Divergent pathogenetic outcomes in BALB/c mice following Omicron subvariant infection

Virus Research, Год журнала: 2024, Номер 341, С. 199319 - 199319

Опубликована: Янв. 21, 2024

Following the emergence of B.1.1.529 Omicron, SARS-CoV-2 virus evolved into a significant number sublineage variants that possessed numerous mutations throughout genome, but particularly within spike glycoprotein (S) gene. For example, BQ.1.1 and XBB.1 XBB.1.5 subvariants contained 34 41 in S, respectively. However, these elicited largely replication only or mild disease phenotypes mice. To better model pathogenic outcomes measure countermeasure performance, we developed mouse adapted versions (BQ.1.1 MA; MA) reflect more acute phase pulmonary symptoms SARS-CoV-2, as well derivative strains expressing nano-luciferase (nLuc) place ORF7 nLuc; nLuc). Amongst (MA) viruses, wide range were observed including mortality, weight loss, lung dysfunction, tissue viral loads nasal turbinates. Intriguingly, MA strains, which identical except at position F486S/P exhibited divergent mice (Ao et al., 2023). infection was associated with loss ∼45 % mortality across two independent studies, while infected animals suffered from 10 same studies. Additionally, development use nanoluciferase provided moderate throughput for live neutralization assays. The availability small animal models assessment Omicron VOC potential will enable refined capacity to evaluate efficacy on market pre-clinical therapeutics interventions.

Язык: Английский

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In Silico Design of miniACE2 Decoys with In Vitro Enhanced Neutralization Activity against SARS-CoV-2, Encompassing Omicron Subvariants

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(19), С. 10802 - 10802

Опубликована: Окт. 8, 2024

The COVID-19 pandemic has overwhelmed healthcare systems and triggered global economic downturns. While vaccines have reduced the lethality rate of SARS-CoV-2 to 0.9% as October 2024, continuous evolution variants remains a significant public health challenge. Next-generation medical therapies offer hope in addressing this threat, especially for immunocompromised individuals who experience prolonged infections severe illnesses, contributing viral evolution. These cases increase risk new emerging. This study explores miniACE2 decoys novel strategy counteract variants. Using silico design molecular dynamics, blocking proteins (BPs) were developed with stronger binding affinity receptor-binding domain multiple than naturally soluble human ACE2. BPs expressed E. coli tested vitro, showing promising neutralizing effects. Notably, BP9 exhibited an average IC50 4.9 µg/mL across several variants, including Wuhan strain, Mu, Omicron BA.1, BA.2 low demonstrates potent ability BP9, indicating its efficacy at concentrations.Based on these findings, emerged therapeutic candidate combating evolving thereby positioning it potential emergency biopharmaceutical.

Язык: Английский

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Pseudotyped virus infection of multiplexed ACE2 libraries reveals SARS-CoV-2 variant shifts in receptor usage

PLoS Pathogens, Год журнала: 2024, Номер 20(5), С. e1012044 - e1012044

Опубликована: Май 20, 2024

Pairwise compatibility between virus and host proteins can dictate the outcome of infection. During transmission, both inter- intraspecies variabilities in receptor protein sequences impact cell susceptibility. Many viruses possess mutable viral entry patterns shift as sequence changes. This combinatorial space is poorly understood, traditional experimental approaches lack throughput to simultaneously test all possible combinations sequences. Here, we created a pseudotyped infection assay where multiplexed target-cell library variants be assayed using DNA barcode sequencing readout. We applied this panel 30 ACE2 orthologs or human mutants for infectability by original SARS-CoV-2 spike Alpha, Beta, Gamma, Delta, Omicron BA1 variant spikes. compared these results an analysis structural shifts that occurred each spike’s interface with ACE2. Mutated residues were directly involved largest shifts, although there also widespread indirect effects altering structure. The N501Y substitution conferred large interaction ACE2, which was partially recreated distal substitutions does not harbor N501Y. from greatly influenced set animal capable interacting with. Out thirteen non-human orthologs, ten exhibited unique variant-specific compatibility, demonstrating changes during transmission toggle potential susceptibility other species, cumulatively increase overall compatibilities new emerge. These experiments provide blueprint similar large-scale assessments diverse viruses. dataset demonstrates complex relationships occur variable proteins.

Язык: Английский

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Efficacies of S-nitrosoglutathione (GSNO) and GSNO reductase inhibitor in SARS-CoV-2 spike protein induced acute lung disease in mice

Frontiers in Pharmacology, Год журнала: 2023, Номер 14

Опубликована: Дек. 8, 2023

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which initially surfaced in late 2019, often triggers pulmonary complications, encompassing various disease mechanisms such as intense lung inflammation, vascular dysfunction, and embolism. Currently, however, there’s no drug addressing all these simultaneously. This study explored the multi-targeting potential of S-nitrosoglutathione (GSNO) N6022, an inhibitor GSNO reductase (GSNOR) on markers inflammatory, vascular, thrombotic diseases related to COVID-19-induced disease. For this, was induced C57BL/6 mice through intranasal administration recombinant SARS-CoV-2 spike protein S1 domain (SP-S1). exhibited fever, body weight loss, increased blood levels expression proinflammatory cytokines (e.g., TNF-α IL-6) well inflammation mediated by ICAM-1 VCAM-1 infiltration immune cells neutrophils, monocytes, activated cytotoxic helper T cells). Further, hyperpermeability (lung Evans blue extravasation) leading edema development elevated coagulation factors fibrinogen, thrombin, platelets, von Willebrand factor) fibrin deposition. Similar patients with COVID-19, male showed more than female mice, along higher GSNOR lungs. Optimization treatment exogenous or inhibition N6022 (or knockout) protects against SP-S1-induced both genders. These findings provide evidence for efficacies inhibitors multi-mechanistic nature SP-associated acute-lung

