AiM  Vol.6 No.4 , April 2016
The Role of Circulating MicroRNAs as Markers of Disease Progression in Hepatitis C Virus Infected Egyptian Patients
Abstract: Background: The discovery of miRNAs circulating in the peripheral blood has opened new directions of research to identify new non-invasive markers for diagnosis of diseases. Aim: The aim of the study was to evaluate the expression levels of circulating plasma miRNAs (miRNA-21 & miRNA-122) in Egyptian patients with chronic uncomplicated and complicated HCV. Patients & Methods: This study was conducted on 60 Chronic HCV infected patients. Patients were divided into three groups (20 patients each): uncomplicated HCV, cirrhosis, and hepatocellular carcinoma (HCC). All patients were subjected to laboratory investigations including complete blood picture, liver function tests. Expression levels of miRNA-21 and -122 in plasma using RT-PCR were determined. Results: MiRNA-21 showed significant fold increase in chronic uncomplicated HCV while significant fold decrease in cirrhotic and HCC groups (P = 0.036). On the other hand, miRNA-122 showed significant fold elevation in both chronic uncomplicated and cirrhotic groups and significant fold decrease in HCC group (P = 0.005). ROC curve analysis for miRNA-122 yielded 68.4% sensitivity and 100% specificity for the differentiation of HCC patients from non-HCC at a cutoff 0.184. Neither miRNA-21 nor miRNA-122 was a successful predictor for HCC diagnosis. Conclusion: MiRNA-122 can be used as novel non-invasive biomarker for monitoring HCV related disease progression.
Cite this paper: Harfoush, R. , Meheissen, M. , Elwafa, R. and Elwazzan, D. (2016) The Role of Circulating MicroRNAs as Markers of Disease Progression in Hepatitis C Virus Infected Egyptian Patients. Advances in Microbiology, 6, 320-331. doi: 10.4236/aim.2016.64031.

[1]   Rehermann, B. (2009) Hepatitis C: Virus versus Innate and Adaptive Immune Responses—A Tale of Coevolution and Coexistence. Journal of Clinical Investigations, 119, 1745-1754.

[2]   Ray, S.C., Arthur, R.R., Carella, A., Bukh, J. and Thomas, D.L. (2000) Genetic Epidemiology of Hepatitis C Virus throughout Egypt. Journal of Infectious Diseases, 182, 698-700.

[3]   El-Zayadi, A.R., Badran, H.M., Barakat, E.M., Mel-D, A., Shawky, S., Mohamed, M.K., Selim, O. and Saeid, A. (2005) Hepatocellular Carcinoma in Egypt: A Single Center Study over a Decade. World Journal of Gastroenterology, 11, 5193-5198.

[4]   Zinkin, N.T., Grall, F., Bhaskar, K., Otu, H.H. and Spentzos, D.D. (2008) Serum Proteomics and Biomarkers in Hepatocellular Carcinoma and Chronic Liver Disease. Clinical Cancer Research, 14, 470-477.

[5]   Du, T. and Zamore, P.D. (2005) Microprimer: The Biogenesis and Function of Micro-RNA. Development, 132, 4645-4652.

[6]   Volinia, S., Calin, G.A., Liu, C.G., Ambs, S. and Cimmino, A. (2006) A Micro-RNA Expression Signature of Human Solid Tumors Defines Cancer Gene Targets. Proceedings of the National Academy of Sciences of the United States of America, 103, 2257-2261.

[7]   Mitchell, P.S., Parkin, R.K., Kroh, E.M., Fritz, B.R. and Wyman, S.K. (2008) Circulating Micro-RNAs as Stable Blood-Based Markers for Cancer Detection. Proceedings of the National Academy of Sciences of the United States of America, 105, 10513-10518.

[8]   Johannes, H., Nicolas, Z. and Mathias, H. (2011) The Parallel Universe: Micro-RNAs and Their Role in Chronic Hepatitis, Liver Tissue Damage and Hepatocarcinogenesis. European Journal of Medical Sciences, 141, Article ID: w13287.

[9]   Matsuzaki, K., Murata, M., Yoshida, K., Sekimoto, G., Uemura, Y., Sakaida, N., Kaibori, M., Kamiyama, Y., Nishizawa, M., Fujisawa, J., Okazaki, K. and Seki, T. (2007) Chronic Inflammation Associated with Hepatitis C Virus Infection Perturbs Hepatic Transforming Growth Factor Beta Signaling, Promoting Cirrhosis and Hepatocellular Carcinoma. Hepatology, 46, 48-57.

[10]   Walter, H.T., Duverlie, G. and Bengrine, A. (2012) Micro-RNAs and Hepatitis C Virus: Toward the End of MiRNA-122 Supremacy. Journal of Virology, 9, 109.

