Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration

Yang, Qi; Hughes, Thomas A.; Kelkar, Anju; Yu, Xinheng; Cheng, Kai; Park, Sheldon J.; Huang, Wei; Lovell, Jonathan F.; Neelamegham, Sriram

doi:10.7554/elife.61552

Cited by 200 publications

(234 citation statements)

References 54 publications

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“…This maturation process is key for proper viral protein functions. For instance, Spike N-linked glycosylation is required for virus entry into the host cells impacting directly on Spike stability during its synthesis instead of its binding ability to the ACE2 receptor (10). These modifications might be also key for antibody recognition.…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV2 envelop proteins reshape the serological responses of COVID-19 patients

Martin

Heslan²,

Jégou

et al. 2021

Preprint

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The SARS-CoV-2 pandemic has elicited a unique international mobilization of the scientific community to better understand this coronavirus and its associated disease and to develop efficient tools to combat infection. Similar to other coronavirae, SARS-CoV-2 hijacks the host cell complex secretory machinery to produce properly folded viral proteins that will compose the nascent virions; including Spike, Envelope and Membrane proteins, the most exposed membrane viral proteins to the host immune system. Antibody response is part of the anti-viral immune arsenal that infected patients develop to fight viral particles in the body. Herein, we investigate the immunogenic potential of Spike (S), Envelope (E) and Membrane (M) proteins using a human cell-based system to mimic membrane insertion and N-glycosylation. We show that both S and M proteins elicit the production of specific IgG, IgM and IgA in SARS-CoV-2 infected patients. Elevated Ig responses were observed in COVID+ patients with moderate and severe forms of the disease. Finally, when SARS-CoV-2 Spike D614 and G614 variants were compared, reduced Ig binding was observed with the Spike G614 variant. Altogether, this study underlines the needs for including topological features in envelop proteins to better characterize the serological status of COVID+ patients, points towards an unexpected immune response against the M protein and shows that our assay could represent a powerful tool to test humoral responses against actively evolving SARS-CoV-2 variants and vaccine effectiveness.

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Section: Introductionmentioning

confidence: 99%

SARS-CoV2 envelop proteins reshape the serological responses of COVID-19 patients

Martin

Heslan²,

Jégou

et al. 2021

Preprint

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“…The SARS-CoV-2 S protein possesses several documented O -glycosylation sites located at the hinge between the RBD and NTD domains and surrounding the furin cleavage site ( Antonopoulos et al, 2020 ; Gao et al, 2020 ; Sanda et al, 2020 ; Shajahan et al, 2020 ; Zhao P. et al, 2020 ). Blocking of O -glycosylation resulted in partial inhibition of SARS-CoV-2 cell entry in vitro , indicating the functional importance of O -glycans in the infection process ( Yang et al, 2020 ). Epithelial cells of the upper respiratory tract, nasopharynx, trachea and large bronchi, appear to be the main producers of virions involved in inter-individual transmission ( Wolfel et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Low Levels of Natural Anti-α-N-Acetylgalactosamine (Tn) Antibodies Are Associated With COVID-19

et al. 2021

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Human serum contains large amounts of anti-carbohydrate antibodies, some of which may recognize epitopes on viral glycans. Here, we tested the hypothesis that such antibodies may confer protection against COVID-19 so that patients would be preferentially found among people with low amounts of specific anti-carbohydrate antibodies since individual repertoires vary considerably. After selecting glycan epitopes commonly represented in the human anti-carbohydrate antibody repertoire that may also be expressed on viral glycans, plasma levels of the corresponding antibodies were determined by ELISA in 88 SARS-CoV-2 infected individuals, including 13 asymptomatic, and in 82 non-infected controls. We observed that anti-Tn antibodies levels were significantly lower in patients as compared to non-infected individuals. This was not observed for any of the other tested carbohydrate epitopes, including anti-αGal antibodies used as a negative control since the epitope cannot be synthesized by humans. Owing to structural homologies with blood groups A and B antigens, we also observed that anti-Tn and anti-αGal antibodies levels were lower in blood group A and B, respectively. Analyses of correlations between anti-Tn and the other anti-carbohydrates tested revealed divergent patterns of correlations between patients and controls, suggesting qualitative differences in addition to the quantitative difference. Furthermore, anti-Tn levels correlated with anti-S protein levels in the patients’ group, suggesting that anti-Tn might contribute to the development of the specific antiviral response. Overall, this first analysis allows to hypothesize that natural anti-Tn antibodies might be protective against COVID-19.

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“…Previous studies have predicted that spike N- glycans may modulate protein folding and multimerization 8 , conformational changes, binding to the ACE-2 human receptor and immunological shielding 5 . Also, it has been shown using recombinant proteins and pseudotyped viruses that mutations in certain N- glycosylation sites of the spike lead to reduced infectivity 10 , and that treatment with kifunensine or MGAT1 knockout to prevent the formation of complex N- glycans reduces pseudovirus infection 11 . However, the effects of altering N- glycosylation in SARS-CoV-2 virions remain undetermined.…”

Section: Discussionmentioning

confidence: 99%

“…By mutating N- glycosylation sites, it has been shown that the absence of glycans on Asn 331 and Asn 343 , both within the RBD, resulted in a drastic reduction of pseudovirus infection 10 . In addition, knocking out MGAT1 gene, which encodes for N -acetylglucosaminyl-transferase, an essential enzyme in the formation of hybrid and complex N -glycans, also reduced invasion of pseudotyped VSV suggesting that hybrid and complex spike N- glycans may be of importance for this process 11 . Although SARS-CoV-2 cDNA clones have been available for months 12,13 , no study has yet used real virions to provide direct evidence.…”

Section: Introductionmentioning

confidence: 99%

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Protein N-glycosylation is essential for SARS-CoV-2 infection

Casas-Sánchez

Romero-Ramirez

Hargreaves

et al. 2021

Preprint

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SARS-CoV-2 extensively N-glycosylates its surface spike (S) proteins. This post-translational modification is essential to modulate protein conformation and host cell invasion. Each S monomer can be modified with up to 22 N-glycans. To meet the high demand of protein glycosylation during virus replication, SARS-CoV-2 upregulates the expression of host N-glycosylation genes. Although a substantial amount of detail is known about the structure of S protein N-glycans, the role of N-glycosylation in SARS-CoV-2 infection remains largely undetermined. Here, we investigated the essentiality of the host N-glycosylation pathway and viral N-glycans for SARS-CoV-2 infection. When either monkey or human cells were preincubated with glycosylation inhibitors, including FDA-approved iminosugars, virus infection was significantly reduced. This infection phenotype was confirmed after RNAi knockdown of several glycosylation genes. In addition, enzymatic deglycosylation of whole viral particles confirmed that accessible oligosaccharides on the SARS-CoV-2 surface are essential for host cell infection. Altogether, we show evidence that the normal functioning of the host N-glycosylation machinery is essential not only for SARS-CoV-2 to infect, but also to produce new functional virions. These findings open the door for developing new approaches targeting N-glycosylation against COVID-19.

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Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration

Cited by 200 publications

References 54 publications

SARS-CoV2 envelop proteins reshape the serological responses of COVID-19 patients

SARS-CoV2 envelop proteins reshape the serological responses of COVID-19 patients

Low Levels of Natural Anti-α-N-Acetylgalactosamine (Tn) Antibodies Are Associated With COVID-19

Protein N-glycosylation is essential for SARS-CoV-2 infection

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