EVOLVING VIRUS-SPECIFIC sACE2 MIMICS FOR COMPETITIVE INHIBITION OF SARS-CoV-2

项目总结 高致病性的新型SARS冠状病毒2(SARS-CoV-2)的快速传播已造成全球 卫生紧急情况。因此，迫切需要有效的抗病毒疗法来对抗这种病毒。 SARS-CoV-2病毒通过ACE2受体1进入细胞，ACE2受体1与病毒刺突蛋白2结合。在ITS中 可溶性ACE2(SACE2)有可能作为一种稳定的、非免疫原性的竞争性抗体 SARS-CoV-2的抑制剂，目前正在临床试验中探索3。由于潜在的负面影响 抗尖峰mAbs18的作用，以及ACE2表现出包括整合素在内的其他生物作用的事实 信号调节7，8，刺激性受体模拟物将产生治疗SARS-CoV-2和 潜在的其他高度传染性疾病。 这一提议寻求使用机器学习和定向进化来发展高亲和力，但 内源性非活性的sACE2模拟物，以创造快速实施的治疗方法来对抗 SARS-CoV-2和潜在的冠状病毒。这种方法将允许生成可伸缩和 可翻译的生物制品，并提供了一个平台，以快速纠正可能出现的潜在突变 在未来。利用UniRep49的深度学习，将设计和生成紧密绑定的sACE2变体 SARS-CoV2-2刺突蛋白，但不与整合素类等内源性靶标交叉作用[目标1]。 同时，我们将执行定向进化来优化尖峰结合，并针对 结合内源蛋白[目标2]。最后，我们将确定领先候选人并评估容忍度和 基因工程sACE2变异体在小鼠中的免疫原性[目标3]。总体而言，这项提案将制定 高度特异的ACE2受体模拟，以便及时创造免疫原性最低的新型抗病毒药物 以拯救生命并防止未来的疫情。 10

PROJECT SUMMARY The rapid spread of the highly-pathogenic, novel SARS-coronavirus 2 (SARS-CoV-2) has caused a global health emergency. Thus, there is a desperate need for effective antiviral therapeutics to counteract this virus. The SARS-CoV-2 virus enters cells using the ACE2 receptor1 which binds the viral spike protein2. In its soluble form, ACE2 (sACE2) has the potential to be used as a stable and non-immunogenic competitive inhibitor to SARS-CoV-2 and is presently being explored in clinical trials3. Due to the potential negative side effects of anti-spike mAbs18, and the fact that ACE2 exhibits other biological roles4–6 including integrin signaling regulation7,8, spike-specific receptor mimics would yield novel therapeutics for SARS-CoV-2 and potentially other highly infectious diseases. This proposal seeks to use machine learning and directed evolution to develop high affinity, yet endogenously-inactive mimics of sACE2 in order to create rapidly implementable therapeutics to combat SARS-CoV-2 and potential corona-like viruses. This approach would allow for the generation of scalable and translatable biologics, and provide a platform to rapidly course-correct for potential mutations that may arise in the future. Utilizing deep-learning with UniRep49, will design and generate sACE2 variants that tightly bind the SARS-CoV2-2 spike protein but do not cross-interact with endogenous targets such as integrins [Aim 1]. Simultaneously, we will perform directed evolution to optimize spike-binding and select against variants that bind endogenous proteins [Aim 2]. Finally, we will identify lead candidates and evaluate the tolerance and immunogenicity of engineered sACE2 variants in mice [Aim 3]. Collectively, this proposal will develop highly-specific ACE2 receptor mimics in order to create novel antivirals with minimal immunogenicity in time to save lives and prevent future outbreaks. 10