Dissecting Structural Details of Hepadnavirus Subviral Particles

剖析嗜肝DNA病毒亚病毒颗粒的结构细节

基本信息

批准号：
10407642
负责人：
Che-Yen Wang
金额：
$ 20.27万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-19 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10407642
关键词：
Animal Experimentation Antibodies Architecture B-Lymphocytes Biological Models Capsid Cell Line Cell physiology Cells Chronic Hepatitis B Clinical Trials Cryo-electron tomography Cryoelectron Microscopy Disease Electron Microscope Endoplasmic Reticulum Epitopes Experimental Models Filament Foundations Future Genome Goals Golgi Apparatus Hepadnaviridae Hepatitis B Hepatitis B Infection Hepatitis B Surface Antigens Hepatitis B Virus Human Immune Immune Evasion Immune response Integral Membrane Protein Label Lead Learning Malaria Vaccines Masks Medical Membrane Membrane Biology Membrane Lipids Modeling Morphogenesis Morphology N-terminal Natural Immunity Pathogenesis Pathway interactions Patients Persons Pharmaceutical Preparations Proteins Public Health Reporting Resolution Role Serum Structure Study models Surface Antigens System T-Lymphocyte Testing Time Vaccines Viral Virion Virus Diseases Woodchuck Woodchuck Hepatitis B Virus anti-hepatitis B base clinically relevant drug testing hepatoma cell insight neutralizing antibody novel therapeutic intervention particle pre-clinical preclinical evaluation prevent serological marker

项目摘要

Hepatitis B virus (HBV) infection is a global public health concern; however, current approval therapies rarely lead to a complete cure. Hepatitis B surface antigen (HBsAg) is the serological marker for HBV infection. It is characterized by three transmembrane proteins having the same C-termini with variable in-frame extension at the N-terminal ends. Naturally, HBsAg proteins are incorporated in the viral envelope or assembled into genome-free and capsid-free subviral particles (SVPs) with the lipid membrane derived from the host cells. Recently, a new therapeutic approach by blocking assembly and release of SVPs has shown promising results in rapidly reducing HBsAg level in chronic HBV- infected patients. Despite its potential clinically relevant applications, there is no structural information available for HBsAg and yet different models of SVPs have been proposed. In this proposal we focus on obtaining the high resolution structures of SVPs from different model systems using cryo-electron microscope (cryo-EM). We will test the central hypothesis that changes in the structural organizations of SVPs will have important implications on their secretion pathway. In Aim 1, we will determine the structures of SVP from woodchuck hepatitis virus (WHV). Woodchuck infected with WHV is a critical experimental model for studying pathogenesis of HBV infection. In Aim 2, we will elucidate the structural details of HBV SVP from carrier's sera and hepatoma cell line. We will use antibodies to spatially label different regions of HBsAg. Finally, all structures determined from this proposal will be compared and used to build a complete picture of SVP with the detailed structural information of Hepadnavirus surface antigen. Successful completion of the proposed project will reveal new structural information of SVPs and will deepen our understanding of HBV particles morphogenesis to facilitate current efforts in finding a cure for HBV infection.

丙型肝炎病毒（HBV）感染是全球公共卫生问题；但是，当前的批准疗法很少导致完全治愈。丙型肝炎表面抗原（HBSAG）是HBV感染的血清学标记。它的特征是三个具有相同C末端的跨膜蛋白，在N末端末端具有可变的框架延伸。自然地，HBSAG蛋白被掺入病毒包膜中，或组装成无基因组和无衣壳亚皮下颗粒（SVP），并带有源自宿主细胞的脂质膜。最近，通过阻止组装和释放SVP的一种新的治疗方法，显示出令人鼓舞的结果，可迅速降低慢性HBV感染患者的HBSAG水平。尽管它具有潜在的临床相关应用，但仍未提出HBSAG的结构信息，但已提出了不同的SVP模型。在此提案中，我们着重于使用低温电子显微镜（Cryo-EM）从不同模型系统中获得SVP的高分辨率结构。我们将检验一个中心假设，即SVP的结构组织的变化将对其分泌途径具有重要意义。在AIM 1中，我们将确定Woodchuck肝炎病毒（WHV）中SVP的结构。感染WHV的Woodchuck是研究HBV感染发病机理的关键实验模型。在AIM 2中，我们将阐明从载体血清和肝癌细胞系中HBV SVP的结构细节。我们将使用抗体在空间上标记HBSAG的不同区域。最后，将比较从该提案中确定的所有结构，并用于构建SVP的完整图片，并具有肝表面抗原的详细结构信息。拟议项目的成功完成将揭示SVP的新结构信息，并将加深我们对HBV颗粒形态发生的理解，以促进当前的努力，以寻求治愈HBV感染的方法。