(058) Detection and Transcriptional Profiling of Mesenchymal Stem Cells in Peyronie’s Plaque Explants

Abstract

Abstract Introduction The mechanisms behind the pathogenesis of Peyronie’s disease largely remain a mystery. Plaques have largely been thought to be derived from epithelial to mesenchymal transformation of fibroblasts to myofibroblasts. Objective We sought to transcriptomically profile explanted cells from Peyronie’s plaques and evaluate their profiles with transforming growth factor B (TGF-B) stimulation. Methods Peyronie’s plaque were excised from 3 patients undergoing penile incision and grafting surgery. Cells were explanted from biopsy tissue and propagated. Cells were profiled using single cell RNA sequencing (scRNAseq). These cells were then exposed to TGF-□ to simulate the active phase of Peyronie’s disease, and profiled with scRNAseq at baseline and 24 hours following TGF-□□ stimulation. We used principle component analysis (PCA) to reduce the dimension of data and built the nearest-neighbors graph using PCA weights. We performed clustering over the nearest-neighbor graph and visualized the clusters using UMAP algorithm. Differential gene expression and pathway analyses were performed using the top 50 genes with increased expression between 0- and 24-hour time points. Multi-channel flow cytometry and immunofluorescence were used for mesenchymal stem cell (MSC) phenotypic validation. Results Assessment of cell clustering identified profiles of fibroblasts, myofibroblasts, smooth muscle, and mesenchymal stem-like cells. We further confirmed rare presence of mesenchymal stem cells using multi-channel flow cytometry in our cultured cells and plaque biopsies. Differential gene expression of the 50 genes showing the greatest induction showed that genes involved in cellular binding and catalytic activity were most commonly upregulated following introduction of TGF-B. Conclusions This analysis provides unique insight into the expression profiles of Peyronie’s plaque cell populations explanted in culture and stimulated via TGF-B to activate myofibroblasts akin to the active phase of Peyronie’s disease. Using the information gained from this single-cell RNA sequencing analysis ongoing efforts can be performed to identify therapeutic targets for active phase Peyronie’s disease. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Coloplast; Boston; Teumo Health Technologies.