颅内动脉瘤环状RNA差异表达鉴定及生物信息学分析

Identification of differentially expressed circular RNA in intracranial aneurysm and bioinformatics analysis

摘要:

目的 拟筛选颅内动脉瘤（IA）差异表达的环状RNA（DECs），并分析其生物学意义。方法 纳入16例临床样本（IA组织10例，颞浅动脉对照6例），利用illumina高通量测序技术筛选DECs来源基因；结合火山图及热图可视化表达差异，应用基因本体论（GO）功能注释及京都基因和基因组数据库（KEGG）联合富集分析挖掘DECs的功能关联；采用实时荧光定量聚合酶链式反应（qRT-PCR）验证候选circRNA的表达水平。结果 共鉴定28 850个DECs（上调10 081个，下调18 769个）；KEGG分析表明，IA相关DECs富集于昼夜节律、AMPK及Hedgehog信号通路（P<0.001）；qRT-PCR验证显示，hsa_circ_0003967、hsa_circ_0063305在IA组织中表达上调（P<0.01）；而hsa_circ_0003091、hsa_circ_0001742表达下调（P<0.01）。结论 circRNA差异表达是IA发生的重要分子特征，hsa_circ_0003967等候选分子可能通过干预昼夜节律、Hedgehog信号通路等信号通路驱动IA形成，这为IA的机制研究与靶向治疗提供新线索。

Abstract:

Objective The development of intracranial aneurysm （IA） is closely associated with genetic factors； however， studies have shown that circular RNA （circRNA） is involved in vascular remodeling and inflammatory response regulation， and its role in the pathogenesis of IA remains unclear. Therefore， this study aims to identify differentially expressed circRNAs （DECs） in IA and analyze their biological significance.Methods A total of 16 clinical samples （10 IA tissue samples and 6 superficial temporal artery samples as controls） were included， and Illumina high-throughput sequencing was used to identify DECs. Volcano plots and heat maps were used to visualize expression differences； the gene ontology （GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were used to explore the functional associations of DECs； qRT-PCR was used to further validate the expression levels of candidate circRNAs.Results A total of 28 850 DECs were identified， among which 10 081 were upregulated and 18 769 were downregulated. The KEGG analysis showed that IA-related DECs were significantly enriched in the circadian rhythm， AMPK， and Hedgehog signaling pathways （P<0.001）. The results of qRT-PCR validation showed that hsa_circ_0003967 and hsa_circ_0063305 were significantly upregulated in IA tissue （P<0.01）， while hsa_circ_0003091 and hsa_circ_0001742 were significantly downregulated （P<0.01）.Conclusions Differential expression of circRNAs is an important molecular feature in the pathogenesis of IA， and candidate molecules such as hsa_circ_0003967 may drive the formation of IA by intervening in the signaling pathways such as circadian rhythm and Hedgehog， which provides new clues for the mechanism research and targeted therapy for IA.