Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro

作者：Hsiang‐Chi Huang, Yun‐Ju Lai, Chun‐Che Liao, Feng-Yang Wang, Ke-Bin Huang, I‐Jung Lee, Wen‐Cheng Chou, Shih‐Han Wang, Linghui Wang, Jung-Mao Hsu, Cheng‐Pu Sun, Chun‐Tse Kuo, Jyun Wang, Tzu‐Chun Hsiao, Po-Jiun Yang, Te-An Lee, Wilson Huang, Fu‐An Li, Chen‐Yang Shen, Yi‐Ling Lin, Mi‐Hua Tao, Chia-Wei Li · 发表于：EBioMedicine · 年份：2021 · DOI：10.1016/j.ebiom.2021.103712 · 被引用次数：65 · 研究领域：SARS-CoV-2 and COVID-19 Research、vaccines and immunoinformatics approaches、Immune responses and vaccinations

BACKGROUND: Despite clinical success with anti-spike vaccines, the effectiveness of neutralizing antibodies and vaccines has been compromised by rapidly spreading SARS-CoV-2 variants. Viruses can hijack the glycosylation machinery of host cells to shield themselves from the host's immune response and attenuate antibody efficiency. However, it remains unclear if targeting glycosylation on viral spike protein can impair infectivity of SARS-CoV-2 and its variants. METHODS: We adopted flow cytometry, ELISA, and BioLayer interferometry approaches to assess binding of glycosylated or deglycosylated spike with ACE2. Viral entry was determined by luciferase, immunoblotting, and immunofluorescence assays. Genome-wide association study (GWAS) revealed a significant relationship between STT3A and COVID-19 severity. NF-κB/STT3A-regulated N-glycosylation was investigated by gene knockdown, chromatin immunoprecipitation, and promoter assay. We developed an antibody-drug conjugate (ADC) that couples non-neutralization anti-spike antibody with NGI-1 (4G10-ADC) to specifically target SARS-CoV-2-infected cells. FINDINGS: The receptor binding domain and three distinct SARS-CoV-2 surface N-glycosylation sites among 57,311 spike proteins retrieved from the NCBI-Virus-database are highly evolutionarily conserved (99.67%) and are involved in ACE2 interaction. STT3A is a key glycosyltransferase catalyzing spike glycosylation and is positively correlated with COVID-19 severity. We found that inhibiti...