Extracellular vesicle-derived microRNA signature in chronic hepatitis C patients with different stages of liver fibrosis

CAIROLI VICTORIA; DANIEL VALLE-MILLARES; PABLO RYAN; LUZ MARTÍN-CARBONERO; IGNACIO DE LOS SANTOS; ELENA DE MATTEO; AMEIGEIRAS BEATRIZ; VERÓNICA BRIZ; MARIA VICTORIA PRECIADO; VALVA PAMELA; FERNANDEZ RODRIGUEZ AMANDA

Viena

Congreso; EASL Congress 2023; 2023

EASL

Background and Aims: In the context of chronic hepatitis C (CHC), co-infection with HIV has a negative impact on liver damage progression. Recently, extracellular vesicles (EVs) have gained relevance in liver diseases as intercellular communicators thus mediating pathological processes. HCV and HIV have been described to modify the microRNA content of EV, but little is known about their impact on pathogenesis or their possible role as liver damage biomarkers. We aimed to characterize the plasma derived-EVs miRNA signature of HCV+ and HCV+/HIV+ patients and to explore their expression regarding the severity of liver fibrosis, to assess its impact on pathogenesis. Method: EVs were purified and characterized from 50 CHC plasma samples [36 % significant fibrosis (F ≥ 2)] [21 HCV+ (52.4 % F ≥ 2) and 29 HCV+/HIV+ (22.6 % F ≥ 2)]. EV-derived total RNA was extracted and massively sequenced. Then, miRNA identification, significant differential expression (SDE) analysis [fold change (FC) ≥ 1.5; adjusted p-value (p.adj) ≤ 0.2], target gene prediction and pathway enrichment analysis (p.adj ≤ 0.5) were performed. The diagnostic value of SDE miRNAs was assessed by ROC curves analysis. Results: Differential expression analysis of plasma EV-miRNAs according to severity of liver fibrosis demonstrated 2 SDE miRNAs (miR-122-5p and miR-92a-3p, up- and down-regulated in the F ≥ 2 group, respectively) which in silico regulate genes involved in cytoskeleton organization. Within HCV+ subgroup, 4 up- (miR-122-5p, miR-320c, miR-3615, miR-320a-3p), and 4 downregulated-miRNAs (miR-374b-5p, let-7a-3p, miR-199a-5p, miR-142-5p) were found in F ≥ 2 patients. Together, they regulate genes involved in macrophage activity and cell growth/death regulation. In turn, the HCV+/HIV+ subgroup displayed 11 up- (miR-4508, miR-122-5p, miR-451a, miR-1290, miR-1246, miR-107, miR-15b-5p, miR-194-5p, miR-22-5p, miR-20b-5p, miR-142-5p) and 7 downregulated-miRNAs (miR-328-3p, miR-335-3p, miR-125a-5p, miR-423-3p, let-7d-3p, miR-128-3p, miR-10a-5p). These miRNAs regulate the expression of genes involved the RNA silencing machinery.Regarding diagnostic performance of SDE miRNAs to discriminate F ≥ 2 cases (AUROC ≥ 0. 800) in each group different miRNAs were identified: A) HCV+: miR-3615 miR-374b-5p, let-7a-3p, miR-142-5p, miR-320a-3p and miR-320c and; B) HCV+/HIV+: miR-423-3p, miR-128-3p, miR-194-5p, miR-10a-5p, miR-328-3p, miR-22-5p, miR-125a-5p, let-7d-3p, miR-335-3p, miR-451a, miR-122-5p and miR-1246.Conclusion: chronic HCV+ and HCV+/HIV+ patients with different stages of liver fibrosis present a differential profile of extracellular vesicle-derived miRNA. This specific miRNA signature would allow elucidation of possible mechanisms involved in clinical evolution and identification of biomarkers of unfavorable progression specific for each group plausible to be used in a diagnostic panel.