Stromal fibroblasts from perimenopausal endometrium exhibit a different transcriptome than those from the premenopausal endometrium†

Abstract

Human endometrium undergoes extensive regeneration on a cyclic basis in premenopausal women and likely occurs through the contribution of stem/progenitor cells. Menopause results in the permanent cessation of menstrual cycles and is preceded by perimenopause, a period of several years in which endocrine and biological changes occur and is a period of risk for endometrial proliferative disorders. The objectives of this study were to identify endometrial mesenchymal stem cells (eMSC) and endometrial stromal fibroblasts (eSF) in endometrium of perimenopausal women and perform expression profile analysis of perimenopausal eMSC and eSF to gain insight into the biology of stem/progenitor and lineage cell populations during the transition to menopause. Endometrial tissue was collected from perimenopausal and premenopausal women (n = 9 each). Microarray analysis was performed on fluorescence-activated cell sorting-isolated eSF and eMSC, and data were validated by quantitative real-time PCR. Principal component analysis showed that cells clustered into three distinct groups in 3-dimensional space: perimenopausal eMSC and premenopausal eMSC clustered together, while perimenopausal eSF and premenopausal eSF formed two discrete clusters separate from eMSC. Hierarchical clustering revealed a branching pattern consistent with principle clustering analysis results, indicating that eMSC from premenopausal and perimenopausal women exhibit similar transcriptomic signatures. Pathway analysis revealed dysregulation of cytoskeleton, proliferation, and survival pathways in perimenopausal vs. Erikson et al., 2017, Vol. 97, No. 3 premenopausal eSF. These data demonstrate that cell populations have altered gene expression in perimenopausal vs. premenopausal endometrium, and that perimenopausal eSF had altered pathway activation when compared to premenopausal eSF. This study provides insight into aging endometrium with relevance to function in reproductively older women. Summary SentenceThe hormonal milieu during the transition to menopause has an effect on endometrial stromal fibroblast gene expression and a minimal effect on the endometrial mesenchymal stem cell population, offering insight into the mechanisms by which the endometrium remains functional after menopause.

Whole-Tissue Deconvolution and scRNAseq Analysis Identify Altered Endometrial Cellular Compositions and Functionality Associated With Endometriosis

“…Finally, we validated that xCell enrichment analysis could be applied to microarray-based profiles of endometrial transcriptomes by applying the approach to artificial mixtures of microarray profiles of sorted cells. Microarray expression data from four FACS-purified endometrial cell types, endothelial cells (n=11 samples), epithelial cells (n=7), mesenchymal stem cells (n=28), and eSF (n=31) (9,(33)(34)(35), were median -summarized per gene, combined together into 20 different artificial mixtures, then analyzed with xCell (Figures 2C, D). The original method has a built-in compensation step to reduce spillover between closely related cell types.…”

“…Microarray data from purified human endometrial cell populations (stromal fibroblasts, endothelial, epithelial, and mesenchymal stem cells) isolated by fluorescence-activated cell sorting (FACS) were from previous studies: GSE73622, GSE31152, GSE48301, and GSE97163 (9,(33)(34)(35). These were used in artificial mixes of pure cell types in determining the signature specificity of the xCell signatures (see below).…”

Unique Molecular Features in High-Risk Histology Endometrial Cancers

“…Endometrial stem cells have not been studied in atrophic endometrium. However, studies suggest that mesenchymal stem cell populations change during the perimenopausal period [27].…”

Whole-tissue deconvolution and scRNAseq analysis identify altered endometrial cellular compositions and functionality associated with endometriosis

“…Finally, we validated that xCell enrichment analysis could be applied to microarray-based profiles of endometrial transcriptomes by applying the approach to artificial mixtures of microarray profiles of sorted cells. Microarray expression data from four FACS-purified endometrial cell types, endothelial cells (n=11 samples), epithelial cells (n=7), mesenchymal stem cells (n=28), and eSF (n=31) ( 8, 32–34 ) were median summarized per gene, combined together into 20 different artificial mixtures, then analyzed with xCell ( Fig. 2C, D ).…”

“…Microarray data from purified human endometrial cell populations (stromal fibroblasts, endothelial, epithelial, and mesenchymal stem cells) isolated by fluorescence-activated cell sorting (FACS) were from previous studies: GSE73622, GSE31152, GSE48301, GSE97163 ( 8, 32–34 ). These were used in artificial mixes of pure cell types in determining signature specificity of the xCell signatures (see below).…”