Volume 10, Issue 2 (Vol.10 No.2 Jul 2021)                   rbmb.net 2021, 10(2): 233-242 | Back to browse issues page


XML Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Lashgarian H E, Valibeik A, Marzban A, Karkhane M, Shahzamani K. The Relationship Between HCV-NS5A Gene Mutations and Resistance to Combination Therapy in Patients with HCV- Genotype 1-B. rbmb.net. 2021; 10 (2) :233-242
URL: http://rbmb.net/article-1-555-en.html
Department of Medical Biotechnology, School of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
Abstract:   (396 Views)
Background: Hepatitis C virus (HCV) is one of the major causes of chronic liver disease, as it holds a significant role in developing liver cirrhosis and hepatocellular carcinoma. Combination therapy with Pegaferon and Ribavirin leads to viral clearance of only 50% of patients. During the host antiviral response, protein kinase R (PKR) interacts with eukaryotic translation initiation factor 2 alpha (eIF2α), that leads to the inhibition of viral protein synthesis. The viral NS5A protein appears to interfere with this antiviral action, evading the host immune response. However, mutations in the NS5A gene have been observed to render HCV more susceptible to treatment. The aim of this study was to determine the mutations present in the IFN Sensitivity Determining Region (ISDR) and NS5A-PKRbinding domain regions in chronic HCV infected patients before and after therapy.

Methods: Viral RNA was isolated from the plasma of 52 chronic HCV infected patients before and after treatment. RT-Nested PCR reaction was used to reverse transcription and amplification of target fragment using the specific primers.

Results: Sequence analysis revealed no relationship between NS5A mutations and response to treatment. No significant difference was found between the mutations before and 3 months after treatment among responders and non-responders.

Conclusions: This study showed that the number of mutations in NS5A did not significantly differ between the patients who responded to treatment and the patients that did not. Therefore, sequencing of these regions does not appear to be a suitable tool for predicting treatment outcomes.
Full-Text [PDF 229 kb]   (198 Downloads)    
Type of Article: Original Article | Subject: Microbiology
Received: 2020/08/24 | Accepted: 2020/09/8 | Published: 2021/08/26

