Circulating ECV-Associated miRNAs as Potential Clinical Biomarkers in Early Stage HBV and HCV Induced Liver Fibrosis

Abstract

Introduction: Chronic hepatitis B (HBV) and C (HCV) virus infection is associated with the activation of hepatic stellate cells (HSCs) toward a myofibroblastic phenotype, resulting in excessive deposition of extracellular matrix, the development of liver fibrosis, and its progression toward cirrhosis. The gold standard for the detection and staging of liver fibrosis remains the liver biopsy, which is, however, associated with some mild and severe drawbacks. Other non-invasive techniques evade these drawbacks, but lack inter-stage specificity and are unable to detect early stages of fibrosis. We investigated whether circulating vesicle-associated miRNAs can be used in the diagnosis and staging of liver fibrosis in HBV and HCV patients.Methods: Plasma samples were obtained from 14 healthy individuals and 39 early stage fibrotic patients (F0–F2) with chronic HBV or HCV infection who underwent transient elastography (Fibroscan). Extracellular vesicles were extracted from the plasma and the level of miRNA-122, -150, -192, -21, -200b, and -92a was analyzed by qRT-PCR in total plasma and circulating vesicles. Finally, these same miRNAs were also quantified in vesicles extracted from in vitro activating primary HSCs.Results: In total plasma samples, only miRNA-200b (HBV: p = 0.0384; HCV: p = 0.0069) and miRNA-122 (HBV: p < 0.0001; HCV: p = 0.0007) were significantly up-regulated during early fibrosis. In circulating vesicles, miRNA-192 (HBV: p < 0.0001; HCV: p < 0.0001), -200b (HBV: p < 0.0001; HCV: p < 0.0001), -92a (HBV: p < 0.0001; HCV: p < 0.0001), and -150 (HBV: p = 0.0016; HCV: p = 0.004) displayed a significant down-regulation in both HBV and HCV patients. MiRNA expression profiles in vesicles isolated from in vitro activating primary mouse HSCs resembled the miRNA expression profile in circulating vesicles.Conclusion: Our analysis revealed a distinct miRNA expression pattern in total plasma and its circulating vesicles. The expression profile of miRNAs in circulating vesicles of fibrotic patients suggests the potential use of these vesicle-associated miRNAs as markers for early stages of liver fibrosis.

Predictors for advanced liver fibrosis in chronic hepatitis B virus infection with persistently normal or mildly elevated alanine aminotransferase

“…The miRNAs identified were involved in 100 signal transduction pathways, the majority of which affected liver fibrosis via the transforming growth factor-β/Smad, Wnt, mitogen-activated protein kinase, Janus kinase/signal transducers and activators of transcription and vascular endothelial growth factor pathways. A study conducted by Lambrecht et al ( 24 ) reported that miRNA-200b and miRNA-122 were significantly upregulated during early liver fibrosis and that miRNA-192, −92a and −150 were significantly downregulated in patients with HBV. Zheng et al ( 25 ) identified that levels of serum miR-125a-5p correlated with liver fibrosis and suggested that serum miR-125a-5p may be used as a non-invasive biomarker to monitor the progression of liver disease.…”

Circulating miRNAs as footprints for liver fibrosis grading in schistosomiasis

“…As attractive targets for novel diagnosis, circulating miRNAs have been proposed as biomarkers for different diseases and disorders including cancers, and infectious and inflammatory diseases [ 12 , 25 , 31 , 32 ]. Furthermore, circulating miRNAs have been proposed as having potential as predictors of fibrosis progression [ 1 , 14 , 26 , 27 , 30 ]. However, as there was limited information available, we aimed to determine the potential of measuring the levels of circulating miRNAs to trace the progression of hepatic fibrosis due to schistosomiasis.…”

Biomarker-Based Targeted Therapeutics

“…A similar study based on identification of miRNA levels, found different miRNAs in the EVs isolated from the plasma of the patients infected with HBV or HCV infection. Expression profile of miRNAs isolated from circulating vesicles showed reduced miR-192, miR-200b, miR-92a and miR-150a levels [32]. Thus, miRNAs expression levels can be correlated to early stage liver fibrosis identification.…”