Single cell RNA sequencing and lineage tracing confirm mesenchyme to epithelial transformation (MET) contributes to repair of the endometrium at menstruation
- University of Edinburgh, United Kingdom
Abstract
The human endometrium experiences repetitive cycles of tissue wounding characterised by piecemeal shedding of the surface epithelium and rapid restoration of tissue homeostasis. In this study we used a mouse model of endometrial repair and three transgenic lines of mice to investigate whether epithelial cells that become incorporated into the newly formed luminal epithelium have their origins in one or more of the mesenchymal cell types present in the stromal compartment of the endometrium. Using scRNAseq we identified a novel population of PDGFRb+ mesenchymal stromal cells that developed a unique transcriptomic signature in response to endometrial breakdown/repair. These cells expressed genes usually considered specific to epithelial cells and in silico trajectory analysis suggested they were stromal fibroblasts in transition to becoming epithelial cells. To confirm our hypothesis we used a lineage tracing strategy to compare the fate of stromal fibroblasts (PDGFRa+) and stromal perivascular cells (NG2/CSPG4+). We demonstrated that stromal fibroblasts can undergo a mesenchyme to epithelial transformation and become incorporated into the re-epithelialised luminal surface of the repaired tissue. This study is the first to discover a novel population of wound-responsive, plastic endometrial stromal fibroblasts that contribute to the rapid restoration of an intact luminal epithelium during endometrial repair. These findings form a platform for comparisons both to endometrial pathologies which involve a fibrotic response (Asherman’s syndrome, endometriosis) as well as other mucosal tissues which have a variable response to wounding.
Data availability
Single cell RNAseq datasets have been deposited in GEO under accession codes GSE198556
Article and author information
Author details
Philippa TK Saunders
Funding
Medical Research Council (MR/N013166/1)
- Phoebe M Kirkwood
Medical Research Council (MR/N024524/1)
- Phoebe M Kirkwood
- Douglas A Gibson
- Isaac Shaw
Wellcome Trust (219542/Z/19/Z)
- Neil C Henderson
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Ricardo Azziz, University at Albany, SUNY, United States
Ethics
Animal experimentation: All animal experiments were performed under a license granted by the UK Home Office (PPL 70/8945) and were approved by the University of Edinburgh Animal Welfare and Ethical Review Body.
Version history
- Preprint posted: December 21, 2021 (view preprint)
- Received: February 7, 2022
- Accepted: December 15, 2022
- Accepted Manuscript published: December 16, 2022 (version 1)
- Version of Record published: January 24, 2023 (version 2)
Copyright
© 2022, Kirkwood 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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Single cell RNA sequencing and lineage tracing confirm mesenchyme to epithelial transformation (MET) contributes to repair of the endometrium at menstruation
eLife 11:e77663.
https://doi.org/10.7554/eLife.77663
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