Abstract
The frequently emerging SARS-CoV-2 variants have weakened the effectiveness of existing COVID-19 vaccines and neutralizing antibody therapy. Nevertheless, the infections of SARS-CoV-2 variants still depend on angiotensin-converting enzyme 2 (ACE2) receptor-mediated cell entry, and thus the soluble human ACE2 (shACE2) is a potential decoy for broadly blocking SARS-CoV-2 variants. In this study, we firstly generated the recombinant AAVrh10-vectored shACE2 constructs, a kind of adeno-associated virus (AAV) serotype with pulmonary tissue tropism, and then validated its inhibition capacity against SARS-CoV-2 infection. To further optimize the minimized ACE2 functional domain candidates, a comprehensive analysis was performed to clarify the interactions between the ACE2 orthologs from various species and the receptor binding domain (RBD) of SARS-CoV-2 spike (S) protein. Based on the key interface amino acids, we designed a series of truncated ACE2 orthologs, and then assessed their potential affinity to bind to SARS-CoV-2 variants RBD in silico. Of note, we found that the 24-83aa fragment of dog ACE2 (dACE2 24-83 ) had a higher affinity to the RBD of SARS-CoV-2 variants than that of human ACE2. Importantly, AAVrh10-vectored shACE2 or dACE2 24-83 constructs exhibited a broadly blockage breadth against SARS-CoV-2 prototype and variants in vitro and ex vivo. Collectively, these data highlighted a promising therapeutic strategy against SARS-CoV-2 variants.
Keywords: Adeno-associated virus (AAV) vector; COVID-19; SARS-CoV-2; Variants; sACE2.
【저자키워드】 COVID-19, SARS-CoV-2, variants, Adeno-associated virus (AAV) vector, sACE2., 【초록키워드】 neutralizing antibody, ACE2, COVID-19 vaccine, therapy, SARS-COV-2 infection, variant, SARS-CoV-2 variant, Infection, in vitro, in silico, virus, angiotensin-converting enzyme 2, variants, Receptor binding domain, human ACE2, Protein, SARS-CoV-2 variants, RBD, Effectiveness, amino acids, therapeutic strategy, breadth, SARS-CoV-2 spike, Angiotensin-converting enzyme, Interaction, angiotensin, adeno-associated virus, tissue tropism, fragment of, Ex vivo, Candidates, serotype, comprehensive analysis, These data, ACE2 orthologs, domain, cell entry, higher affinity, sACE2, weakened, was performed, exhibited, functional, the RBD, ACE2 ortholog, 【제목키워드】 ACE2, SARS-CoV-2 variant, domain, cell entry, functional,