Hepatitis C Virus Quasispecies in the Resistance to Antiviral Therapy

丙型肝炎病毒准种对抗病毒治疗的耐药性

• 批准号: 8242874
• 负责人: XIAOFENG FAN
• 金额: $ 28.81万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8242874
• 关键词: Acute Hepatitis C; Address; American; Antiviral Agents; Antiviral Therapy; Appearance; Behavior; Biology; Cessation of life; Chronic; Chronic Hepatitis C; Cirrhosis; Clinical; Cloning; Consensus Sequence; Data; Detection; Drug resistance; Experimental Models; Extinction (Psychology); Future; Genetic; Genetic Recombination; Genome; Genotype; Hepatitis C; Hepatitis C Antiviral; Hepatitis C virus; Infection; Integration Host Factors; Interferons; Length; Mutation; Nature; Patients; Pattern; Pegylated Interferon Alfa; Play; Population; Population Study; Public Health; Relapse; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Ribavirin; Role; Sampling; Serum; Structure; Techniques; Technology; Testing; Therapeutic; Therapeutic Agents; Time; United States; Variant; Viral; Viral Drug Resistance; Viral Genome; Virus; Yin; comparative; design; drug withdrawal; innovation; liver transplantation; new technology; novel; research study; resistance mechanism; response; success; viral resistance

Project Summary Hepatitis C virus (HCV) infection is one of the major concerns in public health. Currently, optimal antiviral therapy with pegylated interferon-alpha plus ribavirin, cures ~50% of patients infected with HCV genotype 1 and ~80% of patients infected with HCV genotypes 2 and 3. One of the difficulties regarding our understanding on treatment resistance is related to the nature of current antiviral agents. Both interferon and ribavirin have long been known for their broad-spectrum antiviral activity by creating a non-specific antiviral status rather than the direct interaction with viruses. Consequently, no explicit targets on HCV genome have been documented. Studies on drug resistance with these agents have generated very controversial data through conventional approaches that frequently focus on short domains of the HCV rather than the entire viral genome. Using a novel long RT-PCR and cloning technology and well- characterized serum samples from the Hepatitis C Antiviral Long-term Treatment against Cirrhosis trial (HALT-C), we propose a viral sequencing project through which viral mechanisms for the resistance to antiviral therapy will be exhaustively examined at multiple levels. HYPOTHESIS: HCV resistance to antiviral therapy is associated with region-dependent mutations of viral quasispecies at either single variants or the population level. Aim 1: To explore genetic signatures at both HCV isolate and quasispecies levels in null responders infected with HCV genotype 1a. The full-length HCV quasispecies profiles at the baseline will be generated from patients with either null or sustained virological responses (SVR), followed by comparative analyses to identify potential genetic signatures that are associated with the treatment resistance. Aim 2: To demonstrate if there are distinct quasispecies structures of HCV genotype 2 in terms of the high response rate to the antiviral therapy. The full-length HCV quasispecies profiles will be generated from twenty SVRs with HCV genotype 2a, followed by comparative analyses with those derived from HCV genotype 1a. Aim 3: To characterize mutational patterns associated with HCV re-emergence in patients with relapse after initial response to antiviral therapy. The relapse indicates the survival of HCV from a putative population bottleneck formed under antiviral therapy. How does HCV respond to such ¿in vivo¿ population bottlenecks? This issue will be addressed through a sequential comparative analysis of full-length HCV quasispecies profiles. Data from these studies will have immediate applications for rational design of future HCV antiviral therapy in which PegIFN-? and ribavirin are still the core components. Project Narrative Hepatitis C virus infection is one of the major concerns in public health and current antiviral therapy has very differential success within and among HCV genotypes. We will examine viral determinants responsible for the treatment resistance through the application of novel technology. This may help to design more effective antiviral therapy to chronic HCV infection.

