Single-cell RNA sequencing redefines the mesenchymal cell landscape of mouse endometrium

FASEB J. 2021 Apr;35(4):e21285. doi: 10.1096/fj.202002123R.

Abstract

The endometrium is a dynamic tissue that exhibits remarkable resilience to repeated episodes of differentiation, breakdown, regeneration, and remodeling. Endometrial physiology relies on a complex interplay between the stromal and epithelial compartments with the former containing a mixture of fibroblasts, vascular, and immune cells. There is evidence for rare populations of putative mesenchymal progenitor cells located in the perivascular niche of human endometrium, but the existence of an equivalent cell population in mouse is unclear. We used the Pdgfrb-BAC-eGFP transgenic reporter mouse in combination with bulk and single-cell RNA sequencing to redefine the endometrial mesenchyme. In contrast to previous reports we show that CD146 is expressed in both PDGFRβ + perivascular cells and CD31 + endothelial cells. Bulk RNAseq revealed cells in the perivascular niche which express the high levels of Pdgfrb as well as genes previously identified in pericytes and/or vascular smooth muscle cells (Acta2, Myh11, Olfr78, Cspg4, Rgs4, Rgs5, Kcnj8, and Abcc9). scRNA-seq identified five subpopulations of cells including closely related pericytes/vascular smooth muscle cells and three subpopulations of fibroblasts. All three fibroblast populations were PDGFRα+/CD34 + but were distinct in their expression of Ngfr/Spon2/Angptl7 (F1), Cxcl14/Smoc2/Rgs2 (F2), and Clec3b/Col14a1/Mmp3 (F3), with potential functions in the regulation of immune responses, response to wounding, and organization of extracellular matrix, respectively. Immunohistochemistry was used to investigate the spatial distribution of these populations revealing F1/NGFR + cells in most abundance beside epithelial cells. We provide the first definitive analysis of mesenchymal cells in the adult mouse endometrium identifying five subpopulations providing a platform for comparisons between mesenchymal cells in endometrium and other adult tissues which are prone to fibrosis.

Keywords: CD146; chondroitin sulfate proteoglycan 4; endometrium; fibroblast; pericyte; platelet-derived growth factor beta.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Endometrium / cytology*
  • Female
  • Gene Expression Regulation
  • Green Fluorescent Proteins
  • Homeostasis
  • Mesenchymal Stem Cells / physiology*
  • Mice
  • Receptor, Platelet-Derived Growth Factor beta / metabolism
  • Sequence Analysis, RNA
  • Single-Cell Analysis
  • Transcriptome

Substances

  • Biomarkers
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Receptor, Platelet-Derived Growth Factor beta