Abstract
Inflammation is a protective response to infection or injury and an essentialprocess in the maintenance of tissue homeostasis. It constitutes a highly complex
and tightly regulated sequence of events in which macrophages play a central
role, by eliciting cellular pathways that produce inflammatory mediators to
neutralize and eliminate invading pathogens and repair tissues damaged by
trauma. The focus of this thesis was to define the global lipid and protein
responses of cultured murine macrophages (RAW 264.7 cells) subjected to
inflammatory stimuli. Cells were treated with immune modulators that either
modelled bacterial infection (Kdo2-Lipid A) or reflected the signals produced by
damaged or dying cells during clearance (ATP). In-depth proteomic analyses
identified significant metabolic reprogramming along with a promotion of
macrophage differentiation and polarisation, and lipidomic analysis identified
significant regulation of arachidonic acid and glycerophospholipid metabolism.
Furthermore, a novel stable isotope labelling methodology was developed using
deuterium oxide (2H2O) in conjunction with high resolution mass spectrometry for the simultaneous measurement of the rates of synthesis of individual lipids and proteins on a cellular-wide scale. The results revealed altered dynamics of
ribosomal proteins and triglycerides following macrophage activation. Ultimately
advanced multi-omic approaches can be utilised to investigate the molecular
pathways of macrophages and provide additional perspectives on the
mechanisms associated with inflammatory responses.
| Date of Award | 9 Jul 2021 |
|---|---|
| Original language | English |
| Awarding Institution |
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| Supervisor | Ingrid Mainland (Supervisor) & Ian Megson (Supervisor) |