Single cell RNA-seq study of wild type and Hox9,10,11 mutant developing uterus

Abstract

The uterus is a remarkable organ that must guard against infections while maintaining the ability to support growth of a fetus without rejection. The Hoxa10 and Hoxa11 genes have previously been shown to play essential roles in uterus development and function. In this report we show that the Hoxa9,10,11, Hoxc9,10,11, Hoxd9,10,11 genes play a redundant role in the formation of uterine glands. In addition, we use single cell RNA-seq to create a high resolution gene expression atlas of the developing wild type mouse uterus. Cell types and subtypes are defined, for example dividing endothelial cells into arterial, venous, capillary, and lymphatic, while epithelial cells separate into luminal and glandular subtypes. Further, a surprising heterogeneity of stromal and myocyte cell types are identified. Transcription factor codes and ligand/receptor interactions are characterized. We also used single cell RNA-seq to globally define the altered gene expression patterns in all developing uterus cell types for two Hox mutants, with 8 or 9 mutant Hox genes. The mutants show a striking disruption of Wnt signaling as well as the Cxcl12/Cxcr4 ligand/receptor axis.

Identification of the lymphangioleiomyomatosis cell and its uterine origin

“…An understanding of the source of cells that infiltrate the lung has remained elusive, however, as have the mechanisms of tumorigenesis, metastasis, invasion, and cystic remodeling Krymskaya and McCormack, 2017;McCormack et al, 2012). In the present study, scRNA-seq analysis of LAM lesions identified unique, non-pulmonary mesenchymal cells, which we termed LAM CORE cells, that share a high degree of transcriptomic similarity to the LAM CORE cells in uterus of an S-LAM patient and with myometrial and stromal cells (express high level of MME, FN1, COL3A1, HOXA10) from normal human and mouse uteri (Mucenski et al, 2019;Wu et al, 2018) . A prominent neural crest signature was also identified in pulmonary and uterine LAM CORE cells and in normal uterus.…”

“…In addition, uterine myocyte, epithelial, immune, endothelial and several stromal cell subtypes were identified (Figure 6a; Figure S15). Endometrial epithelia (expressing high level of KRT8, KRT18, EPCAM, CLDN3) and stromal cells (expressing high level of MME, FN1, COL3A1, HOXA10, VIM ) were defined based on the gene markers from the human and mouse uterus altas (Mucenski et al, 2019;Wu et al, 2018) . Hierarchical clustering of major cell types from LAM lung and LAM uterus demonstrated close correlation of LAM CORE cells from both tissues (Figure 6b).…”

“…Hierarchical clustering of major cell types from LAM lung and LAM uterus demonstrated close correlation of LAM CORE cells from both tissues (Figure 6b). Uterine and neural crest associated genes identified in pulmonary LAM CORE cells were compared using scRNA-seq data from the human LAM uterus and normal human/mouse uterus (Mucenski et al, 2019) . Most neural crest and uterine associated genes selectively expressed in pulmonary LAM CORE cells were expressed in the LAM cell cluster from the human LAM uterus (Figure 6a; Figure S15).…”

Uterine Patterning, Endometrial Gland Development, and Implantation Failure in Mice Exposed Neonatally to Genistein

“…AP differentiation is driven by Hoxa9 in the oviduct, Hoxa10 in the uterus, Hoxa11 in the lower uterus and cervix, and Hoxa13 in the cervix and upper vagina (Du and Taylor 2015;Taylor et al 1997). Hoxc9-11 and Hoxd9-11 have some functional redundancy with Hoxa9-11 (Du and Taylor 2015;Mucenski et al 2019;Raines et al 2013). Disruption of Hoxa10 or combined disruption of Hoxa9-11 causes posteriorization of the reproductive tract with the anterior movement of stratified epithelium, including basal cells and extracellular matrix, from the cervix and upper vagina up into the uterus (Benson et al 1996;Mucenski et al 2019;Raines et al 2013).…”

“…Hoxc9-11 and Hoxd9-11 have some functional redundancy with Hoxa9-11 (Du and Taylor 2015;Mucenski et al 2019;Raines et al 2013). Disruption of Hoxa10 or combined disruption of Hoxa9-11 causes posteriorization of the reproductive tract with the anterior movement of stratified epithelium, including basal cells and extracellular matrix, from the cervix and upper vagina up into the uterus (Benson et al 1996;Mucenski et al 2019;Raines et al 2013). Neonatal GEN-exposed mice exhibited all of these characteristics and, in the uterus, expressed genes normally restricted to the cervix and upper vagina, including Trp63, Krt14, and Six1 (Suen et al 2016.…”