Neutralizing Aptamers Block S/RBD‐ACE2 Interactions and Prevent Host Cell Infection

Liu, Xiaohui; Wang, Yi ling; Wu, Jacky; Qi, Jianjun; Zeng, Zihua; Wan, Quanyuan; Chen, Zhenghu; Manandhar, Pragya; Cavener, Victoria S.; Boyle, Nina R.; Fu, Xinping; Salazar, Eric; Kuchipudi, Suresh V.; Hudson, Peter J.; Zhang, Xiaoliu; Umetani, Michihisa; Şen, Mehmet; Willson, Richard C.; Chen, Shu hsia; Zu, Youli

doi:10.1002/anie.202100345

Cited by 95 publications

(105 citation statements)

References 33 publications

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“…Inhibiting the RBD-ACE2 interaction, thereby preventing viral attachment to target cells, is used as an approach to develop vaccines and therapeutics. The latter include antibodies, aptamers, peptides, and specific small-molecule inhibitors (SMIs) [ 22 , 23 , 24 , 25 , 26 , 27 , 28 ]. One effective and rapid strategy for discovering inhibitors is by screening compound libraries, as was recently applied to find new therapeutics for COVID-19.…”

Section: Discussionmentioning

confidence: 99%

Identification of SARS-CoV-2 Receptor Binding Inhibitors by In Vitro Screening of Drug Libraries

et al. 2021

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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the coronavirus disease 2019 (COVID-19) global pandemic. The first step of viral infection is cell attachment, which is mediated by the binding of the SARS-CoV-2 receptor binding domain (RBD), part of the virus spike protein, to human angiotensin-converting enzyme 2 (ACE2). Therefore, drug repurposing to discover RBD-ACE2 binding inhibitors may provide a rapid and safe approach for COVID-19 therapy. Here, we describe the development of an in vitro RBD-ACE2 binding assay and its application to identify inhibitors of the interaction of the SARS-CoV-2 RBD to ACE2 by the high-throughput screening of two compound libraries (LOPAC®1280 and DiscoveryProbeTM). Three compounds, heparin sodium, aurintricarboxylic acid (ATA), and ellagic acid, were found to exert an effective binding inhibition, with IC50 values ranging from 0.6 to 5.5 µg/mL. A plaque reduction assay in Vero E6 cells infected with a SARS-CoV-2 surrogate virus confirmed the inhibition efficacy of heparin sodium and ATA. Molecular docking analysis located potential binding sites of these compounds in the RBD. In light of these findings, the screening system described herein can be applied to other drug libraries to discover potent SARS-CoV-2 inhibitors.

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Section: Discussionmentioning

confidence: 99%

Identification of SARS-CoV-2 Receptor Binding Inhibitors by In Vitro Screening of Drug Libraries

et al. 2021

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“…Several potential treatments were suggested for COVID-19 involving aptamers during the past year. At the beginning of this year, two publications demonstrated the power of aptamers selected against the receptor binding domain of the SARS-CoV-2 spike protein as diagnostic and therapeutic tools [ 51 , 127 ]. In the first study, initial screening selected aptamer CoV2-6C3 ( Figure 3 (3)), which was further modified to improve overall performance.…”

Section: Aptamers As Therapeutics In Covid-19 Diseasementioning

confidence: 99%

“…Post-modification, the virus was blocked in the sub-nanomolar range, binding affinity increased to K D = 0.13 nM, and the aptamer was stable in serum and at room temperature for prolonged periods [ 51 ]. Soon after, another group published enrichment of potent ssDNA aptamers capable of blocking ligand–receptor interaction, not only preventing host cell infection but also neutralizing the virus in cell culture [ 127 ]. Both groups proceeded to in vivo and/or preclinical studies to test the aptamers’ translational potential.…”

Section: Aptamers As Therapeutics In Covid-19 Diseasementioning

confidence: 99%

Aptamer Applications in Emerging Viral Diseases

et al. 2021

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Aptamers are single-stranded DNA or RNA molecules which are submitted to a process denominated SELEX. SELEX uses reiterative screening of a random oligonucleotide library to identify high-affinity binders to a chosen target, which may be a peptide, protein, or entire cells or viral particles. Aptamers can rival antibodies in target recognition, and benefit from their non-proteic nature, ease of modification, increased stability, and pharmacokinetic properties. This turns them into ideal candidates for diagnostic as well as therapeutic applications. Here, we review the recent accomplishments in the development of aptamers targeting emerging viral diseases, with emphasis on recent findings of aptamers binding to coronaviruses. We focus on aptamer development for diagnosis, including biosensors, in addition to aptamer modifications for stabilization in body fluids and tissue penetration. Such aptamers are aimed at in vivo diagnosis and treatment, such as quantification of viral load and blocking host cell invasion, virus assembly, or replication, respectively. Although there are currently no in vivo applications of aptamers in combating viral diseases, such strategies are promising for therapy development in the future.

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“…Live virus neutralization (VN) for variant of concern B.1.1.7 and WA/1 were performed in an orthogonal assay. The ability of the monoclonal antibodies to neutralize SARS-CoV-2 was determined with a traditional VN assay using SARS-CoV-2 strain USA-WA1/2020 (NR-52281-BEI resources), as previously described 40,41,42,43 . All experiments with SARS-CoV-2 were performed in the Eva J Pell BSL-3 laboratory at Penn State and were approved by the Penn State Institutional Biosafety Committee (IBC # 48625).…”

Section: Live Virus Neutralization Assaysmentioning

confidence: 99%

Synthetic repertoires derived from convalescent COVID-19 patients enable discovery of SARS-CoV-2 neutralizing antibodies and a novel quaternary binding modality

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Hsieh

Horton

et al. 2021

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The ongoing evolution of SARS-CoV-2 into more easily transmissible and infectious variants has sparked concern over the continued effectiveness of existing therapeutic antibodies and vaccines. Hence, together with increased genomic surveillance, methods to rapidly develop and assess effective interventions are critically needed. Here we report the discovery of SARS-CoV-2 neutralizing antibodies isolated from COVID-19 patients using a high-throughput platform. Antibodies were identified from unpaired donor B-cell and serum repertoires using yeast surface display, proteomics, and public light chain screening. Cryo-EM and functional characterization of the antibodies identified N3-1, an antibody that binds avidly (Kd,app = 68 pM) to the receptor binding domain (RBD) of the spike protein and robustly neutralizes the virus in vitro. This antibody likely binds all three RBDs of the trimeric spike protein with a single IgG. Importantly, N3-1 equivalently binds spike proteins from emerging SARS-CoV-2 variants of concern, neutralizes UK variant B.1.1.7, and binds SARS-CoV spike with nanomolar affinity. Taken together, the strategies described herein will prove broadly applicable in interrogating adaptive immunity and developing rapid response biological countermeasures to emerging pathogens.

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Neutralizing Aptamers Block S/RBD‐ACE2 Interactions and Prevent Host Cell Infection

Cited by 95 publications

References 33 publications

Identification of SARS-CoV-2 Receptor Binding Inhibitors by In Vitro Screening of Drug Libraries

Identification of SARS-CoV-2 Receptor Binding Inhibitors by In Vitro Screening of Drug Libraries

Aptamer Applications in Emerging Viral Diseases

Synthetic repertoires derived from convalescent COVID-19 patients enable discovery of SARS-CoV-2 neutralizing antibodies and a novel quaternary binding modality

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