Medical Scientist-Programm 2021

Thema: "Dissecting the differential roles of pre-clinical adenosine ligands on adenosine receptor signaling using optical approaches"

Zusammenfassung:
Adenosine is the natural agonist for A1, A2A, A2B and A3 family of GPCRs and is implicated in inflammation, cancer and sepsis. To this end, several synthetic ligands have been developed to target these receptors and influence ensuing cell phenotypes such as cytokine production, maturation or motility in pre-clinical models. However, characterization of these ligands at the primary level of receptor signaling is lacking. Over the previous years, several optic and biochemical tools have been developed to dissect this complex signaling network and also discover new phenomena such as intracellular signaling of receptors. Such findings have shed new light on possible efficacies of agonist stimulation. With these preliminary findings in mind, I want to characterize selected pre-clinical adenosine ligands with respect to their receptor activation, G-protein coupling, GRK/arrestin mediated desensitization and internalization as well as intracellular signaling. I plan to use combinations of parental and CRISPR/Cas9-treated HEK293 cells to unravel this complexity and ultimately transfer the findings to cancer cell lines and investigate differences in signaling using endogenous receptors and signaling machinery. In cancer cell lines, I plan to investigate gene expression and connect these findings with the observed cell phenotypes.