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 我们程序的长期目标是提高对病毒宿主细胞的了解 通过表征对丙型肝炎病毒（HCV）感染必不可少的细胞辅助因子的相互作用。 HCV在全球范围内感染了1.7亿人，是肝细胞癌的主要原因。 了解该病毒与宿主的相互作用是至关重要的 诊断，抗病毒药物和预防性疫苗的诊断。我们以前的研究表明 培养的肝癌细胞的HCV感染严重取决于细胞蛋白， 环磷脂A（CYPA），并且消融该蛋白质的功能不仅可以防止新的 感染但也抑制了现有的感染。此外，CYPA是 HCV对环孢素A（CSA）及其衍生物的抗性，它抑制HCV复制 一种不确定的机制，目前正在临床试验中评估作为候选抗 HCV药物。这里提出的实验将研究分子机制 确定CYPA的必需辅因子函数，CSA的作用和相关CSA 反抗。将进行以下实验：（1）CSA和小干扰RNA 针对CYPA mRNA的针对HCV复制酶的特定功能将使用 需要CYPA作为辅助因子。对病毒需要以及在哪里需要的详细理解 生存的细胞伴侣将为HCV的复制策略提供新的观点。 （2） 生化测定和诱变将用于表征 CYPA和病毒复制酶，它代表病毒宿主细胞的新结构界面 可能出于治疗目的而破坏的互动。 （3）大分子相互作用和 将采用反向遗传学来剖析CSA的作用方式和分子基础 CSA耐药性的最终目标是预测和规避这种抗性。这 拟议研究的结果不仅将提供有关高度高度的重要见解 成功的寄生虫，例如人类病毒劫持宿主细胞机器以有效复制 但也揭示了抗病毒干预的新治疗靶标。

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