Complete mapping of the functional and antigenic effects of mutations to Lassa virus glycoprotein

拉沙病毒糖蛋白突变的功能和抗原效应的完整图谱

• 批准号: 9906444
• 负责人: Katharine Holt Dusenbury Crawford
• 金额: $ 4.04万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9906444
• 关键词: Address; Affect; Africa; Amino Acid Sequence; Amino Acids; Antibodies; Antibody Response; Automobile Driving; Bar Codes; Binding; Case Fatality Rates; Cells; Cessation of life; Clinical; Development; Disease Outbreaks; Ebola virus; Epitopes; Etiology; Evolution; Genetic Variation; Glycoproteins; Growth; HIV; Health; Human; Immune; Immune response; Immunity; Immunotherapy; Infection; Influenza; Knowledge; Lassa Fever; Lassa virus; Length; Libraries; Link; Maps; Measures; Mediating; Medical; Medicine; Methods; Middle East Respiratory Syndrome Coronavirus; Monoclonal Antibodies; Mutation; Nucleotides; Proteins; Resistance; Scheme; System; Techniques; Therapeutic; Therapeutic antibodies; Time; Vaccines; Variant; Viral; Viral Pathogenesis; Virion; Virus; Virus Diseases; Work; World Health Organization; antibody immunotherapy; base; drug development; env Gene Products; experimental study; human disease; human monoclonal antibodies; improved; insight; mutant; neutralizing antibody; neutralizing monoclonal antibodies; pressure; prevent; priority pathogen; protein function; receptor; research and development; therapeutic development; tool; vaccine development; virus envelope

Project Summary/Abstract Despite many medical advances, viruses continue to cause significant human disease. Viral infections are difficult to prevent or treat in large part due to the rapid evolution of viral entry proteins. Mutations in these proteins can permit viruses to infect new hosts (including humans), better spread between hosts, and evade immune responses and some therapeutics. The Bloom lab has developed high-throughput tools to completely characterize the effects of all single amino-acid mutations to the viral entry proteins from influenza and HIV. These tools have proven very powerful for better understanding how these viruses evolve and how they escape from antibodies proposed for clinical use. However, current tools cannot be easily applied to other viruses, including most emerging viruses. To better understand the effects of mutations to viral entry proteins from emerging viruses, I am developing a general platform for the high-throughput characterization of mutations to viral entry proteins. Based on a pseudotyped lentiviral system, this platform allows me to study the viral entry proteins from most enveloped viruses. I will leverage this system, along with the high-throughput tools already developed in the Bloom lab, to measure the effects of all amino-acid mutations to the viral entry proteins from emerging viruses. Specifically, I will use my platform to characterize the effects of mutations to the Lassa virus entry protein from several strains of Lassa virus. Understanding the effects of mutations to the viral entry protein from divergent lineages of Lassa virus will be important for determining how the diversity of this virus affects the development of a broadly-protective treatment or vaccine. To further address the question of antibody therapeutic development, I will completely characterize the ability of mutations to the Lassa virus entry protein to mediate antibody escape from three human monoclonal antibodies currently undergoing therapeutic development. These complete maps of antibody resistance will determine from which antibody it is most difficult for the virus to escape and help evaluate and refine potential antibody immunotherapies. Overall, I will develop a general method to characterize the effects of mutations to viral entry proteins, including those from emerging viruses. I will then leverage this approach to study the Lassa virus entry protein, gaining actionable insight into Lassa virus entry protein function and antibody escape.

项目总结/摘要 尽管有许多医学进步，但病毒继续引起重大的人类疾病。病毒感染 在很大程度上由于病毒进入蛋白的快速进化而难以预防或治疗。这些基因突变 蛋白质可以使病毒感染新的宿主（包括人类），更好地在宿主之间传播， 免疫反应和一些治疗方法。Bloom实验室已经开发出高通量工具， 表征所有单个氨基酸突变对流感和HIV病毒进入蛋白的影响。 事实证明，这些工具非常强大，可以更好地了解这些病毒如何进化，以及它们如何 逃避临床使用的抗体。然而，目前的工具不能轻易地应用于其他领域。 病毒，包括大多数新兴病毒。 为了更好地了解突变对新出现病毒的病毒进入蛋白的影响，我 开发一个通用平台，用于病毒进入蛋白突变的高通量表征。 基于假型慢病毒系统，这个平台使我能够研究大多数病毒的病毒进入蛋白， 包膜病毒我将利用这个系统，沿着已经在 Bloom实验室，以测量所有氨基酸突变对新出现病毒的病毒进入蛋白的影响。 具体来说，我将使用我的平台来描述突变对拉沙病毒条目的影响， 几种拉沙病毒的蛋白质。了解突变对病毒进入蛋白的影响 从拉沙病毒的不同谱系中分离出的病毒对于确定这种病毒的多样性如何影响 广泛保护性治疗或疫苗的开发。为了进一步解决抗体问题， 治疗发展，我将完全表征突变的能力，以拉沙病毒进入蛋白 介导目前正在进行治疗的三种人单克隆抗体的抗体逃逸 发展这些完整的抗体耐药性图谱将决定它最 病毒难以逃脱，并有助于评估和改进潜在的抗体免疫疗法。 总之，我将开发一种通用方法来表征突变对病毒进入蛋白的影响， 包括新出现的病毒。然后我将利用这种方法来研究拉沙病毒的进入蛋白， 获得对拉沙病毒进入蛋白功能和抗体逃逸的可行见解。