Circulating microRNAs as non-invasive biomarkers for hepatitis B virus liver fibrosis

Table 1 Circulating and intrahepatic microRNA regulation and target processes involved in liver fibrogenesis in hepatitis B virus infection

MicroRNA	Plasma level	Liver tissue	Mediated processes involved in liver fibrosis	Ref.
miR-34a	↑	↑	Cell-cycle regulator (cell differentiation, proliferation, metabolism, apoptosis); HSC activation	Guo et al[45]; Singh et al[42]
miR-221 miR-222	↑	↑	Collagen synthesis; HSC proliferation; Liver fibrosis; Oncogenesis	Singh et al[42]
miR 27a/b	↑	↑	HSC activation, differentiation and proliferation	Zhang et al[131]
miR 181a/b	↑	↑	Cell cycle regulator; HSC activation and proliferation	Yu et al[132]
miR 199a/b	↓	↑	HSC activation	Murakami et al[91]
miR-223	↓; ↓ EVs	↑	Inflammatory response	Akamatsu et al[133]; Bao et al[43]; Ye et al[59]; Wang et al [106]
miR-125 (-125a-5p/ -125b)	↑	↑	HSC activation, proliferation	Zheng et al[134]; You et al[49]
miR 21-5p	↑in total plasma; ↓ in EVs	↑	Collagen synthesis; Oncogenesis	Bao et al[43]; Wei et al[68]; Wang et al[135]

↑ means upregulation. ↓ means downregulation. EVs: Extracellular vesicles; HSC: Hepatic stellate cell; MiR: MicroRNA.

Table 2 Circulating microRNAs in hepatitis B virus infection and their significance for the staging of liver fibrosis

Circulating microRNAs, plasma or serum	Significance for liver disease	Ref.
Upregulated microRNAs
miR-185	↑ in advanced (F3-F4) vs early liver fibrosis (F1-F2); and ↑ in early liver fibrosis vs healthy volunteers; No increase with HBV plasma DNA	Li et al[136]
miR-2861, miR-345-3p, miR-3620, miR-3656, miR-371a-5p, miR-4646-5p, miR-4651, miR-4695, miR-4800-5p, miR-638	Individually ↑ in F1-F4 vs F0; Plasma expression differs between each stage of liver fibrosis	Zhang et al[137]
miR-1, mR-10b-5p, miR-20b-5p, miR-96b-5p, miR-133b, miR-455-ep, miR-671-5p	Increase in the serum in F3-F4 liver fibrosis	Singh et al[42]
miR-499-5p	Increases in the serum in F1-F2 stages	Singh et al[42]
miR-106b, miR-181b	Panel for the diagnosis of liver cirrhosis due both HBV and non-HBV associated infection	Chen et al44]
Downregulated microRNAs
miR-29	↓ in liver cirrhosis vs healthy controls	Xing et al[138]; Wang et al[106]
miR-223	↓ with the progression of liver fibrosis from F0-F2 to F3-F4	Bao et al[43]; Wang et al[106]
miR-21
miR-143
miR-374
miR-486-3p, miR-497-5p	Individually ↓ in F1-F4 vs F0; Plasma expression differs between each stage of liver fibrosis	Zhang et al[137]
miR-1227-3p	↓ in the serum in F1-F2 stages	Singh et al[42]

↑ means upregulation. ↓ means downregulation. HBV: Hepatitis B virus; miR: MicroRNA.

Table 3 Overview of the major studies on the use of microRNAs in hepatitis B virus infected patients for the staging of liver fibrosis

