Whole Genome CRISPR Screening Strategy to Identify Genes Contributing to SARS-CoV-2 Spike and VSV-G Mediated Entry. DOI Open Access
Adam D. Hoppe, Md. Saddam Hossain,

Jason G. Kerkvliet

et al.

Authorea (Authorea), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 12, 2023

Background Understanding the cellular host factors that promote and inhibit viral entry is important for identifying countermeasures. CRISPR whole genome screens can be used to rapidly discover contribute or impair entry. However, when using live viruses lethality selection, these identify an overwhelming number of genes without specificity stage infection cycle. New screening methods are needed machinery contributing specific steps infection. Here, we developed a screen counter strategy based on pseudoviral platform allowed identification SARS-CoV-2 spike vesicular stomatitis virus glycoprotein VSV-G mediated Methods To focus onto step, non-lytic fluorescent reporters in combination with comparative distinguish affecting reporter from those unique envelope-mediated Screening same lentiviral pseudovirus entry-specific relative associated retro-transcription, integration, expression pseudovirus. Second, Cre-Gag fusion protein was bypass retro-transcription integration by directly activating floxed GFP upon reduce gene hits increase Results Our approach correctly identified receptors ACE2 LDLR, respectively distinguished retroviral Moreover, CRE-Gag fusion/flox increased genes. Validation few demonstrates this distinguishes envelope-specific expression. Conclusion Overall, provides new influencing confounding complexity live-viral which produce long lists all aspects pathogenesis replication. This pathway increasing

Language: Английский

Integrated multi-omics analyses identify anti-viral host factors and pathways controlling SARS-CoV-2 infection DOI Creative Commons
Jiakai Hou, Yanjun Wei, Jing Zou

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Jan. 2, 2024

Host anti-viral factors are essential for controlling SARS-CoV-2 infection but remain largely unknown due to the biases of previous large-scale studies toward pro-viral host factors. To fill in this knowledge gap, we perform a genome-wide CRISPR dropout screen and integrate analyses multi-omics data screen, association studies, single-cell RNA-Seq, host-virus proteins or protein/RNA interactome. This study uncovers many that currently underappreciated, including components V-ATPases, ESCRT, N-glycosylation pathways modulate viral entry and/or replication. The cohesin complex is also identified as an pathway, suggesting important role three-dimensional chromatin organization mediating host-viral interaction. Furthermore, discover another regulator KLF5, transcriptional factor involved sphingolipid metabolism, which up-regulated, harbors genetic variations linked COVID-19 patients with severe symptoms. Anti-viral effects three candidates (DAZAP2/VTA1/KLF5) confirmed individually. Molecular characterization DAZAP2/VTA1/KLF5-knockout cells highlights involvement genes related coagulation system determining severity COVID-19. Together, our results provide further resources understanding network during may help develop new countermeasure strategies.

Language: Английский

Citations

19
Molecular basis of convergent evolution of ACE2 receptor utilization among HKU5 coronaviruses DOI
Young‐Jun Park, Chen Liu, Jimin Lee

et al.

Cell, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 1, 2025

Language: Английский

Citations

5
Understanding Long COVID; Mitochondrial Health and Adaptation—Old Pathways, New Problems DOI Creative Commons
Alistair V.W. Nunn,

Geoffrey W. Guy,

Wolfgang Brysch

et al.

Biomedicines, Journal Year: 2022, Volume and Issue: 10(12), P. 3113 - 3113

Published: Dec. 2, 2022

Many people infected with the SARS-CoV-2 suffer long-term symptoms, such as "brain fog", fatigue and clotting problems. Explanations for "long COVID" include immune imbalance, incomplete viral clearance potentially, mitochondrial dysfunction. As conditions sub-optimal function are associated initial severity of disease, their prior health could be key in resistance to long COVID recovery. The SARs virus redirects host metabolism towards replication; response, can metabolically react control virus. Resolution is normally achieved after stress activates a hormetic negative feedback mechanism. It therefore possible that, some individuals function, "tip" into chronic inflammatory cycle. This might explain main including platelet Long thus described virally induced self-perpetuating imbalanced non-resolving state characterised by dysfunction, where reactive oxygen species continually drive inflammation shift glycolysis. would suggest that sufferer's needs "tipped" back using stimulus, physical activity, calorie restriction, or chemical compounds mimic these enhancing perhaps combination inhibitors quell response.

