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Late Breaking Posters
Day 1, June 22（Sun.）
Room P (Maesato East, Foyer, Ocean Wing)
- 1P-LB-07
Live Single-Cell Mass Spectrometry to Study the Metabolic Mechanisms Behind Triple Negative Breast Cancer Cell Migration.
(Leiden Univeristy)
oXiaoyue Huang, Sylvia Le Devedec, Thomas Hankemeier, Ahmed Ali
TNBC metastasis is associated with altered lipid metabolism, yet lipid profiling in the context of migration has largely been overlooked. Single-cell lipidomics offers a powerful tool to investigate lipidomic changes and their link to migratory behavior in rare leader cells.
After 18 hours of post-wounding time-lapse imaging, live single cells were sampled using a 10 μm capillary from an HBSS buffer spiked with the internal standard PC(15:0/15:0) under 37°C and 5% CO₂ conditions. After backfilling with 3 μL of ionization solvent spiked with the internal standard EquiSPLASH, the single-cell samples were directly infused into a Thermo Orbitrap Exploris™ 480 for 10 min. Full-scan and tandem MS with electrospray ionization (ESI) were employed for molecular ion detection. Lipidomic data from leader cells, follower cells, and less-migrated cells were analyzed and compared against our MS/MS feature list to identify lipid metabolite differences associated with collective cell migration.
Through annotation using LipidMaps, 266 distinct m/z values were identified in single cells based on MS1 data. A comprehensive MS² feature list was generated from mass spectrometry spectra obtained from serially diluted cell lysates, with 45 lipid species confirmed at the MS² level in negative ionization mode. By comparing the highest intensities to the highest intensity of PC(15:0/15:0) in each single-cell sample, our data showed that PC(34:1) levels were relatively higher (Log2FC = 1.13, p = 0.0306) in leader cells compared to follower cells.