Study group	Fibrosis staging/validation method	microRNA detection method/sample1	Data normalization	microRNA regulation depending on fibrosis staging			microRNA panel for liver fibrosis	Ref.
102 treatment naïve CHB	58 pts F0-F2; 44 pts F3-F4 / liver biopsy and laboratory data	RT-qPCR / serum samples	Spiked-in cel-miR-39	F3-F4 vs F0-F2	miR-122, -27b	↑	miR-122, -222, PLT, ALP	Appourchaux et al[107], 2016
					miR-222, -224	↓
330 CHB, 165 HC	165 pts F0-F3; 165 pts F4 / clinical and laboratory data	RT-qPCR / serum samples	Exogenous control using cel-miR-67	CHB: F4 vs F0-F3	miR-18a-5p, -21-5p, -29c-3p, -122-5p, -106b-5p, 185-5p	↓	Three panels: F4 vs other stages: miR-18a-5p, -21-5p -29c-3p, -122-5p, -106b-5p, 185-5p; F4 vs HC: miR-1, -146a-5p, -451a; CHB vs HC: miR-21-5p, -27a-3p -122-5p, -146a-5p	Jin et al[139], 2015
				CHB F4 vs HC	miR-1, -146a-5p	↑
					miR-451a	↓
				CHB vs HC	miR-21-5p, -27a-3p, -122-5p, -146a-5p	↑
123 treatment naïve CHB	69 pts F0-F2 vs 54 staged F3-F4 / liver biopsy	RT-qPCR; Serum samples	Spiked-in cel-miR-39	F3-F4 vs F0-F2	miR-29a, -143, -223, -21, -374	↓	miR-29a, -143, -223, PLT	Bao et al[43], 2017
8 ASC, 8 AVH, 7 HC, 49 treatment naïve CHB	49 CHB patients: 16 pts F0, 19 pts F1-F2, 14 pts F3-F4 / liver biopsy, clinical and laboratory data	RT-qPCR and microarray analysis; Serum samples	U6 RNA control relative miRNA	F1-F2	miR-499-5p	↑	Analysis of miRNA networks and of individual MiRNA s	Singh et al[42], 2018
					miR-1227-3p	↓
				F3-F4	miR-34b-3p, -1224-3p, -1, -10b-5p, -20b-5p, -96b-5p, -133, -455-3p, -671-5p	↑
19 CHB, 14 HC	19 CHB pts with F0-F2	RT-qPCR total plasma EVs/liver stiffness	Spiked-in cel-miR-39	Plasma (F0-F2)	miR-192, -200b	↑	Expression pattern of each individual miRNA in EVs vs total plasma	Lambrecht et al[93], 2017
				EVs (F0-F2)	miR-192, -200b, -92a, -150	↓
207 CHB, 47 non-HBV-LC, 7 non-CHB, 137 HC	207 CHB of which: 127 pts F4; 79 pts F0-F3 / liver biopsy	RT-qPCR / plasma samples	miR-1228 control with a log-2 scale transformation	CHB F4 and non-CHB F4; vs other groups; (panel for F4 diagnosis)	miR-106b	↓	Panel composed of miR-106 and miR-181b	Chen et al[44], 2013
					miR-181b	↑
50 treatment naïve CHB	10 pts F0, 10 pts F1, 10 pts F2, 10 pts F3, 10 pts F4 / liver biopsy	Microarray analysis / plasma samples	Log standardization of MiRNA s whose target gene expression levels > 2 folds and FDR < 0.05	F4 vs F0	miR-2861, -345-3p, -3620-3p, -3656, -371a-5p, -4646-5p, -4651, 4695-5p, -4800-5p, -638,	↑	Detailed differential expression of individual MiRNA s for each stage of liver fibrosis F0-F4	Zhang et al[137], 2015
					miR-497-5p, -486-3p	↓
92 CHB	11 pts F0, 16 pts F1, 12 pts F2, 13 pts F3, 40 pts F4 / liver biopsy and laboratory data	RT-qPCR; Plasma samples	Quanto, EC1, EC2 controls; relative miRNA expression was assessed using 2−ΔΔCq calculation	≥ F2	miR-122-5p	↑	miR-122-5p, -21-5p, -146a-5p, -223, -29c-3p, -22-3p, -381-3p	Wang et al[106], 2018
					miR-223, -29c-3p	↓
				≥ F3	miR-122-5p	↑
				F4 vs F0	miR-122-5p, -29c-3p, -146a-5p, -223	-/↓

¹miRNA sample refers only to the samples collected from the serum/plasma in each of the mentioned studies. ↑ means upregulation. ↓ means downregulation. ALP: Alkaline phosphatase; ASC: Asymptomatic carriers; AVH: Acute viral hepatitis; CHB: Chronic hepatitis B; EVs: Extracellular vesicles; FDR: False discovery rate; HC: Healthy controls; LC: Liver cirrhosis; miR: MicroRNA; PLT: Platelet; Pts: Patients; RT-qPCR: Real time quantitative polymerase chain reaction.

Table 4 Circulating microRNAs in hepatitis B virus infection and their role in necroinflammation vs fibrosis

Circulating microRNA	microRNA regulation	Clinical significance in HBV infection	Ref.
miR-122	↑	Correlates with the necroinflammatory activity, HBsAg and HBV DNA; Also correlated with ≥ F2 stage of liver fibrosis	Waidmann et al[103], Ji et al[109], Wang et al[106]
miR-210	↑	Marker of necroinflammation; Varies with the severity of HBV hepatitis	Song et al[140]
miR-125 (-125a-5p/ -125b)	↑	Correlates with HBV intrahepatic replication and necroinflammatory activity	Li et al[141], Zheng et al[134]
miR-124	↑	Marker of HBV-associated necroinflammation	Wang et al[142]
miR-29	↓	Marker of liver fibrosis irrespective of aetiology	Xing et al[139]
miR-223	↓	Marker of liver fibrosis, decreases with the progression to cirrhosis	Bao et al[43]
miR-185	↑	Increases in advanced HBV fibrosis; Could play a therapeutic role in HBV gene suppression in tumoral cells	Li et al[136], Fan et al[143]

↑ means upregulation. ↓ means downregulation. HBsAg: Hepatitis B surface antigen; HBV: Hepatitis B virus; miR: MicroRNA.