The expression levels and significance of miR-92a-3p in exosomes, serum and liver tissue of patients with hepatitis B-related liver fibrosis

Abstract

Abstract: Objective To investigate the expression levels of microRNA miR-92a-3p in exosomes, serum and liver tissue samples of patients with HBV-related hepatic fibrosis and to analyze the difference and significance. Methods Real-time fluorescent quantitative PCR technology was used to detect the expression levels of miR-92a-3p in exosomes, serum, and liver tissue of patients with hepatitis B-related liver fibrosis at different stages. The Mann-Whitney U test or Kruskal-Wallis test was used to compare the differences in miR-92a-3p expression levels during different stages of liver fibrosis. Spearman’s correlation coefficient method was employed to analyze the correlation between the level of miR-92a-3p in exosomes and that in serum and liver tissue. The receiver operating characteristic curve (ROC) was applied to evaluate the predictive efficacy of exosomal and serum miR-92a-3p levels for significant liver fibrosis in hepatitis B. Results The expression level of miR-92a-3p in exosomes showed significant differences among different stages of liver fibrosis (P<0.05). The expression level of miR-92a-3p in exosomes from patients in S2-4 stages was significantly higher than that in S0-1 stages (P<0.0001). The relative expression levels of serum miR-92a-3p in S0-1, S2, S3 and S4 stages were 0.69±0.20, 1.01±1.35, 0.74±0.88 and 0.87±1.33, respectively. There was no statistically significant difference in the expression level of miR-92a-3p in serum among different stages of liver fibrosis (P=0.3817). The relative expression levels of liver tissue miR-92a-3p in S0-1, S2, S3 and S4 stages were 1.69±1.31, 1.54±1.26, 1.05±0.61 and 0.68±0.54, respectively. Similarly, no statistically significant difference was observed in the expression level of miR-92a-3p in liver tissue among different stages of liver fibrosis (P=0.1040). The expression level of miR-92a-3p in exosomes did not show a significant correlation with its expression level in serum (r=0.1787, P=0.1083). Similarly, there was no significant correlation between the expression levels of miR-92a-3p in exosomes and liver tissue (r=0.051, P=0.7311). When the cut-off value was 0.245, the AUROC for serum miR-92a-3p was 0.61 (95%CI: 0.48~0.75), with a sensitivity and specificity for diagnosing significant liver fibrosis of 63.46% and 60%, respectively. For exosomal miR-92a-3p, at a cut-off value of 0.7459, the AUROC was highest at 0.87 (95%CI: 0.80~0.94), with a sensitivity and specificity for diagnosing significant liver fibrosis of 84.13% and 72.09%, respectively. Conclusion The expression levels of miR-92a-3p in exosomes, serum, and liver tissue showed no significant correlation. The concentration of miR-92a-3p in exosomes is significantly higher than in serum and liver tissue, indicating that circulating miR-92a-3p is enriched in exosomes. Compared to serum miR-92a-3p, exosomal miR-92a-3p demonstrated higher diagnostic efficacy for significant liver fibrosis in hepatitis B patients.

Key words: miR-92a-3p, Exosomes, Serum, Liver tissue, Liver fibrosis

WANG Qian-qian, JIANG Xu-hua, LI Xin-yan, SONG Jie, ZHANG Yi, JI Yuan-yuan, REN Xiao-jing, QI Xun, CHEN Liang, ZHANG Jian, HUANG Yu-xian. The expression levels and significance of miR-92a-3p in exosomes, serum and liver tissue of patients with hepatitis B-related liver fibrosis[J]. Chinese Hepatolgy, 2024, 29(11): 1325-1329.

