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

项目摘要/摘要 尽管医学上取得了许多进步，但病毒仍在继续导致重大的人类疾病。病毒感染 很大程度上是由于病毒进入蛋白的快速进化而难以预防或治疗。这些基因的突变 蛋白质可以允许病毒感染新的宿主(包括人类)，更好地在宿主之间传播，并逃避 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，以完全 描述所有单一氨基酸突变对流感和艾滋病毒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是如何演变的以及它们是如何 逃避建议用于临床的抗体。然而，当前的工具不能很容易地应用于其他 病毒，包括大多数新出现的病毒。 为了更好地了解来自新兴病毒的突变对病毒进入蛋白的影响，我正在 开发高通量表征病毒进入蛋白突变的通用平台。 基于一个假型慢病毒系统，这个平台允许我研究大多数 被包裹的病毒。我将利用这个系统，以及在 Bloom实验室，以测量所有氨基酸突变对来自新兴病毒的病毒进入蛋白的影响。 具体地说，我将使用我的平台来描述突变对Lassa病毒进入的影响 来自几种拉萨病毒株的蛋白质。了解突变对病毒进入蛋白的影响 从拉萨病毒不同的谱系中分离出来将对确定该病毒的多样性如何影响很重要 广泛保护性治疗或疫苗的发展。进一步解决抗体问题 随着治疗的发展，我将完全表征突变对拉萨病毒进入蛋白的能力 介导人抗体逃逸目前正在接受治疗的三种人单抗 发展。这些完整的抗体抵抗力图谱将确定它最多来自哪种抗体 病毒很难逃脱，并帮助评估和改进潜在的抗体免疫疗法。 总体而言，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， 包括那些来自新兴病毒的病毒。然后我将利用这种方法来研究拉萨病毒进入蛋白， 对拉萨病毒进入蛋白功能和抗体逃逸获得可操作的洞察力。

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.