key: cord-0755286-3ubithu4 authors: Marchetti, Carlo; Mould, Kara; Tengesdal, Isak W.; Janssen, William J; Dinarello, Charles A. title: Targeting of the NLRP3 Inflammasome for early COVID-19 date: 2021-02-24 journal: bioRxiv DOI: 10.1101/2021.02.24.432734 sha: 4ee9a74da55c7d95bf4cda8f4acc29a9372fc2c5 doc_id: 755286 cord_uid: 3ubithu4 Following entry and replication of Severe Acute Respiratory Syndrome-coronavirus-2 (SARS-CoV-2) into ACE2 expressing cells, the infected cells undergo lysis releasing more virus but also cell contents. In the lung, constitutive cytokines such as IL-1α are released together with other cell contents. A cascade of inflammatory cytokines ensues, including chemokines and IL-1β, triggering both local as well as systemic inflammation. This cascade of inflammatory cytokines in patients with COVID-19 is termed “Cytokine Release Syndrome” (CRS), and is associated with poor outcomes and death. Many studies reveal that blocking IL-1 activities in COVID-19 patients reduces disease severity and deaths. Here we report highly significant circulating levels of IL-1β, IL-1 Receptor antagonist, IL-6, TNFα, IL-10 and soluble urokinase plasminogen activator receptor in COVID-19 patients with mild or no symptoms. We also report that in circulating myeloid cells from the same patients, there is increased expression of the NOD-, LRR- and pyrin domain-containing 3 (NLRP3) early in the infection. We observed increased NLRP3 gene expression in myeloid cells correlated with IL-1β gene expression and also with elevated circulating IL-1β levels. We conclude that early in SARS-CoV-2 infection, NLRP3 activation takes place and initiates the CRS. Thus, NLRP3 is a target to reduce the organ damage of inflammatory cytokines of the CRS. Emerging from these reports is the concept that targeting of IL-1 result in improved outcomes, 46 including deaths. For example, high doses of anakinra reduces deaths as well as number of days 47 in the hospital (Cauchois et al. 2020, Cavalli et al. 2020, Huet et al. 2020 ). Anakinra has also been 48 administered in less severe hospitalized patients and resulted in similar reduction in disease 49 (Kyriazopoulou et al. 2020 ). Since anakinra blocks the IL-1 Receptor, the efficacy of anakinra may 50 be due to reducing IL-1a or IL-1b. Other studies report that specifically targeting IL-1b with the 51 neutralizing monoclonal antibody canakinumab also reduces outcomes. 52 The intracellular processing of IL-1β into its biologically active form is largely governed 53 by cytosolic macromolecular complexes termed inflammasomes (Franchi et al. 2009 As shown in Figure 1a influenza or sepsis. The use of anakinra or canakinumab in COVID-19 disease is associated with 118 recovery and survival. However, it is possible to reduce the detrimental properties of IL-1b and its 119 downstream cytokines by first preventing the processing and release of active IL-1b. The present 120 data provide a rationale to treat patients infected with SARS-CoV-2 early in the course of the 121 disease using a specific NLRP3 inhibitor in order to arrest the progression of IL-1b-mediated CRS. 122 Such a treatment offers an opportunity to reduce hospitalization and the need for supplemental 123 oxygen, particularly in subjects with high risk co-morbidities. in patients with COVID-19, acute respiratory distress syndrome, and 198 hyperinflammation: a retrospective cohort study Severe Acute Respiratory 200 Syndrome Coronavirus Viroporin 3a Activates the NLRP3 Inflammasome An 207 inflammatory cytokine signature predicts COVID-19 severity and survival The inflammasome: a caspase-210 1-activation platform that regulates immune responses and disease pathogenesis Inhibitory effects of colchicine on inflammasomes Anakinra for severe forms of COVID-19: 217 a cohort study Therapy for Early COVID-19: A Critical Need Dapansutrile, an oral selective NLRP3 inflammasome inhibitor, for 223 treatment of gout flares: an open-label, dose-adaptive, proof-of-concept, phase 2a trial Giamarellos-Bourboulis (2020) Anakinra To 232 Prevent Respiratory Failure In COVID-19 Omer & A. Iwasaki (2020) 239 Longitudinal analyses reveal immunological misfiring in severe COVID-19 The NLRP3 Inflammasome as a Pharmacological Target OLT1177, a beta-sulfonyl nitrile compound, safe in humans, inhibits the NLRP3 248 inflammasome and reverses the metabolic cost of inflammation /IL-18BP in patients with secondary hemophagocytic syndrome Colchicine in Patients with Chronic Coronary 259 Disease Inflammasomes are activated in response to SARS-CoV-2 infection and 269 are associated with COVID-19 severity in patients Frame-8b triggers intracellular stress pathways and activates NLRP3 inflammasomes Severe acute respiratory syndrome 275 coronavirus ORF3a protein activates the NLRP3 inflammasome by promoting TRAF3-276 dependent ubiquitination of ASC Efficacy of Colchicine in Non-Hospitalized Patients with 284 COVID-19. medRxiv Host and 290 Environmental Factors Influencing Individual Human Cytokine Responses Inflammasome formation in the 294 lungs of patients with fatal COVID-19 Repeat 299 Dose Safety and Pharmacodynamics Study of the Oral NLRP3 Inhibitor Dapansutrile in 300 Subjects With NYHA II-III Systolic Heart Failure SARS-CoV-2 viroporin triggers the NLRP3 inflammatory pathway. 303 bioRxiv