Presented by: Rei Unno MD,PhD
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences

Introduction

Benign prostatic hyperplasia (BPH) affects the majority of aging men and leads to lower urinary tract symptoms. Based on histological characteristics and bulk sequencing data, the pathophysiology and underlying mechanisms of BPH are poorly understood. The aim of this study was to investigate the transcriptional profile of BPH at single cell resolution to improve our understanding of the molecular features of BPH.

 

Materials

Fifteen BPH specimens were collected from patients undergoing holmium laser enucleation of the prostate. Cells from each specimen were isolated, captured, and sequenced using a bead-based single-cell RNA sequencing platform (Seq-well). Transcriptomic analysis of the gene expression matrices was performed. The absence of malignant pathologic diagnoses was independently confirmed.

Results

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16,234 cells from 15 BPH specimens were analyzed. By canonical cell type markers, we annotated three major cell types (epithelial, stromal, and immune cells). Using significantly upregulated genes in BPH compared to normal prostates previously derived from a bulk RNA sequencing comparison, bulk sequencing highlighted only the fibroblast populations. After performing a sub-clustering analysis of the 6249 epithelial cells, we identified four luminal cell subclusters showing similar levels of luminal epithelial markers KLK3, KLK2, and NKX3-1, as well as one distinct basal and one club cell population. Analysis of the differentially expressed genes among the four luminal subclusters revealed that luminal subcluster 4 was enriched in a specific signature gene set, including ribosomal genes and KLK4, which has been shown to be abundant in BPH, compared to the other subclusters. Gene ontology analysis using this gene set demonstrated that luminal subcluster 4 was associated with pathways such as oxidative phosphorylation and adipogenesis. Analyses of the BPH immune microenvironment showed a larger proportion of pro-inflammatory cells, such as M1 macrophages, compared to anti-inflammatory cells within the myeloid cell population.

Conclusion

Single cell profiling identified epithelial cells, stromal cells, and immune cells. We confirmed the upregulation of previously derived BPH-associated genes in the fibroblast population and the enrichment of pro-inflammatory myeloid cells in the immune microenvironment. Within the epithelial cell population, we identified a luminal epithelial subcluster that may represent a cell state associated with BPH.

Funding

None

Co-Authors

Heiko Yang,
Department of Urology, University of California, San Francisco

Kazumi Taguchi,
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences

Shuzo Hamamoto,
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences

Atsushi Okada,
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences

Takahiro Yasui,
Department of Nephro-urology, Nagoya City University Graduate School of Medical Sciences

Thomas Chi,
Department of Urology, University of California, San Francisco

Franklin Huang,
Department of Urology, University of California, San Francisco