Document text
Principal Investigator: Jason Elinoff
Organization: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
Fiscal Year: 2024
Award: $194,017
Funding agency: National Heart Lung and Blood Institute
Primary human pulmonary artery and lung microvascular ECs express low levels of ACE2, TMPRSS2, and CD209L but high levels of NRP1. In contrast to lentiviral particles pseudotyped with the wild-type Wuhan variant, SARS-CoV-2 pseudovirus expressing the D614G spike protein variant successfully entered PAECs (P<0.05 for MOI 2.5 and 5 compared to MOI 0; P<0.05 for the fold increase compared to the Wuhan strain at the same MOI), while alternative variants (N439K and N501Y) did not. Likewise, using immunofluorescent microscopy, recombinant spike protein from the SARS-CoV-2 Delta variant (contains D614G mutation) dose-dependently entered pulmonary artery ECs while the Wuhan variant recombinant spike protein did not. Yet following inoculation with either the USA/WA1 strain, an early pandemic isolate carrying the D614G mutation, or the Delta variant of SARS-CoV-2 variant, there was no evidence of viral replication as determined by viral quantification (TCID50) of PAEC culture supernatants.
In FY24, SARS-CoV-2 nucleocapsid RNA transcript was detected by digital droplet PCR of pulmonary artery and lung microvascular whole cell lysates 1 hour, 24 and 72 hours following inoculation with different SARS-CoV-2 variants (MOI 0.1). Cell lysates from HEK293 cells served as a positive control since this cell type is known to be permissive to SARS-CoV-2. Notably the amount of viral RNA detected in pulmonary artery and lung microvascular ECs following inoculation was much lower than in HEK293 cells. Therefore, while SARS-CoV-2 appears to enter lung ECs, the virus does not appear to effectively replicate in vitro. Further experiments are planned to confirm these results and explore potential mechanisms that explain lack of viral replication.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><2019-nCoV variant><2019-nCoV variant forms><2019-nCoV variant strains><ACE2><Acute><Apoplexy><Apoptosis><Apoptosis Pathway><Autopsy><B.1.617.2><Blood Coagulation Disorders><Blood Vessels><Brain Vascular Accident><COVID associated pneumonia><COVID induced pneumonia><COVID infected patient><COVID patient><COVID pneumonia><COVID positive patient><COVID related pneumonia><COVID-19 S protein><COVID-19 associated pneumonia><COVID-19 induced pneumonia><COVID-19 infected patient><COVID-19 infection><COVID-19 patient><COVID-19 pneumonia><COVID-19 positive patient><COVID-19 related pneumonia><COVID-19 spike><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 variant><COVID-19 variant forms><COVID-19 variant strains><COVID-19 viral pneumonia><COVID-19 virus><COVID-19 virus infection><COVID19 infection><COVID19 patient><COVID19 positive patient><COVID19 virus><Cardiovascular><Cardiovascular Body System><Cardiovascular Manifestation><Cardiovascular Organ System><Cardiovascular system><Cell Body><Cells><Cerebral Stroke><Cerebrovascular Apoplexy><Cerebrovascular Stroke><Cessation of life><Circulatory Collapse><CoV emergence><CoV-2><CoV2><Coagulation Disorder><Coagulopathy><Death><Delta variant><Disease><Disorder><Distal><Dose><Electron Microscopy><Endothelial Cells><Endothelium><Epitheliasin Gene><Genetic Alteration><Genetic Change><Genetic defect><Heart Vascular><Hour><Human><Hypoxemic Respiratory Failure><Hypoxic Respiratory Failure><In Situ Hybridization><In Vitro><Infection><Inflammatory Infiltrate><Injury><Lung><Lung Respiratory System><MIS-C><Microscopy><Modern Man><Molecular><Multiorgan Inflammatory Syndrome in Children><Multisystem Inflammatory Syndrome in Children><Mutation><NRP1><NRP1 gene><Non-Polyadenylated RNA><Nucleocapsid><Organ><PRSS10><Programmed Cell Death><Proteins><Pulmonary Artery><Pulmonary Hypertension><Pulmonary artery structure><RNA><RNA Gene Products><Recombinants><Ribonucleic Acid><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 B.1.617.2><SARS-CoV-2 S><SARS-CoV-2 S protein><SARS-CoV-2 associated pneumonia><SARS-CoV-2 