The circulating microRNA (miRNA) profile has been widely used for identifying potential biomarkers against viral infections. However, data on circulating microRNA expression patterns in dengue patients are scanty. Considering the impact of severity caused by dengue infection, circulating miRNA profiles in plasma of dengue patients may prove to be valuable for developing early prognostic markers for the disease severity. Here, we described an in-depth analytical study of small RNA sequencing data obtained from the plasma of 39 dengue patients. Integrating bioinformatics and in vitro studies, we identified differentially expressed miRNAs (DEMs) (log₂ fold change ≥1.5, P < 0.05) associated with dengue disease progression. In comparing miRNA expression pattern with the follow-up samples, nine miRNAs were found to exhibit an altered expression that could distinguish between severe dengue and the convalescent patients. To understand the abundance and specificity of the DEMs in the context of dengue infection and disease progression, eight top-hit DEMs were further validated in the dengue virus-infected cell lines as well as in the patient’s plasma and peripheral blood mononuclear cells (PBMCs) using the quantitative reverse transcription-PCR (qRT‐PCR) method. Importantly, receiver operating curve analysis further confirmed that the plasma expression pattern of hsa-miR-122-5p could differentiate between different stages of dengue infection (area under the concentration-time curve [AUC] = 0.792), and dengue-negative patients with other febrile illnesses (AUC = 0.984). The in silico analysis of DEM target genes suggested an enrichment of the pathways associated with metabolism and inflammation. Our study gives a global view of miRNA expression in the plasma from dengue patients and provides a precious resource of candidate miRNAs involved in dengue infection and disease progression.

中文翻译：

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血浆样本的高通量 RNA 测序分析揭示了在登革热病进展中具有生物标志物潜力的循环 microRNA 特征。

循环 microRNA (miRNA) 谱已被广泛用于识别针对病毒感染的潜在生物标志物。然而，关于登革热患者循环 microRNA 表达模式的数据很少。考虑到登革热感染引起的严重程度的影响，登革热患者血浆中的循环 miRNA 谱可能被证明对于开发疾病严重程度的早期预后标志物是有价值的。在这里，我们描述了从 39 名登革热患者血浆中获得的小 RNA 测序数据的深入分析研究。结合生物信息学和体外研究，我们确定了差异表达的 miRNA (DEM)（log ₂倍变化≥1.5，P< 0.05) 与登革热疾病进展相关。在将 miRNA 表达模式与后续样本进行比较时，发现 9 种 miRNA 表现出改变的表达，可以区分严重的登革热患者和恢复期患者。为了了解登革热感染和疾病进展背景下 DEM 的丰度和特异性，在登革热病毒感染的细胞系以及患者的血浆和外周血单核细胞 (PBMC) 中进一步验证了八个热门 DEM使用定量逆转录-PCR（qRT-PCR）方法。重要的是，受试者工作曲线分析进一步证实了 hsa-miR-122-5p 的血浆表达模式可以区分登革热感染的不同阶段（浓度-时间曲线下面积 [AUC] = 0.792），以及患有其他发热性疾病的登革热阴性患者（AUC = 0.984）。这DEM 靶基因的计算机模拟分析表明与代谢和炎症相关的通路的富集。我们的研究提供了登革热患者血浆中 miRNA 表达的全局视图，并提供了参与登革热感染和疾病进展的候选 miRNA 的宝贵资源。