急性病毒性心肌炎血浆外泌体Circular RNA的转录及功能分析

Transcriptional and Functional Analysis of Plasma Exosomal circRNA in Acute Viral Myocarditis

摘要: 目的：评估急性病毒性心肌炎(AVMC)患者外泌体circular RNA (circRNA)的表达差异并揭示其潜在功能。方法：于2021年7月至2023年3月在青岛大学附属医院采集9例临床确诊为AVMC的患者和9例健康对照(HC)的外周血。采用RNA高通量测序检测外泌体circRNA表达。我们进行了GO及KEGG富集分析预测circRNA在外泌体中的潜在分子、生物学功能和相关信号通路。我们选取了3个外泌体中差异性表达的circRNA进行了实时荧光定量聚合酶链反应(qRT-PCR)对测序结果进行了验证。结果：以P < 0.05和Fold Change绝对值 > 2为截断值筛选具有差异性表达的circRNA。本研究共发现了外泌体中有84个差异性表达的circRNA，其中83个下调及1个上调。GO和KEGG富集分析结果表明，差异表达的circRNA可能与ATP酶活动、GTP酶结合、溶酶体膜、微管、内质网蛋白质加工等有关。PCR结果表明hsa_circ_0001546在AVMC患者中下调，与测序结果一致。结论：差异性表达的hsa_circ_0001546可能是AVMC的潜在诊断标志物；差异性表达的circRNA在AVMC中具有一定生物功能，可能参与AVMC的发病机制。

Abstract: Objective: To evaluate the differential expression profile of exosomal circular RNA (circRNA) in pa-tients with acute viral myocarditis (AVMC) and to reveal its potential biological function. Methods: From July 2021 to March 2023, the peripheral blood of 9 patients clinically diagnosed with AVMC and 9 healthy controls (HC) were collected in the Affiliated Hospital of Qingdao University. CircRNA expression in exosomes was detected by high throughput RNA sequencing. We performed GO and KEGG functional analyses to predict the potential molecules, biological functions, and associated signaling pathways in exosomal circRNA. We selected three differentially expressed circRNAs in exosomes and performed quantitative real time polymerase chain reaction (qRT-PCR) to verify the sequencing results. Results: circRNA with differential expression was screened with P < 0.05 and Fold Change absolute value > 2 as truncation values. In this study, 84 differentially expressed circRNAs were found in exosomes, of which 83 were down-regulated and 1 was up-regulated. GO and KEGG analysis showed that differentially expressed circRNA may be related to ATPase activity, GTPase binding, lysosomal membrane, microtubules and protein processing in endoplasmic reticu-lum. PCR results showed that hsa_circ_0001546 was down-regulated in AVMC patients, which is consistent with the sequencing results. Conclusion: The differentially expressed hsa_circ_0001546 may be potential diagnostic markers for AVMC. Differentially expressed circRNAs may have certain biological functions in AVMC and be involved in the pathogenesis of AVMC.