A genome-wide CRISPR screen identifies N-acetylglucosamine-1-phosphate transferase as a potential antiviral target for Ebola virus

Flint, Mike; Chatterjee, Payel; Lin, David L.; McMullan, Laura K.; Shrivastava-Ranjan, Punya; Bergeron, Éric; Lo, Michael K.; Welch, Stephen R.; Nichol, Stuart T.; Tai, Andrew W.; Spiropoulou, Christina F.

doi:10.1038/s41467-018-08135-4

Cited by 58 publications

(53 citation statements)

References 67 publications

(74 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Genome-wide knockout screens have been widely used to identify host factors for various viruses (Flint et al, 2019; but the only coronavirus for which genome-wide knockout screens have been performed to date is SARS-CoV-2. These screens were performed either in the African green monkey cell line Vero E6, or in human cells that were engineered to overexpress ACE2.…”

Section: Cov-2mentioning

confidence: 99%

Identification of TMEM106B as proviral host factor for SARS-CoV-2

Baggen

Persoons

Jansen

et al. 2020

Preprint

View full text Add to dashboard Cite

SUMMARYThe ongoing COVID-19 pandemic is responsible for worldwide economic damage and nearly one million deaths. Potent drugs for the treatment of severe SARS-CoV-2 infections are not yet available. To identify host factors that support coronavirus infection, we performed genome-wide functional genetic screens with SARS-CoV-2 and the common cold virus HCoV-229E in non-transgenic human cells. These screens identified PI3K type 3 as a potential drug target against multiple coronaviruses. We discovered that the lysosomal protein TMEM106B is an important host factor for SARS-CoV-2 infection. Furthermore, we show that TMEM106B is required for replication in multiple human cell lines derived from liver and lung and is expressed in relevant cell types in the human airways. Our results identify new coronavirus host factors that may potentially serve as drug targets against SARS-CoV-2 or to quickly combat future zoonotic coronavirus outbreaks.

show abstract

Section: Cov-2mentioning

confidence: 99%

Identification of TMEM106B as proviral host factor for SARS-CoV-2

Baggen

Persoons

Jansen

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…XKR8 is required for apoptosis-induced PS externalization in HAP1 cells. HAP1 cells have been used to screen and identify cellular genes critical for EBOV entry, as these nearly haploid cells are straightforward to knock out genes and are susceptible to EBOV infection (13,62). In order to investigate the specific contributions of TMEM16F and XKR8 scramblases to EBOV replication, ΔTMEM16F and ΔXKR8 knockout HAP1 cell lines were first tested for scramblase activity.…”

Section: Resultsmentioning

confidence: 99%

Ebola virus requires phosphatidylserine scrambling activity for efficient budding and optimal infectivity

Acciani

Lay-Mendoza

Havranek

et al. 2020

Preprint

View full text Add to dashboard Cite

word count: 250 AbstractEbola virus (EBOV) interacts with cells using two categories of cell surface receptors, C-type lectins and phosphatidylserine (PS) receptors. PS receptors typically bind to apoptotic cell membrane PS and orchestrate the uptake and clearance of apoptotic bodies. Many viruses coated with PS-containing lipid envelopes, acquired during budding from host cells, can also exploit these receptors for internalization. PS is restricted to the inner leaflet of the plasma membrane in homeostatic cells, an orientation that would be unfavorable for PS receptor-mediated uptake if conserved on the viral envelope. Therefore, it is theorized that viral infection induces host cell PS externalization to the outer leaflet during replication. Cells have several membrane scramblase enzymes that enrich outer leaflet PS when activated. Here, we investigate two scramblases, TMEM16F and XKR8, as possible mediators of cellular and viral envelope surface PS levels during recombinant VSV/EBOV-GP replication and EBOV virus-like particle (VLP) production. We found that rVSV/EBOV-GP and EBOV VLPs produced in XKR8 knockout cells contain decreased levels of PS in their outer leaflets. ΔXKR8-made rVSV/EBOV-GP is 70% less efficient at infecting cells through apoptotic mimicry compared to viruses made in parental cells. Our data suggest that virion surface PS acquisition requires XKR8 activity, whereas TMEM16F activity is not essential. Unexpectedly, we observed defective rVSV/G, rVSV/EBOV-GP, and EBOV VLP budding in ΔXKR8 cells, suggesting that phospholipid scrambling via XKR8 enhances both Ebola infectivity and budding efficiency. Overexpression of XKR8 dramatically increased budding activity, suggesting outer leaflet PS is required for both particle production and increased infectivity. ImportanceThe Democratic Republic of the Congo experienced its deadliest Ebola outbreak from 2018 to 2020, with 3,444 confirmed cases and 2,264 deaths (as of March 12, 2020). Owing to the extensive damage that these outbreaks have caused in Africa, as well as its future epidemic potential, Ebola virus (EBOV) ranks among the top eight priority pathogens outlined by the WHO in 2018. A comprehensive understanding of Ebola entry pathways into target cells is critical for antiviral development and outbreak control. Thus far, host-cell scramblases TMEM16F and XKR8 have each been named as the sole mediator of Ebola envelope surface phosphatidylserine (PS). We assessed the contributions of these proteins using CRISPR knockout cells and two EBOV models: rVSV/EBOV-GP and EBOV VLPs. We observed that XKR8 is required for optimal EBOV envelope PS levels, PS receptor engagement, and particle budding across all viral models, whereas TMEM16F did not play a major role.

