Functional and Pharmacological Characterization of Endogenous PAR2 A Study on Ligand-mediated Modulation in Diverse Human Cancer Cell Models

Protease-activated receptor 2 (PAR2) is a G protein-coupled receptor, activated by serine proteases, of which the activity has been proven to be involved in inflammatory signaling and cancer biology. While the main signaling pathways of this receptor have been studied, its broader physiological and pathological function is still only partially understood, and the current choice of pharmacological agents capable of selectively engaging or inhibiting PAR2, remains relatively limited. In this project, to assess the activity of this specific receptor, three cell lines were selected to evaluate the presence of the endogenous expression of the PAR2 and its function, by employing three distinct assays. RT-qPCR analysis was conducted to detect and quantify endogenous PAR2 mRNA levels, while a complementary label-free approach and a measurement of intracellular calcium mobilization were chosen to defy the functionality of the receptor when in contact with specific compounds, selected based on previously selected studies. By testing already known agonists and antagonists, these approaches provided a quantitative and precise insight on the expression of the PAR2 and the activity related to different compounds, which provide a foundation for further investigation of the receptor as a therapeutic target.