Urology Research Laboratory, Department of Clinical Research, University of Bern, Bern, Switzerland.
Am J Pathol. 2013 Feb;182(2):431-48. doi: 10.1016/j.ajpath.2012.10.020. Epub 2012 Nov 29.
Defects in urothelial integrity resulting in leakage and activation of underlying sensory nerves are potential causative factors of bladder pain syndrome, a clinical syndrome of pelvic pain and urinary urgency/frequency in the absence of a specific cause. Herein, we identified the microRNA miR-199a-5p as an important regulator of intercellular junctions. On overexpression in urothelial cells, it impairs correct tight junction formation and leads to increased permeability. miR-199a-5p directly targets mRNAs encoding LIN7C, ARHGAP12, PALS1, RND1, and PVRL1 and attenuates their expression levels to a similar extent. Using laser microdissection, we showed that miR-199a-5p is predominantly expressed in bladder smooth muscle but that it is also detected in mature bladder urothelium and primary urothelial cultures. In the urothelium, its expression can be up-regulated after activation of cAMP signaling pathways. While validating miR-199a-5p targets, we delineated novel functions of LIN7C and ARHGAP12 in urothelial integrity and confirmed the essential role of PALS1 in establishing and maintaining urothelial polarity and junction assembly. The present results point to a possible link between miR-199a-5p expression and the control of urothelial permeability in bladder pain syndrome. Up-regulation of miR-199a-5p and concomitant down-regulation of its multiple targets might be detrimental to the establishment of a tight urothelial barrier, leading to chronic pain.
尿路上皮完整性的缺陷导致渗漏和潜在感觉神经的激活,这可能是膀胱疼痛综合征的致病因素,这是一种在没有特定原因的情况下出现骨盆疼痛和尿急/尿频的临床综合征。在此,我们确定 microRNA miR-199a-5p 是细胞间连接的重要调节剂。在尿路上皮细胞中过表达时,它会损害正确的紧密连接形成,并导致通透性增加。miR-199a-5p 直接靶向编码 LIN7C、ARHGAP12、PALS1、RND1 和 PVRL1 的 mRNA,并在相似程度上降低它们的表达水平。使用激光微切割,我们表明 miR-199a-5p 主要在膀胱平滑肌中表达,但也在成熟的膀胱尿路上皮和原代尿路上皮培养物中检测到。在尿路上皮中,cAMP 信号通路的激活可以上调其表达。在验证 miR-199a-5p 靶标时,我们描绘了 LIN7C 和 ARHGAP12 在尿路上皮完整性中的新功能,并证实了 PALS1 在建立和维持尿路上皮极性和连接组装中的重要作用。这些结果表明 miR-199a-5p 表达与膀胱疼痛综合征中尿路上皮通透性的控制之间可能存在联系。miR-199a-5p 的上调和其多个靶标的下调可能对建立紧密的尿路上皮屏障有害,导致慢性疼痛。