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

• 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
• 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

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.

项目摘要 从动物水库到人类的病毒溢出会使公共卫生系统降低并削弱 世界经济，与当前的SARS-COV-2大流行相同。继续野生动植物栖息地破坏， 人类的扩张和常规的全球旅行不断增加另一个病毒大流行的可能性 在接下来的几十年中再次发生。 β冠状病毒是一个非常多样化的病毒家族 在亚洲，欧洲和非洲都观察到，这些人都能够像他们一样涌向人类 在过去的二十年中，全世界爆发了多次爆发。我们仍然缺乏基本 了解决定了决定分子兼容性的分子和遗传因素 哪些β冠状病毒最有可能在未来跳入人类。 SARS样β冠状病毒利用ACE2作为细胞进入的受体的能力是一个主要因素 确定跨物种或生物组织内的冠状病毒的程度。尽管 SARS-COV-2已经进行了大量研究，该病毒的其他大多数成员几乎一无所知 家庭。传统研究只能一次测试少数条件 相关的实验条件，特别是对于数百个未表征的β冠状病毒。大的- 规模，最小偏见的基于细胞的入口测定需要建模这些因素如何收敛以决定 感染的可能性。 我们将将细胞工程和合成生物学的新方法与启用DNA测序配对 多重遗传测定法以执行一系列大规模感染测定，揭示了决定因素 对β冠状病毒进入的敏感性。这些大规模实验将揭示ACE2序列和 细胞表面密度影响病毒进入的效率。通过测试受体结合域的库 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拥有 足够与人ACE2的亲和力，以增强跨物种的传输，并创建一个描述所有内容的目录 这些病毒在不同的方式中演变了序列以参与ACE2。通过建模 尖峰和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和其他类似SARS的BAT冠状病毒的潜在动物储藏室，并预测哪种 病毒与人ACE2具有足够的结合，可能会激发下一个大流行。