[11]   Shiha, G. and Zalata, K. (2001) Ishak versus METAVIR: Terminology, Convertibility and Correlation with Laboratory Changes in Chronic Hepatitis C. In: Takahashi, H., Ed., Liver Biopsy, InTech, Rijeka, 155-170.

[12]   Bruix, J. and Sherman, M. (2011) American Association for the Study of Liver Diseases (AASLD) Practice Guidelines. Management of Hepatocellular Carcinoma: An Update. Hepatology, 53, 1020-1022.

[13]   Varnholt, H., Drebber, U., Schulze, F., Wedemeyer, I., Schirmacher, P., Dienes, H.P. and Odenthal, M. (2008) MicroRNA Gene Expression Profile of Hepatitis C Virus-Associated Hepatocellular Carcinoma. Hepatology, 47, 12123-12132.

[14]   Ladeiro, Y., Couchy, G., Balabaud, C., Bioulac-Sage, P., Pelletier, L., Rebouissou, S. and Zucman-Rossi, J. (2008) MicroRNA Profiling in Hepatocellular Tumors Is Associated with Clinical Features and Oncogene/Tumor Suppressor Gene Mutations. Hepatology, 47, 1955-1963.

[15]   Chan, J.A., Krichevsky, A.M. and Kosik, K.S. (2005) MicroRNA-21 Is an Antiapoptotic Factor in Human Glioblastoma Cells. Cancer Research, 65, 6029-6033.

[16]   Fulci, V., Chiaretti, S., Goldoni, M., Azzalin, G., Carucci, N., Tavolaro, S., Castellano, L., Magrelli, A., Citarella, F., Messina, M., Maggio, R., Peragine, N., Santangelo, S., Mauro, F.R., Landgraf, P., Tuschl, T., Weir, D.B., Chien, M., Russo, J.J., Ju, J., Sheridan, R., Sander, C., Zavolan, M., Guarini, A., Foà, R. and Macino, G. (2007) Quantitative Technologies Establish a Novel microRNA Profile of Chronic Lymphocytic Leukemia. Blood, 109, 4944-4951.

[17]   Iorio, M.V., Ferracin, M., Liu, C.G., Veronese, A., Spizzo, A., Sabbioni, S., Magri, E., Pedriali, M., Fabbri, M., Campiglio, M., Ménard, S., Palazzo, J.P., Rosenberg, A., Musiani, P., Volinia, S., Nenci, I., Calin, G.A., Querzoli, P., Negrini, M. and Croce, C.M. (2005) MicroRNA Gene Expression Deregulation in Human Breast Cancer. Cancer Research, 65, 7065-7070.

[18]   Lee, E.J., Gusev, Y., Jiang, J., Nuovo, G.J., Lerner, M.R., Frankel, W.L., Morgan, D.L., Postier, R.G., Brackett, D.G. and Schmittgen, T.D. (2007) Expression profiling Identifies microRNA Signature in Pancreatic Cancer. International Journal of Cancer, 120, 1046-1054.

[19]   Huang, Y.S., Dai, Y., Yu, X.F., Bao, S.Y., Yin, Y.B., Tang, M. and Hu, C.X. (2008) Microarray Analysis of microRNA Expression in Hepatocellular Carcinoma and Non-Tumorous Tissues without Viral Hepatitis. Journal of Gastroenterology and Hepatology, 23, 87-94.

[20]   Bihrer, V., Waidmann, O., Friedrich-Rust, M., Forestier, N., Susser, S. and Haupenthal, J. (2011) Serum microRNA-21 as Marker for Necroinflammation in Hepatitis C Patients with and without Hepatocellular Carcinoma. PLoS ONE, 6, e26971.

[21]   Marquez, R.T., Bandyopadhyay, S., Wendlandt, E.B., Keck, K., Hoffer, B.A., Icardi, M.S., Christensen, R.N., Schmidt, W.N. and McCaffrey, A.P. (2010) Correlation between microRNA Expression Levels and Clinical Parameters Associated with Chronic Hepatitis C Viral Infection in Humans. Laboratory Investigations, 90, 1727-1736.

[22]   Bihrer, V., Friedrich-Rust, M., Kronenberger, B., Forestier, N., Haupenthal, J., Shi, Y., Peveling-Oberhag, J., Radeke, H.H., Sarrazin, C., Herrmann, E., Zeuzem, S., Waidmann, O. and Piiper, A. (2011) Serum miR-122 as a Biomarker of Necroinflammation in Patients with Chronic Hepatitis C Virus Infection. American Journal of Gastroenterology, 106, 1663-1669.

[23]   Trebicka, J., Anadol, E., Elfimova, N., Strack, I., Roggendorf, M., Viazov, S., Wedemeyer, I., Drebber, U., Rockstroh, J., Sauerbruch, T., Dienes, H.P. and Odenthal, M. (2013) Hepatic and Serum Levels of miR-122 after Chronic HCV-Induced Fibrosis. Journal of Hepatology, 58, 234-239.

