Identification of Cellular Factors Required for SARS-CoV-2 Replication

Synowiec, Aleksandra; Jedrysik, Malwina; Branicki, Wojciech; Klajmon, Adrianna; Lei, Jing; Owczarek, Katarzyna; Suo, Chen; Szczepański, Artur; Wang, Jingru; Zhang, Pengyan; Łabaj, Paweł P.; Pyrć, Krzysztof

doi:10.3390/cells10113159

Cited by 14 publications

(16 citation statements)

References 46 publications

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“…This is also the reason why, in the current work, further studies were focused on the activity of P. galeata CCNP1313 metabolites against SARS-CoV-2. For this purpose, the human lung epithelial cell line A549 ACE2/TMPRSS2 overexpressing the angiotensin-converting enzyme 2 (ACE2), and the transmembrane serine proteases TMPRSS2, crucial for the SARS-CoV-2 spike protein priming was used [ 42 , 43 ]. A549 ACE2/TMPRSS2 cells recapitulate the natural route of entry for the coronavirus, which enters through fusion directly on the cell surface.…”

Section: Discussionmentioning

confidence: 99%

“…A549 ACE2/TMPRSS2 cells recapitulate the natural route of entry for the coronavirus, which enters through fusion directly on the cell surface. As a result, this makes the virus independent of the endocytic machinery and cellular cathepsins [ 43 ]. Similarly to cytotoxic effects, the antiviral activity was recorded in fractions eluted with solvent of different MeOH content.…”

Section: Discussionmentioning

confidence: 99%

“…Cell Culture. Vero E6 (ATCC: CRL-1586), HCT-8 (ATCC: CCL-244), primary human dermal fibroblasts, A549 (ATCC: CCL-185; ATCC, Manassas, VA, USA) overexpressing ACE2 and TMPRSS2 (A549 ACE2/TMPRSS2 in-house generated [ 43 ]) were maintained in DMEM medium (Gibco, Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 5% v/v heat-inactivated fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific Inc., Waltham, MA, USA) and penicillin-streptomycin solution (100 U mL −1 and 100 µg mL −1 , respectively) (PAN Biotech GmbH, Aidenbach, Bayern, Germany). Medium for A549 ACE2/TMPRSS2 cells was additionally supplemented with blasticidin S (10 µg mL −1 ; Sigma-Aldrich, Merck, Warsaw, Poland) and puromycin (0.5 µg mL −1 ; Sigma-Aldrich, Merck, Warsaw, Poland).…”

Section: Methodsmentioning

confidence: 99%

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Pseudanabaena galeata CCNP1313—Biological Activity and Peptides Production

Cegłowska

Szubert

Grygier

et al. 2022

Toxins

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Even cyanobacteria from ecosystems of low biodiversity, such as the Baltic Sea, can constitute a rich source of bioactive metabolites. Potent toxins, enzyme inhibitors, and anticancer and antifungal agents were detected in both bloom-forming species and less commonly occurring cyanobacteria. In previous work on the Baltic Pseudanabaena galeata CCNP1313, the induction of apoptosis in the breast cancer cell line MCF-7 was documented. Here, the activity of the strain was further explored using human dermal fibroblasts, African green monkey kidney, cancer cell lines (T47D, HCT-8, and A549ACE2/TMPRSS2) and viruses (SARS-CoV-2, HCoV-OC43, and WNV). In the tests, extracts, chromatographic fractions, and the main components of the P. galeata CCNP1313 fractions were used. The LC-MS/MS analyses of the tested samples led to the detection of forty-five peptides. For fourteen of the new peptides, putative structures were proposed based on MS/MS spectra. Although the complex samples (i.e., extracts and chromatographic fractions) showed potent cytotoxic and antiviral activities, the effects of the isolated compounds were minor. The study confirmed the significance of P. galeata CCNP1313 as a source of metabolites with potent activity. It also illustrated the difficulties in assigning the observed biological effects to specific metabolites, especially when they are produced in minute amounts.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Pseudanabaena galeata CCNP1313—Biological Activity and Peptides Production

