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HypoSUMOylation in embryonic stem cells generate head-trunk embryo-like structures [scRNAseq]

ABSTRACT: HypoSUMOylation in embryonic stem cells generate head-trunk embryo-like structures [scRNAseq]

PROVIDER: PRJNA749070 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15218300_1.fastq.gz	Fastqsanger.gz
	SRR15218300_2.fastq.gz	Fastqsanger.gz
	SRR15218400_1.fastq.gz	Fastqsanger.gz
	SRR15218400_2.fastq.gz	Fastqsanger.gz
	SRR15218301_1.fastq.gz	Fastqsanger.gz

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HypoSUMOylation in embryonic stem cells generate head-trunk embryo-like structures [ChIP-seq]

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Project description:Numerous models of synthetic embryos have recently been established to simulate mammalian development. Two main strategies have been developed to build mouse or human embryo-like structures (ELS): by assembling embryonic and extraembryonic stem cells or by challenging embryonic stem cells (ESCs) with a pulse of WNT agonist. However, both models did not fully recapitulate early organogenesis, particularly the emergence of brain derivatives. SUMOylation was recently identified as a general barrier to cell fate transitions. Here, we show that mouse ESCs exposed to a small-molecule inhibitor of SUMOylation alone generate adherent spheroids which, once in suspension, self-organize in gastrulating structures containing cell types spatially and functionally-related to embryonic and extraembryonic compartments. Alternatively, spheroids cultured in an optimized droplet-microfluidic device form elongated ELS characterized by a multi-axial organization of the body plan reminiscent of natural embryo morphogenesis. Single-cell transcriptomics further revealed various cell types including anterior neuronal cell types, Schwann cell precursors and somites. Mechanistically, transient SUMOylation repression gradually increases the global level of DNA methylation, which in turn represses transcription of Nanog and other pluripotency-associated genes, enhancing cellular plasticity of ESCs. Our morphogen-free protocol constitutes a new facet to study regulative mechanisms of early development by targeting reprogramming roadblocks to shape multicellular architecture.

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HypoSUMOylation in embryonic stem cells generates head-and-trunk embryo-like structure [Methyl-seq]

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HypoSUMOylation in embryonic stem cells generates head-and-trunk embryo-like structure [RNA-Seq]

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Project description:Recent advances in synthetic mammalian embryo models have opened new avenues to understand the complex events controlling lineage decisions and morphogenetic processes occurring during peri-implantation and early organogenesis. Two main strategies have been developed to build embryo-like structures (ELS): by assembling extraembryonic and embryonic stem cells (ESCs) or by subjecting ESCs to various morphogens. Here, we show that mouse ESCs solely exposed to chemical inhibition of SUMOylation, a posttranslational modification which acts as a general barrier to cell-fate transitions, generates early ELSs comprising both anterior neural and trunk-associated regions. HypoSUMOylated ESCs give rise to adherent spheroids which, once in suspension, self-organize into gastrulating structures containing cell types spatially and functionally related to embryonic and extraembryonic compartments. Alternatively, spheroids cultured in an optimized droplet-microfluidic device form elongated structures characterized by a multi-axial organization reminiscent of natural embryo morphogenesis. Single-cell RNA-sequencing further revealed a variety of cellular lineages including properly positioned anterior neuronal cell types, Schwann cell precursors and paraxial mesoderm segmented into somite-like structures. Mechanistically, transient SUMOylation suppression gradually increases DNA methylation genome-wide and repressive marks deposition at Nanog, enhancing ESC plasticity. Our approach provides a proof of principle for a potential new strategy to study early embryogenesis by targeting reprogramming roadblocks to shape multicellular architecture.

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