MP38-11 ROLE OF MICRORNAS, INHIBITED BY TNF-?, IN SMOOTH MUSCLE REMODELING DURING OUTLET OBSTRUCTION-INDUCED LOWER URINARY TRACT DYSFUNCTION

Abstract

You have accessJournal of UrologyUrodynamics/Lower Urinary Tract Dysfunction/Female Pelvic Medicine: Basic Research & Pathophysiology II1 Apr 2018MP38-11 ROLE OF MICRORNAS, INHIBITED BY TNF-?, IN SMOOTH MUSCLE REMODELING DURING OUTLET OBSTRUCTION-INDUCED LOWER URINARY TRACT DYSFUNCTION Ali Hashemi Gheinani, Ivonne Koeck, Fiona C. Burkhard, and Katia Monastyrskaya Ali Hashemi GheinaniAli Hashemi Gheinani More articles by this author , Ivonne KoeckIvonne Koeck More articles by this author , Fiona C. BurkhardFiona C. Burkhard More articles by this author , and Katia MonastyrskayaKatia Monastyrskaya More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1238AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Dysregulation of microRNAs (miRNAs) contributes to molecular changes in the bladder wall during bladder outlet obstruction (BOO). Using next generation sequencing (NGS) of mRNAs and miRNAs in human patients′ biopsies and cell based systems we identified and validated TNF-α as the top upstream regulator of signaling in obstructed bladders. In the patients′ biopsies, the up-regulated targets of the inhibited miRNAs contributed to the majority of activated signaling pathways. Here we sought to investigate the specific contribution of TNF-α-induced and -inhibited miRNAs to aberrant signaling in the smooth muscle cells (SMC), leading to cell proliferation and bladder fibrosis. METHODS SMC cells were treated with TNF-α and RNA isolated. Transcriptome analysis performed by NGS, NanoString nCounter miRNA assays were used to profile miRNAs. miRNAs and anti-miRs were expressed in SMCs using lentiviral vectors. NFkB-luciferase assays were performed to determine the effect of miRNAs on TNF-α signaling. RESULTS Compensatory overexpression of SM-specific, TNF-α-inhibited miR-199a-5p significantly reduced NFkB-driven luciferase gene expression via down-regulation of its direct targets IKKB, MAP3K5, NFKB1 and PIK3CD. siRNA-mediated knockdown of IKKB or MAP3K5 mimicked the effect of miR-199a-5p. Transcriptome analysis of TNF-treated cells showed that 629 targets of 15 down-regulated miRs were up-regulated, whereas only 352 targets of 13 elevated miRs were decreased, indicating the higher impact of down-regulated miRNAs. 269 elevated mRNA targets of down-regulated miRNAs were implicated in regulation of 80% TNF-induced pathways. Top pathway elements were targeted specifically by miR-199a-5p, miR-424-5p and miR-149-5p. These miRNAs are reduced in BOO patients’ biopsies, making them candidates for therapeutic intervention. CONCLUSIONS Compensatory up-regulation of miR-199a-5p and other miRNAs, inhibited by TNF-α might have a beneficial effect by reducing the proliferative and fibrotic changes in BOO bladders. The importance of the targets of down-regulated miRNAs for signaling might mean that the therapeutic efforts should concentrate on finding the pharmacological ways to enhance their expression to counter-balance fibrotic bladder remodeling. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e504 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Ali Hashemi Gheinani More articles by this author Ivonne Koeck More articles by this author Fiona C. Burkhard More articles by this author Katia Monastyrskaya More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...