Abstract
MicroRNAs (miRNA) have been implicated in HCV infection. The present study analyzed the effects of let-7g on HCV infection in vitro, in clinical tissue and serum samples. Here, we show that the expression of let-7g in serum and liver tissue is significantly higher in patients with sustained virologic response (SVR). We show that interferon (IFN)/ribavirin (RBV) induces let-7g expression through p38/AP-1 signaling. Overexpression of let-7g reduced HCV gene or core protein level and inhibited the HCV viral load. The let-7g and IFN/RBV have additively inhibitory effect on HCV replication. These data implicate let-7g as a new therapeutic drug to additively cooperate with IFN/RBV to repress HCV replication.
Key messages
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let-7g expression is increased in serum and liver tissue of patients with SVR.
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Interferon/ribavirin induces let-7g expression through p38/AP-1 signaling.
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Overexpression of let-7g can repress HCV replication.
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Let-7g additively cooperates with interferon/ribavirin to repress HCV replication.
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Lin28B silencing can reverse let-7g expression and repress HCV replication.
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References
Williams R (2006) Global challenges in liver disease. Hepatology 44:521–526
Global Burden Of Hepatitis CWG (2004) Global burden of disease (GBD) for hepatitis C. J Clin Pharmacol 44:20–29
Wang JH, Lu SN, Wu JC, Huang JF, Yu ML, Chen SC, Chuang WL (1999) A hyperendemic community of hepatitis B virus and hepatitis C virus infection in Taiwan. Trans R Soc Trop Med Hyg 93:253–254
Yang JF, Lin YY, Hsieh MH, Tsai CH, Liu SF, Yu ML, Dai CY, Huang JF, Lin WY, Lin ZY et al (2011) Performance characteristics of a combined hepatitis C virus core antigen and anti-hepatitis C virus antibody test in different patient groups. Kaohsiung J Med Sci 27:258–263
Yu ML, Dai CY, Huang JF, Chiu CF, Yang YH, Hou NJ, Lee LP, Hsieh MY, Lin ZY, Chen SC et al (2008) Rapid virological response and treatment duration for chronic hepatitis C genotype 1 patients: a randomized trial. Hepatology 47:1884–1893
Yu ML, Dai CY, Huang JF, Hou NJ, Lee LP, Hsieh MY, Chiu CF, Lin ZY, Chen SC, Hsieh MY et al (2007) A randomised study of peginterferon and ribavirin for 16 versus 24 weeks in patients with genotype 2 chronic hepatitis C. Gut 56:553–559
Huang CF, Huang JF, Yang JF, Hsieh MY, Lin ZY, Chen SC, Wang LY, Juo SH, Chen KC, Chuang WL et al (2012) Interleukin-28B genetic variants in identification of hepatitis C virus genotype 1 patients responding to 24 weeks peginterferon/ribavirin. J Hepatol 56:34–40
Huang CF, Yeh ML, Huang JF, Yang JF, Hsieh MY, Lin ZY, Chen SC, Wang LY, Hsi E, Juo SH et al (2012) Host interleukin-28B genetic variants versus viral kinetics in determining responses to standard-of-care for Asians with hepatitis C genotype 1. Antiviral Res 93:239–244
Omata MKT, Yu M-L, Yokosuka O, Lim S-G, Jafri W, Tateishi R et al (2012) APASL consensus statements and management algorithms for hepatitis C virus infection. Hepatol Int 6:409–435
Lindow M, Kauppinen S (2012) Discovering the first microRNA-targeted drug. J Cell Biol 199:407–412
Hildebrandt-Eriksen ES, Aarup V, Persson R, Hansen HF, Munk ME, Orum H (2012) A locked nucleic acid oligonucleotide targeting microRNA 122 is well-tolerated in cynomolgus monkeys. Nucleic Acid Ther 22:152–161
Pedersen IM, Cheng G, Wieland S, Volinia S, Croce CM, Chisari FV, David M (2007) Interferon modulation of cellular microRNAs as an antiviral mechanism. Nature 449:919–922