show abstract

“…Genes required for uptake of Ebola, influenza A, and Zika viruses were obtained from published CRISPR screens [31][32][33] . Since the Ebola data were also analysed with MAGeCK, we defined genes required for viral uptake using an unadjusted p-value threshold of 0.01; this information was not available for the influenza A and Zika studies, so we defined genes required for viral uptake as having an adjusted p-value < 0.05.…”

Section: Discussionmentioning

confidence: 99%

“…The host factors or genes that regulate the entry and life cycle of several viruses have been comprehensively studied using RNAi and CRISPR screens. These include the neurotropic Zika virus 31 and Influenza A 32 , which enter cells via receptor-mediated endocytosis, and Ebola 33 , which uses micropinocytosis.…”

Section: Neuronal Uptake Of Extracellular Tau Shares Functional Similmentioning

confidence: 99%

Whole genome CRISPR screens identify a LRRK2-regulated pathway for extracellular tau uptake by human neurons

Evans¹,

Strano²,

Campbell³

et al. 2020

Preprint

View full text Add to dashboard Cite

Pathological protein aggregation in Alzheimer's disease and other dementias is proposed to spread through the nervous system by a process of intercellular transfer of pathogenic forms of tau protein. Defining the cellular mechanisms of tau entry to human neurons is essential for understanding dementia pathogenesis and the rational design of disease-modifying therapeutics. Using whole genome CRISPR knockout screens in human iPSC-derived excitatory neurons, the primary cell type affected in these diseases, we identified genes and pathways required specifically for uptake of monomeric and aggregated tau. Monomeric and aggregated tau are both taken up by human neurons by receptor-mediated endocytosis, with the low-density lipoprotein LRP1 a significant surface receptor for both forms of tau. Perturbations of the endolysosome and autophagy systems at many levels, and specifically endosome sorting and receptor recycling, greatly reduced tau uptake. Of particular therapeutic interest is that loss of function of the endocytosis and autophagy regulator LRRK2, as well as acute inhibition of its kinase activity, reduced neuronal uptake of monomeric and aggregated tau. Kinase-activating mutations in LRRK2 are a cause of Parkinson's disease accompanied by neuronal tau aggregation, suggesting that LRRK2 mediates tau spreading in vivo and that LRRK2 inhibition has the potential to inhibit interneuronal spread of tau pathology, slowing disease progression. Overall, pathways for tau entry share significant similarity with those required for virus entry by receptor-mediated endocytosis, suggesting that tau spreading is a quasi-infectious process.

show abstract

A genome-wide CRISPR screen identifies N-acetylglucosamine-1-phosphate transferase as a potential antiviral target for Ebola virus

Cited by 58 publications

References 67 publications

Identification of TMEM106B as proviral host factor for SARS-CoV-2

Identification of TMEM106B as proviral host factor for SARS-CoV-2

Ebola virus requires phosphatidylserine scrambling activity for efficient budding and optimal infectivity

Whole genome CRISPR screens identify a LRRK2-regulated pathway for extracellular tau uptake by human neurons

Contact Info

Product

Resources

About