References
1. Thrift AP, El-Serag HB, Kanwal F. Global epidemiology and burden of HCV infection and HCV-related disease. Nat Rev Gastroenterol Hepatol. 2017;14(2):122-132. [DOI:10.1038/nrgastro.2016.176] [PMID]
2. Karkhane M, Mohebbi SR, Azimzadeh P, Niasar MS, Sarbazi MR, Sharifian A, et al. Lack of association between interleukin 28B gene polymorphisms (rs8099917G/T, rs12979860 C/T) and susceptibility to chronic hepatitis C virus infection, Tehran, Iran. Gastroenterol Hepatol Bed Bench. 2016;9(Suppl1):S29-S35.
3. Edlin BR, Eckhardt BJ, Shu MA, Holmberg SD, Swan T. Toward a more accurate estimate of the prevalence of hepatitis C in the United States. Hepatology. 2015;62(5):1353-63. [DOI:10.1002/hep.27978] [PMID] [PMCID]
4. Ip PP, Nijman HW, Wilschut J, Daemen T. Therapeutic vaccination against chronic hepatitis C virus infection. Antiviral research. 2012;96(1):36-50. [DOI:10.1016/j.antiviral.2012.07.006] [PMID]
5. Bartenschlager R, Lohmann V. The Hepatitis C Virus Replicon System and Its Role in Drug Development. 2019:69-96. [DOI:10.1007/7355_2018_34]
6. Susser S, Flinders M, Reesink HW, Zeuzem S, Lawyer G, Ghys A, et al. Evolution of hepatitis C virus quasispecies during repeated treatment with the NS3/4A protease inhibitor telaprevir. Antimicrob Agents Chemother. 2015;59(5):2746-55. [DOI:10.1128/AAC.04911-14] [PMID] [PMCID]
7. Moradpour D, Penin F, Rice C. Chapter 7- Replication of Hepatitis C Virus. In: Boyer TD, Manns MP, Sanyal AJ, editors. Zakim and Boyer's Hepatology (Sixth Edition). Saint Louis: W.B. Saunders; 2012. p. 97-110. [DOI:10.1016/B978-1-4377-0881-3.00007-3] [PMID]
8. Gupta SP. Chapter 3 - Inhibition of Viruses: Promising Targets and Their Importance. In: Gupta SP, editor. Studies on Hepatitis Viruses: Academic Press; 2018. p. 35-65. [DOI:10.1016/B978-0-12-813330-9.00003-X]
9. Pawlotsky J-M, Germanidis G, Neumann AU, Pellerin M, Frainais P-O, Dhumeaux D. Interferon resistance of hepatitis C virus genotype 1b: relationship to nonstructural 5A gene quasispecies mutations. J Virol. 1998;72(4):2795-2805 [DOI:10.1128/JVI.72.4.2795-2805.1998] [PMID]
10. Veillon P, Payan C, Le Guillou-Guillemette H, Gaudy C, Lunel F. Quasispecies evolution in NS5A region of hepatitis C virus genotype 1b during interferon or combined interferon-ribavirin therapy. World J Gastroenterol. 2007;13(8):1195-1203. [DOI:10.3748/wjg.v13.i8.1195] [PMID] [PMCID]
11. Holysz M, Bialas K, Migdalski P, Kmieciak D, Trzeciak W. Identification of mutations in the HVR1 and PKR-BD regions in HCV-infected patients resistant to PEG-IFNα/RBV therapy. J Appl Genet. 2015;56(3):403-9. [DOI:10.1007/s13353-014-0267-0] [PMID] [PMCID]
12. Jarret A, McFarland AP, Horner SM, Kell A, Schwerk J, Hong M, et al. Hepatitis-C-virus-induced microRNAs dampen interferon-mediated antiviral signaling. Nat Med. 2016;22(12):1475-1481. [DOI:10.1038/nm.4211] [PMID] [PMCID]
13. Yokozaki S, Katano Y, Hayashi K, Ishigami M, Itoh A, Hirooka Y, et al. Mutations in two PKR-binding domains in chronic hepatitis C of genotype 3a and correlation with viral loads and interferon responsiveness. J Med Virol. 2011;83(10):1727-32. [DOI:10.1002/jmv.21959] [PMID]
14. García-Sastre A. Ten strategies of interferon evasion by viruses. Cell Host Microbe. 2017;22(2):176-184. [DOI:10.1016/j.chom.2017.07.012] [PMID] [PMCID]
15. de Rueda PM, Rodríguez JMF, Pérez RQ, Medina AG, Álvarez ABM, Ruíz JC, et al. Hepatitis C virus NS5A region mutation in chronic hepatitis C genotype 1 patients who are non-responders to two or more treatments and its relationship with response to a new treatment. World J Gastroenterol. 2017;23(25):4538-4547. [DOI:10.3748/wjg.v23.i25.4538] [PMID] [PMCID]
16. Nousbaum J-B, Polyak S, Ray S, Sullivan D, Larson A, Carithers R, et al. Prospective characterization of full-length hepatitis C virus NS5A quasispecies during induction and combination antiviral therapy. J Virol. 2000;74(19):9028-38. [DOI:10.1128/JVI.74.19.9028-9038.2000] [PMID] [PMCID]
17. Sayan M, Yıldırım FS, Akhan S, Yıldırım AA, Şirin G, Cabalak M, et al. NS5A resistance - associated substitutions in chronic hepatitis C patients with direct acting antiviral treatment failure in Turkey. Int J Infect Dis. 2020;95:84-89. [DOI:10.1016/j.ijid.2020.03.061] [PMID]
18. Aldunate F, Echeverría N, Chiodi D, López P, Sánchez-Cicerón A, Fajardo A, et al. Pretreatment Hepatitis C Virus NS5A/NS5B Resistance-Associated Substitutions in Genotype 1 Uruguayan Infected Patients. Dis Markers. 2018;2018:2514901. [DOI:10.1155/2018/2514901] [PMID] [PMCID]
19. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997;25(24):4876-82. [DOI:10.1093/nar/25.24.4876] [PMID] [PMCID]
