TNF-a initiates miRNA-mRNA signaling cascades leading to loss of contractility in obstruction-induced bladder dysfunction

Bladder outlet obstruction (BOO), and clinical lower urinary tract (LUT) dysfunction are common in elderly patients. BOO is accompanied by urodynamic changes in bladder function and leads to organ fibrosis and loss of contractility. Comprehensive transcriptome analysis of bladder samples from human patients with different urodynamically-defined states of BOO revealed TNF- a as top upstream signaling pathway regulator. Here we validated the next-generation sequencing and pathway analysis in cell-based models using bladder smooth muscle (SM) and urothelial cells exposed to TNF-a. MicroRNA profiling and transcriptome analysis of TNF-treated bladder smooth muscle cells (SMCs) revealed striking similarities with human BOO. Using the comparative approach, we delineated the TNF-specific and -independent pathways in the human BOO samples. Concomitant down-regulation of SMC-specific miRNAs and SM markers after TNF-a treatment is in accordance with the loss of contractility in human patients in advanced obstruction-induced organ remodeling. We modulated the expression levels of four abundant TNF-regulated miRNAs, and showed that compensatory up-regulation of miR-199a-5p reduced NF-?B signaling. Important hubs of TNF signaling pathways MAP3K5 (ASK1), and IKBKB (IKKß) were targeted by miR-199a-5p. MiR-199a-5p might be part of a negative feedback loop, reducing the impact of TNF while its down-regulation in acontractile bladders of BOO patients advances the disease. The compensatory up-regulation of miR-199a-5p together with TNF-a inhibition might bring therapeutic benefits.