Structure-guided and epitope-based design of potent and broadly neutralizing nanobodies for COVID-19 mucosal immunotherapy

项目摘要 2019冠状病毒病（COVID-19）在人群中迅速传播 并成为全球性的流行病。在开发疫苗或 基于SARS-CoV-2刺突糖蛋白的抗体疗法。其中一个挑战是 策略是识别刺突上的保守表位，并预测可能 降低疫苗和免疫疗法的有效性。到目前为止，超过20种SARS变种- 已经报道了CoV-2基因组序列。因此，结构指导和基于表位的 设计对于生产针对当前和未来SARS-CoV爆发的有效药物至关重要- 2或相关的冠状病毒。纳米抗体可以识别高变蛋白上的保守表位 病原体在这里，我们提出抗刺突纳米抗体可用于快速识别 保护性表位为疫苗和治疗剂的设计提供信息。此外，我们假设， 针对SARS-CoV-2的有效和广泛保护性纳米抗体可以被开发为吸入剂， 预防或治疗药物。 在这项建议中，我们会透过一个跨专业的 结合粘膜免疫学、结构生物学和病毒学的方法， 一组不同的纳米抗体对刺突的分子相互作用和差异特异性 SARS-CoV-2和Betacoronaviruese家族的其他成员（Aim 1）。全面 保守和非保守表位的列表（目的2）将用于基于结构的设计， 用于体内表征的有效纳米抗体多聚体（目的3）。

Project Summary The coronavirus disease 2019 (COVID-19) rapidly disseminated through the human population and became a global pandemic. Significant efforts have been put into developing vaccines or antibody therapies based on the spike glycoprotein of SARS-CoV-2. One challenge of such strategies is to identify conserved epitopes on the Spike and predict viral mutations that could diminish the effectiveness of the vaccine and immunotherapy. To date, over 20 variant of SARS- CoV-2 genome sequences have been reported. Therefore, structure guided and epitope based design are crucial to generate effective medicines for current and future outbreaks of SARS-CoV- 2 or related coronavirus. Nanobodies can recognize conserved epitopes on hypervariable pathogens. Here, we propose that anti-spike nanobodies can be utilized for rapid identification of protective epitopes to inform design of vaccine and therapeutics. Further, we hypothesize that potent and broadly protective nanobodies against SARS-CoV-2 can be developed as an inhaled prophylactic or therapeutic medicine. In this proposal, we will leverage our complementary strengths through a multi-disciplinary approach combining mucosal immunology, structural biology, and virology, to characterize the molecular interactions and differential specificities of a diverse panel of nanobodies against spike of SARS-CoV-2 and other members of the Betacoronaviruese family (Aim 1). A comprehensive list of conserved and non-conserved epitopes (Aim 2) will be used for structure-based design of potent nanobody multimers for in vivo characterization (Aim 3).