Immature neutrophils and the vasculature: friends or foes?

PROJECT OVERVIEW

Vascular pathologies underline devastating diseases ranging from auto-immune vasculitis to the recent COVID-19 pandemic (1). Neutrophils, as the most abundant immune cells, have been reported to intimately interact with the vascular system either via direct cell-cell contact or indirectly through release of inflammatory cytokines or cellular substances.  Fully functional mature neutrophils patrol the circulation and tissues to exert anti-microbial activity through several mechanisms including release of cytotoxic products, reactive oxygen species (ROS), neutrophil extracellular traps (NETs) and pore-forming molecules. These activities can cause vascular tissue damage if poorly controlled.

Inflammatory responses trigger the release of functionally distinct immature neutrophils into the circulation and tissues in different diseases, including severe COVID-19, where we, and others, identify the presence of neutrophil progenitors (2). Our recent work on giant cell arteritis (GCA) has shown that immature neutrophils can interact with endothelium, generate ROS and cause vascular leakage and damage that may lead to systemic vascular pathology (3).

This project will experimentally test a possible direct involvement of immature neutrophils in endothelial damage in a mouse system for imaging of neutrophil subset interactions with vascular walls in vivo (4). We will exploit our recently unravelled cell-intrinsic molecular regulators of neutrophil maturation and function (5) to genetically manipulate neutrophil phenotypes (i.e. cells and mouse strains deficient in specific regulators). In parallel, we will expand our analysis of neutrophil- and oxidative tissue damage-associated biomarkers in human biopsies (biobanked and fresh) of large (GCA) and small (ANCA, Lupus) vessel vasculitis patients, using the state-of-the-art spatial biology approaches, such as multi-parameter confocal microscopy and single cell spatial transcriptomics. Correlations between molecular signatures of vascular damage associated with immature neutrophils, endpoints and treatment outcomes will be assessed in a clinically well-defined cohorts. 

The outcome of this study is expected to contribute significantly to development of new targets for therapeutic interventions to prevent detrimental vascular damage that is implicated in many diseases such as auto-immune vasculitis.

KEYWORDS

Neutrophils, Vasculitis, Multiplex Imaging, Spatial transcriptomics, Vascular pathologies 

TRAINING OPPORTUNITIES

The Kennedy Institute is a world-renowned research centre and is housed in a brand new state-of-the-art research facility. Training will be provided in techniques in a wide range of immunological tool kits (cell isolation, FACS, ELISA, primary cell culture); imaging (immunofluorescence on tissue sections, confocal intravital microscopy) and genomic (single cell and spatial transcriptomics) approaches.  Multiplex assays such as the Luminex assay will be used for patient plasma profiling to identify key signaling molecules that modulate neutrophil-vasculature interaction. The candidate can benefit from the hands-on experience with these techniques in the Udalova and Clarke labs, and from access to clinical samples in the Luqmani group.

A core curriculum of lectures will be taken in the first term to provide a solid foundation in a broad range of subjects including inflammation, genomics, epigenetics, translational immunology and data analysis. Students will attend weekly seminars within the department and those relevant in the wider University. Students will be expected to present data regularly to the department, the Genomics of Inflammation lab and to attend external conferences to present their research globally. Students will also have the opportunity to work closely with both internal and external collaborators on live imaging and spatial omic analyses. 

KEY PUBLICATIONS

(1)        Wang L, Luqmani R, Udalova IA. The role of neutrophils in rheumatic disease-associated vascular inflammation. Nature Review Rheumatology. 2022 Mar;18(3):158-170.

(2)        Oxford Covid-19 Immunology Consortium. A blood atlas of COVID-19 defines hallmarks of disease severity and specificity. Cell. 2022 Mar 3;185(5):916-938.e58.

(3)        Wang L, Ai Z, Khoyratty T, Zec K, Eames HL, van Grinsven E, Hudak A, Morris S, Ahern D, Monaco C, Eruslanov EB, Luqmani R, Udalova IA. ROS producing immature neutrophils are linked to GCA vascular pathologies. Journal of Clinical Investigations Insight. 2020 Oct 15;5(20):e139163

(4)        Finsterbusch M, Voisin MB, Beyrau M, Williams TJ, Nourshargh S. Neutrophils recruited by chemoattractants in vivo induce microvascular plasma protein leakage through secretion of TNF. J Exp Med. 2014 Jun 30;211(7):1307-14.

(5)        Khoyratty T*, Ai Z*, Ballesteros I, Mathie S, Eames HL, Martín-Salamanca S, Wang L, Hemmings A, Willemsen N, von Werz V, Zehrer A, Walzog B, van Grinsven E, Hidalgo A, Udalova IA. Distinct transcription factor networks control neutrophil-driven inflammation. Nature Immunology, 2021 Sep;22(9):1093-1106.

CONTACT INFORMATION OF ALL SUPERVISORS

Professor Irina Udalova Irina.udalova@kennedy.ox.ac.uk

Professor Alex Clarke alexander.clarke@kennedy.ox.ac.uk

Professor Raashid Luqmani raashid.luqmani@ndorms.ox.ac.uk

Dr Kristina Zec Kristina.zec@kennedy.ox.ac.uk