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

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 respon...

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Autores principales:	Hsiang-Chi Huang, Yun-Ju Lai, Chun-Che Liao, Wang-Feng Yang, Ke-Bin Huang, I-Jung Lee, Wen-Cheng Chou, Shih-Han Wang, Ling-Hui 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
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	SARS-CoV-2 variant STT3A NGI-1 ADC Deglycosylation Medicine R Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/8fed32a6141e421c81852255ebc22345
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spelling	oai:doaj.org-article:8fed32a6141e421c81852255ebc223452021-11-26T04:31:39ZTargeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro2352-396410.1016/j.ebiom.2021.103712https://doaj.org/article/8fed32a6141e421c81852255ebc223452021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2352396421005065https://doaj.org/toc/2352-3964Background: 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 inhibiting STT3A using N-linked glycosylation inhibitor-1 (NGI-1) impaired SARS-CoV-2 infectivity and that of its variants [Alpha (B.1.1.7) and Beta (B.1.351)]. Most importantly, 4G10-ADC enters SARS-CoV-2-infected cells and NGI-1 is subsequently released to deglycosylate spike protein, thereby reinforcing the neutralizing abilities of antibodies, vaccines, or convalescent sera and reducing SARS-CoV-2 variant infectivity. Interpretation: Our results indicate that targeting evolutionarily-conserved STT3A-mediated glycosylation via an ADC can exert profound impacts on SARS-CoV-2 variant infectivity. Thus, we have identified a novel deglycosylation method suitable for eradicating SARS-CoV-2 variant infection in vitro. Funding: A full list of funding bodies that contributed to this study can be found in the Acknowledgements sectionHsiang-Chi HuangYun-Ju LaiChun-Che LiaoWang-Feng YangKe-Bin HuangI-Jung LeeWen-Cheng ChouShih-Han WangLing-Hui WangJung-Mao HsuCheng-Pu SunChun-Tse KuoJyun WangTzu-Chun HsiaoPo-Jiun YangTe-An LeeWilson HuangFu-An LiChen-Yang ShenYi-Ling LinMi-Hua TaoChia-Wei LiElsevierarticleSARS-CoV-2 variantSTT3ANGI-1ADCDeglycosylationMedicineRMedicine (General)R5-920ENEBioMedicine, Vol 74, Iss , Pp 103712- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	SARS-CoV-2 variant STT3A NGI-1 ADC Deglycosylation Medicine R Medicine (General) R5-920
spellingShingle	SARS-CoV-2 variant STT3A NGI-1 ADC Deglycosylation Medicine R Medicine (General) R5-920 Hsiang-Chi Huang Yun-Ju Lai Chun-Che Liao Wang-Feng Yang Ke-Bin Huang I-Jung Lee Wen-Cheng Chou Shih-Han Wang Ling-Hui 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 Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
description	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 inhibiting STT3A using N-linked glycosylation inhibitor-1 (NGI-1) impaired SARS-CoV-2 infectivity and that of its variants [Alpha (B.1.1.7) and Beta (B.1.351)]. Most importantly, 4G10-ADC enters SARS-CoV-2-infected cells and NGI-1 is subsequently released to deglycosylate spike protein, thereby reinforcing the neutralizing abilities of antibodies, vaccines, or convalescent sera and reducing SARS-CoV-2 variant infectivity. Interpretation: Our results indicate that targeting evolutionarily-conserved STT3A-mediated glycosylation via an ADC can exert profound impacts on SARS-CoV-2 variant infectivity. Thus, we have identified a novel deglycosylation method suitable for eradicating SARS-CoV-2 variant infection in vitro. Funding: A full list of funding bodies that contributed to this study can be found in the Acknowledgements section
format	article
author	Hsiang-Chi Huang Yun-Ju Lai Chun-Che Liao Wang-Feng Yang Ke-Bin Huang I-Jung Lee Wen-Cheng Chou Shih-Han Wang Ling-Hui 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
author_facet	Hsiang-Chi Huang Yun-Ju Lai Chun-Che Liao Wang-Feng Yang Ke-Bin Huang I-Jung Lee Wen-Cheng Chou Shih-Han Wang Ling-Hui 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
author_sort	Hsiang-Chi Huang
title	Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
title_short	Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
title_full	Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
title_fullStr	Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
title_full_unstemmed	Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro
title_sort	targeting conserved n-glycosylation blocks sars-cov-2 variant infection in vitro
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/8fed32a6141e421c81852255ebc22345
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Targeting conserved N-glycosylation blocks SARS-CoV-2 variant infection in vitro

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