microRNAs from plasmatic Extracellular vesicles as biomarkers of liver fibrosis in cases with Chronic Hepatitis C

CAIROLI, V; VALLE-MILLARES, D; FERNÁNDEZ RODRÍGUEZ, A; PRECIADO, MV; VALVA, P

Conferencia; SNEV International Virtual Conference (virtual); 2023

SNEV

Introduction: Extracellular vesicles (EVs) have gained significance as a component of liquid biopsies for evaluating liver damage in chronic hepatitis C (CHC) patients due to their molecular content and stability. Among EV-cargo, microRNAs (miRNAs) play a vital role as fine-tuning regulators of gene expression. Consequently, miRNAs may modulate fibrogenic signalling pathways, making them a promising biomarker tool for assessing liver damage. We aimed to characterize the miRNA signature of plasma-derived EVs with respect to the severity of liver fibrosis and to evaluate their diagnostic accuracy to discriminate liver damage stages.Method: From plasma samples of 50 CHC patients [36% significant fibrosis (F≥2)] EVs were purified (exoRNeasy QIAGEN) and then characterized by transmission and cryo electron-microscopy (EM) along with nanoparticle tracking analysis (NTA). miRNA-enriched RNA was extracted and high throughput sequenced. Then, miRNA identification and significant differential expression (SDE) analysis [fold change (FC) ≥ 1.5; adjusted p-value (p.adj) ≤ 0.2] according to fibrosis severity were performed. SDE miRNAs diagnostic value was assessed by ROC curves analysis. Finally, a new score to discriminate F≥2 including miRNAs with SDE was generated by multiple ordered logistic regression analysis. Results: Regarding EV characterization, TEM and cryo-EM revealed the presence of spherical bilayered particles with a mean diameter of ~130 nm. No significant differences in particle size [F