Development of a mass spectrometric workflow for the analysis of stable isotope labelled metabolites in cancer cells
File(s)
Author(s)
Turkseven, Seyma
Type
Thesis
Abstract
Altered lipid metabolism is one of the prominent features observed in cancer, and its underlying mechanisms and impact on the lipidomic composition remain incompletely understood due to the diverse range of lipids and the complexity of cancer cells. Metabolomics studies play a crucial role in understanding these changes and reflect as metabolite ion abundance, facilitating the identification of potential biomarkers and therapeutic targets in cancer. However, to uncover the underlying metabolite changes, isotopically labelled substrates have been increasingly integrated into the experimental designs. By incorporating labelled substrates, it becomes possible to track the metabolic fate of specific molecules and elucidate the altered pathways involved in lipid metabolism.
The novel ambient ionisation technique known as rapid evaporative ionisation mass spectrometry (REIMS) is one promising technique for investigating lipid metabolism in cancer cells. Initially developed for tissue classification, REIMS offers distinct advantages over traditional mass spectrometric approaches, such as its ability to perform rapid analysis without extensive sample preparation. These advantages make REIMS a valuable analytical tool for wide-ranging studies.
The primary objective of this study was to trace the altered lipid metabolism in breast cancer cells using labelled palmitate and analyse the cells using the REIMS approach. To achieve this, a mass spectrometric workflow utilising REIMS in the tracer study was developed. Several factors affecting data quality were examined, and preliminary findings indicated that the REIMS spectra effectively characterise the lipid composition of cancer cells and detect labelled metabolites belonging to various lipid classes, including PE, PC, PI, FA and Cer...
The novel ambient ionisation technique known as rapid evaporative ionisation mass spectrometry (REIMS) is one promising technique for investigating lipid metabolism in cancer cells. Initially developed for tissue classification, REIMS offers distinct advantages over traditional mass spectrometric approaches, such as its ability to perform rapid analysis without extensive sample preparation. These advantages make REIMS a valuable analytical tool for wide-ranging studies.
The primary objective of this study was to trace the altered lipid metabolism in breast cancer cells using labelled palmitate and analyse the cells using the REIMS approach. To achieve this, a mass spectrometric workflow utilising REIMS in the tracer study was developed. Several factors affecting data quality were examined, and preliminary findings indicated that the REIMS spectra effectively characterise the lipid composition of cancer cells and detect labelled metabolites belonging to various lipid classes, including PE, PC, PI, FA and Cer...
Version
Open Access
Date Issued
2023-06-01
Date Awarded
2023-09-01
Copyright Statement
Attribution-NonCommercial 4.0 International Licence (CC BY-NC)
License URL
Advisor
Takats, Zoltan
Poulogiannis, George
Sponsor
British Council
Türkiye Bilimsel ve Teknolojik Araştırma Kurumu
Publisher Department
Department of Metabolism, Digestion and Reproduction
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)