Breast cancer organelle atlas : combining ESPRESSO with spatial proteomics to investigate tumor heterogeneity

Lorenzo Scipioni & Giulia Tedeschi

Breast cancer is a highly heterogeneous disease, composed by numerous subpopulations that differ in metabolic profile, metastatic potential, resistance to therapies and tumor initiation capability. To investigate this heterogeneity and the possible transition between these phenotypes, we couple ESPRESSO phenotyping with highly-multiplexed spatial proteomics, generating datasets connecting organelle properties with protein expression in 2D cells and 3D spheroids. We monitor the expression of proteins reporting on glycolysis, oxidative phosophorylation, proliferation, lipid metabolism, migration, stemness, drug resistance, chromating organization and breast cancer classification, to unravel the biological role of organelle properties. This unique spatiotemporal multiomics approach allows us to identify organelle and protein expression signature, their temporal evolution and their implication in chemoresistance and relapse.

 Illustration :

Triple negative breast cancer cells (MDA-MB-231) labeled with seven fluorescent dyes targeting DNA, mitochondria, lysosomes, endosomes, lipid droplets, Golgi apparatus and tubulin, imaged in hyperspectral confocal microscopy.