Structural studies of Zika virus for vaccine design

项目摘要/摘要 寨卡病毒(Zika Virus，ZIKV)是一种由节肢动物传播的黄病毒家族的病原体，包括西尼罗河、黄色 发烧和登革热病毒。虽然ZIKV和其他黄病毒一样是通过蚊子传播的，但ZIKV也可以 寨卡病毒通过性传播，并在2007年首次意识到寨卡病毒能够引起流行病。感染通常是 轻微，产生发烧、结膜炎、头痛、皮疹、结膜炎和关节痛的病例约占20% 罕见的脑膜脑炎或格林-巴利综合征。然而，寨卡病毒感染的主要担忧是 在怀孕期间，这可能导致严重的胎儿后果，包括神经发育异常 例如小头畸形和其他显著的眼科和神经学变化。考虑到大规模的 传播、症状的严重性，以及目前没有治疗或预防措施，一种安全有效的ZIKV 目前迫切需要疫苗。针对寨卡病毒的疫苗正在开发中，但人们严重担心 使用全病毒或大部分ZIKV包膜(E)糖蛋白的疫苗可能导致非中和 通过抗体依赖与随后的黄病毒感染交叉反应并增强的抗体 增强(ADE)。尤其令人担忧的是，在以后的疾病中通过ADE感染严重出血热的风险 感染登革热病毒(DENV1)。相比之下，安全的疫苗将选择性地诱导抗体中和表位 而不是非中和的，因此可能会增强感染的抗体。 设计ZIKV疫苗需要了解抗体如何识别ZIKV包膜蛋白的结构知识。在……里面 帕梅拉·比约克曼教授在加州理工学院的实验室，这个项目将使用结构生物学，特别是最近 低温电子显微镜(Cryo-EM)在了解ZIKV抗体反应方面的进展，以及2)使用酵母 设计和测试免疫原的展示，基于结构分析，产生中和反应和 最大限度地减少非保护性抗体的产生，因此可用于开发安全的疫苗。这将是 通过与查尔斯·赖斯、玛格丽特·麦克唐纳和米歇尔教授的合作完成 洛克菲勒大学的努森茨韦格。我将准备病毒材料，以便在教授的实验室进行结构分析。 莱斯和麦克唐纳教授，以及我将在小鼠模型中测试候选免疫原的有效性和安全性 Nussenzweig教授的实验室，希望候选免疫原将适合向 临床测试。 培训计划包括在比约克曼教授的专业知识和指导下完成博士学位 加州理工学院，在赖斯教授的实验室呆了三个月，在赖斯教授的实验室呆了六个月。 纽森维格在洛克菲勒确保有很高的训练潜力和可行性。此培训，在此之前 回到加州大学洛杉矶分校的医学院，将提供成为独立医生的经验和技能 内科医生--学术医学的科学家。

Project Summary/Abstract Zika virus (ZIKV) is a pathogen from a family of arthropod-borne flaviviruses that includes West Nile, yellow fever, and dengue viruses. Though ZIKV is transmitted by mosquitos like other flaviviruses, ZIKV can also be transmitted sexually, and the ability of ZIKV to cause an epidemic was first realized in 2007. Infection is usually mild, producing fever, conjunctivitis, headache, rash, conjunctivitis and arthralgia in ~20% of cases and meningoencephalitis or Guillain-Barré Syndrome in rare cases. However, the major concern of ZIKV infection is during pregnancy, which can cause severe fetal consequences, including neurodevelopmental abnormalities such as microcephaly and other significant ophthalmologic and neurologic changes. Given the large-scale spread, severity of symptoms, and that there is no current treatment or prophylaxis, a safe and effective ZIKV vaccine is urgently needed. Vaccines against ZIKV are being development but there is serious concern that vaccines using whole virus or large portions of the ZIKV envelope (E) glycoprotein may induce non-neutralizing antibodies (Abs) that cross-react with and enhance subsequent flavivirus infections trough Ab-dependent enhancement (ADE). Of particular concern is the risk of severe hemorrhagic fever through ADE upon subsequent infection by dengue virus (DENV1). In contrast, a safe vaccine would selectively elicit Abs to neutralizing epitopes rather than non-neutralizing and therefore potentially infection-enhancing Abs. Design of a ZIKV vaccine requires structural knowledge of how Abs recognize ZIKV envelope proteins. In Prof. Pamela Bjorkman’s laboratory at Caltech, this project will 1) use structural biology, particularly recent advances in cryo-electron microscopy (cryo-EM), to understand the Ab response to ZIKV, and 2) use yeast display to design and test immunogens that, based on structural analysis, yield a neutralizing response and minimize production of non-protective Abs, and therefore may be used to develop a safe vaccine. This will be accomplished through collaboration with Professors Charles Rice, Margaret MacDonald, and Michel Nussenzweig at Rockefeller University. I will prepare viral material for structural analysis in the laboratory of Prof. Rice and Prof. MacDonald, and I will test candidate immunogens for efficacy and safety in mouse models in the laboratory of Prof. Nussenzweig with the aspiration that candidate immunogens will be suitable to move towards clinical testing. The training plan involves completion of the PhD under the expertise and guidance of Prof. Bjorkman at Caltech, with three-months spent in the laboratory of Prof. Rice and six-months in the laboratory of Prof. Nussenzweig at Rockefeller to ensure there is high training potential and feasibility. This training, before transitioning back to medical school at UCLA, will provide the experience and skillset to become an independent physician-scientist in academic medicine.