RNA-Seq Profiling of Circular RNAs During Development of Hindgut in Rat Embryos With Ethylenethiourea-Induced Anorectal Malformations.
RNA-Seq Profiling of Circular RNAs During Development of Hindgut in Rat Embryos With Ethylenethiourea-Induced Anorectal Malformations.
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亚乙基硫脲诱导的肛门直肠畸形大鼠胚胎后肠发育过程中环状 RNA 的 RNA 测序分析
DOI：
10.3389/fgene.2021.605015
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发表时间：
2021
影响因子：
3.7
通讯作者：
Bai YZ
中科院分区：
文献类型：
--
作者：
Li SY;Wang CY;Xiao YX;Tang XB;Yuan ZW;Bai YZ
肛门直肠畸形是最常见的先天性末端消化道畸形之一。Circular RNA（circRNA）是一类新型的内源性非编码RNA，在消化系统发育中发挥重要作用，但其在ARMs发病机制中的作用尚不明确。在这项研究中，我们通过RNA-seq分析circRNA表达并构建调控circRNA-miRNA-mRNA网络来探索乙烯硫醚（ETU）诱导ARMs的机制。孕10 d（GD 10），9只孕鼠单次灌胃1%ETU（ARM组）125 mg/kg，另9只孕鼠灌胃等量生理盐水（正常组）作为对照。在肛门直肠发育的关键时间点（GD 14、GD 15和GD 16）通过剖宫产获得胚胎。通过高通量测序评估从胎儿分离的后肠样品，并通过逆转录-定量聚合酶链反应、琼脂糖凝胶电泳和桑格克隆和测序验证差异表达的circRNA。在正常组和ARM组中总共鉴定了18295个circRNA。基于425个差异表达的circRNA（|FC|> 2，p < 0.05），使用miREAP、米兰达和TargetScan预测circRNA-miRNA和miRNA-mRNA对。预计总共有55种circRNA（与正常组相比，ARM组中有14种上调，41种下调）与195种miRNA和947种mRNA结合。竞争性内源RNA网络和京都基因和基因组百科全书分析显示，novel_circ_001042具有最大的连接性，并且与ARM相关的信号通路密切相关，例如无翼型MMTV整合位点家族、促分裂原活化蛋白激酶和转化生长因子β通路。这些结果为circRNA在ARMs中的作用提供了原始见解，并为进一步分析分子机制和信号网络提供了宝贵的资源。
Anorectal malformations (ARMs) are among the most common congenital terminal digestive tract malformations. Circular RNAs (circRNAs), a novel type of endogenous non-coding RNAs, play roles in the development of the digestive system; however, their contributions to the pathogenesis of ARMs are not well-established. In this study, we explored the mechanism underlying ethylenethiourea (ETU)-induced ARMs by profiling circRNA expression via RNA-seq and constructing a regulatory circRNA-miRNA-mRNA network. Nine pregnant rats were gavage-fed a single dose of 125 mg/kg 1% ETU (ARM group) on gestational day 10 (GD10), and another 9 pregnant rats received a similar dose of saline (normal group) as a control. Embryos were obtained by cesarean section on the key time-points of anorectal development (GD14, GD15, and GD16). Hindgut samples isolated from the fetuses were evaluated by high-throughput sequencing and differentially expressed circRNAs were validated by reverse transcription-quantitative polymerase chain reaction, agarose gel electrophoresis, and Sanger cloning and sequencing. A total of 18295 circRNAs were identified in the normal and ARM groups. Based on the 425 differentially expressed circRNAs (|Fc| > 2, p < 0.05), circRNA-miRNA and miRNA-mRNA pairs were predicted using miREAP, miRanda, and TargetScan. A total of 55 circRNAs (14 up- and 41 downregulated in the ARM group compared to the normal group) were predicted to bind to 195 miRNAs and 947 mRNAs. Competing endogenous RNA networks and a Kyoto Encyclopedia of Genes and Genomes analysis revealed that novel_circ_001042 had the greatest connectivity and was closely related to ARM-associated signaling pathways, such as the Wingless Type MMTV integration site family, mitogen-activated protein kinase, and transforming growth factor-β pathways. These results provide original insight into the roles of circRNAs in ARMs and provide a valuable resource for further analyses of molecular mechanisms and signaling networks.
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影响因子：
14.9
作者：
Danan M;Schwartz S;Edelheit S;Sorek R
通讯作者：
Sorek R
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
14.9
作者：
Kanehisa M;Araki M;Goto S;Hattori M;Hirakawa M;Itoh M;Katayama T;Kawashima S;Okuda S;Tokimatsu T;Yamanishi Y
通讯作者：
Yamanishi Y
影响因子：
4.6
作者：
Bachmayr-Heyda A;Reiner AT;Auer K;Sukhbaatar N;Aust S;Bachleitner-Hofmann T;Mesteri I;Grunt TW;Zeillinger R;Pils D
通讯作者：
Pils D
DOI：
10.1002/ajmg.1532
发表时间：
2001-10-15
期刊：
AMERICAN JOURNAL OF MEDICAL GENETICS
影响因子：
--
作者：
Cuschieri, A
通讯作者：
Cuschieri, A