About the Project
Research project outline :
Obesity, insulin resistance, type 2 diabetes (T2D), and associated cardiovascular disease (CVD) often coexist and are associated with inflammation of metabolic, vascular and endocrine tissues. Rheumatoid arthritis (RA) is strongly associated with metabolic and vascular comorbidity. Pattern recognition receptors (PRR) such as toll-like receptors (TLR), NOD-like receptors and C-type lectins are microbial and tissue damage sensors that strongly dictate the outcome of an inflammatory response. PRRs are apt at sensing microenvironmental cues that are fundamental in shaping organ- and tissue-specific inflammatory responses. Cell surface TLR2, TLR4, and TLR5 recognize components of the bacterial cell wall including lipopolysaccharide (LPS). TLR3, TLR7, TLR8 and TLR9 are located on the membranes of the endoplasmic reticulum, endosomes and lysosomes, where they detect nucleic acids of bacterial or viral origin. The therapeutic potential of interfering with or exploiting PRR signaling in immuno-metabolic diseases is so far unexplored.
Our previous work allowed the elegant characterization of the inflammatory and synthetic properties of human atherosclerosis. Using this system and ex vivo biochemical approaches we demonstrated that the dysregulated inflammatory response in human atherosclerosis is driven by the extracellular sensor TLR2 and NFkB activation. We then showed that the endosomal receptors TLR3 and TLR7 mediate atheroprotection by exerting an unexpected anti-inflammatory effect. Hence, pattern recognition by TLRs could elicit both protective and detrimental effects in atherosclerosis and the outcome of TLR signaling depends on sensing patterns (extracellular vs. endosomal) and specific ligands present in the tissue. There is initial evidence suggesting that these effects might be cell type specific but the information of the specific repertoire and function of TLRs of the various cell subsets involved in atherosclerosis is scant.
This DPhil project will map the TLR and CLR expression repertoire in myeloid and resident vascular cell types and their relevance to atherosclerosis and vascular injury, fluorescence activated cell sorting of live cells, quantitation of gene expression and single cell genomics. Recent advances in immune-monitoring including multi-parameter flow and mass cytometry will also be applied. Mass cytometry is a significant innovation that takes advantage of the measurement resolution of Inductively Coupled Plasma (ICP) mass spectrometry and applies it to single-cell analysis. In mass cytometry stable isotopes of non-biological rare earth metals are conjugated to the same clones of antibody used in conventional flow cytometry, overcoming the interference caused by spectral overlap between fluorescent agents and autofluorescence (https://www.kennedy.ox.ac.uk/facilities/cytof-mass-cytometry/overview).
Moreover the DPhil Candidate will dissect the crosstalk between endosomal and extracellular TLRs in models of atherosclerosis. We recently screened transcriptomics changes in human atherosclerotic tissues and obtained a shortlist of potential molecular mediators of the TLR crosstalk we observed in vivo. The role of candidate genes will be validated using techniques including siRNA knockdown, cell activation and proliferation, phagocytic and efferocytosis function in genetically modified cells. The consequences of PRR-mediated signalling in resident vs. inflammatory cell types will be evaluated in state-of-the-art models atherosclerosis, arterial injury and plaque rupture with genetic deletions and bone marrow chimeras (including mixed bone marrow chimeras).
Details of the research group (e.g. how many students, post-docs, website, etc.)
The DPhil Candidate will join a highly committed research team focused on understanding the role of inflammation in atherosclerosis and identifying new therapeutic pathways for cardiovascular disease. The Cardiovascular Inflammation Group headed by Claudia is currently formed by 4 post-doctoral research fellows, 2 DPhil students and 3 senior research technicians (https://www.kennedy.ox.ac.uk/research/inflammation-in-atherosclerosis).
Training opportunities:
The project will be carried out at the Kennedy Institute of Rheumatology, which is a world-renowned research center housed in a new state-of-the-art facility in Oxford. The Kennedy Institute, headed by Professor Fiona Powrie, is an exciting research institute focusing on inflammatory diseases involving the musculoskeletal system, the vascular system and the gut. The Institute houses a vibrant community of basic and clinician scientists working on diverse aspects of immunology, inflammation and tissue repair. The PhD programme includes a core curriculum of 20 lectures in the first term of year 1 to provide a solid foundation in musculoskeletal sciences, immunology and data analysis. Full training will be provided in a range of immunology, cell and molecular biology techniques, as well as opportunities for debate between peers and senior academics. Students will attend weekly group meetings and journal clubs, and will be expected to attend seminars in the department and relevant seminars in the wider University. Students will attend national and international meetings, at which they will present their data.
Further information:
Contact: Professor Claudia Monaco, Kennedy Institute of Rheumatology, University of Oxford
E-mail: [Email Address Removed]
References
Relevant publications
1. Cole JE, Astola N, Cribbs AP, Goddard ME, Park I, Green P, Davies AH, Williams RO, Feldmann M, Monaco C. Indoleamine 2,3-dioxygenase-1 is protective in atherosclerosis and its metabolites provide new opportunities for drug development. Proc Natl Acad Sci U S A. 2015 Oct 20;112(42):13033-8. doi: 10.1073/pnas.1517820112. Epub 2015 Oct 5.
2. Strom AC, Cross AJ, Cole JE, Blair PA, Leib C, Goddard ME, Rosser EC, Park I, Hultgårdh Nilsson, Nilsson J, Mauri C, Monaco C. B regulatory cells are increased in hypercholesterolemic mice and protect from lesion development via IL-10. Thromb Haemost. 2015 Oct;114(4):835-47.
3. Cole JE, Navin TJ, Cross AJ, Goddard ME, Alexopoulou L, Mitra AT, Davies AH, Flavell R, Feldmann M, Monaco C. An unexpected protective role for TLR3 in the arterial wall. Proc Natl Acad Sci U S A. 2011 Feb 8;108(6):2372-7. Comment on PNAS USA 2011; 108 (7): 2637-8.
4. Monaco C, Gregan S, Navin T, Davies AH, Feldmann M. TLR-2 drives inflammation and matrix degradation in human atherosclerosis Circulation 2009; 120: 2462-2469.
5. Monaco C., Andreakos E., Kiriakidis S., Mauri C., Bicknell C., Foxwell B., Cheshire N., Paleolog E., Feldmann M. The canonical pathway of NFκB activation selectively regulates pro-inflammatory and pro-thrombotic responses in human atherosclerosis. PNAS USA 2004, 2004 13;101:5634-9.