Control of embryonic stem cell metastability by L-proline catabolism

Laura Casalino, Stefania Comes, Giuseppina Lambazzi, Benedetta De Stefano, Stefania Filosa, Sandro De Falco, Dario De Cesare, Gabriella Minchiotti, Eduardo Jorge Patriarca

Research output: Contribution to journalArticle

Abstract

The molecular mechanisms controlling mouse embryonic stem cell (ESC) metastability, i.e. their capacity to fluctuate between different states of pluripotency, are not fully resolved. We developed and used a novel automation platform, the Cellmaker, to screen a library of metabolites on two ESC-based phenotypic assays (i.e. proliferation and colony phenotype) and identified two metabolically related amino acids, namely L-proline (L-Pro) and L-ornithine (L-Orn), as key regulators of ESC metastability. Both compounds, but mainly L-Pro, force ESCs toward a novel epiblast stem cell (EpiSC)-like state, in a dose- and time-dependent manner. Unlike EpiSCs, L-Pro-induced cells (PiCs) contribute to chimeric embryos and rely on leukemia inhibitor factor (LIF) to self-renew. Furthermore, PiCs revert to ESCs or differentiate randomly upon removal of either L-Pro or LIF, respectively. Remarkably, PiC generation depends on both L-Pro metabolism (uptake and oxidation) and Fgf5 induction, and is strongly counteracted by antioxidants, mainly L-ascorbic acid (vitamin C, Vc). ESCs ↔ PiCs phenotypic transition thus represents a previously undefined dynamic equilibrium between pluripotent states, which can be unbalanced either toward an EpiSC-like or an ESC phenotype by L-Pro/L-Orn or Vc treatments, respectively. All together, our data provide evidence that ESC metastability can be regulated at a metabolic level.

Original languageEnglish
Pages (from-to)108-122
Number of pages15
JournalJournal of Molecular Cell Biology
Volume3
Issue number2
DOIs
Publication statusPublished - Apr 2011

Keywords

  • colony phenotype
  • embryonic stem cells
  • L-proline
  • metastability
  • pluripotent states
  • vitamin C

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Genetics

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  • Cite this

    Casalino, L., Comes, S., Lambazzi, G., De Stefano, B., Filosa, S., De Falco, S., De Cesare, D., Minchiotti, G., & Patriarca, E. J. (2011). Control of embryonic stem cell metastability by L-proline catabolism. Journal of Molecular Cell Biology, 3(2), 108-122. https://doi.org/10.1093/jmcb/mjr001