20. Choo Q, Richman K, Han J, Berger K, Lee C, Dong C, et al. Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A. 1991;88(6):2451-2455. [DOI:10.1073/pnas.88.6.2451] [PMID] [PMCID]
21. Jabari H, Bayatian A, Sharifi AH, Zaer RH, Fakharzadeh E, Asadi R, et al. Safety and efficacy of locally manufactured pegylated interferon in hepatitis C patients. 2010;13(4):306-12.
22. Pascu M, Martus P, Höhne M, Wiedenmann B, Hopf U, Schreier E, et al. Sustained virological response in hepatitis C virus type 1b infected patients is predicted by the number of mutations within the NS5A-ISDR: a meta-analysis focused on geographical differences. Gut. 2004;53(9):1345-51. [DOI:10.1136/gut.2003.031336] [PMID] [PMCID]
23. Enomoto N, Sakuma I, Asahina Y, Kurosaki M, Murakami T, Yamamoto C, et al. Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection. N Engl J Med. 1996;11;334(2):77-81. [DOI:10.1056/NEJM199601113340203] [PMID]
24. Khorrami S, Mohammadpour H, Shahzamani K, Zarif MN, Sharifi AH, Merat S, et al. The relationship between HLA-G and viral loads in non-responder HCV-infected patients after combined therapy with IFN-α2α and ribavirin. Hum Immunol. 2015;76(2-3):181-6. [DOI:10.1016/j.humimm.2014.12.012] [PMID]
25. Sepahi S, Pasdar A, Gerayli S, Rostami S, Gholoobi A, Meshkat Z. CTLA-4 Gene Haplotypes and the Risk of Chronic Hepatitis C Infection; a Case Control Study. Rep Biochem Mol Biol. 2017;6(1):51-58.
26. McHutchison JG, Gordon SC, Schiff ER, Shiffman ML, Lee WM, Rustgi VK, et al. Interferon alfa-2b alone or in combination with ribavirin as initial treatment for chronic hepatitis C. Hepatitis Interventional Therapy Group. N Engl J Med. 1998;339(21):1485-92. [DOI:10.1056/NEJM199811193392101] [PMID]
27. Tokumoto Y, Hiasa Y, Horiike N, Michitaka K, Matsuura B, Chung RT, et al. Hepatitis C virus expression and interferon antiviral action is dependent on PKR expression. Journal of medical virology. 2007;79(8):1120-1127. [DOI:10.1002/jmv.20902] [PMID]
28. El-Dahshan D, Bahy D, Wahid A, Ahmed AE, Hanora A. Two novel SNPs in the promoter region of PKR gene in hepatitis C patients and their impact on disease outcome and response to treatment. Arab J Gastroenterol. 2018;19(3):106-115. [DOI:10.1016/j.ajg.2018.06.002] [PMID]
29. von vorgelegt, Gisa A, Berlin aus. Pathophysiology of Hepatitis E Virus Infection: Viral Evolution During Antiviral Therapy and Virus-specific T Cell Responses. Medizinische Hochschule Hannover. 2015.
30. Arase Y, Ikeda K, Chayama K, Murashima N, Tsubota A, Suzuki Y, et al. Efficacy and changes of the nonstructural 5A GENE by prolonged interferon therapy for patients with hepatitis C virus genotype 1b and a high level of serum HCV-RNA. Intern Med. 1999;38(6):461-6. [DOI:10.2169/internalmedicine.38.461] [PMID]
31. Aslan N, Bozdayi AM, Cetinkaya H, Sarioglu M, Turkay C, Bozkaya H, et al. The mutations in ISDR of NS5A gene are not associated with response to interferon treatment in Turkish patients with chronic hepatitis C virus genotype 1b infection. Turk J Gastroenterol. 2004;15(1):21-6.
32. Murayama M, Katano Y, Nakano I, Ishigami M, Hayashi K, Honda T, et al. A mutation in the interferon sensitivity‐determining region is associated with responsiveness to interferon‐ribavirin combination therapy in chronic hepatitis patients infected with a Japan‐specific subtype of hepatitis C virus genotype 1B. J Med Virol. 2007;79(1):35-40. [DOI:10.1002/jmv.20766] [PMID]
33. Jardim ACG, Yamasaki LHT, de Queiróz ATL, Bittar C, Pinho JRR, Carareto CMA, et al. Quasispecies of hepatitis C virus genotype 1 and treatment outcome with peginterferon and ribavirin. Infect Genet Evol. 2009;9(4):689-98. [DOI:10.1016/j.meegid.2008.11.001] [PMID]
34. Zhang L, Han F, Zhang D, Dou X-G. Mutations in different regions of the genome of hepatitis C virus genotype 1b and association with response to interferon therapy. Int J Mol Med. 2012;30(6):1438-42. [DOI:10.3892/ijmm.2012.1155] [PMID]
35. Puig-Basagoiti F, Sáiz J-C, Forns X, Ampurdanès S, Giménez-Barcons M, Franco S, et al. Influence of the genetic heterogeneity of the ISDR and PePHD regions of hepatitis C virus on the response to interferon therapy in chronic hepatitis C. J Med Virol. 2001;65(1):35-44. [DOI:10.1002/jmv.1098] [PMID]
36. Duverlie G, Khorsi H, Castelain S, Jaillon O. Izopet J, Lunel F, Eb F, Penin F, Wychows C. Sequence analysis of the NS5A protein of European hepatitis Cvirus 1b isolates and relation to interferon sensitivity. J Gen Virol. 1998;79( Pt 6):1373-81. [DOI:10.1099/0022-1317-79-6-1373] [PMID]

Add your comments about this article : Your username or Email:
CAPTCHA

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2015 All Rights Reserved | Reports of Biochemistry and Molecular Biology

Designed & Developed by : Yektaweb