Document text
Principal Investigator: PETER D. SUN
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2022
Award: $171,318
Funding agency: National Institute of Allergy and Infectious Diseases
To characterize the SARS-CoV-2 spike protein interaction with lectin receptors, we have expressed recombinant SARS-CoV-2 spike prefusion trimer using 293F freestyle expression system. Given the nanomolar high affinity of the spike protein binding to human ACE2, the entry receptor for SARS-CoV-2, and the physiological importance of ACE2 in balancing blood pressure, we investigated if this interaction would affect the enzymatic activity of ACE2. Surprisingly, SARS-CoV-2 trimeric spike protein increased ACE2 proteolytic activity 3-10 fold when fluorogenic caspase-1 substrate and Bradykinin-analog peptides were used to characterize ACE2 activity. In addition, the enhancement was mediated by ACE2 binding of RBD domain of SARS-CoV-2 spike. These results highlighted the altered activity of ACE2 during SARS-CoV-2 infection and would shed new lights on the pathogenesis of COVID-19 and its complications for better treatments. A manuscript describing this finding is under revision for publication in Journal of Biological Chemistry.
COVID-induced lung fibrosis is a major confounding factor associated with the current pandemic related death. Understanding the mechanism of COVID-associated lung fibrosis and developing a therapeutic treatment are an urgent public health need to reduce the current pandemic-associated death. Through proteomic analyses of bronchoalveolar lavage fluid (BALF) from COVID patients, we found dramatically elevated levels of fibrinogen and prothrombin in the acute COVID lung fluid compared to the healthy donors. Using primary human bronchoalveolar epithelial cells as models, we discovered that SARS-CoV-2 infected primary lung cells but not other SARS-CoV-2 susceptible cells, including Vero and 293T cells, induced fibrin to clot in the absence of many plasma coagulation factors. Many fibrin fibers induced by SARS-CoV-2 infection originated from lung cells. These infected lung cell induced fibrin clotting occurred with all strains of SARS-CoV-2 pseudoviruses, including omicron, as well as the circulating replication competent SARS-CoV-2 strains tested. They are indistinguishable from those thrombin clots and can be inhibited by pharmacological direct thrombin inhibitors. Importantly, infected lung cells triggered fibrin clotting in 3 of 4 acute but not recovered COVID nor healthy BALF. We suggest the viral infection activated members of cell surface expressed transmembrane serine proteases to directly activate prothrombin for fibrin clotting, as evidenced from the shedding of one of the transmembrane proteases, ST14, in response to the infection, and both recombinant catalytic matriptase and human airway trypsin-like protease activated prothrombin for fibrin clot formation. Our findings revealed a lung cell-mediated fibrosis triggered by SARS-CoV-2 infection that is independent of plasma coagulation, and suggest the need to focus therapeutic treatments to respiratory airway rather than blood vessels to mitigate COVID-induced lung fibrosis.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><ACE2><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Acute><Acylneuraminyl hydrolase><Affect><Affinity><Apoptosis-Related Cysteine Protease Caspase 1><Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg><Binding><Binding Proteins><Biochemistry><Biological Chemistry><Blood Coagulation Factor><Blood Coagulation Factor I><Blood Coagulation Factor II><Blood Coagulation Factor One><Blood Factor One><Blood Plasma><Blood Pressure><Blood Vessels><Bradykinin><Bronchoalveolar Lavage Fluid><CASP-1><CASP1><CASP1 gene><COVID><COVID disease severity><COVID infected patient><COVID patient><COVID positive patient><COVID severity><COVID-19><COVID-19 S protein><COVID-19 disease severity><COVID-19 infected patient><COVID-19 infection><COVID-19 pathogenesis><COVID-19 patient><COVID-19 positive patient><COVID-19 severity><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 virus><COVID19><COVID19 S protein><COVID19 disease severity><COVID19 infection><COVID19 pathogenesis><COVID19 patient><COVID19 positive patient><COVID19 severity><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 virus><CV-19><CV19><Caspase-1><Caspase-1 Gene><Cell Body><Cell surface><Cells><Cessation of life><Clinical><Clotting><CoV S