MicroRNA-27a Modulates HCV Infection in Differentiated Hepatocyte-Like Cells from Adipose Tissue-Derived Mesenchymal Stem Cells

Abstract

Background and AimsDespite the discovery of hepatitis C virus (HCV) entry factor, the mechanism by which it is regulated by miRNAs remains unclear. Adipose tissue-derived human mesenchymal stem cells (AT-hMSCs) have been widely used for differentiated hepatocyte-like cells (DHCs). Here, we established an in vitro HCV infection model using DHCs from AT-hMSCs and identified miRNAs that modulate HCV infectivity.MethodsAT-hMSCs were differentiated into DHCs using the conditional media, and evaluated for hepatocyte characteristics using RT-PCR, immunocytochemistry, periodic acid-Schiff staining, and a urea synthesis assay. The expression of HCV candidate receptors was also verified using immunocytochemistry. The levels of candidate miRNAs targeting HCV receptors were then determined by relative quantitative RT-PCR (rqRT-PCR). Finally, DHCs were infected using HCVcc and serum from HCV-infected patients, and infectivity of the virus was measured by rqRT-PCR and transmission electron microscopy (TEM).ResultsThe expected changes in morphology, function and hepatic gene expression were observed during hepatic differentiation. Moreover, the expression of candidate HCV entry factors and miR-27a were altered during hepatic differentiation. The infection and replication of HCV occurred efficiently in DHCs treated with HCVcc or infected with serum from HCV-infected patients. In addition, HCV infectivity was suppressed in miR-27a-transfected DHCs, due to the inhibition of LDLR expression by miR-27a.ConclusionsOur results demonstrate that AT-hMSCs are a good source of DHCs, which are suitable for in vitro cultivation of HCV. Furthermore, these results suggest that miR-27a modulates HCV infectivity by regulating LDLR expression.

Exosomal MicroRNAs Derived From Umbilical Mesenchymal Stem Cells Inhibit Hepatitis C Virus Infection

“…Together, the combination of the four miRNAs showed a potent inhibitory effect on HCV replication. A recent study revealed the role of miR-27a in suppressing HCV [58]. However, because the amount and fold changes of miR27a were not among the top in sequencing data, it may not be critical for the antiviral ability of uMSC-Exo.…”

MicroRNA-27a decreases the level and efficiency of the LDL receptor and contributes to the dysregulation of cholesterol homeostasis

“…Choi et al [37] previously reported that miR-27a decreases hepatitis C virus (HCV) infectivity by downregulating LDLR --a candidate HCV entry receptor-- in differenciated hepatocyte-like cells (DHCs). Furthermore, these authors confirmed that, as the expression of miR-27a decreases during hepatic differentiation, the levels of LDLR increased.…”

“…Furthermore, these authors confirmed that, as the expression of miR-27a decreases during hepatic differentiation, the levels of LDLR increased. In contrast, miR-27b --the other member of the miR-27 family that shares the same seed region and 20 out of 21 nucleotides with 27a— increases during hepatic differentiation [37] suggesting that miR-27a but not miR-27b, modulates LDLR in DHCs. Although Choi et al [37] have clearly demonstrated that miR-27a downregulates LDLR, they did not characterize the underlying mechanism and whether this observed effect was direct or indirect.…”

Mesenchymal stem cells and infectious diseases: Smarter than drugs

“…Regarding infectivity of MSCs by HCV Choi et al found that hepatocyte-like cells differentiated from AMSCs were suitable for in vitro cultivation of this virus [41].…”