Closely related reovirus lab strains induce opposite expression of rig-i/ifn-dependent versus -independent host genes, via mechanisms of slow replication versus polymorphisms in dsrna binding σ3 respectively

Adil Mohamed; Prathyusha Konda; Heather E. Eaton; Shashi Gujar; James R. Smiley; Maya Shmulevitz

doi:10.1371/journal.ppat.1008803

Closely related reovirus lab strains induce opposite expression of rig-i/ifn-dependent versus -independent host genes, via mechanisms of slow replication versus polymorphisms in dsrna binding σ3 respectively

Adil Mohamed, Prathyusha Konda, Heather E. Eaton, Shashi Gujar, James R. Smiley, Maya Shmulevitz

Medicine

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

20 Citas (Scopus)

Resumen

The Dearing isolate of Mammalian orthoreovirus (T3D) is a prominent model of virus-host relationships and a candidate oncolytic virotherapy. Closely related laboratory strains of T3D, originating from the same ancestral T3D isolate, were recently found to exhibit significantly different oncolytic properties. Specifically, the T3D^PL strain had faster replication kinetics in a panel of cancer cells and improved tumor regression in an in vivo melanoma model, relative to T3D^TD. In this study, we discover that T3D^PL and T3D^TD also differentially activate host signalling pathways and downstream gene transcription. At equivalent infectious dose, T3D^TD induces higher IRF3 phosphorylation and expression of type I IFNs and IFN-stimulated genes (ISGs) than T3D^PL. Using mono-reassortants with intermediate replication kinetics and pharmacological inhibitors of reovirus replication, IFN responses were found to inversely correlate with kinetics of virus replication. In other words, slow-replicating T3D strains induce more IFN signalling than fast-replicating T3D strains. Paradoxically, during co-infections by T3D^PL and T3D^TD, there was still high IRF3 phosphorylation indicating a phenodominant effect by the slow-replicating T3D^TD. Using silencing and knock-out of RIG-I to impede IFN, we found that IFN induction does not affect the first round of reovirus replication but does prevent cell-cell spread in a paracrine fashion. Accordingly, during co-infections, T3D^PL continues to replicate robustly despite activation of IFN by T3D^TD. Using gene expression analysis, we discovered that reovirus can also induce a subset of genes in a RIG-I and IFN-independent manner; these genes were induced more by T3D^PL than T3D^TD. Polymorphisms in reovirus σ3 viral protein were found to control activation of RIG-I/ IFN-independent genes. Altogether, the study reveals that single amino acid polymorphisms in reovirus genomes can have large impact on host gene expression, by both changing replication kinetics and by modifying viral protein activity, such that two closely related T3D strains can induce opposite cytokine landscapes.

Idioma original	English
Número de artículo	e1008803
Publicación	PLoS Pathogens
Volumen	16
N.º	9
DOI	https://doi.org/10.1371/journal.ppat.1008803
Estado	Published - sep. 2020

Nota bibliográfica

Funding Information:
This work was funded by project grants to M.S. from the Canadian Institutes of Health Research (CIHR), Li Ka Shing Institute of Virology (LKSIoV), and Cancer Research Society (CRS), a salary award to M.S. from the Canada Research Chairs (CRC) and infrastructure support from Canada Foundation for Innovation (CFI). A.M. received scholarships from an Alberta Cancer Foundation Graduate Studentship (ACF), University of Alberta Faculty of Medicine and Dentistry/Alberta Health Services Graduate Recruitment Studentship, and University of Alberta Doctoral Recruitment Award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Kevin Coombs at the University of Manitoba and Terrence Dermody at University of Pittsburgh for generously sharing their laboratory reovirus T3D virus lysates, Aja Reiger at the Faculty of Medicine and Dentistry flow cytometry facility, Rob Maranchuk at the Li Ka Shing Institute of Virology RNAi screening facility and Stephen Ogg at the Faculty of Medicine & Dentistry cell imaging centre, for valuable technical advice, training and support. We appreciate all the helpful discussions and suggestions by members of the Maya Shmulevitz, David Evans, Mary Hitt, Ronald Moore and Patrick Lee laboratories.

