Single Cell Insights on Inflammation-Induced Transitional States in Benign Prostate Hyperplasia 2253526

Abstract Introduction Benign prostatic hyperplasia (BPH) is an age-related prostate enlargement involving epithelial and stromal expansion. Inflammation drives BPH, but its impact on epithelial lineage hierarchies, especially transit-amplifying (TA) states with basal and luminal features, is unclear. We hypothesized inflammation induces epithelial transitions that promote basal-to-luminal differentiation contributing to prostate growth. Methods Using the POET3 inducible prostatitis murine model, inflammation was triggered by OT-1 T cells. Benign prostate stem cells (bPSCs, n = 11) from naïve and inflamed prostates (3 technical replicates/condition) were isolated by FACS and profiled by single-cell RNA sequencing (scRNA-seq, SMARTer, Illumina). Data underwent QC, Seurat integration, cluster annotation, marker analysis, and gene set enrichment. Integrated analysis with an AIRE_KO chronic inflammation dataset assessed conserved versus model-specific responses. Results We identified basal and luminal epithelial populations, with inflammation promoting intermediate states expressing basal (Krt5, Krt14, Trp63) and luminal (Krt8, Krt18) markers. Inflamed cells showed increased proliferation, altered androgen receptor signaling, and elevated pro-inflammatory cytokines (Il1a, Il1b, Ccl2, CxCl2) with EMT-related changes. Integration with chronic inflammation data revealed both conserved and model-specific transcriptional programs, highlighting epithelial heterogeneity. Conclusion This study reveals inflammation-induced intermediate epithelial states and disrupted lineage hierarchies contributing to BPH. These findings could guide therapeutic targeting of inflammatory signaling and epithelial remodeling in BPH and prostate diseases. Funding Source NIH R01DK126478 (PI: Dr. Ratliff) NIH R21ActfAI151723 (PI: Dr. Ratliff) Society for Basic Urologic Research (SBUR) Topic Categories Translational and Interventional Immunology (TI)