Finnur Freyr Eiriksson (Presenter)
University of Iceland
Bio: I have a M.Sc. Degree in pharmaceutical sciences from the University of Iceland Curretly pursuing a Ph.D. degree in Biomedical Sciences from the University of Iceland. I work as laboratory manager at ArcticMass Ltd. a bio-analytical spin-off company. I regularly supervise M.Sc. students from the faculty of Pharmaceutical Sciences, University of Iceland in the field of analytical chemistry with a focus on lipid analysis with mass spectrometry.
Authorship: Finnur Freyr Eiriksson (1, 2), Manuela Magnusdottir (1), Skarphedinn Halldorsson (1), Ottar Rolfsson (1), Helga M. Ogmundsdottir (1), Margret Thorsteinsdottir (1, 2)
(1) Universtity of Iceland, Reykjavík, Iceland; (2) ArcticMass, Reykjavík, Iceland
It is now realized that lipids exhibit a wide variety of physiological functions, structural as well as regulatory. Evidence links carcinogenesis to metabolic control and indicates risk association of cancer with obesity. The aim of this project is to establish a lipidomic-based UPLC-QToF method for evaluation of lipid composition in cultured cells of normal and cancerous origin. PCA analyses revealed differences in the lipidomes of several cancer cell lines. Phosphocholins were significantly elevated in Sk-Br-3 (overexpresses fatty acid synthase) compared to other cell lines. The lipidomes of D492 and its subline D492M, which has undergone epithelial-to-mesenchymal transition, were significantly different.
Lipids were first proposed to be only structural building blocks and energy storage in living organisms but have been shown to have numerous functions in biological processes. It is now realized that lipids exhibit a much wider variety of physiological functions, structural as well as regulatory. Accumulating evidence links carcinogenesis to metabolic control and indicates risk association of several types of cancer with obesity. The importance of metabolic changes in cancer cell phenotypes has been demonstrated by experimental data. Cancer cells show differences from healthy normal cells in their metabolism, including lipid metabolism and this difference can contribute to their ability to survive and grow at a faster rate. Such metabolic changes and how they could be affected by drugs can be studied with a lipidomic approach. Overexpression of fatty acid synthase (FASN) is commonly observed in cancer cells and there are also suggestions for other lipid pathways being changed in tumor cells. The general aim of this project is to establish a lipidomic-based UPLC-QToF method for evaluation of lipid composition in cultured cells of normal and cancerous origin.
An Ultra Performance Liquid Chromatography (UPLC) coupled to a Synapt G1 QToF mass spectrometer was utilized for lipidomic evaluation of different cultured cancer cell lines and non-malignant cell lines. Cell lines were selected partly based on knowledge of their lipid metabolism, including six breast cancer cell lines: SK-Br-3 (overexpresses FASN and HER-2), T47D (TP53-mutated), MCF7 (estrogen receptor-positive) Cama-1 (luminal type), MDA-MB-231 (highly invasive), MDA-MB-436 (triple-negative); one non-malignant breast epithelial cell line, MCF10a, and one pancreatic cancer cell line, ASPC-1(overexpresses 5-and 12-lipoxygenases). All cells were extracted according to a modified Bligh and Dyer procedure.
Furthermore, we utilized a Synapt G2 with IonKey source for analyzing the immortalized breast epithelial stem cell line D492 along with its subline, D492M, that has undergone epithelial to mesenchymal transition (EMT). Data acquisition was carried out using MassLynx 4.1 and lipids identified by MarkerLynx XS. SIMCA 14.1 was used for Principal component analysis (PCA) and statistical evaluation of the results.
Lipid composition in cultured cancer cell lines and non-malignant cell lines was determined by the UPLC-QToF method. Principal component analyzes (PCA) revealed distinct differences in lipid composition between all cancer cell lines tested, each cell line shows a clear cluster of lipid components and a pattern that was reproducible between experiments. Sk-Br-3 was most clearly separated from the other two breast cancer cell lines. By utilizing orthogonal projections to latent structures discriminant analysis (OPLS-DA) when comparing the breast cancer cell lines, we were able to interpret the data visually using s-plots and detected markers that explain the variance between different groups in the PCA plot. The data revealed significant differences in levels of phosphocholins (PC) that are elevated in Sk-Br-3 breast cancer cell line when compared to other breast cancer cell lines. Further investigations of the raw data and utilizing MSE, specific PCs could be identified that differ between cell lines. With our new lipidomic platform on a Synapt G2 with IonKey we were able to detect significant differences in the lipid composition of the immortalized breast epithelial stem cell line (D492) and its subline D492M, which has undergone EMT.
Conclusions & Discussion
In conclusion, different levels of lipid-synthesizing enzymes are reflected in distinct lipid profiles in breast cancer cells. The observed differences in lipid profiles of the breast epithelial cell line and its subline may provide an insight into membrane changes associated with EMT and help evaluate lipidomes of tumor samples from patients in terms of invasive potential. Finally, by comparing breast cancer cells with different receptor profiles and invasive potential, as well as analyzing cancer vs. non-malignant breast epithelial cells we hope to identify lipidome patterns that could enhance our understanding of aberrations in lipid metabolism in cancer, have diagnostic potential and indicate therapeutic targets.
References & Acknowledgements:
University of Iceland Doctoral Fund
University of Iceland Research Fund
IP Royalty: no
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