Publisher Copyright:
© 2020 Mohamed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

ASJC Scopus Subject Areas

Parasitology
Microbiology
Immunology
Molecular Biology
Genetics
Virology

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Mohamed, A., Konda, P., Eaton, H. E., Gujar, S., Smiley, J. R., & Shmulevitz, M. (2020). Closely related reovirus lab strains induce opposite expression of rig-i/ifn-dependent versus -independent host genes, via mechanisms of slow replication versus polymorphisms in dsrna binding σ3 respectively. PLoS Pathogens, 16(9), Artículo e1008803. https://doi.org/10.1371/journal.ppat.1008803

Producción científica: Contribución a una revista › Artículo › revisión exhaustiva

Mohamed, A, Konda, P, Eaton, HE, Gujar, S, Smiley, JR & Shmulevitz, M 2020, 'Closely related reovirus lab strains induce opposite expression of rig-i/ifn-dependent versus -independent host genes, via mechanisms of slow replication versus polymorphisms in dsrna binding σ3 respectively', PLoS Pathogens, vol. 16, n.º 9, e1008803. https://doi.org/10.1371/journal.ppat.1008803

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abstract = "The Dearing isolate of Mammalian orthoreovirus (T3D) is a prominent model of virus-host relationships and a candidate oncolytic virotherapy. Closely related laboratory strains of T3D, originating from the same ancestral T3D isolate, were recently found to exhibit significantly different oncolytic properties. Specifically, the T3DPL strain had faster replication kinetics in a panel of cancer cells and improved tumor regression in an in vivo melanoma model, relative to T3DTD. In this study, we discover that T3DPL and T3DTD also differentially activate host signalling pathways and downstream gene transcription. At equivalent infectious dose, T3DTD induces higher IRF3 phosphorylation and expression of type I IFNs and IFN-stimulated genes (ISGs) than T3DPL. Using mono-reassortants with intermediate replication kinetics and pharmacological inhibitors of reovirus replication, IFN responses were found to inversely correlate with kinetics of virus replication. In other words, slow-replicating T3D strains induce more IFN signalling than fast-replicating T3D strains. Paradoxically, during co-infections by T3DPL and T3DTD, there was still high IRF3 phosphorylation indicating a phenodominant effect by the slow-replicating T3DTD. Using silencing and knock-out of RIG-I to impede IFN, we found that IFN induction does not affect the first round of reovirus replication but does prevent cell-cell spread in a paracrine fashion. Accordingly, during co-infections, T3DPL continues to replicate robustly despite activation of IFN by T3DTD. Using gene expression analysis, we discovered that reovirus can also induce a subset of genes in a RIG-I and IFN-independent manner; these genes were induced more by T3DPL than T3DTD. Polymorphisms in reovirus σ3 viral protein were found to control activation of RIG-I/ IFN-independent genes. Altogether, the study reveals that single amino acid polymorphisms in reovirus genomes can have large impact on host gene expression, by both changing replication kinetics and by modifying viral protein activity, such that two closely related T3D strains can induce opposite cytokine landscapes.",

author = "Adil Mohamed and Prathyusha Konda and Eaton, {Heather E.} and Shashi Gujar and Smiley, {James R.} and Maya Shmulevitz",

note = "Funding Information: This work was funded by project grants to M.S. from the Canadian Institutes of Health Research (CIHR), Li Ka Shing Institute of Virology (LKSIoV), and Cancer Research Society (CRS), a salary award to M.S. from the Canada Research Chairs (CRC) and infrastructure support from Canada Foundation for Innovation (CFI). A.M. received scholarships from an Alberta Cancer Foundation Graduate Studentship (ACF), University of Alberta Faculty of Medicine and Dentistry/Alberta Health Services Graduate Recruitment Studentship, and University of Alberta Doctoral Recruitment Award. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We would like to thank Kevin Coombs at the University of Manitoba and Terrence Dermody at University of Pittsburgh for generously sharing their laboratory reovirus T3D virus lysates, Aja Reiger at the Faculty of Medicine and Dentistry flow cytometry facility, Rob Maranchuk at the Li Ka Shing Institute of Virology RNAi screening facility and Stephen Ogg at the Faculty of Medicine & Dentistry cell imaging centre, for valuable technical advice, training and support. We appreciate all the helpful discussions and suggestions by members of the Maya Shmulevitz, David Evans, Mary Hitt, Ronald Moore and Patrick Lee laboratories. Publisher Copyright: {\textcopyright} 2020 Mohamed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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