Recapitulating the human segmentation clock with pluripotent stem cells

Nature. 2020 Apr;580(7801):124-129. doi: 10.1038/s41586-020-2144-9. Epub 2020 Apr 1.

Abstract

Pluripotent stem cells are increasingly used to model different aspects of embryogenesis and organ formation1. Despite recent advances in in vitro induction of major mesodermal lineages and cell types2,3, experimental model systems that can recapitulate more complex features of human mesoderm development and patterning are largely missing. Here we used induced pluripotent stem cells for the stepwise in vitro induction of presomitic mesoderm and its derivatives to model distinct aspects of human somitogenesis. We focused initially on modelling the human segmentation clock, a major biological concept believed to underlie the rhythmic and controlled emergence of somites, which give rise to the segmental pattern of the vertebrate axial skeleton. We observed oscillatory expression of core segmentation clock genes, including HES7 and DKK1, determined the period of the human segmentation clock to be around five hours, and demonstrated the presence of dynamic travelling-wave-like gene expression in in vitro-induced human presomitic mesoderm. Furthermore, we identified and compared oscillatory genes in human and mouse presomitic mesoderm derived from pluripotent stem cells, which revealed species-specific and shared molecular components and pathways associated with the putative mouse and human segmentation clocks. Using CRISPR-Cas9-based genome editing technology, we then targeted genes for which mutations in patients with segmentation defects of the vertebrae, such as spondylocostal dysostosis, have been reported (HES7, LFNG, DLL3 and MESP2). Subsequent analysis of patient-like and patient-derived induced pluripotent stem cells revealed gene-specific alterations in oscillation, synchronization or differentiation properties. Our findings provide insights into the human segmentation clock as well as diseases associated with human axial skeletogenesis.

MeSH terms

  • Abnormalities, Multiple / genetics
  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / deficiency
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Biological Clocks / genetics
  • Biological Clocks / physiology*
  • Embryonic Development / genetics
  • Embryonic Development / physiology*
  • Gene Editing
  • Gene Expression Regulation, Developmental / genetics
  • Glycosyltransferases / deficiency
  • Glycosyltransferases / genetics
  • Hernia, Diaphragmatic / genetics
  • Humans
  • In Vitro Techniques
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intracellular Signaling Peptides and Proteins / deficiency
  • Intracellular Signaling Peptides and Proteins / genetics
  • Male
  • Membrane Proteins / deficiency
  • Membrane Proteins / genetics
  • Mice
  • Phenotype
  • Pluripotent Stem Cells / cytology*
  • Somites / cytology*
  • Somites / growth & development*
  • Somites / metabolism
  • Time Factors

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • DKK1 protein, human
  • DLL3 protein, human
  • HES7 protein, human
  • Intercellular Signaling Peptides and Proteins
  • Intracellular Signaling Peptides and Proteins
  • MESP2 protein, human
  • Membrane Proteins
  • Glycosyltransferases
  • LFNG protein, human

Supplementary concepts

  • Jarcho-Levin syndrome