Atherosclerosis is one of the main factors contributing to cardiovascular diseases and remains a leading cause of global mortality. Certain critical aspects of this pathology, such as the role of post-translational modifications (PTMs) in cellular interactions and the extracellular microenvironment, remain unexplored. Investigating these aspects at the subcellular and single-cell levels could represent a significant breakthrough in the fight against this disease.

Therefore, the main objective of this project is to quantitatively describe and study the mechanisms of post-translational regulation in the different cells involved in atherosclerosis: endothelial cells, smooth muscle cells, and immune cells, within the context of disease progression.

To complete this main goal, the study of post-translational regulation at the spatiotemporal level of the global proteome, focusing on its relationship with extracellular matrix remodeling in the three main cell types present in atherosclerotic plaques, and in a co-culture model, is proposed. Additionally, the project involves examining the impact of disease progression on cellular heterogeneity and population diversity at the single-cell level for the studied cell types in the co-culture model. This includes exploring their relationship with the extracellular matrix by integrating protein expression and post-translational modification information at this level.

To achieve the proposed objectives, an in vitro cell culture and co-culture model of the different cell populations will be designed, followed by subcellular fractionation techniques, cell isolation by FACS-sorting, mass spectrometry analysis and bioinformatics analysis.

Upon completion of this project, it is expected to obtain differential proteomic profiles, PTMs, and subcellular localization for each cell type studied in response to extracellular matrix remodeling in atherosclerosis, both in culture in separate media and in co-culture. Additionally, population heterogeneity analysis will enable the generation of a proteomic map of the resulting cellular subpopulations. Finally, the information generated throughout the project will be used to create PTM libraries, which will serve as the basis for mapping proteins and peptidoforms across different cellular subpopulations and correlating them with extracellular matrix types and the progression of atherosclerosis.