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Title: Proteomic and metabolomic signatures of arthropathies
Author: Anderson, J. R.
Awarding Body: University of Liverpool
Current Institution: University of Liverpool
Date of Award: 2019
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Osteoarthritis (OA) and rheumatoid arthritis (RA) are the most common arthropathies in people, causing limited mobility, pain and subsequent reduction in quality of life, resulting in a substantial impact on human society. Conditions affecting articular joints are also common in horses, including OA, osteochondrosis (OC) and synovial sepsis. Despite their high prevalence and clinical relevance, diagnosis, staging, monitoring and determining an accurate prognosis remain a challenge. Thus, there is a need to identify reliable biomarkers of disease. The development of 'omics' technologies, such as metabolomics and proteomics, has provided global analysis of biological tissues and fluids, enabling discovery of disease biomarkers and providing a greater understanding of their underlying pathogeneses. Within this thesis, protocols for collection and processing of synovial fluid (SF) for nuclear magnetic resonance (NMR) metabolomic and liquid chromatography-tandem mass spectrometry (LC-MS/MS) proteomic analysis have been optimised. Lys-C endopeptidase pre-digestion greatly improved on-bead tryptic SF protein digestion when used in conjunction with randomised hexapeptide ProteoMiner™ beads for low abundant protein enrichment. Equine OA was stratified using metabolomic and proteomic SF profiles, identifying a panel of markers which may be applicable to grading OA severity. This is the first study to undertake computational integration of NMR metabolomic and LC-MS/MS proteomic datasets of any biological system. This thesis is also the first study to use a multi 'omics' approach to simultaneously investigate the metabolomic profile of ex-vivo cartilage and metabolomic/proteomic profiles of culture media using the TNF-α/IL-1β ex-vivo cartilage OA model. A panel of metabolites, proteins and neopeptides were identified which were differentially abundant within an early phase of this OA model and may provide further information on the underlying disease pathogenesis. Comparing human OA and RA SF identified that the metabolic pathways that differed most were glycolysis, amino acid biosynthesis and taurine and hypotaurine metabolism. This thesis has also identified a panel of metabolites and proteins within equine SF which can distinguish synovial sepsis from nonseptic joint pathologies, with glucose the principal metabolite discriminator.
Supervisor: Peffers, Mandy ; Clegg, Peter ; Rubio-Martinez, Luis ; Lian, Lu-Yun Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral