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病属的成员引起的。高遗传多样性将lyssavirus成员分为三个门群，而交叉菌群（尤其是到门群2和3）尚未通过当前暴露后预防（PEP）或抗体疗法。为了实现全球狂犬病疾病的更广泛有效的对策，并为新裂解病毒的出现做准备，针对所有系统群的有效治疗剂是必要的。 lyssavirus g糖蛋白是病毒体表面上的唯一抗体靶标，它采用了独特的融合前和融合后状态之间的构象与其他III类病毒融合机器。域-III lyssavirus g糖蛋白包括在天然状态下具有大型中和抗体可访问的表面。对域-III的序列分析揭示了所有天线的几个保守斑块，可以有可能充当泛透析病毒中和器的靶标表位。抗原特异性纳米构造已经被认为是针对各种传染病和非感染疾病的有前途的治疗剂，在绑定紧凑和隐藏的表位上具有优势，这些表位是常规的抗体。我们假设通过将免疫识别集中在lyssavirus g域-III中表位，我们可以鉴定泛透析病毒中和免疫疗法的纳米体。在这项研究中，我们提出了两个具体的目的：（1）使用基于结构的抗体引导的方法设计和表征 lyssavirus糖蛋白结构域-III的抗原性，这是一个广泛针对的病毒脆弱性部位中和抗体，以启示抗体中和机制；（2）识别和表征通过将免疫识别聚焦于lyssavirus g域，泛透析病毒中和纳米疗法用于治疗剂 III保守的表位。