SARS-CoV-2 Omicron efficiently infects human airway, but not alveolar epithelium

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Jan. 20, 2022

Abstract In late 2021, the highly mutated SARS-CoV-2 Omicron variant emerged, raising concerns about its potential extensive immune evasion, increased transmissibility and pathogenicity. Here, we used organoids of human airways alveoli to investigate Omicron’s fitness replicative in comparison with earlier variants. We report that replicates more rapidly has an compared early 614G Delta. contrast, did not replicate productively alveolar type 2 cells. Mechanistically, show does efficiently use TMPRSS2 for entry or spread through cell-cell fusion. Altogether, our data altered tropism protease usage, potentially explaining higher decreased

Language: Английский

---

Choosing a cellular model to study SARS-CoV-2

Frontiers in Cellular and Infection Microbiology, Journal Year: 2022, Volume and Issue: 12

Published: Oct. 21, 2022

As new pathogens emerge, challenges must be faced. This is no different in infectious disease research, where identifying the best tools available laboratories to conduct an investigation can, at least initially, particularly complicated. However, context of emerging virus, such as SARS-CoV-2, which was recently detected China and has become a global threat healthcare systems, developing models infection pathogenesis urgently required. Cell-based approaches are crucial understanding coronavirus biology, growth kinetics, tropism. Usually, laboratory cell lines first line experimental study viral pathogenicity perform assays aimed screening antiviral compounds efficient blocking replication viruses, saving time resources, reducing use animals. determining ideal type can challenging, especially when several researchers have adapt their studies specific requirements. review strives guide scientists who venturing into studying SARS-CoV-2 help them choose right cellular models. It revisits basic concepts virology presents currently

Language: Английский

---

SARS-CoV-2 Omicron entry is type II transmembrane serine protease-mediated in human airway and intestinal organoid models

Journal of Virology, Journal Year: 2023, Volume and Issue: 97(8)

Published: Aug. 9, 2023

SARS-CoV-2 can enter cells after its spike protein is cleaved by either type II transmembrane serine proteases (TTSPs), like TMPRSS2, or cathepsins. It now widely accepted that the Omicron variant uses TMPRSS2 less efficiently and instead enters via cathepsins, but these findings have yet to be verified in more relevant cell models. Although we could confirm efficient cathepsin-mediated entry for a monkey kidney line, experiments with protease inhibitors showed (BA.1 XBB1.5) did not use cathepsins into human airway organoids utilized TTSPs. Likewise, CRISPR-edited intestinal of BA.1 relied on expression cathepsin L B. Together, data force us rethink concept has adapted indicate TTSP should dismissed as prophylactic therapeutic antiviral strategy against SARS-CoV-2. IMPORTANCE Coronavirus relies host activate viral fusion protein, spike. These determine route, tropism, range, attractive drug targets. Whereas earlier studies using lines suggested changed usage, from surface (TTSPs) endosomal report this case organoid models, suggesting inhibition still viable current variants highlighting importance vitro

Language: Английский

---

Glycosylated extracellular mucin domains protect against SARS-CoV-2 infection at the respiratory surface

PLoS Pathogens, Journal Year: 2023, Volume and Issue: 19(8), P. e1011571 - e1011571

Published: Aug. 10, 2023

Mucins play an essential role in protecting the respiratory tract against microbial infections while also acting as binding sites for bacterial and viral adhesins. The heavily O-glycosylated gel-forming mucins MUC5AC MUC5B eliminate pathogens by mucociliary clearance. Transmembrane MUC1, MUC4, MUC16 can restrict invasion at apical surface of epithelium. In this study, we determined impact host mucin glycans on epithelial entry SARS-CoV-2. Human lung Calu-3 cells express SARS-CoV-2 receptor ACE2 high levels glycosylated but not MUC4 MUC16, their cell surface. O-glycan-specific mucinase StcE specifically removed part MUC1 extracellular domain leaving underlying SEA cytoplasmic tail intact. treatment significantly enhanced infection with pseudovirus authentic virus, removal terminal sialic acid fucose from did entry. cells, transmembrane are located to close proximity results purified spike protein. Both expressed human organoid-derived air-liquid interface (ALI) differentiated airway cultures led increased replication. these cultures, was highly non-ciliated enriched goblet cells. conclusion, domains different might have similar protective functions types restricting

Language: Английский

---

Human conjunctiva organoids to study ocular surface homeostasis and disease

Cell stem cell, Journal Year: 2024, Volume and Issue: 31(2), P. 227 - 243.e12

Published: Jan. 11, 2024

The conjunctival epithelium covering the eye contains two main cell types: mucus-producing goblet cells and water-secreting keratinocytes, which present mucins on their apical surface. Here, we describe long-term expanding organoids air-liquid interface representing mouse human conjunctiva. A single-cell RNA expression atlas of primary cultured conjunctiva reveals that keratinocytes express multiple antimicrobial peptides identifies tuft cells. IL-4/-13 exposure increases differentiation drastically modifies secretome. Human NGFR+ basal are identified as bipotent stem Conjunctival cultures can be infected by herpes simplex virus 1 (HSV1), adenovirus 8 (hAdV8), SARS-CoV-2. HSV1 infection was reversed acyclovir addition, whereas hAdV8 infection, lacks an approved drug therapy, inhibited cidofovir. We document transcriptional programs induced hAdV8. Finally, transplanted. Together, organoid enable study (patho)-physiology.

