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

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.