[24]   Su, T.H., Liu, C.H., Liu, C.G., Chen, C.L., Ting, T.T., Tseng, T.C., Chen, P.J., Kao, L.H. and Chen, D.S. (2013) Serum microRNA-122 Level Correlates with Virologic Responses to Pegylated Interferon Therapy in Chronic Hepatitis C. Proceedings of the National Academy of Sciences of the United States of America, 110, 7844-7849.

[25]   Poynard, T., Bedossa, P. and Opolon, P. (1997) Natural History of Liver Fibrosis Progression in Patients with Chronic Hepatitis C. The OBSVIRC, METAVIR, CLINIVIR, and DOSVIRC Groups. Lancet, 349, 825-832.

[26]   Gramantieri, L., Ferracin, M., Fornari, F., Veronese, A., Sabbioni, S., Liu, C.G., Calin, G.A., Giovannini, C., Ferrazzi, E., Grazi, G.L., Croce, C.M., Bolondi, L. and Negrini, M. (2007) Cyclin G1 Is a Target of miR-122a, a microRNA Frequently Down-Regulated in Human Hepatocellular Carcinoma. Cancer Research, 67, 6092-6099.

[27]   Köberle, V., Kronenberger, B., Pleli, T., Trojan, J., Imelmann, E., Peveling-Oberhag, J., Welker, M.W., Elhendawy, M., Zeuzem, S., Piiper, A. and Waidmann, O. (2013) Serum microRNA-1 and microRNA-122 Are Prognostic Markers in Patients with Hepatocellular Carcinoma. European Journal of Cancer, 49, 3442-3449.

[28]   EL-Abd, N.E., Fawzy, A.N., EL-Sheikh, S.M. and Soliman, M.E. (2015) Circulating miRNA-122, miRNA-199a, and miRNA-16 as Biomarkers for Early Detection of Hepatocellular Carcinoma in Egyptian Patients with Chronic Hepatitis C Virus Infection. Molecular Diagnostic Therapy, 19, 213-220.

[29]   Ezzat, H., Lotfy, A.M., Alalfy, M.N., El-Taher, S.M., Mokhtar, A., Mohamed, S.A. and EL-Senosy, F.M. (2014) The Significance of Circulating MicroRNA-122 as a Non Invasive Diagnostic Marker of Liver Injury in Egyptian Chronic Hepatitis C Virus Infected and Cirrhotic Patients with and without Hepatocellular Carcinoma. Clinical Medicine and Diagnostics, 4, 1-8.

[30]   Fattovich, G., Giustina, G., Sanchez-Tapias, J., Quero, C., Mas, A., Olivotto, P.G., Solinas, A., Almasio, P., Hadziyannis, S., Degos, F., de Moura, M.C., Krogsgaard, K., Pantalena, M., Realdi, G., Corrocher, R. and Schalm, S.W. (1998) Delayed Clearance of Serum HBsAg in Compensated Cirrhosis B: Relation to Interferon Alpha Therapy and Disease Prognosis. European Concerted Action on Viral Hepatitis (EUROHEP). American Journal of Gastroenterology, 93, 896-900.

[31]   Parkin, D.M., Bray, F.I. and Deves, S.S. (2001) Cancer Burden in the Year 2000. The Global Picture. European Journal of Cancer, 37, S4-S66.

[32]   Borel, F., Konstantinova, P. and Jansen, P.L. (2012) Diagnostic and Therapeutic Potential of miRNA Signatures in Patients with Hepatocellular Carcinoma. Journal of Hepatology, 56, 1371-1383.

[33]   Bandiera, S., Pfeffer, S., Thomas, F. and Zeisel, M.B. (2015) MiR-122—A Key Factor and Therapeutic Target in Liver Disease. Journal of Hepatology, 62, 448-457.

[34]   Fan, H.-X. and Tang, H. (2014) Complex Interactions between microRNAs and Hepatitis B/C Viruses. World Journal of Gastroenterology, 20, 13477-13492.

[35]   El-Garem, H., Ammer, A., Shehab, H., Shaker, O., Anwer, M., El-Akel, W. and Omar, H. (2014) Circulating microRNA, miR-122 and miR-221 Signature in Egyptian Patients with Chronic Hepatitis C Related Hepatocellular Carcinoma. World Journal of Hepatology, 27, 818-824.

[36]   Motawi, T.K., Shaker, O.G., El-Maraghy, S.A. and Senousy, M.A. (2015) Serum microRNAs as Potential Biomarkers for Early Diagnosis of Hepatitis C Virus-Related Hepatocellular Carcinoma in Egyptian Patients. PLoS ONE, 10, e0137706.

[37]   Yi, R., Qin, Y., Macara, I.G. and Cullen, B.R. (2003) Exportin-5 Mediates the Nuclear Export of Pre-MicroRNAs and Short Hairpin RNAs. Genes & Development, 17, 3011-3016.