delta><SARS-CoV-2 induced pneumonia><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 patient><SARS-CoV-2 pneumonia><SARS-CoV-2 positive patient><SARS-CoV-2 related pneumonia><SARS-CoV-2 spike><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 variant><SARS-CoV-2 variant forms><SARS-CoV-2 variant strains><SARS-CoV-2 viral pneumonia><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Secondary to><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 coronavirus 2><Severe acute respiratory syndrome coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Shock><Stroke><Syndrome><TMPRSS2><TMPRSS2 gene><Thrombus><Transcript><VEGF165R><Variant><Variation><Vascular Endothelium><Viral><Virus><Virus Replication><Wuhan coronavirus><angiotensin converting enzyme 2><angiotensin converting enzyme II><arteriole><bleeding disorder><brain attack><cell type><cerebral vascular accident><cerebrovascular accident><circulatory shock><circulatory system><clotting disorder><corona virus emergence><coronavirus disease 2019 S protein><coronavirus disease 2019 associated pneumonia><coronavirus disease 2019 induced pneumonia><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 patient><coronavirus disease 2019 pneumonia><coronavirus disease 2019 positive patient><coronavirus disease 2019 related pneumonia><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 variant><coronavirus disease 2019 variant forms><coronavirus disease 2019 variant strains><coronavirus disease 2019 virus><coronavirus disease associated pneumonia><coronavirus disease induced pneumonia><coronavirus disease infected patient><coronavirus disease patient><coronavirus disease pneumonia><coronavirus disease positive patient><coronavirus disease related pneumonia><coronavirus disease-19 patient><coronavirus disease-19 pneumonia><coronavirus disease-19 virus><coronavirus emergence><coronavirus patient><digital><emergent CoV><emergent corona virus><emergent coronavirus><emerging CoV><emerging corona virus><emerging coronavirus><experiment><experimental research><experimental study><experiments><genome mutation><hCoV19><in situ Hybridization Genetics><in situ Hybridization Staining Method><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><injuries><lung microvascular endothelial cells><lung vascular endothelial cells><mortality><multisystem inflammatory syndrome of children><nCoV><nCoV2><necropsy><new CoV><new corona virus><new coronavirus><novel CoV><novel corona virus><novel coronavirus><pandemic><pandemic disease><particle><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><pediatric inflammatory multisystem syndrome><permissiveness><pneumonia due to COVID><pneumonia due to COVID-19><pneumonia due to SARS-CoV-2><pneumonia due to coronavirus disease><pneumonia due to coronavirus disease 2019><pneumonia due to severe acute respiratory syndrome coronavirus 2><pneumonia in COVID><pneumonia in COVID-19><pneumonia in SARS-CoV-2><pneumonia in coronavirus disease><pneumonia in coronavirus disease 2019><pneumonia in severe acute respiratory syndrome coronavirus 2><postmortem><pulmonary><pulmonary arterial endothelial cell><pulmonary artery endothelial cell><pulmonary microvascular endothelial cells><pulmonary vascular endothelial cells><severe acute respiratory syndrome coronavirus 2 B.1.617.2><severe acute respiratory syndrome coronavirus 2 associated pneumonia><severe acute respiratory syndrome coronavirus 2 induced pneumonia><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 pneumonia><severe acute respiratory syndrome coronavirus 2 positive patient><severe acute respiratory syndrome coronavirus 2 related pneumonia><severe acute respiratory syndrome coronavirus 2 variant><severe acute respiratory syndrome coronavirus 2 variant forms><severe acute respiratory syndrome coronavirus 2 variant strains><shocks><spike proteins on SARS-CoV-2><stroked><strokes><systemic inflammation><systemic inflammatory response><transcriptomics><vascular><vascular inflammation><venous thromboembolism><viral RNA><viral multiplication><viral replication><virus RNA><virus multiplication>