Language: Английский

Citations

54
Targeting SARS-CoV-2 entry processes: The promising potential and future of host-targeted small-molecule inhibitors DOI

Aijia Wu,

Kunyu Shi,

Jiaxing Wang

et al.

European Journal of Medicinal Chemistry, Journal Year: 2023, Volume and Issue: 263, P. 115923 - 115923

Published: Oct. 31, 2023

Language: Английский

Citations

17
Development of SARS-CoV-2 entry antivirals DOI Creative Commons
Meiyue Dong, Jazmin Galván Achi, Ruikun Du

et al.

Cell Insight, Journal Year: 2024, Volume and Issue: 3(1), P. 100144 - 100144

Published: Jan. 30, 2024

The global outbreak of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) threatened human health and public safety. development anti-SARS-CoV-2 therapies have been essential to curb spread SARS-CoV-2. Particularly, antivirals targeting viral entry become an attractive target for therapies. In this review, we elucidate mechanism SARS-CoV-2 summarize antiviral inhibitors entry. Moreover, speculate upon future directions toward more potent This study is expected provide novel insights efficient discovery promising candidate drugs against SARS-CoV-2, contribute broad-spectrum anti-coronavirus drugs.

Language: Английский

Citations

6
Multicenter, single-blind, randomized controlled study of the efficacy and safety of favipiravir and nafamostat mesilate in patients with COVID-19 pneumonia DOI Creative Commons
Mahoko Ikeda, Shu Okugawa, Kosuke Kashiwabara

et al.

International Journal of Infectious Diseases, Journal Year: 2023, Volume and Issue: 128, P. 355 - 363

Published: Jan. 4, 2023

To evaluate the efficacy and safety of nafamostat combined with favipiravir for treatment COVID-19.We conducted a multicenter, randomized, single-blind, placebo-controlled, parallel assignment study in hospitalized patients mild-to-moderate COVID-19 pneumonia. Patients were randomly assigned to receive alone (n = 24) or 21). The outcomes included changes World Health Organization clinical progression scale score, time improvement body temperature, oxygen saturation (SpO2).There was no significant difference between groups (median, -0.444 vs -0.150, respectively; least-squares mean difference, -0.294; P 0.364). temperature significantly shorter combination group (5.0 days; 95% confidence interval, 4.0-7.0) than (9.0 7.0-18.0; =0.009). SpO2 greater (0.526% -1.304%, 1.831; 0.022). No serious adverse events deaths reported, but phlebitis occurred 57.1% group.Although our showed differences progression, earlier defervescence, recovery observed group.

Language: Английский

Citations

13
Dual spatially resolved transcriptomics for human host–pathogen colocalization studies in FFPE tissue sections DOI Creative Commons
Hailey Sounart, Enikő Lázár, Yuvarani Masarapu

et al.

Genome biology, Journal Year: 2023, Volume and Issue: 24(1)

Published: Oct. 19, 2023

Abstract Technologies to study localized host–pathogen interactions are urgently needed. Here, we present a spatial transcriptomics approach simultaneously capture host and pathogen transcriptome-wide gene expression information from human formalin-fixed paraffin-embedded (FFPE) tissue sections at near single-cell resolution. We demonstrate this methodology in lung samples COVID-19 patients validate our detection of SARS-CoV-2 against RNAScope situ sequencing. Host–pathogen colocalization analysis identified putative modulators infection cells. Our provides new insights into response through the simultaneous, unbiased two transcriptomes FFPE samples.

