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Deterministic Restriction on Pluripotent State Dissolution by Cell-Cycle Pathways.

Kevin Andrew Uy Gonzales1, Hongqing Liang2, Yee-Siang Lim2

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Human embryonic stem cells (hESCs) lose pluripotency during differentiation via pluripotent state dissolution (PSD). Cell cycle progression, particularly S and G2 phases, actively inhibits PSD, revealing a hardwired link between cell cycle and pluripotency.

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Area of Science:

  • Stem cell biology
  • Cell cycle regulation
  • Epigenetics

Background:

  • Human embryonic stem cells (hESCs) possess pluripotency, the ability to differentiate into various cell types.
  • Pluripotency is lost during differentiation through a process termed pluripotent state dissolution (PSD).
  • The regulatory networks governing PSD are not fully understood.

Purpose of the Study:

  • To identify key regulators of pluripotent state dissolution (PSD) in human embryonic stem cells (hESCs).
  • To investigate the role of the cell cycle in regulating pluripotency.
  • To understand how cell cycle progression influences the initiation of differentiation.

Main Methods:

  • High-throughput RNA interference (RNAi) screen to identify genes regulating PSD.
  • Diverse differentiation conditions were employed.
  • Genetic and chemical perturbations were used to study cell cycle phase-specific effects.

Main Results:

  • Identified central and context-dependent regulators of PSD, including histone acetylation, chromatin remodeling, RNA splicing, and signaling pathways.
  • Discovered a significant enrichment of cell-cycle genes, particularly those involved in DNA replication and G2 phase progression.
  • Demonstrated that S and G2 phases attenuate PSD due to an intrinsic propensity for pluripotency, independent of G1 phase.

Conclusions:

  • Pluripotency control is intrinsically linked to the cell cycle machinery.
  • Specific pathways active during S and G2 phases of the cell cycle deterministically restrict pluripotent state dissolution.
  • The absence of these pathways in G1 phase may permit the initiation of differentiation.