SFRP4+ stromal cell subpopulation with IGF1 signaling in human endometrial regeneration

Abstract

Our understanding of full-thickness endometrial regeneration after injury is limited by an incomplete molecular characterization of the cell populations responsible for the organ functions. To help fill this knowledge gap, we characterized 10,551 cells of full-thickness normal human uterine from two menstrual phases (proliferative and secretory phase) using unbiased single cell RNA-sequencing. We dissected cell heterogeneity of main cell types (epithelial, stromal, endothelial, and immune cells) of the full thickness uterine tissues, cell population architectures of human uterus cells across the menstrual cycle. We identified an SFRP4+ stromal cell subpopulation that was highly enriched in the regenerative stage of the human endometria during the menstrual cycle, and the SFRP4+ stromal cells could significantly enhance the proliferation of human endometrial epithelial organoid in vitro, and promote the regeneration of endometrial epithelial glands and full-thickness endometrial injury through IGF1 signaling pathway in vivo. Our cell atlas of full-thickness uterine tissues revealed the cellular heterogeneities, cell population architectures, and their cell–cell communications during the monthly regeneration of the human endometria, which provide insight into the biology of human endometrial regeneration and the development of regenerative medicine treatments against endometrial damage and intrauterine adhesion.

Single-cell RNA sequencing and lineage tracing confirm mesenchyme to epithelial transformation (MET) contributes to repair of the endometrium at menstruation

“…Comparison between the genes that defined each subpopulation of stromal cells (GFBP1, MGP, IL11, SRGN, HSPA6) with our current dataset did not find any that corresponded to F4 which is probably unsurprising as these signatures were based on shed cells not those that remained in situ and therefore able to contribute to the new luminal epithelium. In contrast, we did find some homologies within the dataset published by Wu et al, 2022 whose analysis included single-cell sequencing of full thickness endometrium from the proliferative and secretory phases (no menstrual tissue). Their stromal cell subclusters included one enriched for SFRP4 which they also showed provided a positive stimulus to regeneration in a rat endometrial injury model.…”

“…The generation of these scRNAseq datasets have allowed us to make some preliminary comparisons to results from studies using this technique to explore cell heterogeneity in human endometrium and human endometrial cells ( Wang et al, 2020 ; Queckbörner et al, 2021 ; Cao et al, 2021 ; Garcia-Alonso et al, 2021 ; Shih et al, 2022 ; Wu et al, 2022 ). Queckborner and colleagues focused their analysis cells from 3 donors obtained during in the proliferative phase of their menstrual cycle (6864 cells) using a protocol that enriched for stromal cells ( Queckbörner et al, 2021 ).…”

Bi-potential hPSC-derived Müllerian duct-like cells for full-thickness and functional endometrium regeneration

“…5d ). A total of 4022 human-derived cells were captured, integrated, and comparatively analysed with the normal human endometrium in the secretory phase (NES) dataset (public database of the Genome Sequence Archive for Humans under the accession number HRA000928) 42 . Hierarchical clustering and scRNA-seq data analysis with uniform manifold approximation and projection (UMAP) identified six cell clusters (clusters 0–5) (Fig.…”

“…The cell source of primary endometrial cells-derived organoids was from the non-pathological part of endometrium from hysterectomy due to leiomyoma in patient, which was approved by the ethical committee of the first affiliated hospital, school of medicine, Zhejiang university (ethics approval No. 2018-113) 42 .…”

“…Sequencing reads were aligned to the human reference genome GRCh38 using the Cell Ranger suite with default parameters. And the data (HRA002267) was integrated and comparatively analysed with the human normal endometrium in secretory phase (NES) dataset (public database of the Genome Sequence Archive for Human under the accession of HRA000928) 42 . Hierarchical clustering and violin plots were generated in Seurat ( http://satijalab.org/seurat/ ) 63 , and UMAP was performed to visualise cells in a two-dimensional space.…”