Engineered trimeric ACE2 binds viral spike protein and locks it in “Three-up” conformation to potently inhibit SARS-CoV-2 infection

Guo, Liang; Bi, Wenwen; Wang, Xinling; Xu, Wei; Yan, Renhong; Zhang, Yuanyuan; Zhao, Kai; Li, Yaning; Zhang, Mingfeng; Cai, Xiujun; Jiang, Shibo; Xie, Youhua; Zhou, Qiang; Lu, Lu; Dang, Bobo

doi:10.1038/s41422-020-00438-w

Cited by 82 publications

(83 citation statements)

References 14 publications

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“…One of the mutations from the mutagenesis screening is T27Y, coinciding with our structure-based design. Another study reported a trimeric ACE2 construct with the wild-type sequence 38 , similar to our ACE2 615 -foldon protein. Moreover, de novo protein design based on the ACE2 helix that interacts with the RBD without the catalytic domain has led to picomolar SARS-CoV-2 miniprotein inhibitors 39 .…”

Section: Discussionsupporting

confidence: 68%

A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent

Xiao

Lü

Zhang

et al. 2021

Nat Struct Mol Biol

124

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Effective intervention strategies are urgently needed to control the COVID-19 pandemic. Human angiotensin-converting enzyme 2 (ACE2) is a membrane-bound carboxypeptidase that forms a dimer and serves as the cellular receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). ACE2 is also a key negative regulator of the renin-angiotensin system that modulates vascular functions. We report here the properties of a trimeric ACE2 ectodomain variant, engineered using a structure-based approach. The trimeric ACE2 variant has a binding affinity of ~60 pM for the spike protein of SARS-CoV-2 (compared with 77 nM for monomeric ACE2 and 12-22 nM for dimeric ACE2 constructs), and its peptidase activity and the ability to block activation of angiotensin II receptor type 1 in the renin-angiotensin system are preserved. Moreover, the engineered ACE2 potently inhibits SARS-CoV-2 infection in cell culture. These results suggest that engineered, trimeric ACE2 may be a promising anti-SARS-CoV-2 agent for treating COVID-19.

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Section: Discussionsupporting

confidence: 68%

A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent

Xiao

Lü

Zhang

et al. 2021

Nat Struct Mol Biol

124

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“…As the individual components, sACE2-5xGCN4 and sACE2-scFvGCN4, had similar activity to the untagged sACE2, the pseudovirus neutralization occurred through multimerization of sACE2. Moreover, this result is compatible with the recent studies indicating natural dimerization (28) or engineered trimerization (29) of sACE2 increased the Spike binding affinity and neutralization capacity.…”

supporting

confidence: 92%

“…However, many viruses including SARS-CoV-2 were shown to acquire escape mutations under such evolutionary pressure (27). Therefore, soluble ACE2 and its more potent mutant forms have been proposed as alternatives to neutralizing antibodies (28)(29)(30) as the virus cannot develop resistance without compromising cell entry. Soluble ACE2 was shown to inhibit SARS-CoV-2 infection in cell culture experiments (31), in vitro human organoids (31), and recently on a COVID-19 patient with severe symptoms (32).…”

Section: Introductionmentioning

confidence: 99%

Protein scaffold-based multimerization of soluble ACE2 efficiently blocks SARS-CoV-2 infectionin vitroandin vivo

Kayabölen

Akcan

Özturan

et al. 2021

Preprint

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Soluble ACE2 (sACE2) decoy receptors are promising agents to inhibit SARS-CoV-2 as they are not affected by common escape mutations in viral proteins. However, their success may be limited by their relatively poor potency. To address these challenges, we developed a highly active multimeric sACE2 decoy receptor via a SunTag system that could neutralize both pseudoviruses bearing SARS-CoV-2 spike protein and SARS-CoV-2 clinical isolates. This fusion protein demonstrated a neutralization efficiency nearly 250-fold greater than monomeric sACE2. SunTag in combination with a more potent version of sACE2 achieved near complete neutralization at a sub-nanomolar range, which is comparable with clinical monoclonal antibodies. We demonstrate that this activity is due to greater occupancy of the multimeric decoy receptors on Spike protein as compared to monomeric sACE2. Overall, these highly potent multimeric sACE2 decoy receptors offer a promising treatment approach against SARS-CoV-2 infections including its novel variants.

