RAPID: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)-Prevention: Multiple-Site Binding with Fusing Aptamers to mitigate Coronavirus Disease 2019

RAPID：严重急性呼吸系统综合症冠状病毒 2 (SARS-CoV-2) - 预防：通过融合适体进行多位点结合以减轻 2019 年冠状病毒病

• 批准号: 2028531
• 负责人: Xiaohong Tan
• 金额: $ 20万
• 依托单位: Bowling Green State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2028531&HistoricalAwards=false
• 关键词: RAPID; Severe; Acute; Respiratory; Syndrome

The SARS-CoV-2 causes the novel coronavirus infectious disease 2019 (COVID-19). The binding of the viral spike protein to a receptor protein (called the angiotensin converting enzyme 2, or ACE2) is the first step in the infection of the human host cell. Blocking or inhibiting this interaction could stop the invasion of the human cell by this highly infectious virus. With this award, the Chemistry of Life Processes program is supporting the research of Dr. Xiaohong Tan at Bowling Green University in Bowling Green, Ohio to create DNA aptamers to inhibit this initial step of viral invasion. Aptamers are short sequences of DNA that fold into a shape that matches that of a target molecule, thereby keeping other molecules from binding. These DNA aptamers are “designed” by successive selection of molecules from large mixtures of different sequences, resulting in a few sequences that keep all other DNAs from binding the target. Dr. Tan uses this selection process to find DNA aptamers that bind tightly to the SARS-CoV-2 spike protein. Two of more of the identified DNA aptamers are linked together to create “super binders” that are much better at inhibiting the interaction than would any single aptamer. Constructing DNA aptamers that block the invasion of SARS-CoV-2 into human cells may lead to the design of antiviral agents that help lessen the infectivity that causes the COVID-19 pandemic. Educational and training activities will focus on postdoctoral researchers learning cutting edge techniques in chemical biology on a “front line” research problem. The team is developing ways to communicate their science to the public, to K-12 institutions, and to other research groups using distance learning technologies.The goal of this project is to develop polymeric molecules (fusion DNA aptamers) that significantly interact with the binding domain of SARS-CoV-2. The objective is to effectively block the virus from binding to the specific ACE2 receptor, thereby blocking the entry of the virus into human cells and, as a result, its ability to cause COVID-19 infections. Professor Tan uses SELEX (systematic evolution of ligands by exponential enrichment) to identify individual aptamers that interact with three distinct regions on the cell binding domain of the spike (S) protein of SARS-CoV-2. The affinities of aptamers to their targets are measured through standard binding assays. These individual high affinity DNA aptamers are connected together using nucleic acid or chemical linkers to make fusion aptamers that have affinities comparable to those of antibodies. The ability of these fused DNA aptamer “super binders” to effectively block this interaction is tested against a recombinant SARS-CoV-2 spike protein using standard technologies, as well as live SARS-CoV-2 through collaboration with the University of Toledo Medical School.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

SARS-COV-2导致新型冠状病毒感染疾病2019（Covid-19）。病毒峰值蛋白与受体蛋白的结合（称为血管紧张素转化酶2或ACE2）是感染人类宿主细胞的第一步。阻止或抑制这种相互作用可以阻止这种高度传染病的病毒侵袭人类细胞。通过此奖项，《生命过程的化学计划》计划支持俄亥俄州鲍林格林大学的鲍灵格林大学的小谭博士的研究，以创建DNA Aptamers，以抑制病毒入侵的最初步骤。 apatamers是DNA的短序列，将折叠成与靶分子相匹配的形状，从而保持其他分子的结合。这些DNA适体是通过从不同序列的大型混合物中成功选择分子来“设计”的，从而导致了一些序列，这些序列使所有其他DNA都无法结合靶标。 Tan博士使用此选择过程来找到与SARS-COV-2尖峰蛋白紧密结合的DNA适体。将两个鉴定的DNA适体链接在一起，以创建“超级粘合剂”，它们在抑制相互作用方面要比任何单个适体都更好。构建阻断SARS-COV-2侵袭人类细胞的DNA适体可能会导致设计抗病毒剂，从而减少导致COVID-19大流行的感染。教育和培训活动将集中于博士后研究人员在“前线”研究问题上学习化学生物学的尖端技术。该团队正在开发将科学传达给公众，K-12机构以及使用远程学习技术的其他研究小组的方法。该项目的目的是开发与SARS-COV-2的结合结构域显着相互作用的聚合物分子（融合DNA适体）。目的是有效地阻断病毒与特定ACE2受体的结合，从而阻断病毒进入人类细胞，从而阻止引起COVID-19感染的能力。 TAN教授使用SELEX（通过指数酶对配体的系统进化）来识别与SARS-COV-2的尖峰蛋白上的细胞结合结构域上与三个不同区域相互作用的单个适体。适体对目标的亲和力是通过标准结合测定法测量的。这些单独的高亲和力DNA适体使用核酸或化学接头将其连接在一起，以使具有与抗体相当的亲和力的融合适体。这些融合的DNA适体“超级粘合剂”有效阻止这种相互作用的能力通过使用标准技术的重组SARS-COV-2尖峰蛋白进行测试，以及通过与托莱多医学院的合作进行实时的SARS-COV-2，该奖项通过评估NSF的法规效果而受到评估，这表明了NSF的奖励，并在众多的知识上进行了评估。