References

[1] Shiha G, Ibrahim A, Helmy A, et al. Asian-Pacific Association for the Study of the Liver (APASL) consensus guidelines on invasive and non-invasive assessment of hepatic fibrosis: a 2016 update[J]. Hepatol Int, 2017, 11(1): 1-30.
[2] Esteller M. Non-coding RNAs in human disease[J]. Nat Rev Genet, 2011, 12(12): 861-74.
[3] Grundhoff A, Sullivan CS. Virus-encoded microRNAs[J]. Virology, 2011, 411(2): 325-43.
[4] Wang Q, Hu Q, Ying Y, et al. Using next-generation sequencing to identify novel exosomal miRNAs as biomarkers for significant hepatic fibrosis[J]. Discov Med, 2021, 31(164):147-159.
[5] 中华医学会肝病学分会,中华医学会感染病学分会. 慢性乙型肝炎防治指南(2015 年版)[J]. 临床肝胆病杂志, 2015, 31(12): 1941-1960.
[6] Scheuer PJ. The nomenclature of chronic hepatitis: time for a change[J]. J Hepatol, 1995, 22(1): 112-4.
[7] Arroyo JD, Chevillet JR, Kroh EM, et al. Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma[J]. Proc Natl Acad Sci U S A, 2011, 108(12): 5003-8.
[8] Tabet F, Vickers KC, Cuesta Torres LF, et al. HDL-transferred microRNA-223 regulates ICAM-1 expression in endothelial cells[J]. Nat Commun, 2014, 5: 3292.
[9] Gallo A, Tandon M, Alevizos I, et al. The majority of microRNAs detectable in serum and saliva is concentrated in exosomes[J]. PLoS One, 2012, 7(3): e30679.
[10] Zheng J, Wu C, Xu Z, et al. Hepatic stellate cell is activated by microRNA-181b via PTEN/Akt pathway[J]. Mol Cell Biochem, 2015, 398(1-2): 1-9.
[11] Li G, Li J, Li C, et al. MicroRNA-125a-5p Contributes to hepatic stellate cell activation through targeting FIH1[J]. Cell Physiol Biochem, 2016, 38(4): 1544-52.
[12] Huang CF, Sun CC, Zhao F, et al. miR-33a levels in hepatic and serum after chronic HBV-induced fibrosis[J]. J Gastroenterol, 2015, 50(4): 480-90.
[13] Huang X, Yuan T, Tschannen M, et al. Characterization of human plasma-derived exosomal RNAs by deep sequencing[J]. BMC Genomics, 2013, 14: 319.
[14] Szabo G, Momen-Heravi F. Extracellular vesicles in liver disease and potential as biomarkers and therapeutic targets[J]. Nat Rev Gastroenterol Hepatol, 2017, 14(8): 455-466.
[15] Baker LA, Lee KC, Palacios Jimenez C, et al. Circulating microRNAs reveal time course of organ injury in a porcine model of acetaminophen-induced acute liver failure[J]. PLoS One, 2015, 10(5): e0128076.
[16] Wang H, Hou L, Li A, et al. Expression of serum exosomal microRNA-21 in human hepatocellular carcinoma[J]. Biomed Res Int, 2014, 2014: 864894.
[17] Matsuura K, De Giorgi V, Schechterly C, et al. Circulating let-7 levels in plasma and extracellular vesicles correlate with hepatic fibrosis progression in chronic hepatitis C[J]. Hepatology, 2016, 64(3): 732-745.
[18] Vickers KC, Remaley AT. Lipid-based carriers of microRNAs and intercellular communication[J]. Curr Opin Lipidol, 2012, 23(2): 91-7.
[19] Manier S, Liu CJ, Avet-Loiseau H, et al. Prognostic role of circulating exosomal miRNAs in multiple myeloma[J]. Blood, 2017, 129(17): 2429-2436.
[20] Morsiani C, Collura S, Sevini F, et al. Circulating miR-122-5p, miR-92a-3p, and miR-18a-5p as Potential Biomarkers in Human Liver Transplantation Follow-Up[J]. Int J Mol Sci, 2023, 24(4):3457.
[21] Sorop A, Iacob R, Iacob S, et al. Plasma small extracellular vesicles derived miR-21-5p and miR-92a-3p as potential biomarkers for hepatocellular carcinoma screening[J]. Front Genet,2020, 23(11):712.
[22] Nóbrega DND, Carvalho TL, do ó KP, et al. MicroRNA dysregulation in schistosomiasis-induced hepatic fibrosis: a systematic review[J]. Expert Rev Mol Diagn, 2023,23(3):257-265.