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“…TMPRSS2 entails proteolytical protein cleavage and folding to a post-fusion conformation coupled with host cell–virus membrane fusion and cytosolic release of viral RNA [ 14 , 15 ]. Thus, the susceptibility to SARS-CoV-2 infection primarily depends on the affinity of the Spike receptor‐binding domain (RBD) to ACE2 in target tissues [ 16 ]. The combined analysis of glycomics-informed glycoproteomics and bioinformatics of variants with molecular dynamics simulations highlighted roles for glycosylation in sterically masking polypeptide epitopes [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Glycated ACE2 receptor in diabetes: open door for SARS-COV-2 entry in cardiomyocyte

D’Onofrio

Scisciola

Sardu

et al. 2021

Cardiovasc Diabetol

103

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Rationale About 50% of hospitalized coronavirus disease 2019 (COVID-19) patients with diabetes mellitus (DM) developed myocardial damage. The mechanisms of direct SARS-CoV-2 cardiomyocyte infection include viral invasion via ACE2-Spike glycoprotein-binding. In DM patients, the impact of glycation of ACE2 on cardiomyocyte invasion by SARS-CoV-2 can be of high importance. Objective To evaluate the presence of SARS-CoV-2 in cardiomyocytes from heart autopsy of DM cases compared to Non-DM; to investigate the role of DM in SARS-COV-2 entry in cardiomyocytes. Methods and results We evaluated consecutive autopsy cases, deceased for COVID-19, from Italy between Apr 30, 2020 and Jan 18, 2021. We evaluated SARS-CoV-2 in cardiomyocytes, expression of ACE2 (total and glycosylated form), and transmembrane protease serine protease-2 (TMPRSS2) protein. In order to study the role of diabetes on cardiomyocyte alterations, independently of COVID-19, we investigated ACE2, glycosylated ACE2, and TMPRSS2 proteins in cardiomyocytes from DM and Non-DM explanted-hearts. Finally, to investigate the effects of DM on ACE2 protein modification, an in vitro glycation study of recombinant human ACE2 (hACE2) was performed to evaluate the effects on binding to SARS-CoV-2 Spike protein. The authors included cardiac tissue from 97 autopsies. DM was diagnosed in 37 patients (38%). Fourth-seven out of 97 autopsies (48%) had SARS-CoV-2 RNA in cardiomyocytes. Thirty out of 37 DM autopsy cases (81%) and 17 out of 60 Non-DM autopsy cases (28%) had SARS-CoV-2 RNA in cardiomyocytes. Total ACE2, glycosylated ACE2, and TMPRSS2 protein expressions were higher in cardiomyocytes from autopsied and explanted hearts of DM than Non-DM. In vitro exposure of monomeric hACE2 to 120 mM glucose for 12 days led to non-enzymatic glycation of four lysine residues in the neck domain affecting the protein oligomerization. Conclusions The upregulation of ACE2 expression (total and glycosylated forms) in DM cardiomyocytes, along with non-enzymatic glycation, could increase the susceptibility to COVID-19 infection in DM patients by favouring the cellular entry of SARS-CoV2.

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Engineered trimeric ACE2 binds viral spike protein and locks it in “Three-up” conformation to potently inhibit SARS-CoV-2 infection

Cited by 82 publications

References 14 publications

A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent

A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent

Protein scaffold-based multimerization of soluble ACE2 efficiently blocks SARS-CoV-2 infectionin vitroandin vivo

Glycated ACE2 receptor in diabetes: open door for SARS-COV-2 entry in cardiomyocyte

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