Novel insights into a reputably irreversible process: combined mRNA and miRNA profiling of tissue from vesicourethral anastomotic stenosis after radical prostatectomy
Novel insights into a reputably irreversible process: combined mRNA and miRNA profiling of tissue from vesicourethral anastomotic stenosis after radical prostatectomy
复制标题
DOI：
10.1007/s00345-017-2060-2
复制
发表时间：
2017-11-01
影响因子：
3.4
通讯作者：
Pfalzgraf, D.
中科院分区：
文献类型：
--
作者：
Worst, T. S.;Daskalova, K.;Pfalzgraf, D.
直到最近，组织纤维化最终导致永久性疤痕一直被认为是一个不可逆转的过程。然而，最近的研究结果表明，它可能是可逆的。膀胱尿道吻合口狭窄是膀胱癌根治术后常见的并发症，复发率高，需要侵入性治疗。其病理生理机制尚不清楚。因此，使用nCounter技术对VUAS组织中的mRNA和miRNA转录进行了组合分析，并进行组织化学染色以评估组织形态和纤维组成。使用nCounter gene 1系统上的人miRNA panel v3和mRNA PanCancer pathway panel和qRT-PCR分析12名患者的健康和纤维化组织的RNA表达。使用实现基于R的高级途径分析工具的nSolver软件进行差异表达数据分析。miRWalk2.0用于miRNA靶点预测，VUAS组织结构线性化程度更高，胶原含量增加，弹性纤维减少。23个miRNAs和118个蛋白质编码基因在纤维化组织中差异表达（p < 0.01）。miRNA靶点预测和重叠分析表明，最强的去调控的miRNA与29个去调控的mRNA相互作用。通路分析揭示了DNA修复、细胞周期调控和TGF-β信号传导的改变。qRT-PCR证实了VUAS组织中最高水平的miRNAs和mRNAs的差异表达。在VUAS组织中，发现了mRNA和miRNAs水平的严重变化。这些一致的变化为了解根治性子宫切除术后VUAS的发病机制提供了见解，并为基于转录组学的风险分层和靶向治疗提供了未来的选择。
Until recently, tissue fibrosis-ultimately leading to permanent scaring-has been considered an irreversible process. However, recent findings indicate that it may be reversible after all. Vesicourethral anastomotic stenosis (VUAS) as fibrous narrowing is a frequent complication after radical prostatectomy with high recurrence rates and requires invasive treatment. The pathophysiology is poorly understood. Therefore, a combined mRNA and miRNA transcription profiling in tissue from VUAS was performed using nCounter technology.To assess tissue morphology and fiber composition, histochemical staining was performed. RNA expression of healthy and fibrotic tissue of twelve patients was analyzed using the human miRNA panel v3 and mRNA PanCancer pathway panel on the nCounter gene1 system and qRT-PCR. Differential expression data analysis was performed using the nSolver software implementing the R-based advanced pathway analysis tool. miRWalk2.0 was used for miRNA target prediction.More linearized tissue architecture, increased collagens, and decreased elastic fibers were observed in VUAS samples. 23 miRNAs and 118 protein coding genes were differentially expressed (p < 0.01) in fibrotic tissue. miRNA target prediction and overlap analysis indicated an interaction of the strongest deregulated miRNAs with 29 deregulated mRNAs. Pathway analysis revealed alterations in DNA repair, cell cycle regulation, and TGF-beta signaling. qRT-PCR confirmed differential expression of top deregulated miRNAs and mRNAs.In VUAS tissue, severe alterations on mRNA and miRNA level are found. These consistent changes give insights into the pathogenesis of VUAS after radical prostatectomy and point to future options for transcriptomics-based risk stratification and targeted therapies.