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 journalArticlepeer-review

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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