as a timing-dependent regulator of stem cell fate.
  • Sulfide preserves pluripotency before commitment and subsequently biases mesoderm toward endothelial specification.
  • Sulfide modulation offers a novel strategy for engineering stem cell fate and enhancing vascularization.