MicroRNA Signature at the Time of Clinical HCV Recurrence Associates With Aggressive Fibrosis Progression Post-Liver Transplantation

Abstract

Diagnosis and prediction of the severity of hepatitis C virus recurrence (HCVrec) after liver transplantation (LT) remain a challenge. MicroRNAs have been recently recognized as potential disease biomarkers.Archival liver biopsy samples from 43 HCV+ LT recipients were collected at clinical HCVrec time and at 3 years post-LT. Patients were classified as progressors (P = F0/F1) or nonprogressors (NP = F3/F4) according to the severity of fibrosis on the 3-year biopsy. Training (n = 27) and validation (n = 16) sets were defined. RNA was isolated from all biopsies at clinical HCVrec time, labeled and hybridized to miRNA-arrays. Progressors versus nonprogressors were compared using the twosample t-test. A p-value ≤0.01 was considered significant. The ingenuity pathway analysis tool was used for microRNA and miRNA:mRNA ontology data integration. Nine microRNAs were differentially expressed between groups. A supervised cluster analysis separated samples in two well-defined groups (progressors vs. nonprogressors). Pathway analysis associated those microRNAs with hepatitis, steatosis, fibrosis, cirrhosis and T cell-related immune response. Data integration identified 17 genes from a previous genomic study as 9-microRNAs signature targets. Seven microRNAs were successfully validated in the validation set using QPCR. We have identified a 9-microRNA signature able to identify early post-LT patients at high risk of severe HCVrec during long-term follow-up.

MicroRNA and hepatitis C virus- challenges in investigation and translation: a review of the literature

“…Cermelli et al (2011) used in vitro analysis of Huh 7.5 replicon supernatants to hypothesize that miR-16, 34a and 122 in serum may be useful in predicting liver fibrosis, and then demonstrated using TaqMan qRT-PCR that serum elevations in miR-34a (undetectable in controls versus mean of 44,000 copies/mL in HCV infected patients, p <.0001) and miR-122 (mean fold change 10.8, p <.0001) were significantly correlated with progression to cirrhosis in 53 HCV infected patients as compared to 19 healthy controls. Elevations in miR-34a in liver biopsy tissue (mean fold change not explicitly reported in paper, p = .0105) as measured by an Affymetrix microarray were also associated with progression to fibrosis in 43 HCV infected post-transplant patients in at least one subsequent study (Gehrau et al, 2013). …”

MicroRNAs in Liver Disease: Bench to Bedside

“…150 Of these 9 miRs, three were up-regulated (miR-155, -34a and -222) and six were down-regulated (miR-23b, -361, -455, -30b, -30c, and -27b). MiR-21 has been shown to regulate pro-and antiinflammatory cytokines and has been implicated in the development of fibrosis leading to chronic rejection following liver transplantation.…”

“…75,151 This has been also validated in an independent set of HCV+ LT recipients and demonstrated organ specificity (molecular pathways involved in liver injury, malignancy and hepatic disease), and relationship with immune response (T cell lineages development). 150 Farid et al have shown that circulating miR-122, miR-148, miR-194 are sensitive biomarkers for hepatocyte injury and rejection after liver transplantation. 152 Disease specific miR signatures would be helpful to monitor post-LT transplant success and supersede the need for multiple invasive, dangerous liver biopsies.…”

Long noncoding RNAs in hepatocellular carcinoma: Novel insights into their mechanism

“…It is estimated that proteincoding genes only account for less than 2% of the human genome while nearly 70% of the human genome is transcribed pervasively [13] . Accordingly, ample ncRNAs are transcribed from human genome, for instance lncRNAs, microRNAs (miRNAs), small interfering RNAs and PIWI-interacting RNAs [14][15][16] . NcRNAs were once thought to be body "garbage" or transcriptional "noise".…”