Abstract
How severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infections engage cellular host pathways and innate immunity in infected cells remains largely elusive. We performed an integrative proteo-transcriptomics analysis in SARS-CoV-2 infected Huh7 cells to map the cellular response to the invading virus over time. We identified four pathways, ErbB, HIF-1, mTOR and TNF signaling, among others that were markedly modulated during the course of the SARS-CoV-2 infection in vitro. Western blot validation of the downstream effector molecules of these pathways revealed a dose-dependent activation of Akt, mTOR, S6K1 and 4E-BP1 at 24 hours post infection (hpi). However, we found a significant inhibition of HIF-1α through 24hpi and 48hpi of the infection, suggesting a crosstalk between the SARS-CoV-2 and the Akt/mTOR/HIF-1 signaling pathways. Inhibition of the mTOR signaling pathway using Akt inhibitor MK-2206 showed a significant reduction in virus production. Further investigations are required to better understand the molecular sequelae in order to guide potential therapy in the management of severe coronavirus disease 2019 (COVID-19) patients.
Keywords:
Akt/mTOR/HIF-1; MK-2206; SARS-CoV-2; proteomics; transcriptomics.
MeSH terms
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Betacoronavirus / pathogenicity*
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COVID-19
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Cell Line
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Chromatography, Liquid
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Coronavirus Infections / genetics
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Coronavirus Infections / metabolism
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Coronavirus Infections / virology*
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Gene Expression Profiling / methods*
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Gene Expression Regulation
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High-Throughput Nucleotide Sequencing
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Humans
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Hypoxia-Inducible Factor 1 / genetics
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Hypoxia-Inducible Factor 1 / metabolism
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Pandemics
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Pneumonia, Viral / genetics
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Pneumonia, Viral / metabolism
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Pneumonia, Viral / virology*
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Proteomics / methods*
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Proto-Oncogene Proteins c-akt / genetics
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Proto-Oncogene Proteins c-akt / metabolism
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SARS-CoV-2
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Signal Transduction*
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TOR Serine-Threonine Kinases / genetics
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TOR Serine-Threonine Kinases / metabolism
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Tandem Mass Spectrometry
Substances
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Hypoxia-Inducible Factor 1
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MTOR protein, human
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AKT1 protein, human
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Proto-Oncogene Proteins c-akt
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TOR Serine-Threonine Kinases
Grants and funding
The study is funded by the Swedish Research Council [grant number 2017-01330, UN]. A. M. is supported by the Swedish research Council [2018-05766 and 2017-03126]. J.M.P. is supported by the Canada 150 Research Chair program and CIHR rapid Response COVID-19 grant. K.S. acknowledges the funding from the Office of Research (Early Concept Grant, Bond Life Sciences Center), University of Missouri, Columbia, MO.