Язык: Английский

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The panzootic potential of SARS-CoV-2

BioScience, Год журнала: 2023, Номер 73(11), С. 814 - 829

Опубликована: Ноя. 1, 2023

Each year, SARS-CoV-2 is infecting an increasingly unprecedented number of species. In the present article, we combine mammalian phylogeny with genetic characteristics isolates found in mammals to elaborate on host-range potential SARS-CoV-2. Infections nonhuman mirror those contemporary viral strains circulating humans, although, certain species, extensive circulation has led unique signatures. As other recent studies, that conservation ACE2 receptor cannot be considered sole major determinant susceptibility. However, are able identify clades and families as candidates for increased surveillance. On basis our findings, argue use term panzootic could a more appropriate than pandemic describe ongoing scenario. This better captures magnitude host range would hopefully inspire inclusive policy actions, including systematic screenings, support management this worldwide event.

Язык: Английский

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Cell type-specific adaptation of the SARS-CoV-2 spike

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2023, Номер unknown

Опубликована: Дек. 20, 2023

Abstract SARS-CoV-2 can infect various human tissues and cell types, principally via interaction with its cognate receptor ACE2. However, how the virus evolves in different cellular environments is poorly understood. Here, we used experimental evolution to study adaptation of spike four lines expressing levels key entry factors. After 20 passages, type-specific phenotypic changes were observed. Selected mutations identified functionally characterized terms efficiency, ACE2 affinity, processing, TMPRSS2 usage, pathway syncytia formation. We found that effects these varied across types. Interestingly, two (L48S A372T) emerged cells low increased induction, efficiency under low-ACE2 conditions. Our results demonstrate specific types have implications for understanding tissue tropism evolution.

Язык: Английский

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Pseudotyped virus infection of multiplexed ACE2 libraries reveals SARS-CoV-2 variant shifts in receptor usage

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Фев. 13, 2024

Abstract Pairwise compatibility between virus and host proteins can dictate the outcome of infection. During transmission, both inter- intraspecies variabilities in receptor protein sequences impact cell susceptibility. Many viruses possess mutable viral entry patterns shift as sequence changes. This combinatorial space is poorly understood, traditional experimental approaches lack throughput to simultaneously test all possible combinations sequences. Here, we created a pseudotyped infection assay where multiplexed target-cell library variants be assayed using DNA barcode sequencing readout. We applied this panel 30 ACE2 orthologs or human mutants for infectability by original SARS-CoV-2 spike Alpha, Beta, Gamma, Delta, Omicron BA1 variant spikes. compared these results an analysis structural shifts that occurred each spike’s interface with ACE2. Mutated residues were directly involved largest shifts, although there also widespread indirect effects altering structure. The N501Y substitution conferred large interaction ACE2, which was partially recreated distal substitutions does not harbor N501Y. from greatly influenced set animal capable interacting with. Out thirteen non-human orthologs, ten exhibited unique variant-specific compatibility, demonstrating changes during transmission toggle potential susceptibility other species, cumulatively increase overall compatibilities new emerge. These experiments provide blueprint similar large-scale assessments diverse viruses. dataset demonstrates complex relationships occur variable proteins.

Язык: Английский

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Cell type-specific adaptation of the SARS-CoV-2 spike

Virus Evolution, Год журнала: 2024, Номер 10(1)

Опубликована: Янв. 1, 2024

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) can infect various human tissues and cell types, principally via interaction with its cognate receptor angiotensin-converting enzyme-2 (ACE2). However, how the virus evolves in different cellular environments is poorly understood. Here, we used experimental evolution to study adaptation of SARS-CoV-2 spike four lines expressing levels key entry factors. After twenty passages a spike-expressing recombinant vesicular stomatitis (VSV), cell-type-specific phenotypic changes were observed sequencing allowed identification sixteen adaptive mutations. We VSV pseudotyping measure efficiency, ACE2 affinity, processing, TMPRSS2 usage, pathway usage all mutants, alone or combination. The fusogenicity mutant spikes was assessed cell-cell fusion assay. Finally, VSVs fitness advantage associated selected found that effects these mutations varied across both terms viral replicative fitness. Interestingly, two (L48S A372T) emerged cells low increased syncytia induction, efficiency under low-ACE2 conditions. Our results demonstrate specific types have implications for understanding tissue tropism evolution.

Язык: Английский

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Characterization of the Pathogenic Features of Multiple SARS-CoV-2 Pandemic Strains in Different Mouse Models

Опубликована: Янв. 1, 2024

Язык: Английский

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Human ACE2 Gene Replacement Mice Support SARS-CoV-2 Viral Replication and Nonlethal Disease Progression

ImmunoHorizons, Год журнала: 2024, Номер 8(9), С. 712 - 720

Опубликована: Сен. 1, 2024

Abstract Many mouse models of SARS-CoV-2 infection involve expression the human ACE2 protein, entry receptor for Spike in tissues. However, most these suffer from nonphysiological regulation expression, which can lead to atypically severe infections and aberrant sites viral replication. In this report, we developed characterized an gene replacement (ACE2-GR) strain Ace2 genomic locus was replaced by entire locus, investigated ability animals respond infection. We show that ACE2-GR mice support replication, but, stark contrast widely used K18-hACE2 transgenic model, leads a mild disease with no detectable involvement CNS. Thus, provide novel, our knowledge, model explore immune responses long-term consequences

Язык: Английский

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