The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism

Martin McLaughlin; Malin Pedersen; Victoria Roulstone; Katharina F. Bergerhoff; Henry G. Smith; Harriet Whittock; Joan N. Kyula; Magnus T. Dillon; Hardev S. Pandha; Richard Vile; Alan A. Melcher; Kevin J. Harrington

doi:10.1016/j.omto.2020.01.001

The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism

Martin McLaughlin, Malin Pedersen, Victoria Roulstone, Katharina F. Bergerhoff, Henry G. Smith, Harriet Whittock, Joan N. Kyula, Magnus T. Dillon, Hardev S. Pandha, Richard Vile, Alan A. Melcher, Kevin J. Harrington

Molecular Medicine

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

Reovirus type 3 Dearing (reovirus) is a tumor-selective oncolytic virus currently under evaluation in clinical trials. Here, we report that the therapeutic efficacy of reovirus in head and neck squamous cell cancer can be enhanced by targeting the unfolded protein response (UPR) kinase, protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). PERK inhibition by GSK2606414 increased reovirus efficacy in both 2D and 3D models in vitro, while perturbing the normal host cell response to reovirus-induced endoplasmic reticulum (ER) stress. UPR reporter constructs were used for live-cell 3D spheroid imaging. Profiling of eIF2a-ATF4, IRE1a-XBP1, and ATF6 pathway activity revealed a context-dependent increase in eIF2a-ATF4 signaling due to GSK2606414. GSK2606414 blocked eIF2a-ATF4 signaling because of the canonical ER stress agent thapsigargin. In the context of reovirus infection, GSK2606414 induced eIF2a-ATF4 signaling. Knockdown of eIF2a kinases PERK, GCN2, and PKR revealed eIF2a-ATF4 reporter activity was dependent on either PERK or GCN2. Knockdown of ATF4 abrogated the GSK2606414-induced increase in reovirus protein levels, confirming eIF2a-ATF signaling as key to the observed phenotype. Our work identifies a novel approach to enhance the efficacy and replication of reovirus in a therapeutic setting.

Original language	English (US)
Pages (from-to)	238-249
Number of pages	12
Journal	Molecular Therapy Oncolytics
Volume	16
DOIs	https://doi.org/10.1016/j.omto.2020.01.001
State	Published - Mar 27 2020

Keywords

ATF4
ER stress
PERK
Reolysin
integrated stress response
oncolytic virus
pelareorep
reovirus type 3 Dearing
unfolded protein response

ASJC Scopus subject areas

Molecular Medicine
Oncology
Cancer Research
Pharmacology (medical)

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10.1016/j.omto.2020.01.001

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McLaughlin, M., Pedersen, M., Roulstone, V., Bergerhoff, K. F., Smith, H. G., Whittock, H., Kyula, J. N., Dillon, M. T., Pandha, H. S., Vile, R., Melcher, A. A., & Harrington, K. J. (2020). The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism. Molecular Therapy Oncolytics, 16, 238-249. https://doi.org/10.1016/j.omto.2020.01.001

McLaughlin, M, Pedersen, M, Roulstone, V, Bergerhoff, KF, Smith, HG, Whittock, H, Kyula, JN, Dillon, MT, Pandha, HS, Vile, R, Melcher, AA & Harrington, KJ 2020, 'The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism', Molecular Therapy Oncolytics, vol. 16, pp. 238-249. https://doi.org/10.1016/j.omto.2020.01.001

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title = "The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism",

abstract = "Reovirus type 3 Dearing (reovirus) is a tumor-selective oncolytic virus currently under evaluation in clinical trials. Here, we report that the therapeutic efficacy of reovirus in head and neck squamous cell cancer can be enhanced by targeting the unfolded protein response (UPR) kinase, protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). PERK inhibition by GSK2606414 increased reovirus efficacy in both 2D and 3D models in vitro, while perturbing the normal host cell response to reovirus-induced endoplasmic reticulum (ER) stress. UPR reporter constructs were used for live-cell 3D spheroid imaging. Profiling of eIF2a-ATF4, IRE1a-XBP1, and ATF6 pathway activity revealed a context-dependent increase in eIF2a-ATF4 signaling due to GSK2606414. GSK2606414 blocked eIF2a-ATF4 signaling because of the canonical ER stress agent thapsigargin. In the context of reovirus infection, GSK2606414 induced eIF2a-ATF4 signaling. Knockdown of eIF2a kinases PERK, GCN2, and PKR revealed eIF2a-ATF4 reporter activity was dependent on either PERK or GCN2. Knockdown of ATF4 abrogated the GSK2606414-induced increase in reovirus protein levels, confirming eIF2a-ATF signaling as key to the observed phenotype. Our work identifies a novel approach to enhance the efficacy and replication of reovirus in a therapeutic setting.",

