A genome-wide siRNA screen identifies a druggable host pathway essential for the Ebola virus life cycle

The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging...

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Published inGenome medicine Vol. 10; no. 1; pp. 58 - 14
Main Authors Martin, Scott, Chiramel, Abhilash I., Schmidt, Marie Luisa, Chen, Yu-Chi, Whitt, Nadia, Watt, Ari, Dunham, Eric C., Shifflett, Kyle, Traeger, Shelby, Leske, Anne, Buehler, Eugen, Martellaro, Cynthia, Brandt, Janine, Wendt, Lisa, Müller, Andreas, Peitsch, Stephanie, Best, Sonja M., Stech, Jürgen, Finke, Stefan, Römer-Oberdörfer, Angela, Groseth, Allison, Feldmann, Heinz, Hoenen, Thomas
Format Journal Article
LanguageEnglish
Published England BioMed Central Ltd 07.08.2018
BioMed Central
BMC
Subjects
Animals
Antiviral activity
Antiviral Agents - pharmacology
Biosynthesis
Carbamoyl phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase
Cell cycle
Cell Line, Tumor
Cercopithecus aethiops
Cloning, Molecular
De novo pyrimidine synthesis
Dihydroorotate dehydrogenase
Disease resistance
Ebola virus
Ebolavirus
Ebolavirus - drug effects
Ebolavirus - pathogenicity
Ebolavirus - physiology
Gene Knockdown Techniques
Genes
Genetic aspects
Genetic screening
Genome, Human
Genomes
HEK293 Cells
Host factor
Host-Pathogen Interactions - genetics
Humans
Kinases
Life cycles
Outbreaks
Proteins
Replication
RNA viruses
siRNA
Small interfering RNA
Teriflunomide
Transcription
Vero Cells
Virus Replication
Viruses
West Africa
Sudan
Ebola virus
Non-segmented negative-sense RNA viruses
De novo pyrimidine synthesis
NNSV
Carbamoyl phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase
Teriflunomide
siRNA screen
Host factor
Dihydroorotate dehydrogenase
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Abstract The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4). In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory. Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses. This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
AbstractList Background The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4). Methods In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory. Results Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses. Conclusions This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses. Keywords: Ebola virus, Carbamoyl phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase, De novo pyrimidine synthesis, Dihydroorotate dehydrogenase, Host factor, Teriflunomide, siRNA screen, Non-segmented negative-sense RNA viruses, NNSV
The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4). In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory. Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses. This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4). In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory. Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses. This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
BackgroundThe 2014–2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4).MethodsIn order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory.ResultsAmong others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses.ConclusionsThis study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4).BACKGROUNDThe 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4).In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory.METHODSIn order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory.Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses.RESULTSAmong others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses.This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.CONCLUSIONSThis study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
Abstract Background The 2014–2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host factors/pathways essential for the virus are advantageous because they can potentially target a wide range of viruses, including newly emerging ones and because the development of resistance is less likely than when targeting the virus directly. However, systematic approaches for screening host factors important for EBOV have been hampered by the necessity to work with this virus at biosafety level 4 (BSL4). Methods In order to identify host factors involved in the EBOV life cycle, we performed a genome-wide siRNA screen comprising 64,755 individual siRNAs against 21,566 human genes to assess their activity in EBOV genome replication and transcription. As a screening platform, we used reverse genetics-based life cycle modelling systems that recapitulate these processes without the need for a BSL4 laboratory. Results Among others, we identified the de novo pyrimidine synthesis pathway as an essential host pathway for EBOV genome replication and transcription, and confirmed this using infectious EBOV under BSL4 conditions. An FDA-approved drug targeting this pathway showed antiviral activity against infectious EBOV, as well as other non-segmented negative-sense RNA viruses. Conclusions This study provides a minable data set for every human gene regarding its role in EBOV genome replication and transcription, shows that an FDA-approved drug targeting one of the identified pathways is highly efficacious in vitro, and demonstrates the power of life cycle modelling systems for conducting genome-wide host factor screens for BSL4 viruses.
ArticleNumber 58
Audience Academic
Author Traeger, Shelby
Feldmann, Heinz
Groseth, Allison
Martin, Scott
Dunham, Eric C.
Buehler, Eugen
Whitt, Nadia
Leske, Anne
Watt, Ari
Müller, Andreas
Schmidt, Marie Luisa
Brandt, Janine
Wendt, Lisa
Chiramel, Abhilash I.
Stech, Jürgen
Martellaro, Cynthia
Best, Sonja M.
Shifflett, Kyle
Römer-Oberdörfer, Angela
Chen, Yu-Chi
Peitsch, Stephanie
Hoenen, Thomas
Finke, Stefan
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/30081931$$D View this record in MEDLINE/PubMed
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Issue 1
Keywords Ebola virus
Non-segmented negative-sense RNA viruses
De novo pyrimidine synthesis
NNSV
Carbamoyl phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase
Teriflunomide
siRNA screen
Host factor
Dihydroorotate dehydrogenase
Language English
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Snippet The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches targeting host...
Background The 2014-2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches...
BackgroundThe 2014–2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus. Approaches...
Abstract Background The 2014–2016 Ebola virus (EBOV) outbreak in West Africa highlighted the need for improved therapeutic options against this virus....
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StartPage 58
SubjectTerms Animals
Antiviral activity
Antiviral Agents - pharmacology
Biosynthesis
Carbamoyl phosphate synthetase 2 aspartate transcarbamylase and dihydroorotase
Cell cycle
Cell Line, Tumor
Cercopithecus aethiops
Cloning, Molecular
De novo pyrimidine synthesis
Dihydroorotate dehydrogenase
Disease resistance
Ebola virus
Ebolavirus
Ebolavirus - drug effects
Ebolavirus - pathogenicity
Ebolavirus - physiology
Gene Knockdown Techniques
Genes
Genetic aspects
Genetic screening
Genome, Human
Genomes
HEK293 Cells
Host factor
Host-Pathogen Interactions - genetics
Humans
Kinases
Life cycles
Outbreaks
Proteins
Replication
RNA viruses
siRNA
Small interfering RNA
Teriflunomide
Transcription
Vero Cells
Virus Replication
Viruses
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Title A genome-wide siRNA screen identifies a druggable host pathway essential for the Ebola virus life cycle
URI https://www.ncbi.nlm.nih.gov/pubmed/30081931
https://www.proquest.com/docview/2089764562
https://www.proquest.com/docview/2084914034
https://pubmed.ncbi.nlm.nih.gov/PMC6090742
https://doaj.org/article/de07dea36f0548debf36fe428049f3e0
Volume 10
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