Cairoli Victoria, Valle-Millares Daniel, Ryan Pablo, Dominguez Lourdes, Martín-Carbonero Luz, De Los Santos Ignacio, De Matteo Elena, Ameigeiras Beatriz, De Sousa Marcela, Briz Verónica, Preciado María V, Fernández-Rodriguez Amanda, Valva Pamela
Multidisciplinary Institute for Investigation in Pediatric Pathologies (IMIPP), CONICET-GCBA, Laboratory of Molecular Biology, Pathology Division, Ricardo Gutiérrez Children's Hospital, C1425EFD CABA, Buenos Aires, Argentina.
Centro de Investigación Biomédica en Red de Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, 28222, Madrid, Spain.
Noncoding RNA Res. 2025 Mar 5;12:132-140. doi: 10.1016/j.ncrna.2025.03.004. eCollection 2025 Jun.
Extracellular vesicles (EVs) are an increasingly promising tool for liquid biopsy in liver diseases. Hepatitis C Virus (HCV) infection, alone or together with Human Immunodeficiency Virus (HIV) infection significantly impacts on the microRNA (miRNA) EVs content resembling chronic hepatitis C (CHC) progression. The objective of the study was to delve into the intricate EVs-miRNA profiles in CHC patients with different liver fibrosis stages, aiming to pinpoint non-invasive markers capable of distinguishing significant fibrosis. Plasma EV-miRNAs from 50 CHC patients (HCV+ and HCV+/HIV+) stratified in no significant (F < 2) and significant (F ≥ 2) fibrosis, were massively sequenced. General linear models (GLM) were used to identify significantly differential expressed (SDE) miRNAs according to liver fibrosis stages (F ≥ 2 and F < 2). Dysregulated biological pathways were subsequently analyzed for the following groups: i) all patients; ii) HCV+; and iii) HCV+/HIV+. Multiple-ordered logistic regression analysis was performed to develop a score to identify F ≥ 2 cases. The diagnostic potential of both the SDE miRNAs and the developed score was assessed using ROC curve analysis. With respect to all CHC patients, two SDE miRNAs (hsa-miR-122-5p and hsa-miR-92a-3p) were identified which regulate genes related to cytoskeleton organization. Regarding their diagnostic performance to discriminate F ≥ 2, both miRNAs individually demonstrated acceptable diagnostic values. However, their combined use in a new score enhanced their diagnostic performance (AUROC = 0.833). In the HCV+ subgroup, 8 SDE miRNAs (hsa-miR-122-5p, hsa-miR-320c, hsa-miR-3615, hsa-miR-320a-3p, hsa-miR-374b-5p, hsa-let-7a-3p, hsa-miR-199a-5p, hsa-miR-142-5p), which regulate macrophage activity and cell growth/death regulation, were recognized. Among them, hsa-miR-3615 displayed the highest diagnostic performance to discriminate F ≥ 2 (AUROC = 0.936). With respect to HCV+/HIV+, 18 SDE miRNAs (hsa-miR-4508, hsa-miR-122-5p, hsa-miR-451a, hsa-miR-1290, hsa-miR-1246, hsa-miR-107, hsa-miR-15b-5p, hsa-miR-194-5p, hsa-miR-22-5p, hsa-miR-20b-5p, hsa-miR-142-5p, hsa-miR-328-3p, hsa-miR-335-3p, hsa-miR-125a-5p, hsa-miR-423-3p, hsa-let-7d-3p, hsa-miR-128-3p, hsa-miR-10a-5p) were recognized that regulate RNA silencing processes. In this case, hsa-miR-423-3p and hsa-miR-128-3p showed outstanding diagnostic performances (AUROC > 0.900). Distinct EVs-miRNA profiles were identified in patients with varying liver fibrosis stages, both in the overall CHC cohort and within HCV+ and HCV+/HIV+ subgroups. These specific miRNA signatures would allow the elucidation of potential mechanisms involved in clinical evolution and identification of specific biomarkers of unfavorable progression, plausible to be used in a diagnostic panel. Furthermore, the developed score demonstrates the ability to discriminate within the CHC group those individuals with significant fibrosis regardless of their HIV infection status.
细胞外囊泡（EVs）是肝脏疾病液体活检中一种越来越有前景的工具。丙型肝炎病毒（HCV）感染，单独或与人类免疫缺陷病毒（HIV）感染一起，会显著影响类似于慢性丙型肝炎（CHC）进展的微小RNA（miRNA）细胞外囊泡含量。本研究的目的是深入探究不同肝纤维化阶段的CHC患者复杂的细胞外囊泡 - miRNA谱，旨在确定能够区分显著纤维化的非侵入性标志物。对50例CHC患者（HCV +和HCV + / HIV +）的血浆细胞外囊泡miRNA进行了大规模测序，这些患者被分为无显著纤维化（F < 2）和显著纤维化（F≥2）两组。使用一般线性模型（GLM）根据肝纤维化阶段（F≥2和F < 2）鉴定显著差异表达（SDE）的miRNA。随后对以下几组进行失调生物途径分析：i）所有患者；ii）HCV +；iii）HCV + / HIV +。进行多序逻辑回归分析以建立一个评分来识别F≥2的病例。使用ROC曲线分析评估SDE miRNA和所建立评分的诊断潜力。对于所有CHC患者，鉴定出两种SDE miRNA（hsa - miR - 122 - 5p和hsa - miR - 92a - 3p），它们调节与细胞骨架组织相关的基因。就其区分F≥2的诊断性能而言，这两种miRNA单独都显示出可接受的诊断价值。然而，它们在新评分中的联合使用提高了其诊断性能（AUROC = 0.833）。在HCV +亚组中，识别出8种SDE miRNA（hsa - miR - 122 - 5p、hsa - miR - 320c、hsa - miR - 3615、hsa - miR - 320a - 3p、hsa - miR - 374b - 5p、hsa - let - 7a - 3p、hsa - miR - 199a - 5p、hsa - miR - 142 - 5p），它们调节巨噬细胞活性和细胞生长/死亡调节。其中，hsa - miR - 3615在区分F≥2方面表现出最高的诊断性能（AUROC = 0.936）。对于HCV + / HIV +，识别出18种SDE miRNA（hsa - miR - 4508、hsa - miR - 122 - 5p、hsa - miR - 451a、hsa - miR - 1290、hsa - miR - 1246、hsa - miR - 107、hsa - miR - 15b - 5p、hsa - miR - 194 - 5p、hsa - miR - 22 - 5p、hsa - miR - 20b - 5p、hsa - miR - 142 - 5p、hsa - miR - 328 - 3p、hsa - miR - 335 - 3p、hsa - miR - 125a - 5p、hsa - miR - 423 - 3p、hsa - let - 7d - 3p、hsa - miR - 128 - 3p、hsa - miR - 10a - 5p），它们调节RNA沉默过程。在这种情况下，hsa - miR - 423 - 3p和hsa - miR - 128 - 3p表现出出色的诊断性能（AUROC > 0.900）。在整个CHC队列以及HCV +和HCV + / HIV +亚组中，不同肝纤维化阶段的患者中鉴定出了不同的细胞外囊泡 - miRNA谱。这些特定的miRNA特征将有助于阐明临床进展中涉及的潜在机制，并识别不良进展的特定生物标志物，有可能用于诊断面板。此外，所建立的评分显示出能够在CHC组中区分出那些具有显著纤维化的个体，而不论其HIV感染状态如何。
