UP-143 Finding the cause of the curve: development of an in vitro 3D Peyronie’s disease model
Thursday June 27, 2019 from
Award Winner
Luke Witherspoon, Canada has been granted the

Luke Witherspoon, Canada



The University of British Columbia


Finding the cause of the curve: Development of an in vitro 3D Peyronie’s disease model

Meghan Robinson1, Ryan Flannigan1,3,4, Luke Witherspoon1,2,3.

1Urology, Vancouver Prostate Centre, Vancouver, BC, Canada; 2Urology, Ottawa Hospital, Ottawa, ON, Canada; 3Urology, University of British Columbia, Vancouver, BC, Canada; 4Urology, Weill Cornell Medicine, New York, NY, United States

Vancouver Coastal Health Research Institute (VCHRI). Canadian Urological Association Scholarship Foundation (CUASF). Canadian Institute of Health Research (CIHR). University of British Columbia Department of Urological Sciences.

Introduction: Understanding of the pathophysiology of Peyronie’s disease is limited. A mechanistic Peyronie’s disease model remains elusive, with currently available animal models representing induced penile fibrosis, and thus do not replicate the actual disease state. This study set out to develop a 3-dimensional (3D) in vitro model to elucidate the pathogenesis of Peyronie’s disease. 

Methods : Peyronie’s plaque tissues were placed in explant culture and expanded. Early passage cells were dissociated and placed either in 6-well culture plates for 2D culture, Aggrewell800 plates to form 3D spheroids, or in a collagen-based hydrogel. To induce Peyronie’s disease pathogenesis, Transforming Growth Factor Beta (TGFβ) was added to cultures at 3 ng/mL for 48 hours. Gene expression was analysed by real time polymerase chain reaction (RT-qPCR), and changes in morphology and cell phenotype were assessed by immunocytochemistry. 

Results:  In 2D plaque cell cultures, exposure to TGFβ for 48 hours resulted in upregulation of genes associated with extracellular matrix protein production such as collagen I (COL1A1), collagen III (COL3A1), elastin (ELN) and Connective Tissue Growth Factor (CTGF) (Figure 1D). In contrast, 3D cultures displayed a more modest upregulation of COL3A1 and CTGF, but nevertheless were able to attach and spread after 48 hours, indicating increased deposition of matrix proteins (Figures 1A-C). The smooth muscle gene Alpha Smooth Muscle Actin 2 (aSMA2/ACTA2), a known marker of myofibroblasts and fibrosis, was upregulated in 2D cultures only, indicating a difference in the reactivity of the plaque cells in 3D versus 2D cultures. 

Conclusions:  This study describes two novel 3D culture models of Peyronie’s Disease and shows that the pro-fibrotic response differs in 3D environments compared to standard 2D cell culture, illustrating the importance of creating in vitro models that closely resemble the in vivo environment.

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