Trimerization of the N-terminal Domain of ACE2 for Bifunctional Trapping of Future SARS-CoV-2 Variants

ACE2 N 末端结构域的三聚化，用于未来 SARS-CoV-2 变体的双功能捕获

基本信息

批准号：
10288255
负责人：
Rihe Liu
金额：
$ 22.49万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-09 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10288255
关键词：
2019-nCoV ACE2 Address Affinity Attenuated Binding Biochemical Biological Biological Assay Biological Process Biophysics COVID-19 COVID-19 pandemic Cathepsins Cell Line Cell Surface Receptors Cell membrane Cells Cessation of life Chiroptera Communicable Diseases Coronavirus Country Development Directed Molecular Evolution Disease Endocytosis Future Half-Life Hamsters Human Immune Evasion Immunologic Surveillance Infection Intervention Length Lentivirus Libraries Luciferases Mediating Minor Modeling Molecular N-terminal Pathogenesis Play Process Proprotein Convertases Proteins Reporter Genes Role SARS coronavirus SARS-CoV-2 infection SARS-CoV-2 variant Severe Acute Respiratory Syndrome Site Specificity Surface TMPRSS2 gene Technology Tertiary Protein Structure Vaccines Variant Viral Virus Virus Receptors Work base cartilage matrix protein inhibitor/antagonist mutant novel novel coronavirus receptor binding respiratory self assembly side effect successful intervention translational study

项目摘要

Abstract The Covid-19 pandemic caused by SARS-Cov-2 has resulted in 38.3 million infection cases across 188 countries and territories with more than 1.08 million deaths by October 14 2020. While various vaccines are under expedite development, there is a serious concern that future variants of SARS-COV-2 will evolve and infect humans in the near future. We propose to develop a novel class of universal molecular blockers against SARS-CoV-2 based on the N-terminal three-helix bundle (3HB, residues 20-99) of hACE2, which is recognized by both SARS-CoV and SARS-CoV-2 as cell surface receptor to infect human cells. Based on our previous work in trimerizing three-helical bundle domains, we hypothesize that the N-terminal 3HB domain of hACE2 can be trimerized for highly potent and specific binding with presumably the RBD of all variants of SARS-CoV-2. Two specific aims will be pursued in this project. In specific aim 1, we will develop a trimeric trap based on the N- terminal three-helix bundle (3HB) of hACE2 that trivalently binds to the RBD of SARS-CoV-2 with high specificity and potency. In specific aim 2, we will use directed molecular evolution to identify a human furin inhibitor from a protein domain library and fuse it with the N-terminal 3HB of hACE2 for a bifunctional SARS-CoV-2 trap that highly specifically binds viral RBD while inhibits its furin-mediated preactivation. The resulting novel SARS-CoV- 2 trapping molecules will have the potential to be universally applied to block hACE2-mediated infection by vast majority of, if not all, future variants of SARS-CoV-2.

抽象的由SARS-COV-2引起的COVID-19-导致188年的3830万例感染病例到2020年10月14日，到期108万人死亡的国家和地区。虽然各种疫苗是在快速发展下，人们严重担心SARS-COV-2的未来变体将不断发展，并且在不久的将来感染人类。我们建议开发一类新型的通用分子阻滞剂 SARS-COV-2基于HACE2的N末端三螺旋束（3HB，残基20-99），已识别通过SARS-COV和SARS-COV-2作为细胞表面受体感染人类细胞。根据我们以前的工作在修剪三螺旋束域中，我们假设HACE2的N末端3HB结构域可以是对SARS-COV-2的所有变体的RBD进行了高度有效和特异性结合而进行了修剪。二该项目将追求具体目标。在特定目标1中，我们将基于n-开发一个三聚陷阱 HACE2的端子三螺旋束（3hb），它与具有高特异性的SARS-COV-2的RBD结合和效力。在特定的目标2中，我们将使用定向的分子进化来鉴定人脂蛋白抑制剂蛋白质域库，并将其与HACE2的N末端3HB融合，以用于双功能SARS-COV-2陷阱高度特异性地结合病毒RBD，而抑制其furin介导的预催化。由此产生的小说sars-cov- 2个捕获分子将有可能普遍应用以阻止HACE2介导的感染。 SARS-COV-2的未来变体中的大多数（如果不是全部）。