Language: Английский

Citations

13
Human cytomegalovirus degrades DMXL1 to inhibit autophagy, lysosomal acidification, and viral assembly DOI Creative Commons
Hanqi Li, Alice Fletcher-Etherington, Leah M. Hunter

et al.

Cell Host & Microbe, Journal Year: 2024, Volume and Issue: 32(4), P. 466 - 478.e11

Published: March 12, 2024

Human cytomegalovirus (HCMV) is an important human pathogen that regulates host immunity and hijacks compartments, including lysosomes, to assemble virions. We combined a quantitative proteomic analysis of HCMV infection with database proteins involved in vacuolar acidification, revealing Dmx-like protein-1 (DMXL1) as the only protein acidifies vacuoles yet degraded by HCMV. Systematic comparison viral deletion mutants reveals uncharacterized 7 kDa US33A necessary sufficient for DMXL1 degradation, which occurs via recruitment E3 ubiquitin ligase Kip1 ubiquitination-promoting complex (KPC). US33A-mediated degradation inhibits lysosome acidification autophagic cargo degradation. Formation virion assembly compartment, requires significantly later US33A-expressing virus infection, reduced replication. These data thus identify strategy cellular remodeling, potential employ therapies or rheumatic conditions, inhibition can attenuate disease.

Language: Английский

Citations

4
Exogenous RNA surveillance by proton-sensing TRIM25 DOI
Myeonghwan Kim, Youngjoon Pyo, Seong‐In Hyun

et al.

Science, Journal Year: 2025, Volume and Issue: 388(6742)

Published: April 4, 2025

Exogenous messenger RNAs (mRNAs) require cellular machinery for delivery and translation but also encounter inhibitory factors. To investigate their regulation, we performed genome-wide CRISPR screens with in vitro–transcribed mRNAs lipid nanoparticles (LNPs). Heparan sulfate proteoglycans (HSPGs) vacuolar adenosine triphosphatase (V-ATPase) were identified as mediators of LNP uptake endosomal escape, respectively. TRIM25—an RNA binding E3 ubiquitin ligase—emerged a key suppressor inducing turnover both linear circular mRNAs. The endoribonucleases N4BP1 KHNYN, along the antiviral protein ZAP, act redundantly TRIM25-dependent surveillance. TRIM25 specifically targets delivered by endosomes, its affinity increases at acidic pH, suggesting activation protons released from ruptured endosomes. N 1 -methylpseudouridine modification reduces TRIM25’s binding, helping evade suppressive effect. This study comprehensively maps pathways regulating LNP-mRNAs, offering insights into immunity therapeutics.

Language: Английский

Citations

0
Identification of niclosamide as a novel antiviral agent against porcine epidemic diarrhea virus infection by targeting viral internalization DOI Creative Commons
Yue Wang, Huimin Huang,

Dongliang Li

et al.

Virologica Sinica, Journal Year: 2023, Volume and Issue: 38(2), P. 296 - 308

Published: Jan. 23, 2023

Porcine epidemic diarrhea virus (PEDV), an enteropathogenic coronavirus, has catastrophic impacts on the global pig industry. However, there remain no effective drugs against PEDV infection. In this study, we utilized a recombinant expressing renilla luciferase (PEDV-Rluc) to screen potential anti-PEDV agents from FDA-approved drug library in Vero cells. Four compounds were identified that significantly decreased activity of PEDV-Rluc. Among them, niclosamide was further characterized because it exhibited most potent antiviral with highest selectivity index. It can efficiently inhibit viral RNA synthesis, protein expression and progeny production classical variant strains dose-dependent manner. Time addition assay showed when added simultaneously or after Furthermore, inhibited entry stage infection by affecting internalization rather than attachment addition, combination other small molecule inhibitors endosomal acidification enhanced effect vitro. Taken together, these findings suggested is novel agent might provide basis for development therapies related pathogenic coronavirus infections.

Language: Английский

Citations

10