Multi-omic Approaches to Investigate the Inflammatory Response in Activated Macrophage Cells

Inflammation is a protective response to infection or injury and an essential<br/>process in the maintenance of tissue homeostasis. It constitutes a highly complex<br/>and tightly regulated sequence of events in which macrophages play a central<br/>role, by eliciting cellular pathways that produce inflammatory mediators to<br/>neutralize and eliminate invading pathogens and repair tissues damaged by<br/>trauma. The focus of this thesis was to define the global lipid and protein<br/>responses of cultured murine macrophages (RAW 264.7 cells) subjected to<br/>inflammatory stimuli. Cells were treated with immune modulators that either<br/>modelled bacterial infection (Kdo2-Lipid A) or reflected the signals produced by<br/>damaged or dying cells during clearance (ATP). In-depth proteomic analyses<br/>identified significant metabolic reprogramming along with a promotion of<br/>macrophage differentiation and polarisation, and lipidomic analysis identified<br/>significant regulation of arachidonic acid and glycerophospholipid metabolism.<br/>Furthermore, a novel stable isotope labelling methodology was developed using<br/>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<br/>ribosomal proteins and triglycerides following macrophage activation. Ultimately<br/>advanced multi-omic approaches can be utilised to investigate the molecular<br/>pathways of macrophages and provide additional perspectives on the<br/>mechanisms associated with inflammatory responses.

Name dc:type.qualificationname

Doctor of Philosophy (awarded by UHI)

Level dc:type.qualificationlevel

Doctoral Thesis

Grantor dc:publisher.institution

University of the Highlands and Islands

Year dc:date.issued

2021

Author dc:creator

• Brace, Nicole

Advisor dc:contributor.advisor

• Mainland, Ingrid

Language dc:language

eng

Identifier

oai:pure.atira.dk:studenttheses/f1cdd473-8a17-4e5a-966b-aa561938e354

OAI identifier oai:identifier

oai:pure.atira.dk:studenttheses/f1cdd473-8a17-4e5a-966b-aa561938e354