| Autor: |
Takuya Yamamoto, Megumi Nishio, Hiroyuki Yoshitomi, Tomoko Matsumoto, Makoto Ikeya, Satoko Sakurai, Knut Woltjen, Yoshihiro Yamanaka, Long Guo, Ayako Nagahashi, Shunsuke Kihara, Megumu K. Saito, Shiro Ikegawa, Maya Uemura, Junya Toguchida, Miki Ebisuya, Mitsujiro Osawa, Masahiro Nakamura, Cantas Alev, Mitsuhiro Matsuda |
| Jazyk: |
angličtina |
| Rok vydání: |
2019 |
| Předmět: |
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| DOI: |
10.1101/562447 |
| Popis: |
Pluripotent stem cells (PSCs) have increasingly been used to model different aspects of embryogenesis and organ formation1. Despite recent advances in the in vitro induction of major mesodermal lineages and mesoderm-derived cell types2,3, experimental model systems that can recapitulate more complex biological features of human mesoderm development and patterning are largely missing. Here, we utilized induced pluripotent stem cells (iPSCs) for the stepwise in vitro induction of presomitic mesoderm (PSM) and its derivatives to model distinct aspects of human somitogenesis. We focused initially on modeling 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 succeeded to observe oscillatory expression of core segmentation clock genes, including HES7 and DKK1, and identified novel oscillatory genes in human iPSC-derived PSM. We furthermore determined the period of the human segmentation clock to be around five hours and showed the presence of dynamic traveling wave-like gene expression within in vitro induced human PSM. Utilizing CRISPR/Cas9-based genome editing technology, we then targeted genes, for which mutations in patients with abnormal axial skeletal development such as spondylocostal dysostosis (SCD) (HES7, LFNG and DLL3) or spondylothoracic dysostosis (STD) (MESP2) have been reported. Subsequent analysis of patient-like iPSC knock-out lines as well as patient-derived iPSCs together with their genetically corrected isogenic controls revealed gene-specific alterations in oscillation, synchronization or differentiation properties, validating the overall utility of our model system, to recapitulate not only key features of human somitogenesis but also to provide novel insights into diseases associated with the formation and patterning of the human axial skeleton. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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