Presentation Authors: Gervaise Henry, Alicia Malewska, Dallas, TX, Diya Joseph, Madison, WI, Venkat Malladi, Ryan Mauck, Jeffrey Gahan, Ganesh Raj, Claus Roehrborn, Ryan Hutchinson, Dallas, TX, Chad Vezina, Madison, WI, Douglas Strand*, Dallas, TX
Introduction: The cellular pathogenesis of human benign prostatic hyperplasia is poorly understood. To understand the functional contribution of specific cell types in disease, a cellular atlas of the normal organ was created using single cell RNA sequencing (scRNA-seq) and immunofluorescence. These data were then used as a control to characterize cellular alterations in BPH.
Methods: BPH tissue from three simple prostatectomy patients and normal prostate specimens from three young organ donors were digested into single cell suspensions. scRNA-seq was performed to derive a molecular profile of each cell type. Unsupervised clustering objectively identified known and novel cell types and newly derived molecular markers were used to 1) improve the flow cytometry-based isolation of individual cell types for functional analysis and 2) provide immunohistochemical confirmation of anatomical position in whole mount specimens. Bioinformatics analysis was used to derive differentially expressed genes for each cell type in normal prostate vs. BPH. Immunohistochemical analysis was used to confirm changes in cellular composition.
Results: We identified two stromal cell types (DCN+ fibroblasts and MYH11+ smooth muscle) and five epithelial cell types (KRT14+ basal, KLK3+ luminal, and CHGA+ neuroendocrine, KRT13+ hillock, and SCGB1A1+ club) in silico. Whole mount immunohistochemical analysis of KRT13+ and SCGB1A1+ epithelial cell types revealed their enrichment in the prostatic urethra and proximal prostatic ducts and their expansion in glandular BPH. Fibroblasts were enriched in the anterior fibromuscular stroma and peri-urethral regions of the normal prostate and were expanded around the urethra in BPH with stromal hyperplasia. M2 macrophages, mast cells, and T cells were enriched in BPH.
Conclusions: An unbiased transcriptional identity of each cell type in the normal prostate is required for understanding functional interactions between cell types in homeostasis and disease. Our study provides the resources to identify, localize, and isolate every cell type in the human prostate. Our identification of novel epithelial cell types concentrated in the urethra and spread throughout the prostate could lead to a deeper understanding of the zonal anatomy and cellular origin of human prostate diseases. Our cell type-specific transcriptional profiles could be used to design new therapeutic agents in BPH.
Source of Funding: Financial support came from K01 DK098277, R03 DK110497, R01 DK115477, and U54DK104310 pilot program award (D.W.S.); U54DK104310, U01DK110807 and R01DK099328 (C.M.V); V.S.M. is supported by a grant from the Cancer Prevention Research Institute of Texas, or