protein><CoV disease><CoV glycoprotein S><CoV spike glycoprotein><CoV spike protein><CoV-2><CoV2><Coagulation><Coagulation Factor I><Coagulation Factor II><Coagulation Factor One><Coagulation Factors><Coagulation Process><Coronaviridae><Coronavirus><Coronavirus glycoprotein S><Coronavirus spike protein><Death><Differentiation Reversal Factor><Drug Targeting><Epithelial Cells><Epithin><Equilibrium><Esteroproteases><Factor I><Factor II><Factor One><Fiber><Fibrin><Fibrinogen><Fibrosis><Glycans><HAI><HIV><HIV Envelope Glycoprotein gp120><HIV Envelope Protein gp120><HIV env Protein gp120><HTLV-III gp120><Human><Human Immunodeficiency Viruses><ICE Protease><IL-1 beta Convertase><IL-1 beta-Converting Enzyme><IL-1BC><IL-1b Converting Enzyme><IL1B-Convertase><IL1BC><IL1BCE><Infection><Influenza Virus><Interleukin 1-B Converting Enzyme><Interleukin 1-Beta Convertase><Interleukin-1 Beta Converting Enzyme><Interleukin-1 Converting Enzyme><Journals><LAV-HTLV-III><Lectin Receptors><Ligand Binding Protein><Ligand Binding Protein Gene><Liquid substance><Lung><Lung Respiratory System><Lung Tissue Fibrosis><Lymphadenopathy-Associated Virus><MT-SP1><MTSP-1><MTSP1><Magazine><Manuscripts><Mediating><Membrane-Type Serine Protease 1><Metabolic Glycosylation><Metallopeptidases><Metalloproteases><Metalloproteinases><Modeling><Modern Man><Molecular Interaction><N-Acetylneuraminic Acids><N-Acylneuraminate Glycohydrolases><Neuraminidase><Oligosaccharide Sialidase><Oseltamivir><PRSS14><Pathology><Peptidases><Peptide Hydrolases><Pharmacology><Physiologic><Physiological><Plasma><Plasma Serum><Polysaccharides><Prostamin><Protease Gene><Proteases><Protein Binding><Proteinases><Proteins><Proteolytic Enzymes><Proteomics><Prothrombin><Public Health><Publications><Pulmonary Fibrosis><Receptor Inhibition><Receptor Protein><Recombinants><Reticuloendothelial System, Serum, Plasma><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 disease severity><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 inhibitor><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV-2 severity><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 infection><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><SNC19><ST14><ST14 gene><Scientific Publication><Serine Endopeptidases><Serine Protease><Serine Protease TADG-15><Serine Protein Hydrolases><Serine Proteinases><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 inhibitor><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Sialic Acids><Sialidase><Site><Suppression of Tumorigenicity 14><Suppressor of Tumorigenicity 14><System><TADG-15><TADG15><Tamiflu><Testing><Therapeutic><Thrombase><Thrombin><Tripcellim><Trypsin><Tumor Associated Differentially Expressed Gene 15 Protein><Viral><Viral Diseases><Virus Diseases><Virus-HIV><Wuhan coronavirus><angiotensin converting enzyme 2><angiotensin converting enzyme II><balance><balance function><block SARS-CoV-2><block severe acute respiratory syndrome coronavirus 2><bound protein><clotting factor><corona virus><corona virus disease><corona virus disease 2019><coronavirus S protein><coronavirus disease><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 disease severity><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 pathogenesis><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 severity><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 virus><coronavirus disease infected patient><coronavirus disease patient><coronavirus disease positive patient><coronavirus disease severity><coronavirus disease-19><coronavirus disease-19 pathogenesis><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus infectious disease-19><coronavirus patient><coronavirus spike glycoprotein><exo alpha sialidase><fibrinogenase><fibrosis in the lung><fluid><glycosylation><gp120><gp120 ENV Glycoprotein><gp120(HIV)><hCoV19><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><influenzavirus><inhibit SARS-CoV-2><inhibit severe acute respiratory syndrome coronavirus 2><inhibitor><kallidin 9><kallidin I><liquid><lung fibrosis><matriptase><member><nCoV2><nano-molar><nanomolar><pandemic><pandemic disease><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><peptide analog><pulmonary><receptor><respiratory><response><severe acute respiratory syndrome coronavirus 2 disease severity><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><severe acute respiratory syndrome coronavirus 2 severity><vascular><viral infection><virus infection><virus-induced disease>