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 大流行已在 188 个国家/地区导致 3830 万例感染病例截至 2020 年 10 月 14 日，死亡人数超过 108 万人的国家和地区。在加速开发过程中，人们严重担心 SARS-COV-2 的未来变种会进化并在不久的将来感染人类。我们建议开发一类新型通用分子阻断剂 SARS-CoV-2 基于 hACE2 的 N 端三螺旋束（3HB，残基 20-99），已被识别 SARS-CoV和SARS-CoV-2作为细胞表面受体来感染人类细胞。基于我们之前的工作在三聚化三螺旋束结构域中，我们假设 hACE2 的 N 端 3HB 结构域可以是三聚化，可与 SARS-CoV-2 所有变体的 RBD 进行高效且特异性的结合。二该项目将追求具体目标。在具体目标1中，我们将开发基于N-的三聚体陷阱 hACE2 的末端三螺旋束 (3HB) 以三价方式与 SARS-CoV-2 的 RBD 结合，具有高特异性和效力。在具体目标 2 中，我们将利用定向分子进化从蛋白质结构域库，并将其与 hACE2 的 N 端 3HB 融合，形成双功能 SARS-CoV-2 陷阱，高度特异性地结合病毒 RBD，同时抑制其弗林蛋白酶介导的预激活。由此产生的新型 SARS-CoV- 2个捕获分子将有可能普遍应用于阻断hACE2介导的感染大部分（如果不是全部）未来的 SARS-CoV-2 变种。