Transient replication of a hepatitis C virus genotype 1b replicon chimera encoding NS5A-5B from genotype 3a
Transient replication of a hepatitis C virus genotype 1b replicon chimera encoding NS5A-5B from genotype 3a
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DOI：
10.1016/j.jviromet.2013.09.017
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发表时间：
2014-01-01
影响因子：
3.1
通讯作者：
Targett-Adams, Paul
中科院分区：
文献类型：
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作者：
Kylefjord, Helen;Danielsson, Axel;Targett-Adams, Paul
尽管丙型肝炎病毒（HCV）是一种具有全球意义的病原体，但目前临床开发的实验性疗法包括高效的全口服HCV直接作用抗病毒药物（DAAs）组合。如果批准使用，这些新的治疗方案将极大地影响我们在大多数病毒感染患者中根除HCV的能力。然而，最近来自后期临床评估的数据表明，感染HCV基因型（GT） 3的个体对全口服DAA联合治疗的反应不如感染其他HCV GTs的患者。鉴于这些观察结果，本研究试图扩大可用的分子工具的数量，以研究临床前组织培养系统中HCV GT3 NS5A和NS5B蛋白的小分子介导抑制。因此，利用GT1b主链修饰，从欧洲丙型肝炎病毒数据库在线存储的HCV GT3基因组序列生成的一致序列中产生NS5A和NS5B蛋白，创建了一个新的亚基因组HCV复制子嵌合体。这种方法避免了从HCV感染患者的血清中分离和扩增HCV基因组的需要。复制子嵌合体，连同一个表达编码Y93H突变的NS5A的版本，在短暂的检测中显示出足够强大的复制水平，可以准确评估代表不同HCV DAA类别的抑制剂的抗病毒活性。因此，复制子嵌合体代表了一种新的简单分子工具，适用于药物发现计划，旨在研究、理解和提高靶向NS5A或NS5B的HCV DAAs的GT3a活性。(C) 2013 Elsevier B.V.版权所有
Although hepatitis C virus (HCV) is a pathogen of global significance, experimental therapies in current clinical development include highly efficacious all-oral combinations of HCV direct-acting antivirals (DAAs). If approved for use, these new treatment regimens will impact dramatically upon our capacity to eradicate HCV in the majority of virus-infected patients. However, recent data from late-stage clinical evaluations demonstrated that individuals infected with HCV genotype (GT) 3 responded less well to all-oral DAA combinations than patients infected with other HCV GTs. In light of these observations, the present study sought to expand the number of molecular tools available to investigate small molecule-mediated inhibition of HCV GT3 NS5A and NS5B proteins in preclinical tissue-culture systems. Accordingly, a novel subgenomic HCV replicon chimera was created by utilizing a GT1b backbone modified to produce NS5A and NS5B proteins from a consensus sequence generated from HCV GT3 a genomic sequences deposited online at the European Hepatitis C Virus database. This approach avoided the need to isolate and amplify HCV genomes from sera derived from HCV-infected patients. The replicon chimera, together with a version engineered to express NS5A encoding a Y93H mutation, demonstrated levels of replication in transient assays robust enough to assess accurate antiviral activities of inhibitors representing different HCV DAA classes. Thus, the replicon chimera represents a new simple molecular tool suitable for drug discovery programmes aimed at investigating, understanding, and improving GT3a activities of HCV DAAs targeting NS5A or NS5B. (C) 2013 Elsevier B.V. All rights reserved.