丙型肝炎病毒（HCV）与人类免疫缺陷病毒（HIV）的共感染对疾病进展有不利影响。越来越多的证据表明，细胞外囊泡（EVs）是宿主与病毒之间重要的交流介质。作为EVs载体的重要组成部分，微型RNA（miRNAs）是调控正常细胞过程并促进病毒复制、病毒发病和疾病进展的关键调节因子。我们的目标是对慢性HCV感染和HIV共感染患者的血浆衍生EVs miRNA特征进行表征，以揭示共感染的分子机制。从50个血浆样本（21个HCV单感染和29个HCV/HIV共感染）中纯化和表征EVs。通过大规模测序分离和分析衍生小RNA。使用miRDeep2鉴定已知和新生miRNAs。使用广义线性模型进行显著差异表达（SDE）miRNA鉴定，并评估其疑似失调的生物途径。大多数临床和流行病学特征的研究组相似。组间在EVs的大小或浓度上没有观察到差异。因此，HCV/HIV共感染状态没有影响EVs的浓度或大小，但造成了血浆衍生EVs miRNA载体的混乱。因此，共鸣鉴定到了149个miRNAs（其中143个已知，6个新生），其中37个SDE miRNAs在HCV/HIV共感染患者中上调了15个，下调了22个。SDE miRNAs调控与HCV和HIV发病机制相关的炎症、纤维化和癌症相关基因。这些发现有助于开发新一代生物标志物和治疗策略，以及揭示病毒与宿主相互作用的机制。关键信息：HCV和HCV/HIV显示了相似的血浆EV大小和浓度。使用NGS对EVs派生的miRNA进行了特征分析。观察到HCV和HCV/HIV之间的37个SDE miRNAs。SDE miRNAs调控与炎症、纤维化和癌症相关的基因。© 2023作者。

Hepatitis C virus (HCV) coinfection with human immunodeficiency virus (HIV) has a detrimental impact on disease progression. Increasing evidence points to extracellular vesicles (EVs) as important players of the host-viral cross-talk. The microRNAs (miRNAs), as essential components of EVs cargo, are key regulators of normal cellular processes and also promote viral replication, viral pathogenesis, and disease progression. We aimed to characterize the plasma-derived EVs miRNA signature of chronic HCV infected and HIV coinfected patients to unravel the molecular mechanisms of coinfection. EVs were purified and characterized from 50 plasma samples (21 HCV mono- and 29 HCV/HIV co-infected). EV-derived small RNAs were isolated and analyzed by massive sequencing. Known and de novo miRNAs were identified with miRDeep2. Significant differentially expressed (SDE) miRNA identification was performed with generalized linear models and their putative dysregulated biological pathways were evaluated. Study groups were similar for most clinical and epidemiological characteristics. No differences were observed in EVs size or concentration between groups. Therefore, HCV/HIV co-infection condition did not affect the concentration or size of EVs but produced a disturbance in plasma-derived EVs miRNA cargo. Thus, a total of 149 miRNAs were identified (143 known and 6 de novo) leading to 37 SDE miRNAs of which 15 were upregulated and 22 downregulated in HCV/HIV co-infected patients. SDE miRNAs regulate genes involved in inflammation, fibrosis, and cancer, modulating different biological pathways related to HCV and HIV pathogenesis. These findings may help to develop new generation biomarkers and treatment strategies, in addition to elucidate the mechanisms underlying virus-host interaction. KEY MESSAGES: HCV and HCV/HIV displayed similar plasma-EV size and concentration. EVs- derived miRNA profile was characterized by NGS. 37 SDE miRNAs between HCV and HCV/HIV were observed. SDE miRNAs regulate genes involved in inflammation, fibrosis and cancer.© 2023. The Author(s).