Cheng M, Si Y, Niu Y, Liu X, Li X, Zhao J, Jin Q, Yang W (2013) High-throughput profiling of alpha interferon- and interleukin-28B-regulated microRNAs and identification of let-7s with anti-hepatitis C virus activity by targeting IGF2BP1. J Virol 87:9707–9718
Cheng JC, Yeh YJ, Tseng CP, Hsu SD, Chang YL, Sakamoto N, Huang HD (2012) Let-7b is a novel regulator of hepatitis C virus replication. Cell Mol Life Sci 69:2621–2633
Piskounova E, Viswanathan SR, Janas M, LaPierre RJ, Daley GQ, Sliz P, Gregory RI (2008) Determinants of microRNA processing inhibition by the developmentally regulated RNA-binding protein Lin28. J Biol Chem 283:21310–21314
Viswanathan SR, Daley GQ, Gregory RI (2008) Selective blockade of microRNA processing by Lin28. Science 320:97–100
Heo I, Joo C, Cho J, Ha M, Han J, Kim VN (2008) Lin28 mediates the terminal uridylation of let-7 precursor MicroRNA. Mol Cell 32:276–284
Guo Y, Chen Y, Ito H, Watanabe A, Ge X, Kodama T, Aburatani H (2006) Identification and characterization of lin-28 homolog B (LIN28B) in human hepatocellular carcinoma. Gene 384:51–61
Ji J, Wang XW (2010) A Yin-Yang balancing act of the lin28/let-7 link in tumorigenesis. J Hepatol 53:974–975
Viswanathan SR, Powers JT, Einhorn W, Hoshida Y, Ng TL, Toffanin S, O’Sullivan M, Lu J, Phillips LA, Lockhart VL et al (2009) Lin28 promotes transformation and is associated with advanced human malignancies. Nat Genet 41:843–848
Wang YC, Chen YL, Yuan RH, Pan HW, Yang WC, Hsu HC, Jeng YM (2010) Lin-28B expression promotes transformation and invasion in human hepatocellular carcinoma. Carcinogenesis 31:1516–1522
Lan FF, Wang H, Chen YC, Chan CY, Ng SS, Li K, Xie D, He ML, Lin MC, Kung HF (2011) Hsa-let-7g inhibits proliferation of hepatocellular carcinoma cells by downregulation of c-Myc and upregulation of p16(INK4A). Int J Cancer 128:319–331
Ji J, Zhao L, Budhu A, Forgues M, Jia HL, Qin LX, Ye QH, Yu J, Shi X, Tang ZY et al (2010) Let-7g targets collagen type I alpha2 and inhibits cell migration in hepatocellular carcinoma. J Hepatol 52:690–697
Kato T, Furusaka A, Miyamoto M, Date T, Yasui K, Hiramoto J, Nagayama K, Tanaka T, Wakita T (2001) Sequence analysis of hepatitis C virus isolated from a fulminant hepatitis patient. J Med Virol 64:334–339
Kato T, Date T, Murayama A, Morikawa K, Akazawa D, Wakita T (2006) Cell culture and infection system for hepatitis C virus. Nat Protoc 1:2334–2339
Chen KC, Hsieh IC, Hsi E, Wang YS, Dai CY, Chou WW, Juo SH (2011) Negative feedback regulation between microRNA let-7g and the oxLDL receptor LOX-1. J Cell Sci 124:4115–4124
Vermehren J, Yu ML, Monto A, Yao JD, Anderson C, Bertuzis R, Schneider G, Sarrazin C (2011) Multi-center evaluation of the Abbott RealTime HCV assay for monitoring patients undergoing antiviral therapy for chronic hepatitis C. J Clin Virol 52:133–137
Luciani MG, Seok J, Sayeed A, Champion S, Goodson WH, Jeffrey SS, Xiao W, Mindrinos M, Davis RW, Dairkee SH (2011) Distinctive responsiveness to stromal signaling accompanies histologic grade programming of cancer cells. PLoS One 6:e20016
Heinemeyer T, Wingender E, Reuter I, Hermjakob H, Kel AE, Kel OV, Ignatieva EV, Ananko EA, Podkolodnaya OA, Kolpakov FA et al (1998) Databases on transcriptional regulation: TRANSFAC, TRRD and COMPEL. Nucleic Acids Res 26:362–367
Miyanari Y, Atsuzawa K, Usuda N, Watashi K, Hishiki T, Zayas M, Bartenschlager R, Wakita T, Hijikata M, Shimotohno K (2007) The lipid droplet is an important organelle for hepatitis C virus production. Nat Cell Biol 9:1089–1097
Filipowicz W, Grosshans H (2011) The liver-specific microRNA miR-122: biology and therapeutic potential. Prog Drug Res 67:221–238
Koberle V, Waidmann O, Kronenberger B, Andrei A, Susser S, Fuller C, Perner D, Zeuzem S, Sarrazin C, Piiper A (2013) Serum microRNA-122 kinetics in patients with chronic hepatitis C virus infection during antiviral therapy. J Viral Hepat 20:530–535