Language: Английский

---

SARS-CoV-2 variants reveal features critical for replication in primary human cells

PLoS Biology, Journal Year: 2021, Volume and Issue: 19(3), P. e3001006 - e3001006

Published: March 24, 2021

Since entering the human population, Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2; causative agent of Disease 2019 [COVID-19]) has spread worldwide, causing >100 million infections and >2 deaths. While large-scale sequencing efforts have identified numerous genetic variants in SARS-CoV-2 during its circulation, it remains largely unclear whether many these changes impact adaptation, replication, or transmission virus. Here, we characterized 14 different low-passage replication-competent isolates representing all major European clades observed first pandemic wave early 2020. By integrating viral data from patient material, virus stocks, passaging experiments, together with kinetic replication nonhuman Vero-CCL81 cells primary differentiated bronchial epithelial (BEpCs), several features that associate distinct phenotypes. Notably, naturally occurring Orf3a (Q57H) nsp2 (T85I) were associated poor but not BEpCs, while expressing Spike D614G variant generally exhibited enhanced abilities BEpCs. Strikingly, Vero-derived stock preparation 3 selected for substitutions at positions 5/6 E highly attenuated revealing a key cell-specific function to this region. Rare isolate-specific deletions also furin cleavage site passage, rapidly against underscoring importance cells. Overall, our study uncovers sequence determine highlights need monitor stocks carefully when phenotyping newly emerging potential concern.

Language: Английский

---

Advancing lung organoids for COVID-19 research

Disease Models & Mechanisms, Journal Year: 2021, Volume and Issue: 14(6)

Published: June 1, 2021

ABSTRACT The COVID-19 pandemic has emphasised the need to develop effective treatments combat emerging viruses. Model systems that poorly represent a virus' cellular environment, however, may impede research and waste resources. Collaborations between cell biologists virologists have led rapid development of representative organoid model study severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We believe lung organoids, in particular, advanced our understanding SARS-CoV-2 pathogenesis, laid foundation future viruses treatments.

Language: Английский

---

Air-Liquid-Interface Differentiated Human Nose Epithelium: A Robust Primary Tissue Culture Model of SARS-CoV-2 Infection

International Journal of Molecular Sciences, Journal Year: 2022, Volume and Issue: 23(2), P. 835 - 835

Published: Jan. 13, 2022

The global urgency to uncover medical countermeasures combat the COVID-19 pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) has revealed an unmet need for robust tissue culture models that faithfully recapitulate key features of human tissues and disease. Infection nose is considered dominant initial site SARS-CoV-2 infection replicate this entry portal offer greatest potential examining demonstrating effectiveness designed prevent or manage highly communicable Here, we test air–liquid-interface (ALI) differentiated nasal epithelium (HNE) system as a model authentic infection. Progenitor cells (basal cells) were isolated from turbinate brushings, expanded under conditionally reprogrammed cell (CRC) conditions at ALI. Differentiated inoculated with different clinical isolates. Infectious virus release into apical washes was determined TCID50, while infected visualized immunofluorescence confocal microscopy. We demonstrate robust, reproducible ALI-HNE established donors. Viral occurred surface, primarily observed in ciliated cells. In contrast ancestral isolate, Delta variant considerable damage. Successful establishment donor dependent. can serve pre-clinical without invasive collection samples.

Language: Английский

---

Host susceptibility and structural and immunological insight of S proteins of two SARS-CoV-2 closely related bat coronaviruses

Cell Discovery, Journal Year: 2023, Volume and Issue: 9(1)

Published: July 28, 2023

Abstract The bat coronaviruses (CoV) BANAL-20-52 and BANAL-20-236 are two newly identified severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) closely related (SC2r-CoV) the genome of shares highest homology with SARS-CoV-2. However, risk their potential zoonotic transmission has not been fully evaluated. Here, we determined host susceptibility among 13 different species 26 animal species, found that both might have extensive ranges, indicating high potential. We also cryo-EM structures S proteins at pH 5.5 complex S1 Rhinolophus affinis ACE2, trimeric adopt all three receptor binding domains (RBDs) in “closed” conformation more compact than Strikingly, unique sugar moiety N370 SC2r-CoVs acts like a “bolt” crosses over neighboring subunits, facilitating locked underpinning architecture stability. Removal glycosylation by T372A substitution substantially enhances virus infectivity but becomes highly sensitive to trypsin digestion 5.5, condition roughly mimicking insectivorous bat’s stomach digestion. In contrast, WT showed considerable resistance conserved T372 CoVs result from selective advantages stability during fecal-oral A372. Moreover, results cross-immunogenicity SARS-CoV-2, BANAL-20-52, A372 pseudoviruses anti-S sera T372, immune evasion play role natural selection evolution. Finally, residues 493 498 protein affect susceptibility, residue influences immunogenicity protein. Together, our findings aid better understanding molecular basis CoV entry, evolution, highlight importance surveillance susceptible hosts these viruses prevent outbreaks.

Language: Английский

---

Highly Efficient SARS-CoV-2 Infection of Human Cardiomyocytes: Spike Protein-Mediated Cell Fusion and Its Inhibition

Journal of Virology, Journal Year: 2021, Volume and Issue: 95(24)

Published: Oct. 6, 2021

Cardiac complications frequently observed in COVID-19 patients are tentatively attributed to systemic inflammation and thrombosis, but viral replication has occasionally been confirmed cardiac tissue autopsy materials. We developed an vitro model of SARS-CoV-2 spread myocardium using induced pluripotent stem cell-derived cardiomyocytes.

Language: Английский

---