Cyclosporine, Cyclophilins and HCV Replication

环孢素、亲环素和 HCV 复制

• 批准号: 8309405
• 负责人: HENGLI TANG
• 金额: $ 32.13万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-04 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8309405
• 关键词: Acute; Address; Affinity; Antiviral Agents; Binding; Biochemical; Biological Assay; Biological Models; C-terminal; Cell Communication; Cell Culture Techniques; Cells; Clinical; Clinical Treatment; Clinical Trials; Complex; Cyclophilin A; Cyclophilins; Cyclosporine; DNA-Directed RNA Polymerase; Data; Dependence; Development; Diagnostic; Drug resistance; Ensure; Face; Fibrosis; Genotype; Goals; Health system; Hepatitis C; Hepatitis C virus; Human; Human Virus; In Vitro; Infection; Infection prevention; Interferons; Intervention; Knowledge; Lead; Liver Cirrhosis; Maps; Mediator of activation protein; Messenger RNA; Molecular; Molecular Biology Techniques; Molecular Chaperones; Mutagenesis; Mutation; Nonstructural Protein; Parasites; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Population; Primary carcinoma of the liver cells; Proteins; RNA Binding; Research; Resistance; Role; Small Interfering RNA; Surface; Testing; Therapeutic; Thumb structure; Vaccines; Variant; Viral; Viral Genome; Virus; Virus Replication; Work; anti-hepatitis C; base; chronic liver disease; cofactor; drug candidate; global health; hepatoma cell; in vivo; insight; interferon therapy; mutant; new therapeutic target; novel; pathogen; positional cloning; prevent; programs; prophylactic; protein function; replicase; research study; resistant strain; viral RNA; viral resistance

Project Summary The broad, long-term goal of our program is to advance the knowledge of virus-host cell interactions by characterizing cellular cofactors essential for hepatitis C virus (HCV) infection. HCV infects 170 million people worldwide and is major cause of hepatocellular carcinoma. Understanding how this virus interacts with the host is of critical importance to the development of diagnostics, antiviral drugs, and a prophylactic vaccine. Our previous studies demonstrated that HCV infection of cultured hepatoma cells is critically dependent on a cellular protein, cyclophilin A (CyPA), and that ablating the function of this protein can not only prevent new infections but also suppress an existing infection. In addition, CyPA is a principal mediator of HCV resistance to cyclosporine A (CsA) and its derivatives, which inhibit HCV replication with an undefined mechanism and are currently being evaluated in clinical trials as candidate anti- HCV drugs. The experiments proposed here will investigate the molecular mechanisms that determine the essential cofactor function of CyPA, the action of CsA, and the related CsA resistance. The following experiments will be performed: (1) CsA and small interfering RNA directed at CyPA mRNA will be used to identify the specific function of HCV replicase that requires CyPA as a cofactor. A detailed understanding of why and where the virus needs this cellular chaperone to survive will offer new perspectives on the replication strategy of HCV. (2) Biochemical assay and mutagenesis will be used to characterize the critical interaction between CyPA and the viral replicase, which represents a novel structural interface of virus-host cell interaction that maybe disrupted for therapeutic purposes. (3) Macromolecular interaction and reverse genetics will be employed to dissect the mode of action for CsA and molecular basis of CsA resistance with the ultimate goal of predicting and circumventing that resistance. The results of the proposed studies will not only provide significant insights into how highly successful parasites such as human viruses hijack host cell machineries to replicate efficiently but also reveal new therapeutic targets for antiviral intervention.

项目摘要 我们的计划的广泛的，长期的目标是推进病毒宿主细胞的知识， 通过表征丙型肝炎病毒（HCV）感染所必需的细胞辅因子来研究相互作用。 HCV感染全球1.7亿人，是肝细胞癌的主要原因。 了解这种病毒如何与宿主相互作用对于开发 诊断、抗病毒药物和预防性疫苗。我们之前的研究表明 培养的肝癌细胞的HCV感染严重依赖于细胞蛋白， 亲环素A（CyPA），并且消除这种蛋白质的功能不仅可以防止新的 感染，但也抑制现有的感染。此外，CyPA是一个主要的调解人， HCV对环孢菌素A（CsA）及其衍生物的耐药性，其抑制HCV复制， 目前正在临床试验中作为候选抗- HCV药物这里提出的实验将研究分子机制， 确定CyPA的基本辅因子功能、CsA的作用和相关CsA 阻力将进行以下实验：（1）CsA和小干扰RNA 针对CyPA mRNA的基因将用于鉴定HCV复制酶的特异性功能， 需要CyPA作为辅助因子。详细了解病毒为什么以及在哪里需要这个 细胞伴侣的存活将提供新的视角对丙型肝炎病毒的复制策略。（二） 将使用生化测定和诱变来表征 CyPA与病毒复制酶的相互作用，代表了病毒-宿主细胞的一种新的结构界面 为了治疗的目的而中断的相互作用。(3)大分子相互作用和 反向遗传学将被用来剖析CsA的作用模式和 CsA耐药的最终目标是预测和规避这种耐药。的 拟议研究的结果不仅将提供重要的见解， 成功的寄生虫，如人类病毒，劫持宿主细胞机制，以有效地复制 而且还揭示了抗病毒干预的新治疗靶点。

项目成果

Cyclophilins as modulators of viral replication.

• DOI: 10.3390/v5071684
• 发表时间: 2013-07-11
• 期刊: Viruses
• 作者: Frausto SD;Lee E;Tang H
• 通讯作者: Tang H

A major determinant of cyclophilin dependence and cyclosporine susceptibility of hepatitis C virus identified by a genetic approach.

• DOI: 10.1371/journal.ppat.1001118
• 发表时间: 2010-09-23
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Yang F;Robotham JM;Grise H;Frausto S;Madan V;Zayas M;Bartenschlager R;Robinson M;Greenstein AE;Nag A;Logan TM;Bienkiewicz E;Tang H
• 通讯作者: Tang H

Cyclophilin inhibitors as a novel HCV therapy.

亲环蛋白抑制剂作为一种新型 HCV 疗法。

• DOI: 10.3390/v2081621
• 发表时间: 2010
• 期刊: Viruses
• 作者: Tang,Hengli