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M. Ilg, M. Mateus, W. Stebbeds, A. Raheem, M. Capece, A. Parnham, G. Garaffa, A. Muneer, N. Christopher, S. Cellek, D. Ralph, HP-01-001 Development of Secondary Assays to Validate Hits from Primary Phenotypic Screen for Anti-Myofibroblast Activity in Peyronie's Disease, The Journal of Sexual Medicine, Volume 14, Issue Supplement_4a, April 2017, Page e142, https://doi.org/10.1016/j.jsxm.2017.03.024
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Objective
Peyronie’s disease (PD) is a fibrotic disorder characterised by the formation of localised fibrous plaques in the tunica albuginea (TA) of the penis, which can result in pain during erection, deformities and erectile dysfunction. PD has been affecting a growing number of men worldwide, however medical treatment for PD is currently lacking efficiency. Following the validation of our primary high-throughput screen for the detection of compounds with anti-myofibroblast activity in cells established from TA, this study aims to develop secondary screening assays to confirm the anti-myofibroblast activity.
Methods
Fibroblasts derived from patients with and without PD were exposed to TGF-β1 to induce myofibroblast transformation and cells were stained for various types of collagen and fibronectin. A modified In Cell Western (ICW) was used to measure intra- and extracellular type V collagen (Col V) staining and cell numbers, in non-PD TA cells exposed to control conditions, TGF-β1 and two FDA approved drugs. Fibroblast-populated collagen lattices (FPCLs) were used to measure differences in contraction of fibroblasts treated with drugs and under control conditions. MTT assay was performed to show the effects of the drugs on cell viability.