A Cellular Anatomy of the Normal Adult Human Prostate and Prostatic Urethra

Abstract

SUMMARYA comprehensive cellular anatomy of normal human prostate is essential for solving the cellular origins of benign prostatic hyperplasia and prostate cancer. The tools used to analyze the contribution of individual cell types are not robust. We provide a cellular atlas of the young adult human prostate and prostatic urethra using an iterative process of single-cell RNA sequencing (scRNA-seq) and flow cytometry on ~98,000 cells taken from different anatomical regions. Immunohistochemistry with newly derived cell type-specific markers revealed the distribution of each epithelial and stromal cell type on whole mounts, revising our understanding of zonal anatomy. Based on discovered cell surface markers, flow cytometry antibody panels were designed to improve the purification of each cell type, with each gate confirmed by scRNA-seq. The molecular classification, anatomical distribution, and purification tools for each cell type in the human prostate create a powerful resource for experimental design in human prostate disease.

Expression ofACE2, the SARS-CoV-2 receptor, andTMPRSS2in prostate epithelial cells

“…Using a publicly-available single-cell RNA sequencing dataset, we analyzed 24,519 epithelial cells from normal human prostates for expression of ACE2 and TMPRSS2 [5].…”

“…18.65% of all epithelial cells (4,573 of 24,519), 41.74% of all stromal cells (882 of 2,113), 16.71% of endothelial cells (265 of 1,586) and 52.07% of leukocytes (239 of 459) expressed TMPRSS2. Moreover, we found 30 cells that co- Prostate club cells were found to have the greatest proportion of double-positive cells in the human prostate, and these cells bear a strong resemblance to lung club cells [5]. To characterize ACE2 and TMPRSS2 expression in prostate club cells, we compared the expression levels of these genes in lung club cells from one mouse lung [6] and four human lung [7][8][9][10] single-cell datasets ( Supplementary Table 1).…”

“…Since both of these proteins are considered to be required for infection, these double-positive cells could potentially serve as reservoirs for SARS-CoV-2 infiltration and damage to the prostate; however, they constitute approximately 0.07% of all prostate epithelial cells. Interestingly, Henry and colleagues recently characterized the prostate club cell [5] as sharing a genetic signature similar to lung club cells [6]. Therefore, in order to better understand the prostate's susceptibility to infection, we compared ACE2 and TMPRSS2 expression in both prostate and lung club cells.…”

“…Datasets were acquired using the raw count matrices made available by previous studies. For the human prostate, mouse, the Reyfman et al and the Habermann et al human lung datasets, cell types were annotated by respective cell type annotation metadata provided in the original studies and validated by top gene markers [5,6,8,9]. The cell type of the epithelial cell population in the Vieira Braga et al, dataset [7] was annotated by the top differentially expressed markers and validated by another previous study [15].…”

An immunohistochemical prostate cell identification key indicates that aging shifts procollagen 1A1 production from myofibroblasts to fibroblasts in dogs prone to prostate-related urinary dysfunction

“…[12] Careful histological and molecular analyses of human prostate have revolutionized our understanding of the cell types of the prostate, revealing a gland that contains at least five epithelial cell types and three fibroblast-like stromal cell types that are organized into three zones. [13, 14] Canine prostate cell types and their spatial distribution across the gland have not been extensively examined using modern molecular and multiplex immunohistochemical methods. This void of knowledge hinders the translational potential of canine prostate findings to human medicine in a model species that bolsters great benefit to the prostate research field.…”