keywords = "ATF4, ER stress, PERK, Reolysin, integrated stress response, oncolytic virus, pelareorep, reovirus type 3 Dearing, unfolded protein response",

author = "Martin McLaughlin and Malin Pedersen and Victoria Roulstone and Bergerhoff, {Katharina F.} and Smith, {Henry G.} and Harriet Whittock and Kyula, {Joan N.} and Dillon, {Magnus T.} and Pandha, {Hardev S.} and Richard Vile and Melcher, {Alan A.} and Harrington, {Kevin J.}",

note = "Publisher Copyright: {\textcopyright} 2020 The Authors",

year = "2020",

month = mar,

day = "27",

doi = "10.1016/j.omto.2020.01.001",

language = "English (US)",

volume = "16",

pages = "238--249",

journal = "Molecular Therapy Oncolytics",

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AU - McLaughlin, Martin

AU - Pedersen, Malin

AU - Roulstone, Victoria

AU - Bergerhoff, Katharina F.

AU - Smith, Henry G.

AU - Whittock, Harriet

AU - Kyula, Joan N.

AU - Dillon, Magnus T.

AU - Pandha, Hardev S.

AU - Vile, Richard

AU - Melcher, Alan A.

AU - Harrington, Kevin J.

PY - 2020/3/27

Y1 - 2020/3/27

N2 - Reovirus type 3 Dearing (reovirus) is a tumor-selective oncolytic virus currently under evaluation in clinical trials. Here, we report that the therapeutic efficacy of reovirus in head and neck squamous cell cancer can be enhanced by targeting the unfolded protein response (UPR) kinase, protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). PERK inhibition by GSK2606414 increased reovirus efficacy in both 2D and 3D models in vitro, while perturbing the normal host cell response to reovirus-induced endoplasmic reticulum (ER) stress. UPR reporter constructs were used for live-cell 3D spheroid imaging. Profiling of eIF2a-ATF4, IRE1a-XBP1, and ATF6 pathway activity revealed a context-dependent increase in eIF2a-ATF4 signaling due to GSK2606414. GSK2606414 blocked eIF2a-ATF4 signaling because of the canonical ER stress agent thapsigargin. In the context of reovirus infection, GSK2606414 induced eIF2a-ATF4 signaling. Knockdown of eIF2a kinases PERK, GCN2, and PKR revealed eIF2a-ATF4 reporter activity was dependent on either PERK or GCN2. Knockdown of ATF4 abrogated the GSK2606414-induced increase in reovirus protein levels, confirming eIF2a-ATF signaling as key to the observed phenotype. Our work identifies a novel approach to enhance the efficacy and replication of reovirus in a therapeutic setting.

AB - Reovirus type 3 Dearing (reovirus) is a tumor-selective oncolytic virus currently under evaluation in clinical trials. Here, we report that the therapeutic efficacy of reovirus in head and neck squamous cell cancer can be enhanced by targeting the unfolded protein response (UPR) kinase, protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). PERK inhibition by GSK2606414 increased reovirus efficacy in both 2D and 3D models in vitro, while perturbing the normal host cell response to reovirus-induced endoplasmic reticulum (ER) stress. UPR reporter constructs were used for live-cell 3D spheroid imaging. Profiling of eIF2a-ATF4, IRE1a-XBP1, and ATF6 pathway activity revealed a context-dependent increase in eIF2a-ATF4 signaling due to GSK2606414. GSK2606414 blocked eIF2a-ATF4 signaling because of the canonical ER stress agent thapsigargin. In the context of reovirus infection, GSK2606414 induced eIF2a-ATF4 signaling. Knockdown of eIF2a kinases PERK, GCN2, and PKR revealed eIF2a-ATF4 reporter activity was dependent on either PERK or GCN2. Knockdown of ATF4 abrogated the GSK2606414-induced increase in reovirus protein levels, confirming eIF2a-ATF signaling as key to the observed phenotype. Our work identifies a novel approach to enhance the efficacy and replication of reovirus in a therapeutic setting.

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KW - ER stress

KW - PERK

KW - Reolysin

KW - integrated stress response

KW - oncolytic virus

KW - pelareorep

KW - reovirus type 3 Dearing

KW - unfolded protein response

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U2 - 10.1016/j.omto.2020.01.001

DO - 10.1016/j.omto.2020.01.001

M3 - Article

AN - SCOPUS:85079871678

SN - 2372-7705

VL - 16

SP - 238

EP - 249

JO - Molecular Therapy Oncolytics

JF - Molecular Therapy Oncolytics

ER -

The PERK Inhibitor GSK2606414 Enhances Reovirus Infection in Head and Neck Squamous Cell Carcinoma via an ATF4-Dependent Mechanism

Abstract

Keywords

ASJC Scopus subject areas

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