An exploratory study to find out circulating miRNAs in Hepatitis C virus associated Hepatocellular carcinoma

Background and Aim: HCV-induced hepatocellular carcinoma (HCC) still remains a significant cause of mortality, despite the availability of a successful direct acting anti-viral (DAA) therapy. The underlying mechanism(s) of occurrence of HCC among patients who achieved sustained virological response (SVR) with DAA treatment is still unknown. The aim of this study was to find out different circulating micro-RNAs (miRNAs) in HCV-associated HCC compared to non-HCC clinical samples. Methods:In this pilot study, we performed miRNA expression profiling from once thawed archived plasma samples in 42 HCV infected patients: Group A with HCC (n=15) and Group B without HCC (n=27) by small RNA sequencing (75bp single end) using QIAseq miRNA Library protocol. The quality check, quantification and annotation were done on CLC Genomics Workbench 22 after controlling for confounding variables. The functional enrichment analysis of differentially expressed miRNAs was done using anmiRNA functional enrichment tool TAM 2.0. Results :The median age of patients in group A was higher (58 years) than of group B (32 years).A total of 19 miRNAs (p-value <0.05) were differentially expressed between group A and group B (15 upregulated and 4 downregulated in HCC-positive patients). The differentially regulated miRNAs was found to be enriched in cell proliferation, differentiation and cell-death pathways. In group A, among cirrhotic (20%) vs non-cirrhotic (80%), 25 deregulated miRNAs were detected. Similarly, in group A, patients with SVR attained showed increased expression of miR-141-5p and miR-582-3p (involved in inflammation and immune response) as compared to no expression in non-SVR patients. Conclusions: Occurrence of HCC in HCV infected patients despite treatment and clearance of virus is a significant problem and needs further studies. The diagnostic performances of deregulated miRNAs will be measured using Receiver Operating Characteristic (ROC) curve analyses.