Role of Complement Activation in Severe COVID-19

Project summary/Abstract: In response to Notice of Special Interest (NOSI): Complement in Basic Immunology (CIBI), we propose to examine the causative effect of complements in the pathogenesis of severe COVID-19. The complement system is activated via one of three pathways-classical, alternative, and mannose-binding lectin (MBL)-which converge at C3 cleavage, leading to the formation of C3 and C5 convertases and concluding with assembly of the membrane attack complex (MAC). MAC is a cytolytic macromolecular pore that can insert into host cell membranes under pathological conditions. Extensive evidence obtained from others and us indicates that the complement (C) system, in particular MAC, may participate in mediating endothelial damage, activating the coagulation pathway and platelets, and causing multiple organ damage leading to severe COVID-19. However, the causative roles of C and MAC in severe COVID-19 have not been experimentally investigated. The proposed studies will utilize our newly developed state-of-the-art tools to block or modify the C activation products to investigate the role of C in endothelial cell damage, platelet activation, and thrombosis formation seen in severe COVID-19, including therapeutic paradigms. To address our needs, we have established and characterized an animal model of severe COVID- 19 using SARS-CoV-2-infected K18-hACE2 mice. The mice develop acute respiratory distress syndrome (ARDS), progressive weight loss, and mortality at 7 days that is associated with severe interstitial inflammation, perivascular inflammation, platelet activation, and thrombosis in the lungs. We also observe (i) endothelial cell (EC) dysfunction of the alveolar septa; (ii) increased vascular permeability associated with the extensive activation of immune cells (e.g., lung macrophage cells); and (iii) increased C3 and MAC deposition in pulmonary vasculature. In addition, single-cell RNAseq shows C activation and coagulation in the lungs of this severe COVID-19 model. These results have prompted us to hypothesize that the C in general, and MAC in particular, significantly contribute to the EC damage, platelet activation, and thrombosis formation seen in severe cases of COVID-19. Aim 1 will investigate whether the inhibition of C activation and MAC formation will reduce EC damage and platelet and coagulation pathway activation in SARS-CoV-2-infected K18 mice. Aim 2 will test the hypothesis that the restriction of MAC formation will protect against EC damage and activation of the myeloid cells, leading to reduced platelet and coagulation activation in SARS-CoV-2-infected K18 mice. Aim 3 will investigate the role of C in the pathogenesis of SAR-CoV-2 infection in a clinically relevant paradigm and evaluate site-targeted C inhibition as a treatment for COVID-19. help us better understand the role of C activation and the MAC in pathogenesis of severe COVID-19, open a new avenue to prevent and treat COVID-19, and foster the development of new therapeutic strategies involving modulation of the C system.