项目摘要 丙型肝炎病毒（HCV）感染是公共卫生的主要问题之一。目前，最佳抗病毒药物 聚乙二醇干扰素-α联合利巴韦林治疗，治愈约50%的HCV基因型1感染患者 约80%的患者感染HCV基因型2和3。关于我们的困难之一 对治疗耐药性的理解与目前抗病毒药物的性质有关。干扰素 和利巴韦林长期以来一直以其广谱抗病毒活性而闻名， 抗病毒状态，而不是与病毒的直接相互作用。因此，没有明确的HCV靶点， 基因组已经被记录下来了。对这些药物的耐药性研究已经产生了非常 通过常规方法获得的有争议的数据，这些方法经常关注HCV的短结构域， 而不是整个病毒基因组。采用新型长链RT-PCR和克隆技术， 丙型肝炎抗病毒长期治疗肝硬化试验的特征血清样本 （HALT-C），我们提出了一个病毒测序项目，通过该项目， 抗病毒治疗将在多个水平上进行彻底检查。 假设：HCV对抗病毒治疗的耐药性与HCV基因的区域依赖性突变有关。 病毒准种无论是在单一变异或人口水平。 目的1：探索无效应答者HCV分离株和准种水平的遗传特征 感染HCV基因型1a。将生成基线时的全长HCV准种谱 从无效或持续病毒学应答（SVR）的患者中，随后进行比较分析， 识别与治疗抗性相关的潜在遗传特征。 目的2：证明HCV基因型2在高表达方面是否存在不同的准种结构， 抗病毒治疗的应答率。全长HCV准种谱将由 20个HCV基因型2a的SVRs，随后与来自HCV的SVRs进行比较分析 基因型1a。 目的3：描述HCV感染后复发患者中与HCV再出现相关的突变模式。 抗病毒治疗的初步反应。复发表明HCV从假定人群中存活 在抗病毒治疗下形成瓶颈。HCV如何应对这种体内群体瓶颈？ 这个问题将通过对全长HCV准种的连续比较分析来解决。 数据区. 这些研究的数据将直接应用于未来HCV抗病毒治疗的合理设计 其中聚乙二醇干扰素-？和利巴韦林仍然是核心成分。项目叙述 丙型肝炎病毒感染是公共卫生中的主要关注点之一，目前的抗病毒治疗已经 在HCV基因型内部和之间非常不同的成功。我们将研究病毒的决定因素 通过新技术的应用，这可能有助于设计更多 有效抗病毒治疗慢性HCV感染。

项目成果

Enhanced protocol for determining the 3' terminus of hepatitis C virus.

用于确定丙型肝炎病毒 3 末端的增强方案。

• DOI: 10.1016/j.jviromet.2010.03.030
• 发表时间: 2010
• 期刊: Journal of virological methods
• 影响因子: 3.1
• 作者: Zhang,Xiaoan;Fan,Xiaofeng;Xu,Yanjuan;DiBisceglie,AdrianM
• 通讯作者: DiBisceglie,AdrianM

Efficient amplification and cloning of near full-length hepatitis C virus genome from clinical samples.

从临床样品中有效扩增和克隆几乎全长丙型肝炎病毒基因组。

• DOI: 10.1016/j.bbrc.2006.06.039
• 发表时间: 2006-08-11
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Fan X;Xu Y;Di Bisceglie AM
• 通讯作者: Di Bisceglie AM

Comprehensive cloning of patient-derived 9022-bp amplicons of hepatitis C virus.

全面克隆患者来源的丙型肝炎病毒 9022 bp 扩增子。

• DOI: 10.1016/j.jviromet.2013.04.010
• 发表时间: 2013
• 期刊: Journal of virological methods
• 影响因子: 3.1
• 作者: Lu,Yang;Xu,Yanjuan;DiBisceglie,AdrianM;Fan,Xiaofeng
• 通讯作者: Fan,Xiaofeng

High diversity of hepatitis C viral quasispecies is associated with early virological response in patients undergoing antiviral therapy.

• DOI: 10.1002/hep.23290
• 发表时间: 2009-12
• 期刊: HEPATOLOGY
• 影响因子: 13.5
• 作者: Fan, Xiaofeng;Mao, Qing;Zhou, Donghui;Lu, Yang;Xing, Jianwei;Xu, Yanjuan;Ray, Stuart C.;Di Bisceglie, Adrian M.
• 通讯作者: Di Bisceglie, Adrian M.

Divergent quasispecies evolution in de novo hepatitis C virus infection associated with bone marrow transplantation.

• DOI: 10.1016/j.bbrc.2011.09.041
• 发表时间: 2011-10-14
• 期刊: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
• 影响因子: 3.1
• 作者: Wang, Weihua;Lin, Jianguo;Tan, De;Xu, Yanjuan;Brunt, Elizabeth M.;Fan, Xiaofeng;Di Bisceglie, Adrian M.
• 通讯作者: Di Bisceglie, Adrian M.