EV-derived miRNA signature in patients with HCV and HCV/HIV with different stages of liver fibrosis

CAIROLI VICTORIA; VALLE MILLARES DANIEL; RYAN PABLO; DOMINGUEZ LOURDES; MARTIN-CARBONERO L; AMEIGEIRAS B; BRIZ VERÓNICA; FERNANDEZ RODRIGUEZ AMANDA; PRECIADO MV *EQUAL CONTRIBUTOR; VALVA P*EQUAL CONTRIBUTOR

Gante

Congreso; 28th International Symposium on Hepatitis C Virus, Flaviviruses, and Related Viruses; 2022

Extracellular vesicles (EVs) are essential players in cell communication, and their cargo modulates the receptor cell response. MicroRNAs (miRNAs) proved to modulate the immune response both in physiological and pathological conditions. Hepatitis C (HCV) and Human Immunodeficiency (HIV) viruses’ infection could modify EVs miRNA content and therefore, the immune response. We aimed to analyze the significant differentially expressed (SDE) EVs-derived miRNAs between HCV and HCV/HIV infected patients with different stages of liver fibrosis and to explore the associated molecular pathways.Plasma from 22 chronic HCV and 31 HCV/HIV patients were analyzed. Total EV-containing RNA enriched with small RNAs was isolated and high-throughput sequenced (1 x 50) to characterize the miRNA cargo. Raw reads were analyzed with Fastqc and trimmed with Cutadapt. Human-miRNA identification was performed with miRDeep2. R package edgeR was used to detect SDE miRNAs between groups. In silico miRNA target prediction was performed with DIANA mirPath.First, HCV patients [54 ys (34 - 71), 54,5 % F ≥ 2] showed 36 SDE miRNAs compared with the HCV/HIV group [50 ys (31 - 71), 22,5 % F ≥ 2], which modulate pathways related to fatty acids biosynthesis, extracellular matrix (ECM) interaction, hippo signaling and viral carcinogenesis. Then, patients were stratified by fibrosis status and differential expression analysis was performed (Figure 1). Patients with F < 2 showed downregulation of hsa-miR-122-5p and hsa-miR-194-5p (log2FC = -1,4, p < 0,001, log2FC = -0,6, p < 0,001, respectively), which, in turn, condition the expression of genes involved in ECM-receptor interaction, proteoglycans in cancer and TGF-beta signaling pathways. Finally, SDE miRNAs between patients with F < 2 and F ≥ 2 were studied in each HCV condition. In the HCV F < 2 group, hsa-miR-3615 and hsa-miR-320a-3p were downregulated (log2FC = -0,9, p < 0,001, log2FC = -0,6, p < 0,001, respectively) which modulate genes of the TGF-beta and hippo signaling pathways. On the other hand, HCV/HIV F < 2 group showed 9 SDE miRNAs where hsa-miR-122-5p (downregulated) and hsa-miR-328-3p (upregulated) had the strongest differences (log2FC = -1,3, p < 0,001, log2FC = 1,3, p < 0,001, respectively). Most of these miRNAs regulate genes involved in cancer and fatty acids related pathways.Differentially expressed EVs-derived miRNAs in HCV and HCV/HIV chronic infection and in different stages of liver fibrosis were observed. The specific miRNA signature of each liver fibrosis stage may elucidate potential mechanisms involved in the clinical evolution of these patients and the identification of biomarkers of unfavorable progression.