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bioRxiv · 10.1101/2020.10.27.357731

SARS-CoV-2 viroporin triggers the NLRP3 inflammatory pathway

Xu, H.·Chitre, S. A.·Akinyemi, I. A.·Loeb, J. C.·Lednicky, J. A.·McIntosh, M. T.·Bhaduri-McIntosh, S.
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Abstract

Cytokine storm resulting from a heightened inflammatory response is a prominent feature of severe COVID-19 disease. This inflammatory response results from assembly/activation of a cell-intrinsic defense platform known as the inflammasome. We report that the SARS-CoV-2 viroporin encoded by ORF3a activates the NLRP3 inflammasome, the most promiscuous of known inflammasomes. ORF3a triggers IL-1{beta} expression via NF{kappa}B, thus priming the inflammasome while also activating it via ASC-dependent and -independent modes. ORF3a-mediated inflammasome activation requires efflux of potassium ions and oligomerization between NEK7 and NLRP3. With the selective NLRP3 inhibitor MCC950 able to block ORF3a-mediated inflammasome activation and key ORF3a residues needed for virus release and inflammasome activation conserved in SARS-CoV-2 isolates across continents, ORF3a and NLRP3 present prime targets for intervention. SummaryDevelopment of anti-SARS-CoV-2 therapies is aimed predominantly at blocking infection or halting virus replication. Yet, the inflammatory response is a significant contributor towards disease, especially in those severely affected. In a pared-down system, we investigate the influence of ORF3a, an essential SARS-CoV-2 protein, on the inflammatory machinery and find that it activates NLRP3, the most prominent inflammasome by causing potassium loss across the cell membrane. We also define key amino acid residues on ORF3a needed to activate the inflammatory response, and likely to facilitate virus release, and find that they are conserved in virus isolates across continents. These findings reveal ORF3a and NLRP3 to be attractive targets for therapy.

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Xu, H., Chitre, S. A., Akinyemi, I. A., Loeb, J. C., Lednicky, J. A., McIntosh, M. T., Bhaduri-McIntosh, S.. 2020-10-27. SARS-CoV-2 viroporin triggers the NLRP3 inflammatory pathway. https://doi.org/10.1101/2020.10.27.357731

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