LAUSR

In cancer, G protein-coupled receptors (GPCRs) are involved in tumor progression and metastasis. In this study we particularly examined one GPCR, the adenosine A2B receptor. This receptor is activated by high concentrations of its endogenous ligand adenosine, which suppresses the immune response to fight tumor progression. A series of adenosine A2B receptor mutations were retrieved from the Cancer Genome Atlas harboring data from patient samples with different cancer types. The main goal of this work was to investigate the pharmacology of these mutant receptors using a 'single-GPCR-one-G protein' yeast assay technology. Concentration-growth curves were obtained with the full agonist NECA for the wild-type receptor and 15 mutants. Compared to wild-type receptor, the constitutive activity levels in mutant receptors F141 L4.61, Y202C5.58 and L310 P8.63 were high, while the potency and efficacy of NECA and BAY 60-6583 on Y202C5.58 was lower. A 33- and 26-fold higher constitutive activity on F141 L4.61 and L310 P8.63 was reduced to wild-type levels in response to the inverse agonist ZM241385. These constitutively active mutants may thus be tumor promoting. Mutant receptors F259 S6.60 and Y113 F34.53 showed a more than one log-unit decrease in potency. A complete loss of activation was observed in mutant receptors C29 R1.54, W130C4.50 and P249 L6.50. All mutations were characterized at the structural level, generating hypotheses of their roles on modulating the receptor conformational equilibrium. Taken together, this study is the first to investigate the nature of adenosine A2B receptor cancer mutations and may thus provide insights in mutant receptor function in cancer.