标题：Identification of potential traumatic spinal cord injury related circular RNA-microRNA networks by sequence analysis
作者：Wang W.; Wang S.; Zhang Z.; Li J.; Xie W.; Su Y.; Chen J.; Liu L.
作者机构：[Wang, W] Department of Orthopedics, West China Hospital, Sichuan University, Chengdu Sichuan610041, China;[ Wang, S] Department of Orthopedics, West 更多
来源：Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery
关键词：circular RNA; microRNA; regulatory network; Traumatic spinal cord injury
摘要：目的: 通过高通量测序获取创伤性脊髓损伤（traumatic spinal cord injury，TSCI）后环状非编码 RNA（circular RNA，circRNA）与微小 RNA（microRNA，miRNA）表达谱，预测潜在 circRNA-miRNA 调控网络。. 方法: 取 48 只雄性 C57BL/6 小鼠（体质量 18～22 g）随机均分为两组（ n=24），TSCI 组采用 Allen’s 打击器械制备 TSCI 模型，假手术组（Sham 组）仅切开椎板不损伤脊髓。术后 3 d，两组取材行 HE 染色，观察脊髓组织结构；提取组织总 RNA 建库，高通量测序鉴定 circRNA 和 miRNA 差异表达谱，基因本体分析（gene ontology，GO）注释差异表达的 circRNA 宿主基因功能，筛选显著差异表达的 miRNA，通过 TargetScan 和 miRanda 预测 circRNA-miRNA 靶向结合，筛选关键 circRNA，构建潜在调控网络。. 结果: HE 染色示 Sham 组小鼠脊髓结构完整无破裂，TSCI 组脊髓结构有明显损伤破裂。测序共鉴定出 17 440 个 circRNA、1 228 个 miRNA。差异表达的 circRNA 宿主基因主要富集在细胞质，生物过程中差异基因主要富集在转录的正调控和蛋白磷酸化过程。差异表达最显著的 miRNA 为 mmu-miR-21-5p，筛选出可与其靶向结合的 circRNA6730，以 circRNA6730 为核心构建潜在 circRNA-miRNA 调控网络。. 结论: 通过表达谱分析和功能注释分析，显著差异表达的 circRNA 和 miRNA 有潜在的临床标志物价值，以 circRNA6730 为核心包含 mmu-miR-21-5p 的靶向互作网络可能在 TSCI 的发生发展过程中起重要调控作用，有助于阐明 TSCI 的病理生理进程机制，为临床诊疗提供新思路。.Objective: To systematically profile and characterize the circular RNA (circRNA) and microRNA (miRNA) expression pattern in the lesion epicenter of spinal tissues after traumatic spinal cord injury (TSCI) and predict the structure and potential functions of the regulatory network. Methods: Forty-eight adult male C57BL/6 mice (weighing, 18-22 g) were randomly divided into the TSCI ( n=24) and sham ( n=24) groups. Mice in the TSCI group underwent T 8-10 vertebral laminectomy and Allen's weight-drop spinal cord injury. Mice in the sham group underwent the same laminectomy without TSCI. The spinal tissues were harvested after 3 days. Some tissues were stained with HE staining to observe the structure. The others were used for sequencing. The RNA-Seq, gene ontology (GO) analysis, and circRNA-miRNA network analyses (TargetScan and miRanda) were used to profile the expression and regulation patterns of network of mice models after TSCI. Results: HE staining showed the severe damage to the spinal cord in TSCI group compared with sham group. A total of 17 440 circRNAs and 1 228 miRNAs were identified. The host gene of significant differentially expressed circRNA enriched in the cytoplasm, associated with positive regulation of transcription and protein phosphorylation. mmu-miR-21-5p was the most significant differentially expressed miRNA after TSCI, and circRNA6730 was predicted to be its targeted circRNA. Then a potential regulatory circRNA-miRNA network was constructed. Conclusion: The significant differentially expressed circRNAs and miRNAs may play important roles after TSCI. A targeted interaction network with mmu-miR-21-5p at the core of circRNA6730 could provide basis of pathophysiological mechanism, as well as help guide therapeutic strategies for TSCI.