Recapitulating the human segmentation clock with pluripotent stem cells
| Autor: | Ayako Nagahashi, Mitsujiro Osawa, Long Guo, Satoko Sakurai, Shiro Ikegawa, Makoto Ikeya, Knut Woltjen, Megumi Nishio, Shunsuke Kihara, Maya Uemura, Megumu K. Saito, Miki Ebisuya, Masahiro Nakamura, Takuya Yamamoto, Hiroyuki Yoshitomi, Junya Toguchida, Tomoko Matsumoto, Noriaki Kawakami, Thomas L. Maurissen, Yoshihiro Yamanaka, Mitsuhiro Matsuda, Cantas Alev |
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| Rok vydání: | 2018 |
| Předmět: |
Male
Pluripotent Stem Cells Mesoderm Time Factors Period (gene) Embryonic Development Biology In Vitro Techniques LFNG 03 medical and health sciences Mice 0302 clinical medicine Biological Clocks Somitogenesis Paraxial mesoderm medicine Basic Helix-Loop-Helix Transcription Factors Animals Humans Abnormalities Multiple Induced pluripotent stem cell 030304 developmental biology Regulation of gene expression Gene Editing Hernia Diaphragmatic 0303 health sciences Multidisciplinary Embryogenesis Intracellular Signaling Peptides and Proteins Gene Expression Regulation Developmental Glycosyltransferases Membrane Proteins Cell biology medicine.anatomical_structure Phenotype Somites Intercellular Signaling Peptides and Proteins 030217 neurology & neurosurgery |
| Zdroj: | Nature. 580(7801) |
| ISSN: | 1476-4687 |
| Popis: | 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. A system involving in vitro induction of presomitic mesoderm recapitulates oscillatory expression of core segmentation clock genes and travelling-wave-like gene expression, suggesting that this system can be used to study the human segmentation clock and provide insights into diseases associated with human axial skeletogenesis. |
| Databáze: | OpenAIRE |
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