Development and use of advanced mass spectrometry techniques for the characterization of cellular and mitochondrial lipidomic profiling in control fibroblasts and Parkinson's disease patients

摘要

Lipidomics (a lipid-targeted metabolomics) aims at global analysis of lipids in biological systems. Recently, lipidomics research has received increased attention due to the well- recognized role of lipids in numerous human diseases. For instance, altered lipid pathways in the primary visual cortex and the anterior cingulate have been demonstrated in Parkinson’s disease (PD) by analyzing post-mortem tissues from patients in advanced neuronal degeneration stage. Such an approach, however, hinders the identification of the first neuronal changes. Skin fibroblasts have been recently proposed as a useful model of primary human cells, capable of reflecting the chronological and biological aging of the patients, according to their polygenic predisposition and environmental etiopathology. Here, hydrophilic interaction liquid chromatography coupled to electrospray ionization-Fourier- transform mass spectrometry was developed to characterize polar lipids occurring in human skin fibroblasts. Different lipid extraction protocols were tested, and Bligh Dyer strategy was selected as the most informative in terms of lipid extracted. Thus, single and tandem MS measurements were performed on a hybrid quadrupole-Orbitrap mass spectrometer for the characterization of fibroblast membrane lipids with the aim to apply this strategy for successive biomarker discover in PD patients.