Table2_Multi-Omics Characterization of Circular RNA-Encoded Novel Proteins Associated With Bladder Outlet Obstruction.XLSX

Bladder outlet obstruction (BOO) is a common urologic disease associated with poorly understood molecular mechanisms. This study aimed to investigate the possible involvements of circRNAs (circular RNAs) and circRNA-encoded proteins in BOO development. The rat BOO model was established by the partial bladder outlet obstruction surgery. Differential expression of circRNA and protein profiles were characterized by deep RNA sequencing and iTRAQ quantitative proteomics respectively. Novel proteins encoded by circRNAs were predicted through ORF (open reading frame) selection using the GETORF software and verified by the mass spectrometry in proteomics, combined with the validation of their expressional alterations by quantitative RT-PCR. Totally 3,051 circRNAs were differentially expressed in bladder tissues of rat BOO model with widespread genomic distributions, including 1,414 up-regulated, and 1,637 down-regulated circRNAs. Our following quantitative proteomics revealed significant changes of 85 proteins in rat BOO model, which were enriched in multiple biological processes and signaling pathways such as the PPAR and Wnt pathways. Among them, 21 differentially expressed proteins were predicted to be encoded by circRNAs and showed consistent circRNA and protein levels in rat BOO model. The expression levels of five protein-encoding circRNAs were further validated by quantitative RT-PCR and mass spectrometry. The circRNA and protein profiles were substantially altered in rat BOO model, with great expressional changes of circRNA-encoded novel proteins.

膀胱出口梗阻（BOO）是一种与分子机制理解欠佳相关的常见泌尿系统疾病。本研究旨在探讨环状RNA（circular RNAs）及其编码蛋白在BOO发病过程中的可能作用。通过膀胱出口部分梗阻手术建立了大鼠BOO模型。利用深度RNA测序和iTRAQ定量蛋白质组学分别对环状RNA和蛋白质谱进行特征化分析。通过GETORF软件的ORF（开放阅读框）选择预测了由环状RNA编码的新蛋白，并通过蛋白质组学中的质谱分析进行验证，结合定量RT-PCR对其表达水平变化进行验证。在大鼠BOO模型的膀胱组织中，共鉴定出3,051个差异表达的环状RNA，其基因组分布广泛，包括1,414个上调和1,637个下调的环状RNA。我们的后续定量蛋白质组学研究揭示了85种蛋白质在大鼠BOO模型中的显著变化，这些蛋白质富集于多种生物学过程和信号通路，如PPAR和Wnt通路。其中，预测为环状RNA编码的21种差异表达蛋白在大鼠BOO模型中显示出一致的环状RNA和蛋白水平。五个编码蛋白的环状RNA的表达水平进一步通过定量RT-PCR和质谱分析得到验证。在大鼠BOO模型中，环状RNA和蛋白质谱发生了显著变化，环状RNA编码的新型蛋白表达水平发生了极大的变化。