Studies of HCV Infection, Vaccine Development and HCV-Host interactions

HCV 感染、疫苗开发和 HCV-宿主相互作用的研究

基本信息

批准号：
10697775
负责人：
T. Jake Liang
金额：
$ 56.91万
依托单位：
NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10697775
关键词：
3-Dimensional Animal Model Antibodies Binding Biotin Cell Culture System Cells Classification Complex Cryoelectron Microscopy Crystallization Data Set Dependence Development Diazomethane Dimensions Disease Gene Expression Genetic Glycoproteins Hepatitis C Hepatitis C Therapy Hepatitis C Vaccine Hepatitis C virus Individual Infectious hepatitides Integration Host Factors Life Cycle Stages Mediating Modeling Morbidity - disease rate Negative Staining Pathogenesis Preventive vaccine Process Proteins RNA Interference Receptor Cell Recombinants Resolution Shapes Structural Models Structure Transmembrane Domain Viral Virus Diseases Virus Replication X-Ray Crystallography base crosslink density design in vivo mortality neutralizing antibody particle small molecule stem therapeutic target three-dimensional modeling vaccine development virus host interaction

项目摘要

HCV dependencies on the host machinery are both intricate and extensive. Each of these host dependencies is a potential therapeutic target. Previous efforts have been successful in discovering important steps in HCV replication, yet many fundamental processes in the viral life cycle remain uncharacterized. Using RNAi-based genetics and an infectious HCV cell culture system, we identified many previously unrecognized host factors required for productive HCV infection. Despite the development of highly effective HCV treatments, an effective prophylactic vaccine is still lacking. HCV infection is mediated by its envelope glycoproteins E1 and E2 in the entry process with E2 binding to cell receptors and E1 mediating endosomal fusion. The structure of E1E2 has only been partially resolved by the X-ray crystallography of the core domain of E2 protein (E2c) and its complex with various neutralizing antibodies. Structural understanding of the E1E2 heterodimer in its native form can advance the design of candidates for HCV vaccine development. Here we analyze the structure of recombinant HCV E1E2 heterodimer with the aid of well-defined monoclonal anti-E1 and E2 antibodies as well as a small molecule chlorcyclizine-diazirine-biotin that can target and cross-link the putative E1 fusion domain. 3D models were generated after extensive 2D classification analysis with negative-stain single particles datasets. We modeled the available crystal structures of the E2c and Fabs into the 3D volumes of E1E2-Fab complexes based on the shape and dimension of the domain density. The E1E2 heterodimer exists in monomeric form and consists of a main globular body presumably depicting the E1 and E2 stem/transmembrane domain and a protruding structure representing the E2c based on anti-E2 Fab binding. At low resolution, model generated from negatively stained analysis reveals the unique binding and orientation of individual or double Fabs onto the E1 and E2 components of the complex. Cryo-electron microscopy (cryo-EM) of double Fab complexes resulting in a refined structural model of the E1E2 heterodimer is presented.

HCV 对宿主的依赖性既复杂又广泛。这些宿主依赖性中的每一种都是潜在的治疗靶点。之前的努力已经成功地发现了丙型肝炎病毒复制的重要步骤，但病毒生命周期中的许多基本过程仍然未知。利用基于 RNAi 的遗传学和感染性 HCV 细胞培养系统，我们确定了许多以前未被识别的 HCV 感染所需的宿主因子。尽管开发了高效的 HCV 治疗方法，但仍然缺乏有效的预防性疫苗。 HCV感染在进入过程中由其包膜糖蛋白E1和E2介导，E2与细胞受体结合，E1介导内体融合。 E1E2 的结构仅通过 E2 蛋白 (E2c) 核心结构域及其与各种中和抗体的复合物的 X 射线晶体学分析得到部分解析。对天然形式的 E1E2 异二聚体的结构了解可以促进 HCV 疫苗开发候选物的设计。在这里，我们借助明确的单克隆抗 E1 和 E2 抗体以及可以靶向并交联假定的 E1 融合结构域的小分子氯环嗪-二嗪-生物素来分析重组 HCV E1E2 异二聚体的结构。使用负染色单颗粒数据集进行广泛的 2D 分类分析后生成 3D 模型。我们根据域密度的形状和尺寸，将 E2c 和 Fab 的可用晶体结构建模为 E1E2-Fab 复合物的 3D 体积。 E1E2 异二聚体以单体形式存在，由可能描绘 E1 和 E2 茎/跨膜结构域的主球体和代表基于抗 E2 Fab 结合的 E2c 的突出结构组成。在低分辨率下，负染色分析生成的模型揭示了单个或双 Fab 在复合物的 E1 和 E2 成分上的独特结合和方向。提出了双 Fab 复合物的冷冻电子显微镜 (cryo-EM)，从而产生了 E1E2 异二聚体的精细结构模型。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hepatitis C virus JFH-1 strain infection in chimpanzees is associated with low pathogenicity and emergence of an adaptive mutation.

DOI：
10.1002/hep.22422
发表时间：
2008-09
期刊：
HEPATOLOGY
影响因子：
13.5
作者：
Kato, Takanobu;Choi, Youkyung;Elmowalid, Gamal;Sapp, Ronda K.;Barth, Heidi;Furusaka, Akihiro;Mishiro, Shunji;Wakita, Takaji;Krawczynski, Krzysztof;Liang, T. Jake
通讯作者：
Liang, T. Jake

Cellular microRNA networks regulate host dependency of hepatitis C virus infection.

DOI：
10.1038/s41467-017-01954-x
发表时间：
2017-11-27
期刊：
Nature communications
影响因子：
16.6
作者：
Li Q;Lowey B;Sodroski C;Krishnamurthy S;Alao H;Cha H;Chiu S;El-Diwany R;Ghany MG;Liang TJ
通讯作者：
Liang TJ

Erratum for Lowey et al., "Hepatitis C Virus Infection Induces Hepatic Expression of NF-κB-Inducing Kinase and Lipogenesis by Downregulating miR-122".

Lowey 等人的勘误表，“丙型肝炎病毒感染通过下调 miR-122 诱导 NF-κB 诱导激酶和脂肪生成的肝脏表达”。

DOI：
10.1128/mbio.02771-19
发表时间：
2019
期刊：
mBio
影响因子：
6.4
作者：
Lowey,Brianna;Hertz,Laura;Chiu,Stephan;Valdez,Kristin;Li,Qisheng;Liang,TJake
通讯作者：
Liang,TJake

Settling the "score" with liver cancer.