elucidating the role of Tumour Necrosis Factor Receptor 1 (TNFR1) in regulating acute inflammatory responses of leukocyte trafficking and vascular per
Lead Research Organisation:
Queen Mary University of London
Department Name: William Harvey Research Institute
Abstract
The specific objective of the PhD project will be to utilise novel pharmacological reagents generated by GSK to dissect the molecular contribution of TNFR1-dependent signalling in regulation of neutrophil-vessel wall dynamics and vascular leakage in models of vascular and tissue injury. The project will involve use of numerous in vivo experimental models of inflammation in conjunction with biochemical and cellular assays. In addition the project will employ cutting edge imaging modalities, including confocal intravital microscopy for real-time tracking of microvascular responses, a specialised technique for which the group is internationally respected.
People |
ORCID iD |
| Sussan Nourshargh (Primary Supervisor) | |
| Charlotte Owen-Woods (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/P504439/1 | 01/10/2016 | 30/09/2020 | |||
| 1797224 | Studentship | BB/P504439/1 | 01/10/2016 | 30/09/2020 | Charlotte Owen-Woods |
| Description | Increased microvascular permeability to plasma proteins and neutrophil emigration into the tissue are fundamental hallmarks of innate immunity and key features of numerous inflammatory disorders. Whilst neutrophils can promote blood vessel leakage, the impact of the latter on neutrophil migration into the tissue is unknown. To address this, we developed a microscopy method to allow for the simultaneous quantification of movement of neutrophils through blood vessels into the surrounding tissue and vascular leakage in the mouse cremaster muscle, a thin, transparent tissue surrounding the testes. Here we identified that reactions exhibiting vascular leakage, had an increased frequency of neutrophil reverse transendothelial migration (reverse TEM), whereby neutrophils that have moved through endothelial cell junctions into the tissue then move back into the blood circulation. Additionally, using a mouse model exhibiting a defect in vascular leakage, we observed that the frequency of neutrophil reverse TEM was decreased. Finally, we have developed a novel labelling strategy designed to exclusively track reverse migrating neutrophils. This methodology allowed us to investigate the fate and phenotype of these reversing white blood cells. Our current data provided direct evidence for the dissemination of reverse migrating neutrophils exhibiting a pro-inflammatory activated phenotype. Furthermore, their dissemination into the lungs was aligned with regions of vascular injury. This cascade of events offers a mechanism to explain how local tissue inflammation and vascular leakage can induce downstream pathological effects in remote organs. |
| Exploitation Route | Local organ trauma or inflammatory events associated with both oedema and immune cell infiltration, are usually accompanied with remote organ damage, leading to severe organ dysfunction, or even death of the patients. At present there are no efficacious treatments available in the clinic to treat these pathologies. In this pre-clinical study we provided the first evidence that a local tissue inflammation associated with blood vessel leakage can induce downstream pathological effects via the dissemination of a subpopulation of activated neutrophils in remote organs, and most notably in the lungs. It is believed that controlling the development of this specific population of neutrophils could be used to develop a novel therapeutic approach to protect patients against remote organ damage. Understanding further the characteristics and functions of these cells will highlight key molecules amenable for the development of new drugs to treat these diseases. |
| Sectors | Pharmaceuticals and Medical Biotechnology |
| URL | https://www.jci.org/articles/view/133661 |
| Description | Characterisation of mice deficient in neutrophil tumour necrosis factor (TNF) |
| Organisation | Russian Academy of Sciences |
| Department | Engelhardt Institute of Molecular Biology (EIMB) |
| Country | Russian Federation |
| Sector | Academic/University |
| PI Contribution | The current project aims to assess the role of neutrophil-derived TNF in mouse models of aberrant leukocyte migration and vascular permeability. To address these aims, we have generated a novel genetically-modified mouse model with selective deletion of TNF in neutrophils through crossing of TNF floxed mice with a neutrophil Cre line. Generation of this novel mouse colony required validation of the extent of TNF deficiency in these animals, in order to assess their value in more complex studies. Functionally we have confirmed that these mice are less able to mount physiological responses to stimuli that rely on the downstream effect of neutrophil TNF. We have further characterised these mice and established that TNF-deficient mice are protected from pathophysiological insults, such as ischaemia/reperfusion injury. Thus, our contribution to this collaboration has resulted in the generation, validation and characterisation of a completely novel mouse model. |
| Collaborator Contribution | Our partners have contributed to this collaboration by supplying the required genetically-modified mice. |
| Impact | This collaboration has led to a number of conference Abstracts being submitted, such as the joint meeting of the UK Cell Adhesion Society/British Microcirculation Society (Birmingham, April 2017) and the Eur Vasc Biol Meeting (Geneva, May 2017). The data contributing to these Abstracts is being built on, collated and prepared for a manuscript . This collaboration is not multi-disciplinary. |
| Start Year | 2014 |
| Description | Characterisation of mice deficient in vascular permeability |
| Organisation | Max Planck Society |
| Department | Max Planck Institute for Molecular Biomedicine |
| Country | Germany |
| Sector | Academic/University |
| PI Contribution | This work aimed to assess the interplay between vascular permeability and leukocyte migration using experiment mouse models of acute inflammation. To address these aims, our collaborator has generated a novel genetically-modified mouse model with selective mutation in the VE-Cadherin gene, resulting in the generation of a mouse strain exhibiting reduced vascular permeability upon stimulation with vascoactive agents. Functionally we have confirmed that these mice exhibit a reduced vascular leakage upon inflammation associated with an inhibition of aberrant transendothelial migration of neutrophils in vivo, a response associated with remote organ damage. Thus, our contribution to this collaboration has resulted in the generation of novel data from these GM animals. This collaboration is closely aligned with the work conducted on the impact of vascular permeability on trafficking of chemokines across vessel walls. |
| Collaborator Contribution | Our partners have contributed to this collaboration by providing intellectual knowledge in vascular permeability signalling pathways and by supplying us with their genetically modified mice |
| Impact | This collaboration has contributed to the submission of a conference abstract to the UK Cell Adhesion Meeting (London, September 2018). Additionally, data generated from this collaboration has led to a strong publication in J Clin Invest in 2020. This collaboration is not multi-disciplinary. |
| Start Year | 2018 |
| Description | Partnership with GlaxoSmithKline to investigate a clinically relevant TNFR1 inhibitor |
| Organisation | GlaxoSmithKline (GSK) |
| Department | Research and Development GSK |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | The aim of this project is to elucidate the specific TNF pathway involved in the aberrant neutrophil migration and vascular permeability upon acute inflammation. As part of this partnership, our team is responsible for the experimental design and execution of most of the in vivo and in vitro studies. Specifically, using the inhibitors provided by our partner, we are currently investigating the role of TNFR1 signalling in the recruitment of leukocytes at site of inflammation, the dynamic and nature of their migration through the different components of the vessel wall (most notably the endothelium), as well as the effect of this inhibition for vascular permeability and remote organ damage. Our contribution to date demonstrate the feasibility to target TNFR1 for inhibiting the recruitment of neutrophils during acute inflammation in a mouse model of ischemia-reperfusion injury. |
| Collaborator Contribution | Our partners are involved in financial and intellectual contributions by providing co-supervision to the PhD Student (Charlotte Owen-Woods) and clinically relevant inhibitors to be tested in different inflammatory models developped in our group. |
| Impact | This collaboration has led, to date, to the generation of data outputs contributing to a manuscript in preparation. This collaboration is not multi-disciplinary. |
| Start Year | 2016 |