Transcriptional heterogeneity and cell cycle regulation as central determinants of primitive endoderm priming
- University of Copenhagen, Denmark
Abstract
During embryonic development cells acquire identity at the same time as they are proliferating, implying that an intrinsic facet of cell fate choice requires coupling lineage decisions to rates of cell division. How is the cell cycle regulated to promote or suppress heterogeneity and differentiation? We explore this question combining time lapse imaging with single cell RNA-seq in the contexts of self-renewal, priming and differentiation of mouse embryonic stem cells (ESCs) towards the Primitive Endoderm lineage (PrE). Since ESCs are derived from the Inner Cell Mass of the mammalian blastocyst, ESCs in standard culture conditions are transcriptionally heterogeneous containing subfractions that are primed for either of the two ICM lineages, Epiblast and PrE. These subfractions represent dynamic states that can readily interconvert in culture, and the PrE subfraction is functionally primed for endoderm differentiation. Here we find that differential regulation of cell cycle can tip the balance between these primed populations, such that naïve ESC culture conditions promote Epiblast-like expansion and PrE differentiation stimulates the selective survival and proliferation of PrE-primed cells. In endoderm differentiation, we find that this change is accompanied by a counter-intuitive increase in G1 length that also appears replicated in vivo. While FGF/ERK signalling is a known key regulator of ESCs and PrE differentiation, we find it is not just responsible for ESCs heterogeneity, but also cell cycle synchronisation, required for the inheritance of similar cell cycles between sisters and cousins. Taken together, our results point to a tight relationship between transcriptional heterogeneity and cell cycle regulation in the context of lineage priming, with primed cell populations providing a pool of flexible cell types that can be expanded in a lineage-specific fashion while allowing plasticity during early determination.
Data availability
The sc-RNAseq data used in this study has been deposited in the Gene Expression Omnibus and are available under the accession number GSE200534. Previously published Nowotschin et al., 2019 data that were used here are available under accession number GSE123046.
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Transcriptional Heterogeneity and Cell Cycle Regulation as Central Determinants of Primitive Endoderm PrimingNCBI Gene Expression Omnibus, GSE200534.
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The Emergent Landscape of the Mouse Gut Endoderm at Single-Cell ResolutionNCBI Gene Expression Omnibus, GSE123046.
Article and author information
Author details
Funding
Lundbeckfonden (R198-2015-412)
- Joshua M Brickman
Danish Agency for Science and Higher Education (DFF- 8020-00100B)
- Joshua M Brickman
Danish National Research Foundation (DNRF116)
- Joshua M Brickman
Lundbeckfonden (R286-2018-1534)
- Marta Perera
Lundbeckfonden (R303-2018-2939)
- Rita Soares Monteiro
Danish National Research Foundation (DNRF116)
- Ala Trusina
Novo Nordisk Fonden (NNF21CC0073729)
- Joshua M Brickman
Novo Nordisk Fonden (NNF17CC002785)
- Joshua M Brickman
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2022, Perera et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Transcriptional heterogeneity and cell cycle regulation as central determinants of primitive endoderm priming
eLife 11:e78967.
https://doi.org/10.7554/eLife.78967
