Phenotyping of immune and endometrial epithelial cells in endometrial carcinomas revealed by single-cell RNA sequencing

Abstract

Tumors are complex ecosystems harboring multiple cell types which might play a critical role in tumor progression and treatment response. The endometrial epithelial cell identities and immune microenvironment of endometrial carcinoma (ECC) are poorly characterized. In this study, a cellular map of endometrial carcinoma was generated by profiling 30,780 cells isolated from tumor and paratumor tissues from five patients using single-cell RNA sequencing. 7 cell types in lymphocytes, 7 types in myeloid cells and 3 types in endometrial epithelial cells were identified. Distinct CD8 + T cell states and different monocyte-macrophage populations were discovered, among which exhausted CD8 + T cells and macrophages were preferentially enriched in tumor. Both CD8 + T cells and macrophages comport with continuous activation model. Gene expression patterns examination and gene ontology enrichment analysis of endometrial epithelial cells revealed 3 subtypes: stem-like cells, secretory glandular cells and ciliated cells. Overall, our study presents a view of endometrial carcinoma at single-cell resolution that reveals the characteristics of endometrial epithelial cells in the endometrium, and provides a cellular landscape of the tumor immune microenvironment.

The Clinicopathological Characteristics, Prognosis and Immune Microenvironment Mapping in MSI-H/MMR-D Endometrial Carcinomas

“…Our previous study [15] also indicated high lymphocytes in ltration in an MMR-D cancer tissue whose MSH6 expression is negative (Figure S2A). The existence of different tumor-in ltrating immune cell types in this sample was also assessed by immunohistochemistry (IHC) staining with CD3, CD8, CD20, CD68 and CD163 antibody (Figure 5A).…”

“…Single-cell RNA-seq data of EC5-T in PRJNA650549 were downloaded from the SRA database (https://www.ncbi.nlm.nih.gov/sra/PRJNA650549). We used the Cell Ranger software pipeline (version 2.2.0, 10xGenomics) to process raw sequencing data and Seurat (version 2.3.4) R package for downstream analysis as previously described [15]. Brie y, principle component analysis (PCA) was performed for dimensional reduction.…”

The clinicopathological characteristics, prognosis and immune microenvironment mapping in MSI-H/MMR-D endometrial carcinomas

“…Following resection, a representative tumor fragment was isolated and transferred rapidly to the laboratory for study as previously described [ 20 ]. Briefly, tissue was initially cut into segments and subjected to digestion by collagenase type I/II (Thermo Fisher Scientific, USA) and DNAse I (Sigma, USA).…”

“…Our previous study also indicated higher lymphocytes infiltration in an MMR-D cancer tissue compared to other (MMR-I) samples [20]. To gain deeper insight into the immune cellular composition and phenotypic diversity of MMR-D endometrial cancer, we profiled 3371 cells obtained from an MMR-D EC sample (labeled "EC-MSI") using the single-cell RNA-seq (10X Genomics platform).…”

Analyzing Modern Biomolecules: The Revolution of Nucleic-Acid Sequencing – Review

“…That allows an unprecedented dissection of transcription within millions of individual cells. Both single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) have exciting applications [65][66][67][68][69][70][71][72][73][74][75][76][77], for instance: (i) discovering and characterizing cell type in health and diseases, such as cancer [13,[78][79][80][81][82][83][84][85][86][87][88][89], with implications in immunology [90], immune-mediated diseases [91], immunotherapy [92][93][94][95][96][97][98][99][100] and drug resistance [101]; (ii) deciphering the roles of such specific cell types in health and disease [102], including mitochondrial heteroplasmy [33]; and (iii) analyzing cell emergence, development and plasticity in tissues and organisms. These studies are also applied to study plant biology [103,104].…”