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Interplay between TNFa and TGF-B1 in a cell culture model of Peyronie’s Disease

Milenkovic, U1; Ilg, MM2; van Renterghem, K3; De Ridder, D1; Albersen, M1

1: KU Leuven, Belgium; 2: Anglia Ruskin University, UK; 3: University of Hasselt, Belgium

Objective: Peyronie’s disease (PD) is a sexual debilitating disease caused by an irreversible fibrotic plaque of the penile tunica albuginea (TA). Treatment is limited to surgically restoring anatomical shape. Evidence for reliable medical treatment is lacking. Our study set out to investigate the interplay between the TNFα and TGF- β1 pathways in terms of fibroblast to myofibroblast (MFB) transformation and ECM production in vitro in order elucidate better the pathophysiological pathways involved.

Materials and Methods: Primary cell culture was initiated from surgically obtained TA tissue from PD patients. To confirm fibroblast (FB)-identity cells were stained for Vimentin (Vim) using immunofluorescence (IF). To induce MFB status, cells where stimulated with 3ng/mL TGF- β1 (TBS). Increasing doses of recombinant TNFα where added (0,1-3 ng/mL). RT-qPCR was utilized to assess mRNA expression of alpha-smooth muscle actin (α-SMA), collagen I and elastin after 24h and 72h. IF was able to visualize MFB differentiation by staining for α-SMA after 72h. Resazurin-based assay was performed to assess cell viability in order to ensure anti-MFB effect of the drugs.

Results: Treatment with TNFα was able to significantly suppress the TBS-induced MFB transformation, as seen in the mRNA expression of α-SMA, which returned to non-stimulated levels after 24 and 72h. Moreover, the production of ECM products collagen I and elastin was significantly inhibited in the TNFα treatment group. Using IF, TBS cells were identified as MFB (α-SMA+, Vim+) as opposed to the non-stimulated and TNFα -treated cells (α-SMA-, Vim+). Using resazurin, cell viability remained uncompromised when administering increasing doses of TNF α.

Conclusion: PD is caused by excessive ECM production of dysregulated MFB. In the presence of TGF- β1 TNFα possesses anti-myofibroblast and anti-ECM producing properties on an mRNA and protein level. This reveals a previously undiscovered cross-talk between TNFα and TGF- β1 pathways in PD. These findings can lead to new insights concerning the pathophysiology and novel treatment strategies for PD.

Acknowledgement: Funded by European Society for Sexual Medicine, Functional Urology Fund of UZ Leuven & Fund for Peyronie’s Disease Research of UZ Leuven.


Work supported by industry: no.

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