Silencing histone deacetylase 2 using small hairpin RNA induces regression of fibrotic plaque in a rat model of Peyronie's disease

Abstract

ObjectivesTo examine the therapeutic effect of adenovirus encoding histone deacetylase 2 (HDAC2) small hairpin RNA (Ad-HDAC2 shRNA) in a rat model of Peyronie's disease (PD) and to determine the mechanisms by which HDAC2 knockdown ameliorates fibrotic responses in primary fibroblasts derived from human PD plaque. Materials and MethodsRats were distributed into four groups (n = 6 per group): age-matched controls without treatment; rats in which PD has been induced (PD rats) without treatment; PD rats receiving a single injection of control adenovirus encoding scrambled small hairpin RNA (Ad-shRNA) (day 15; 1 × 10 8 pfu/0.1 mL phosphate-buffered saline [PBS]); and PD rats receiving a single injection of Ad-HDAC2 shRNA (day 15; 1 × 10 8 pfu/0.1 mL PBS) into the lesion. PD-like plaque was induced by repeated intratunical injections of 100 μL each of human fibrin and thrombin solutions on days 0 and 5. On day 30, the penis was harvested for histological examination. Fibroblasts isolated from human PD plaque were pretreated with HDAC2 small interfering (si)RNA (100 pmoL) and then stimulated with transforming growth factor (TGF)-β1 (10 ng/mL) to determine hydroxyproline levels, procollagen mRNA, apoptosis and protein expression of poly(ADP-ribose) polymerase 1 (PARP1) and cyclin D1. ResultsWe observed that Ad-HDAC2 shRNA decreased inflammatory cell infiltration, reduced transnuclear expression of phospho-Smad3 and regressed fibrotic plaque of the tunica albuginea in PD rats in vivo. siRNA-mediated silencing of HDAC2 significantly decreased the TGF-β1-induced transdifferentiation of fibroblasts into myofibroblasts and collagen production, and induced apoptosis by downregulating the expression of PARP1, and decreased the expression of cyclin D1 (a positive cell-cycle regulator) in primary cultured fibroblasts derived from human PD plaque in vitro. ConclusionSpecific inhibition of HDAC2 with RNA interference may represent a novel targeted therapy for PD.

The Genetic Basis of Peyronie Disease: A Review

“…95 The authors observed that PD model rats receiving injections of Ad-HDAC2 shRNA had regression of fibrotic penile plaque, fewer intralesional inflammatory cells, impaired nuclear translocation of phosphorylated Smad3, inhibited differentiation of fibroblasts into myofibroblasts, and reduced collagen production. Furthermore, cultured rat fibroblasts from PD plaques exhibited reduced expression of cyclin D1, which blocked cell cycle entry and a propensity towards apoptosis in both basal and TGFβ-stimulated conditions.…”

Inhibition of penile tunica albuginea myofibroblasts activity by adipose‑derived stem cells

“…Myofibroblasts was the most important cell in PD plaque and was derived from transformation of penile TAFs, which was the largest class of cells that make up the normal TA (42). The persistence activation of myofibroblasts facilitated the progress of fibrosis, which led the penile structural remodeling.…”

A positive readout single transcript reporter for site-specific mRNA cleavage

“…The reporter can also be used to fine-tune the efficacy of synthetic short hairpin RNA (s-shRNA) (Stevenson et al, 2013). RNA interference is a potent technology to repress disease-causing genes by degrading their mRNAs with s-shRNA (Davidson & McCray, 2011; Kwon et al, 2014; Yoo et al, 2007). Both s-shRNAs and en-siRNAs are loaded into Ago2-RISC and can guide specific endonucleolytic cleavage of RNA molecules.…”