Developing Lectins as Inhibitors of Coronavirus Spike Proteins

项目总结/摘要 2019年底，SARS-CoV2病毒成为全球大流行，导致严重的呼吸道疾病 COVID-19导致全球数百万人死亡。作为一种冠状病毒，SARS-CoV2宿主相互作用是 通过包围病毒体的刺突蛋白决定。SARS-CoV 2的刺突蛋白是一种高度保守的蛋白质， 糖基化三聚体，通过受体结合结构域（RBD）与宿主细胞上的ACE2受体相互作用 以促进病毒进入。作为病毒感染中的关键参与者，Spike三聚体和RBD已成为靶标 用于治疗或预防SARS-CoV2的大多数批准的疗法和疫苗。在本提案中， PI使用了一种新的抗病毒凝集素BOA，他已经证明BOA可以结合刺突蛋白上的聚糖， 并抑制SARS-CoV 2病毒进入。为了了解BOA如何抑制病毒进入，在K99阶段， 研究者将使用一种方法确定BOA抑制SARS-CoV2病毒感染的机制， 病毒抑制测定、生物物理测定和电子显微镜的组合。随后，PI将 测试BOA抑制新出现的SARS-CoV2变异的能力，这些变异已经积累了突变， 促进免疫逃逸的刺突蛋白的不同区域。在此基础上， 在本奖项的独立R00阶段，PI将使用在K99阶段开发的技能， 自己的独立研究小组，在那里他们将测试BOA凝集素抑制病毒进入β- 已知会感染人类的冠状病毒。成功地完成这些目标可以提供一个真正的广泛的 谱冠状病毒抑制剂将被进一步开发作为下一种冠状病毒的治疗或诊断 出现了。拟议的研究将为PI提供新的和令人兴奋的病毒学培训， 他打算把这门技术传授给他未来的学员。拟议工作将于 匹兹堡大学的结构生物学系和疫苗研究中心将 提供无与伦比的资源、仪器和专家访问。PI已经召集了一个 令人兴奋的导师团队的病毒学家和生物药理学家谁，他将会见每月接受建议， 讨论结果，设计新的实验，并准备过渡到一个独立的职业生涯。来帮助 在他的职业发展中，大学提供了许多讲习班，以发展诸如撰写赠款等技能。 和指导PI还将参加会议，进行演示并磨练他的科学 沟通技巧。总之，拟议的培训和职业发展计划将使PI做好准备， 领导自己的独立研究小组，并成为该领域的领导者。

Project Summary/Abstract In late 2019, the SARS-CoV2 virus emerged as a global pandemic, causing the severe respiratory disease COVID-19 and resulting in millions of deaths worldwide. As a coronavirus, SARS-CoV2 host interactions are dictated through the Spike proteins that encompass the virion. The Spike protein of SARS-CoV2, is a highly glycosylated trimer that interacts with the ACE2 receptor on host cells via the receptor binding domain (RBD) to facilitate viral entry. As a key player in viral infection, the Spike trimer, and the RBD, have become the target for a majority of approved therapeutics and vaccines used to treat or prevent SARS-CoV2. In this proposal, the PI uses a novel antiviral lectin, BOA, which he has demonstrated that BOA binds glycans on the spike protein, and inhibits SARS-CoV2 viral entry. To understand how BOA inhibits viral entry, during the K99 phase of the award, the PI will determine the mechanism by which BOA inhibits in SARS-CoV2 viral infection using a combination of viral inhibition assays, biophysical assays, and electron microscopy. Subsequently the PI will test the ability of BOA to inhibit emerging SARS-CoV2 variants of concern that have accumulated mutations to various regions of the spike protein that facilitate immune escape. Expanding upon these findings during the independent R00 phase of this award, the PI will use skills developed during the K99 phase to establish his own independent research group, where they will test the ability of the BOA lectin to inhibit viral entry of Beta- Coronaviruses known to infect humans. Successful completion of these aims could provide a bona fide broad spectrum coronavirus inhibitor to be further developed as a therapeutic or diagnostic for the next coronavirus that emerges. The proposed research, will provide the PI with new and exciting training in virology as well as electron microscopy which he intends to pass on to his future trainees. The proposed work will be completed at The University of Pittsburgh, where the Department of Structural Biology and Center for Vaccine Research will provide unmatched access to resources, instrumentation, and experts in the field. The PI has assembled an exciting mentorship team of virologists and biophysicists who he will meet with monthly to receive advice, discuss results, designed new experiments, and prepare for transitioning into an independent career. To aide in his career development, the university offers numerous workshops to develop skills such as grant writing and mentorship. The PI will also attend conferences to give presentations and hone his science communications skills. In Summary, the proposed training and career development plans will prepare the PI to lead his own independent research group and become a leader in his field.