Objective: To investigate the serum microRNA (miRNA) expression and examine the impact of miRNA expression profiles on T helper type 17 (Th17)/regulatory T cells (Treg) imbalance among patients with cystic echinococcosis, so as to provide insights into the illustration of the mechanisms underlying chronic Echinococcus granulosus infections, and long-term pathogenesis.
Methods: Total RNA was extracted from the sera of cystic echinococcosis patients and healthy controls, and subjected to high-throughput sequencing with the Illumina sequencing platform. Known miRNAs were annotated and new miRNAs were predicted using the miRBase database and the miRDeep2 tool, and differentially expressed miRNAs were identified. The target genes of differentially expressed miRNAs were predicted using the software miRanda and TargetScan, and the intersection was selected for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Among the differentially expressed miRNAs with the 20 highest fold changes, miRNAs that targeted genes relating to key transcription factors RORC and FOXP3 that determine the production of Th17 and Treg cells or their important regulatory pathways (PI3K-Akt and mTOR pathways) were matched.
Results: A total of 53 differentially expressed miRNAs were screened in sera of cystic echinococcosis patients and healthy controls, including 47 up-regulated miRNAs and 6 down-regulated miRNAs. GO enrichment analysis showed that these differentially expressed miRNA were involved DNA transcription and translation, cell components, cell morphology, neurodevelopment and metabolic decomposition, and KEGG pathway analysis showed that the differentially expressed miRNA were mainly involved in MAPK, PI3K-Akt and mTOR signaling pathways. Among the differentially expressed miRNAs with the 20 highest fold changes, there were 3 miRNAs that had a potential for target regulation of RORC, and 15 miRNAs that had a potential to target the PI3K-Akt and mTOR signaling pathways.
Conclusions: Significant changes are found in serum miRNA expression profiles among patients with E. granulosus infections, and differentially expressed miRNAs may lead to Th17/Treg imbalance through targeting the key transcription factors of Th17/Treg or PI3K-Akt and mTOR pathways, which facilitates the long-term parasitism of E. granulosus in hosts and causes a chronic disease.
［摘要］ 目的 观察细粒棘球蚴病患者血清微小 RNA (miRNA) 表达水平、探讨 miRNA 对辅助性 T 细胞 17 (Th17)/调节 性 T 细胞 (Treg) 失衡的影响, 以阐明细粒棘球蚴慢性感染并长期致病的机制。方法 提取细粒棘球蚴病患者与健康对照 者血清总 RNA, 采用 Illumina 测序平台进行高通量测序。分别采用 miRBase 数据库和 miRDeep2 工具进行已知 miRNA 注 释和新 miRNA 预测, 并进行差异分析。采用 miRanda 软件和 TargetScan 软件分别预测差异表达 miRNA 靶基因后取交集, 进行基因本体 (GO) 富集分析以及京都基因和基因组百科全书 (KEGG) 通路分析。在差异表达变化倍数居前 20 位的 miRNA 中, 匹配可靶向决定 Th17 细胞和 Treg 细胞生成的关键转录因子 (RORC 和 FOXP3) 或重要调控通路 (PI3K-Akt 和 mTOR 通路) 相关基因的 miRNA。结果 细粒棘球蚴病患者与健康对照血清中共有 53 个差异表达 miRNA, 其中 47 个上 调表达 miRNA、6 个下调表达 miRNA。GO 富集分析显示, 差异表达 miRNA 功能涉及 DNA 转录翻译、细胞成分、细胞形 态、神经发育及代谢分解等过程。KEGG 富集分析显示, 这些差异表达 miRNA 靶基因涉及的主要信号通路包括 MAPK、PI3K-Akt、mTOR 等信号通路。在差异表达变化倍数居前 20 位的 miRNA 中, 有 3 个潜在靶向调控 RORC 的 miRNA、15 个 潜在靶向 PI3K-Akt、mTOR 信号通路的 miRNA。结论 细粒棘球蚴感染后可使患者血清 miRNA 表达谱出现明显改变, 差 异表达 miRNA 可能通过靶向 Th17/Treg 关键转录因子或 PI3K-Akt、mTOR 通路而导致 Th17/Treg 免疫失衡, 进而利于细粒 棘球蚴在宿主体内长期寄生并慢性致病。.
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