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

项目摘要 冠状病毒病2019年(新冠肺炎)在人类人群中迅速传播 并成为一场全球性的大流行。已经投入了大量的努力来开发疫苗或 基于SARS-CoV-2刺突糖蛋白的抗体治疗。其中一个挑战就是 策略是识别尖峰蛋白上的保守表位，并预测可能 降低疫苗和免疫疗法的有效性。到目前为止，有20多种SARS变种- 冠状病毒2型基因组序列已有报道。因此，结构导向和表位为基础 设计对于生产有效的药物来应对目前和未来的SARS-CoV疫情至关重要- 2或相关冠状病毒。纳米抗体可以识别高变区上的保守表位 病原体。在这里，我们提出可以利用抗尖峰纳米体来快速鉴定 保护性表位为疫苗和治疗药物的设计提供信息。此外，我们假设 针对SARS-CoV-2的有效和广泛保护的纳米抗体可以被开发为吸入型 预防或治疗药物。 在这项建议中，我们将通过多学科的合作来利用我们的互补优势 结合粘膜免疫学、结构生物学和病毒学的方法，来表征 一组不同纳米体抗SPAKE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(目标1)。一个全面的 保守和非保守表位列表(目标2)将用于基于结构的设计 用于活体表征的有效纳米多聚体(目标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).