Cegłowska

Szubert

Grygier

et al. 2022

Toxins

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“…To identify the mechanisms through which the symptoms caused by COVID-19 manifest, studies were conducted using cell-based in vitro or animal-based in vivo models [22,23]. It has been found that angiotensin-converting enzyme 2 (ACE2), a cell membrane protein, contributes to the introduction of SARS-CoV-2 into the cell [23].…”

Section: Introductionmentioning

confidence: 99%

Understanding the cellular pathogenesis of COVID-19 symptoms using organoid technology

Hyun

Kim

2022

Organoid

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Patients with coronavirus disease 2019 (COVID-19), which has recently caused a pandemic, have reported symptoms of coronavirus infection that are not well understood by the medical community in general. After severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, several symptoms, including acute clinical signs and possible sequelae, manifest in multiple organs. It is necessary to precisely identify the cells susceptible to SARS-CoV-2 infection in order to comprehend the mechanism of symptom occurrence, identify molecular targets for therapeutic development, and prevent current or future threats. Following the use of cell lines, animal models, and stem cell-derived symptom-relevant cells, recent research on the pathophysiology of human diseases has utilized organoid models. This article provides a summary of recent research on the tissue- or organ-specific cellular targets of SARS-CoV-2 aiming to understand the pathophysiology of COVID-19.

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“…Our screens revealed several pro-viral genes shared by SARS-CoV, SARS-CoV-2, and MERS-CoV in both Vero-E6 and Calu-3 cells 22,23 . Other genome-wide CRISPR/Cas9 screens for SARS-CoV-2 and other related coronaviruses have since identified additional host dependency factors including proteins involved in viral entry, endocytic trafficking and sorting, cholesterol homeostasis, and autophagy [27][28][29][30][31][32] . While reproducibility between screens was excellent when performed on the same cell type, there was otherwise limited overlap across hits in these loss of function screens [27][28][29][30][31][32] .…”

Section: Introductionmentioning

confidence: 99%

Kinase-independent activity of DYRK1A promotes viral entry of highly pathogenic human coronaviruses

Strine

Cai

Jin

et al. 2022

Preprint

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Identifying host genes essential for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has the potential to reveal novel drug targets and further our understanding of coronavirus disease 2019 (COVID-19). We previously performed a genome-wide CRISPR/Cas9 screen to identify pro-viral host factors for highly pathogenic human coronaviruses. Very few host factors were required by diverse coronaviruses across multiple cell types, but DYRK1A was one such exception. Although its role in coronavirus infection was completely unknown, DYRK1A encodes Dual Specificity Tyrosine Phosphorylation Regulated Kinase 1A and regulates cell proliferation, and neuronal development, among other cellular processes. Interestingly, individuals with Down syndrome overexpress DYRK1A 1.5-fold and exhibit 5-10x higher hospitalization and mortality rates from COVID-19 infection. Here, we demonstrate that DYRK1A regulates ACE2 and DPP4 transcription independent of its catalytic kinase function to support SARS-CoV, SARS-CoV-2, and MERS-CoV entry. We show that DYRK1A promotes DNA accessibility at the ACE2 promoter and a putative distal enhancer, facilitating transcription and gene expression. Finally, we validate that the pro-viral activity of DYRK1A is conserved across species using cells of monkey and human origin and an in vivo mouse model. In summary, we report that DYRK1A is a novel regulator of ACE2 and DPP4 expression that may dictate susceptibility to multiple highly pathogenic human coronaviruses. Whether DYRK1A overexpression contributes to heightened COVID-19 severity in individuals with Down syndrome through ACE2 regulation warrants further future investigation.

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Identification of Cellular Factors Required for SARS-CoV-2 Replication

Cited by 14 publications

References 46 publications

Pseudanabaena galeata CCNP1313—Biological Activity and Peptides Production

Pseudanabaena galeata CCNP1313—Biological Activity and Peptides Production

Understanding the cellular pathogenesis of COVID-19 symptoms using organoid technology

Kinase-independent activity of DYRK1A promotes viral entry of highly pathogenic human coronaviruses

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