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

• 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
• 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

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.

摘要 致命的狂犬病脑炎疾病是由人畜共患狂犬病病毒属的成员引起的。高 遗传多样性将狂犬病病毒属成员分成三个不同的组，而交叉组（特别是 （第2组和第3组）保护尚未通过当前的暴露后预防（PEP）建立，或 抗体疗法为了在全球范围内对狂犬病采取更广泛有效的对策， 为新狂犬病病毒的出现做好准备，针对所有狂犬病病毒组的有效治疗剂是 必要Lysavirus G糖蛋白是病毒粒子表面唯一的抗体靶点， 在融合前和融合后状态之间的构象，如其他III类病毒融合机器。域III 狂犬病病毒G糖蛋白在天然状态下包含大的中和抗体可及表面。 对结构域III的序列分析揭示了在所有基因组中的几个保守的补丁， 潜在地用作泛狂犬病毒中和剂的靶表位。抗原特异性纳米抗体已经被 被认为是有希望的抗各种感染性疾病和非感染性疾病的治疗剂， 其在结合常规免疫球蛋白无法达到的紧凑和隐藏的表位方面具有优势 抗体的我们假设，通过将免疫识别集中在狂犬病毒G结构域-III保守的 表位，我们可以鉴定用于免疫治疗的泛狂犬病毒中和纳米抗体。本研究 提出两个具体目标：（1）使用基于结构和抗体指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，该结构域是几种广泛靶向的病毒脆弱性位点， 中和抗体，用于揭示抗体中和机制;（2）鉴定和表征 通过将免疫识别聚焦于狂犬病病毒G结构域用于治疗的泛狂犬病病毒中和纳米抗体 III保守表位。