Skip to main content Skip to navigation

Dissecting the role of endogenous anti-inflammatory agents on COVID-driven immunothrombosis

Primary Supervisor: Professor Dimitris Grammatopoulos, Warwick Medical School

Secondary Supervisor: Professor LY Young

PhD project title: Dissecting the role of endogenous anti-inflammatory agents on COVID-driven  immunothrombosis

University of Registration: University of Warwick

Project outline:

    COVID-19 patients exhibit unexpected presentation and complications. Patients requiring hospital admissions often develop severe disease during the acute phase and suffer long term complications post-recovery. Emerging evidence suggests that endothelial cells are essential contributors to the initiation and development of severe COVID-19 disease. Vascular endothelial dysfunction associated with inflammation (endotheliitis) can be amplified in the presence of a cytokine storm, one of the key features of COVID-19 disease. In severe cases of COVID-19, immunothrombosis (which identifies the role of the innate immune system in coangulation) is attributed to high level of neutrophil extracellular traps (NETs), extracellular webs of chromatin, microbicidal proteins, and oxidant enzymes that when not properly regulated have the potential to propagate inflammation and microvascular thrombosis, through platelet-neutrophil interactions. Activated leukocytes and neutrophil extracellular traps (NETosis) also promote immunothrombotic clots that embed into the target tissues of severe COVID-19 patients. Emerging evidence suggest that contributing factors might include other components of NETosis such as Damage-Associated Molecular Patterns (DAMPs) released by damaged cells to trigger inflammatory responses; development of transient autoimmunity and an anti-phospholipid syndrome (APS)-like disease. The impact of these mechanisms might worsen in the presence of deficient anti-inflammatory defence mechanisms such as vitamin D and oestrogens. The latter might explain why women have reduced risk for severe disease or death during the acute phase of the disease.

    This project will develop preclinical in vitro cellular models of neutrophil-endothelial cell or neutrophil-alveolar macrophages interactions and study effects of vitamin D and oestrogens on NETosis and DAMP release upon exposure to serum from COVID-19 patients. Markers of NETosis will be quantified by immunofluorescence and immunoassays. This model will also provide a test system to evaluate other potential therapeutic interventions.

    In addition, we will establish neutrophil-endothelial cell co-culture experiments using Netwell permeable support, supernatants from NETosis-activated neutrophils will be co-cultured with HUVECs or MH-S murine alveolar macrophage cells or primary alveolar macrophages obtained by bronchoalveolar lavage form normal or COVID-19 patients.

    BBSRC Strategic Research Priority: Understanding the Rules of Life: Immunology & Integrated Understanding of Health: Ageing & Diet and Health

    Techniques that will be undertaken during the project:

    • Cell co-culture
    • Immunoassay and multiplex immunoassays
    • Confocal microscopy
    • Cell stimulation assays and signalling pathways exploration
    • PCR-including digital droplet PCR

    Contact: Professor Dimitris Grammatopoulos, University of Warwick