Large-scale compatibility assessments between ACE2 proteins and diverse sarbecovirus spikes

ACE2 蛋白和多种 sarbecovirus 尖峰之间的大规模兼容性评估

基本信息

批准号：
10722852
负责人：
Kenneth A Matreyek
金额：
$ 24.15万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-23 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10722852
关键词：
2019-nCoV ACE2 Affinity Africa Amino Acid Sequence Amino Acids Animals Arenavirus Asia Bar Codes Binding Biological Assay Biology Biotechnology COVID-19 pandemic Catalogs Cell surface Cells Chiroptera Complex Computer Models Coronavirus Coronavirus spike protein DNA sequencing Data Data Analyses Disease Outbreaks Europe Event Family Filovirus Flow Cytometry Future Genetic Genetic study Goals Habitats Health system High-Throughput Nucleotide Sequencing Human Infection Integration Host Factors Learning Libraries Light Machine Learning Mammals Merbecovirus Methods Modeling Modernization Molecular Organism Orthologous Gene Pattern Plasmids Play Predisposition Probability Proteins Public Health Receptor Cell Research Infrastructure Role Sampling Sarbecovirus Series Severe Acute Respiratory Syndrome Testing Time Tissues Training Travel Tropism Variant Viral Virus Work Zoonoses betacoronavirus cellular engineering cross-species transmission data access density experimental study future pandemic genetic information improved insight interest luminescence member mutant mutation screening pandemic disease prevent receptor receptor binding synthetic biology tool viral pandemic zoonotic spillover

项目摘要

PROJECT SUMMARY Viral spillover from animal reservoirs into humans can decimate public health systems and cripple the world economy, as evident with the current SARS-CoV-2 pandemic. Continued wildlife habitat destruction, human expansion, and routine global travel keep increasing the likelihood that another viral pandemic will occur again within the next few decades. Beta coronaviruses are an incredibly diverse family of viruses observed across Asia, Europe, and Africa, that have proven capable of zoonotic spillover into humans as they have caused multiple worldwide outbreaks over the last two decades. We still lack the fundamental understanding of the molecular and genetic factors that dictate the molecular compatibilities that determine which beta coronaviruses are most likely to jump into humans in the future. The ability of SARS-like beta coronaviruses to utilize ACE2 as a receptor for cell entry is a major factor determining the extent of coronavirus tropism across species or within the tissues of an organism. While SARS-CoV-2 has been heavily studied, almost nothing is known about most other members of this virus family. Traditional studies can only test a handful of conditions at a time, incompletely sampling the vast range of relevant experimental conditions, particularly for the hundreds of uncharacterized beta coronaviruses. Large- scale, minimally-biased, cell-based entry assays are needed to model how these factors converge to dictate the probability of infection. We will pair new methods in cell engineering and synthetic biology with DNA-sequencing enabled multiplex genetic assays to perform a series of large-scale infection assays revealing the factors determining susceptibility to beta coronavirus entry. These large-scale experiments will reveal how ACE2 sequence and cell surface density impact the efficiency of virus entry. By testing a library of receptor binding domain sequences identified from ecological surveillance of bat coronaviruses, we will identify which viruses possess sufficient affinity to human ACE2 to potentiate cross-species transmission, and create a catalog describing all of the different ways these viruses have evolved their sequences to engage ACE2. By modeling the relationship between spike and ACE2 protein sequence, expression level, and efficiency of cell entry, we will identify potential animal reservoirs for SARS-CoV-2 and other SARS-like bat coronaviruses, and predict which viruses have sufficient binding with human ACE2 to potentially spark the next pandemic.

项目概要病毒从动物宿主传播到人类可能会摧毁公共卫生系统并削弱世界经济，当前的 SARS-CoV-2 大流行就证明了这一点。野生动物栖息地持续遭到破坏，人类扩张和日常全球旅行不断增加另一场病毒大流行的可能性在接下来的几十年内再次发生。 β 冠状病毒是一个极其多样化的病毒家族在亚洲、欧洲和非洲观察到，已证明它们能够传播给人类，因为它们过去二十年来在世界范围内多次爆发疫情。我们还缺乏根本了解决定分子相容性的分子和遗传因素哪些β冠状病毒未来最有可能进入人类体内。 SARS样β冠状病毒利用ACE2作为细胞进入受体的能力是一个主要因素确定冠状病毒跨物种或生物体组织内的趋向性程度。尽管 SARS-CoV-2 已被深入研究，但对该病毒的大多数其他成员几乎一无所知家庭。传统研究一次只能测试少数条件，无法对广泛的范围进行完全采样相关实验条件，特别是数百种未表征的β冠状病毒。大的- 需要规模化、偏差最小、基于细胞的进入分析来模拟这些因素如何汇聚到一起决定感染的概率。我们将细胞工程和合成生物学的新方法与 DNA 测序相结合多重基因检测进行一系列大规模感染检测，揭示决定因素对β冠状病毒进入的易感性。这些大规模实验将揭示 ACE2 序列如何细胞表面密度影响病毒进入的效率。通过测试受体结合域库通过对蝙蝠冠状病毒的生态监测确定的序列，我们将确定哪些病毒具有与人类 ACE2 有足够的亲和力以增强跨物种传播，并创建一个描述所有这些病毒以不同的方式进化其序列以接合 ACE2。通过建模为了研究spike和ACE2蛋白序列、表达水平和细胞进入效率之间的关系，我们将识别 SARS-CoV-2 和其他 SARS 样蝙蝠冠状病毒的潜在动物宿主，并预测哪些病毒病毒与人类 ACE2 有足够的结合力，有可能引发下一次大流行。