Functional and antigenic evolution of SARS-related coronaviruses

SARS相关冠状病毒的功能和抗原进化

• 批准号: 10351951
• 负责人: Tyler Nelson Starr
• 金额: $ 12.86万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-10 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10351951
• 关键词: 2019-nCoV; ACE2; Address; Affect; Affinity; Amino Acids; Antibodies; Antigenic Diversity; Binding; Binding Proteins; Biochemical; Biochemistry; Biological Assay; Biophysics; COVID-19 pandemic; COVID-19 treatment; Cells; Chiroptera; Disease Outbreaks; Epidemic; Epitopes; Evolution; Exhibits; Future; Genetic; Geography; Goals; Health; Human; Immunity; Infection; Infection prevention; Maps; Measurement; Measures; Mediating; Mentorship; Methods; Modernization; Molecular; Mutagenesis; Mutation; Orthologous Gene; Phase; Phenotype; Phylogeny; Population; Positioning Attribute; Process; Property; Proteins; Readiness; Reagent; SARS coronavirus; Sarbecovirus; Site; Specificity; Surface; Surveys; System; Techniques; Therapeutic; Therapeutic antibodies; Time; Training; Vaccines; Variant; Viral; Viral Antibodies; Viral Proteins; Virus; Work; Yeasts; Zoonoses; antibody engineering; biophysical analysis; clinical development; combinatorial; design; experimental study; human pathogen; molecular recognition; mutant; mutation screening; neutralizing antibody; next generation; novel; preference; prevent; prospective; receptor; receptor binding; reconstruction; systems research; therapeutic development; tool; trait; vaccine-induced antibodies; virology

PROJECT SUMMARY SARS-related coronaviruses (also known as sarbecoviruses) circulate in bat reservoirs, but two human spillovers in the past 20 years—SARS-CoV-1 in 2002 and SARS-CoV-2 in 2019—have caused globally devastating outbreaks. The emergence of SARS-CoV-2 from a previously unknown bat lineage highlights the pressing need to trace the evolutionary origins of viral properties that enable human spillover and develop therapeutic reagents that can broadly inhibit infection by diverse, potentially unsampled sarbecovirus lineages. Sarbecovirus zoonosis depends in part on the evolution of viruses to interact with ACE2 receptors on the surface of human and potential intermediate hosts’ cells, mediated by the viral spike receptor-binding domain (RBD). The RBD is also a key component to therapeutic control of SARS-CoV-2 and presumably other sarbecoviruses, as it is the target of the most potently neutralizing antibodies, including those in clinical development and in polyclonal human sera. Although I and others have begun surveying the impacts of mutations within SARS-CoV-2 itself on important biochemical phenotypes including binding to ACE2 receptor and antiviral antibodies, these studies have only limited utility in understanding the broader evolution of sarbecovirus RBDs, which exhibit considerable divergence in sequence and function. To characterize sarbecovirus functional and antigenic evolution, I propose to combine high-throughput protein binding experiments with computational evolutionary analyses, functional virology, and biochemistry: 1. I will profile the ACE2-binding specificities of all known sarbecovirus RBDs and their evolutionary precursors. I hypothesize that the ability to bind ACE2, including the human ACE2 ortholog, is more evolutionarily and geographically widespread than previously appreciated. 2. I will systematically characterize how diverse RBD mutations affect ACE2-binding phenotypes. I hypothesize that human ACE2 binding is an easily evolvable trait in sarbecovirus lineages where it has not been previously considered. I also hypothesize that divergence among sarbecovirus lineages shifts the genetic and biochemical determinants of ACE2 binding over evolutionary time. 3. I will identify RBD epitopes that are susceptible to broad pan-sarbecovirus antibody binding. By determining the relationship between breadth and other antibody properties across RBD epitopes, I highlight key features to target in the design of next-generation pan-sarbecovirus vaccines and antibody therapeutics. Identification of the evolutionary and biochemical basis for key sarbecovirus features enhances viral surveillance and therapeutic development, preparing for or even preventing future sarbecovirus spillovers. The new experimental expertise, broader training and mentorship, and research systems developed in this work will enable me to achieve my goal of leading an academic lab studying protein evolution at the host-virus interface.

项目摘要 SARS相关的冠状病毒（也称为sarbecoviruses）在蝙蝠宿主中传播，但两种人类 过去20年的溢出效应--2002年的SARS-CoV-1和2019年的SARS-CoV-2--在全球范围内造成了 毁灭性的爆发SARS-CoV-2从一个以前未知的蝙蝠谱系中出现， 迫切需要追踪病毒特性的进化起源，这些特性使人类能够传播并发展 可以广泛抑制不同的、可能未取样的肉瘤病毒谱系的感染的治疗试剂。 Sarbecovirus人畜共患病部分取决于病毒的进化，以与ACE 2受体相互作用。 病毒刺突受体结合域介导的人和潜在中间宿主细胞表面 （RBD）。RBD也是SARS-CoV-2治疗控制的关键组分，并且可能是其他治疗控制的关键组分。 肉瘤病毒，因为它是最有效的中和抗体的靶标，包括临床上的中和抗体。 发育和多克隆人血清中。尽管我和其他人已经开始调查 SARS-CoV-2本身在重要生化表型上的突变，包括与ACE 2受体的结合 和抗病毒抗体，这些研究在理解更广泛的进化方面只有有限的效用。 肉瘤病毒RBD，其在序列和功能上表现出相当大的差异。 为了表征Sarbecovirus的功能和抗原进化，我建议将联合收割机高通量蛋白 结合计算进化分析、功能病毒学和生物化学的实验： 1.我将分析所有已知的肉瘤病毒RBD的ACE 2结合特异性及其进化 前体我推测，结合ACE 2的能力，包括人类ACE 2的直系同源物， 在进化上和地理上比以前认识到的更广泛。 2.我将系统地描述不同的RBD突变如何影响ACE 2结合表型。我 假设人ACE 2结合在肉瘤病毒谱系中是一种容易进化的特征， 以前考虑过。我还假设，肉瘤病毒谱系之间的分歧改变了遗传 和ACE 2结合的生物化学决定因素。 3.我将确定RBD表位，这些表位对广泛的泛Sarbecovirus抗体结合敏感。通过 为了确定RBD表位的宽度和其他抗体特性之间的关系，我强调 在设计下一代泛肉瘤病毒疫苗和抗体疗法时要瞄准的关键特征。 确定关键Sarbecovirus特征的进化和生化基础， 监测和治疗开发，为将来的Sarbecovirus溢出做好准备甚至预防。的 新的实验专业知识，更广泛的培训和指导，以及在这项工作中开发的研究系统将 使我能够实现我的目标，领导一个学术实验室，研究宿主-病毒界面的蛋白质进化。