Integrated Single-Cell and Bulk RNA Sequencing Reveals Inflammatory Microenvironment and Glycolysis-Driven Epithelial...

Benign prostatic hyperplasia (BPH) is a prevalent condition and a leading cause of urinary dysfunction in middle-aged and elderly men. The hyperplasia of prostate stromal and epithelial cells enlarges the prostate, compressing the bladder outlet and resulting in obstructive symptoms. However, the crosstalk between the inflammatory microenvironment and metabolic alterations in BPH remains poorly understood.

This study integrated single-cell RNA sequencing and spatial transcriptomics datasets to characterize the biological and metabolic properties of BPH. Analyses were conducted using CellChat, CytoTRACE, Monocle, and SCENIC, alongside functional pathway exploration. The CARD algorithm mapped single-cell clusters onto spatial transcriptomics, enabling subtype-specific investigations of cell dependencies and pathway activities. Bulk RNA sequencing of rat prostate tissues and in vivo experiments were performed to validate these findings.

Our analysis revealed a pro-inflammatory microenvironment in BPH, marked by a higher ratio of M1-like macrophages and an enriched cell-cell interaction network that promote disease progression. Significant alterations in epithelial cells were identified, with basal epithelial (BE) cells playing a pivotal role in driving hyperplasia. SCENIC and metabolic pathway analyses demonstrated that glycolysis-mediated BE cell proliferation is a crucial mechanism in BPH progression. Bulk RNA sequencing of rat prostate tissues further confirmed glycolysis pathway enrichment in BPH models. Notably, glycolysis inhibitors demonstrated therapeutic effects from both macroscopic and histological perspectives in BPH rat models.

This study provides a comprehensive analysis of the inflammatory microenvironment and metabolic characteristics of BPH and identifies glycolysis-mediated proliferation of basal epithelial cells as a key driver of disease progression. Pharmacological inhibition of glycolysis with 2-deoxy-D-glucose reduced prostate weight, prostate index, epithelial proliferation and M1-like macrophage infiltration in vivo, indicating that targeting glycolytic metabolism is a promising therapeutic strategy for BPH. These findings offer novel insights into BPH pathogenesis and provide a rationale for developing metabolism-directed, and potentially locally delivered, interventions that could benefit patients who respond poorly to current medical therapy.