Developing Lectins as Inhibitors of Coronavirus Spike Proteins

项目摘要/摘要 2019年末，SARS-CoV2病毒以全球大流行的形式出现，导致严重的呼吸道疾病 新冠肺炎，并导致全球数以百万计的人死亡。作为一种冠状病毒，SARS-CoV2宿主相互作用是 通过包围病毒粒子的刺突蛋白口述。SARS-CoV2的刺突蛋白是一种高度 糖基化三聚体，通过受体结合域(RBD)与宿主细胞上的ACE2受体相互作用 以便于病毒进入。作为病毒感染的关键角色，Spike三聚体和RBD已成为攻击目标 用于治疗或预防SARS-CoV2的大多数已获批准的疗法和疫苗。在这项提案中， Pi使用了一种新型的抗病毒凝集素BOA，他已经证明BOA能将多糖结合到Spike蛋白上， 并抑制SARS-CoV2病毒进入。为了了解BOA是如何抑制病毒进入的，在K99阶段 获奖后，PI将确定BOA通过使用一种 病毒抑制试验、生物物理试验和电子显微镜相结合。随后，PI将 测试BOA抑制新出现的令人担忧的SARS-CoV2变异的能力，这些变异累积到 促进免疫逃逸的尖峰蛋白的不同区域。在这些发现的基础上，在 这个奖项的独立R00阶段，PI将使用在K99阶段培养的技能来建立HIS 自己的独立研究小组，在那里他们将测试BOA凝集素抑制Beta- 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真正广泛的 冠状病毒光谱抑制剂将进一步开发用于治疗或诊断下一种冠状病毒 这就浮现出来了。拟议的研究将为PI提供新的和令人兴奋的病毒学培训以及 电子显微镜，他打算把它传给他未来的实习生。拟议的工作将于 匹兹堡大学，结构生物系和疫苗研究中心将在那里 提供无与伦比的资源、仪器和该领域专家的访问。私家侦探已经组装了一个 由病毒学家和生物物理学家组成的令人兴奋的指导团队，他将每月与他们会面，接受建议， 讨论结果，设计新的实验，为过渡到独立的职业生涯做好准备。协助 在他的职业发展过程中，该大学提供了许多研讨会来培养他的技能，如撰写拨款 和导师关系。PI还将出席会议，发表演讲并磨练他的科学 沟通技巧。总之，拟议的培训和职业发展计划将使专业人员做好准备 领导自己的独立研究小组，成为他所在领域的领导者。

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.