(113) A Novel 3D Cellular Model for Peyronie's Disease Treatment Screening: PD.SPHERE

Abstract

Abstract Introduction Peyronie's disease (PD) is a common issue that affects approximately 10% of males. Currently, researchers primarily study the pathogenesis and potential treatments for PD using monolayer fibroblast cells and a limited number of animal models. However, these models do not accurately represent the complex cellular structure and interactions involved in PD pathogenesis. We have successfully developed a three-dimensional (3D) cellular model, called PD.SPHERE, using a ground-breaking 'micro-gravity' cell culture method. This model is cultivated from extracted human PD tissues and serves as a valuable tool for in vitro treatment screening for PD. However, we have yet to use this model to evaluate the effectiveness of available and potential treatments. Objective Our objective is to determine the efficacy of the novel 3D cellular PD model, PD.SPHERE, as a treatment screening tool. Methods To achieve this, we cultured fibroblasts isolated from human PD tissues using the 'micro-gravity' approach. In this method, cells were seeded in a non-adhesive 96-well plate, placed on an orbital shaker, and incubated at 37°C with 5% CO2 and 95% relative humidity. We confirmed the proper morphology and cellular composition through microscopic assays. This 3D model was then used to assess the effects of collagenase clostridium histolyticum (CCH) and the actinidin enzyme, obtained from kiwi fruits. The PD.SPHERE was treated with actinidin at 10 mg/ml for 48 hours. We subsequently quantified collagen content using a soluble collagen quantification kit and observed viability, morphology, and changes in cellular compositions of the spheroids through immunofluorescent techniques and confocal microscopy. Results Preliminary results from our model have indicated significant changes in cellular morphology. Treatment with actinidin led to a significant decrease in spheroid size and density compared to the control group (P<0.01). The ratio of vimentin and actin filaments to nuclei staining was slightly but significantly reduced (P<0.05) in the spheroids treated with actinidin at 10 mg/ml. The cellular viability of the treated group showed a slight decrease, as revealed by Sytox Green staining. The assessment of collagen contents in the treated spheroids is ongoing. Conclusions Preliminary results suggest that our PD.SPHERE model can be effectively used to assess morphological and cellular component changes in PD tissues after treatment. Actinidin appears to compromise the plasma membrane of fibroblasts in PD.SPHERE. Our model holds promise for personalized treatment planning and potential for developing high-throughput treatment screening platforms. It also demonstrates consistency and reproducibility while closely mimicking the morphology of a PD plaque. Further research is required to collect signature gene expression data, physiological and immunological information, such as collagen production, extracellular matrix organization, and cellular expression of tight junctions, chemokines, and cytokines. These additional measures will help verify our model's pathophysiological, immunological, and treatment response aspects. Disclosure No.