Presentations

Abstract

Introduction:

Benign prostatic hyperplasia (BPH) is a significant health concern in the geriatric male population, primarily due to its effect on the quality of life through urinary symptoms, encompassing increased frequency and hesitancy. Furthermore, BPH introduces the potential for medical complications, including urinary tract infections and nephropathy. BPH pathology is influenced by steroid hormonal imbalance, which is depicted by the testosterone and 17β-estradiol (T+E2) mouse model. This model showcases enlarged bladders, narrowed urethral lumens, and increased prostate mass - all hallmarks of human BPH, making this model a valuable platform for exploring disease mechanisms. Recent studies in our lab have highlighted that macrophages infiltrating the glandular lumen due to steroid hormone imbalance accumulate lipid droplets, adopting a foam cell phenotype. We hypothesized that foam cells secrete cytokines that contribute to BPH. Accordingly, this study focused on validating the expressional alterations of foam-cell derived cytokines, Tgfb1, Ccl6, and Cxcl16, previously identified via single-cell RNA sequencing.

Methods:

Male C57BL/6J mice were subcutaneously implanted with pellets containing 25 mg testosterone (T), and 2.5 mg estradiol (E2), and their prostates were collected after two weeks. Paraffin- embedded tissues from the ventral prostate were subjected to in-situ hybridization (RNAScopeTM) using probes targeting Tgfb1, Ccl6, and Cxcl16. Tissue samples were analyzed using a Mantra II. Pathological Workstation and InForm software. Student's t-test and one-way ANOVA or their non-parametric equivalent were used for statistical analysis.

Results:

We found increased expressional changes of Tgfb1, Ccl6, and Cxcl16 in foam cells in T+E2 tissues. Furthermore, when quantifying the expressional changes in the tissue, Tgfb1 expression was dramatically upregulated by 10.07-fold in T+E2 tissues compared to the control group. Likewise, Ccl6 and Cxcl16 -positive cells, likely macrophages, demonstrated a 3.21 and 1.96-fold increase in T+E2 tissues compared to the controls, respectively.

Conclusion:

This study corroborates prior scRNA sequencing studies, affirming the elevated expression of Tgfb1, Ccl6, and Cxcl16 in foam cells. The pronounced expression of these factors suggests an active role of foam-cell-derived cytokines in stimulating an inflammatory response in the prostate, consequently contributing to BPH.