Targeting glycoprotein (G) domain-III for pan-lyssavirus nanobody therapeutics

靶向糖蛋白 (G) 结构域 III 用于泛狂犬病病毒纳米抗体治疗

基本信息

批准号：
10667756
负责人：
Kai Xu
金额：
$ 22.82万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-07 至 2023-11-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10667756
关键词：
Acute Adopted Affect Affinity Antibodies Antigen-Antibody Complex Antigenic Diversity Antigens Bacteriophages Binding Biochemical Biological Assay Camels Central Nervous System Cessation of life Clinical Communicable Diseases Development Disease Disease Progression Dose Electron Microscopy Encephalitis Epitopes Family Future GTP-Binding Proteins Generations Genetic Variation Glycoproteins Homologous Protein Human Immune Immunize Immunoglobulins Immunotherapeutic agent Libraries Lyssavirus Mammals Maps Modality Modeling Molecular Conformation Monoclonal Antibodies PH Domain Persons Phage Display Prophylactic treatment Proteins Rabies Rabies Vaccines Reporting Rhabdoviridae Sequence Analysis Site Structure Surface Tertiary Protein Structure Therapeutic Therapeutic Agents Therapeutic antibodies Vaccines Viral Virion Virus X-Ray Crystallography Zoonoses design design and construction extracellular flexibility glycoprotein G human monoclonal antibodies improved in vivo member mouse model nanobodies neutralizing antibody novel prevent protective efficacy protein structure screening segregation structural biology therapeutically effective

项目摘要

Abstract The lethal rabies encephalitis diseases are caused by members of the zoonotic lyssavirus genus. The high genetic diversity separates lyssavirus members into three phylogroups, while cross-phylogroup (especially to phylogroups 2 and 3) protection has not been established by current post-exposure prophylaxis (PEP) or antibody therapeutics. To achieve more broadly effective countermeasures to rabies disease worldwide and to prepare for the emergence of new lyssaviruses, effective therapeutic agents against all phylogroups are necessary. Lyssavirus G glycoprotein is the sole antibody target on the virion surface, which adopt distinct conformations between pre- and post- fusion states as other class-III viral fusion machineries. Domain-III of lyssavirus G glycoprotein encompasses a large neutralizing antibody-accessible surface in the native state. Sequence analysis on domain-III revealed several conserved patches across all phylogroups, which could potentially serve as target epitopes for pan-lyssavirus neutralizers. Antigen-specific nanobodies have been considered as promising therapeutic agents against various infectious diseases and non-infectious diseases, which have the advantage in binding compact and hidden epitopes that are out of reach for conventional antibodies. We hypothesized that by focusing immune recognition on the lyssavirus G domain-III conserved epitopes, we can identify pan-lyssavirus neutralizing nanobodies for immunotherapeutics. In this study, we propose two specific aims: (1) to use a structure-based and antibody-guided approach to design and characterize the antigenicity of lyssavirus glycoprotein domain-III, a site of viral vulnerability targeted by several broadly neutralizing antibodies, for revelation of the antibody neutralization mechanism; (2) to identify and characterize pan-lyssavirus neutralizing nanobodies for therapeutics by focusing immune recognition on lyssavirus G domain III conserved epitopes.

抽象的致命的狂犬病脑炎疾病是由人畜共患狂犬病病毒属成员引起的。高遗传多样性将狂犬病病毒属病毒成员分为三个系统群，而跨系统群（尤其是目前的暴露后预防 (PEP) 尚未建立对系统群 2 和 3）的保护，或者抗体疗法。在全世界范围内采取更广泛有效的狂犬病对策并为新的狂犬病病毒的出现做好准备，针对所有系统群的有效治疗剂必要的。狂犬病病毒G糖蛋白是病毒颗粒表面的唯一抗体靶标，它采用独特的与其他 III 类病毒融合机制一样，融合前和融合后状态之间的构象。域 III 狂犬病病毒G糖蛋白在天然状态下包含一个大的中和抗体可接触表面。结构域 III 的序列分析揭示了所有系统群中的几个保守补丁，这可能可能作为泛狂犬病病毒中和剂的目标表位。抗原特异性纳米抗体已被被认为是针对各种感染性疾病和非感染性疾病的有前途的治疗剂，其优点是可以结合传统方法无法达到的紧凑和隐藏表位抗体。我们假设通过将免疫识别集中在狂犬病病毒 G 结构域 III 上，表位，我们可以识别用于免疫治疗的泛狂犬病病毒中和纳米抗体。在这项研究中，我们提出两个具体目标：（1）使用基于结构和抗体引导的方法来设计和表征狂犬病病毒糖蛋白结构域 III 的抗原性，这是多种病毒广泛针对的病毒脆弱性位点中和抗体，揭示抗体中和机制； (2) 识别和表征通过将免疫识别集中在狂犬病病毒G结构域上，用于治疗的泛狂犬病病毒中和纳米抗体 III 保守表位。