Dai CY, Huang JF, Hsieh MY, Hou NJ, Lin ZY, Chen SC, Hsieh MY, Wang LY, Chang WY, Chuang WL et al (2009) Insulin resistance predicts response to peginterferon-alpha/ribavirin combination therapy in chronic hepatitis C patients. J Hepatol 50:712–718
Yu ML, Huang CF, Huang JF, Chang NC, Yang JF, Lin ZY, Chen SC, Hsieh MY, Wang LY, Chang WY et al (2011) Role of interleukin-28B polymorphisms in the treatment of hepatitis C virus genotype 2 infection in Asian patients. Hepatology 53:7–13
Conrad KD, Giering F, Erfurth C, Neumann A, Fehr C, Meister G, Niepmann M (2013) MicroRNA-122 dependent binding of Ago2 protein to hepatitis C virus RNA is associated with enhanced RNA stability and translation stimulation. PLoS One 8:e56272
Janssen HL, Reesink HW, Lawitz EJ, Zeuzem S, Rodriguez-Torres M, Patel K, van der Meer AJ, Patick AK, Chen A, Zhou Y et al (2013) Treatment of HCV infection by targeting microRNA. N Engl J Med 368:1685–1694
Tokumoto Y, Hiasa Y, Uesugi K, Watanabe T, Mashiba T, Abe M, Kumagi T, Ikeda Y, Matsuura B, Onji M (2012) Ribavirin regulates hepatitis C virus replication through enhancing interferon-stimulated genes and interleukin 8. J Infect Dis 205:1121–1130
Felger JC, Cole SW, Pace TW, Hu F, Woolwine BJ, Doho GH, Raison CL, Miller AH (2012) Molecular signatures of peripheral blood mononuclear cells during chronic interferon-alpha treatment: relationship with depression and fatigue. Psychol Med 42:1591–1603
Humar M, Loop T, Schmidt R, Hoetzel A, Roesslein M, Andriopoulos N, Pahl HL, Geiger KK, Pannen BH (2007) The mitogen-activated protein kinase p38 regulates activator protein 1 by direct phosphorylation of c-Jun. Int J Biochem Cell Biol 39:2278–2288
Platanias LC (2003) The p38 mitogen-activated protein kinase pathway and its role in interferon signaling. Pharmacol Ther 98:129–142
Acknowledgments
We would like to thank Dr. Takaji Wakita of Tokyo Metropolitan Institute of Neuroscience for kindly providing the HCV strain JFH-1; Dr. Wei-Lun Tsai of the Kaohsiung Veterans General Hospital for kindly providing the Huh7.5.1 cell line; Apath, LLC for the gift of the Ava.5, J6/JFH and Con1 replicon cells. This work was supported by grants from the National Science Council of Taiwan (grant number NSC101-2314-B-037-063-MY3) and Kaohsiung Medical University Hospital (grant number KMUH101-1R06) and grants partially from Kaohsiung Medical University “Aim for the Top Universities Grant, grant no. KMU-TP103D12, KMU-TP103D13, KMU-TP103D14, and KMU-KMU-TP103E07.” We appreciate the help from the Taiwan Liver Research Function.
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Summary statement
Let-7g reduces the HCV expression, and the level of let-7g in serum and liver tissue is significantly higher in patients with sustained virologic response in HCV infection. Let-7g may be developed as target drug and predictor for HCV therapy.
Author contributions
Conceived and designed the experiments: Wen-Wen Chou, Suh-Hang Hank Juo, Chia-Yen Dai, Ming-Lung Yu. Performed the experiments: Wen-Wen Chou, Yi-Shan Tsai, Chung-Feng Huang, Ming-Lun Yeh, Ming-Yen Hsieh, Ching-I Huang, Jee-Fu Huang. Analyzed the data: Wen-Wen Chou, Pei-Chien Tsai, Edward Hsi. Contributed reagents/materials/analysis tools: Wen-Wen Chou, Chung-Feng Huang, Ming-Lun Yeh, Ming-Yen Hsieh, Ching-I Huang, Jee-Fu Huang, Suh-Hang Hank Juo, Wei-Lun Tsai, Wan-Long Chuang, Chia-Yen Dai, Ming-Lung Yu. Wrote the paper: Wen-Wen Chou, Chia-Yen Dai, Ming-Lung Yu.
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Chou, WW., Huang, CF., Yeh, ML. et al. MicroRNA let-7g cooperates with interferon/ribavirin to repress hepatitis C virus replication. J Mol Med 94, 311–320 (2016). https://doi.org/10.1007/s00109-015-1348-1
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DOI: https://doi.org/10.1007/s00109-015-1348-1