Cancer-Associated Mutations of the Adenosine A2A Receptor Have Diverse Influences on Ligand Binding and Receptor Functions

Abstract

The adenosine A2A receptor (A2AAR) is a class A G-protein-coupled receptor (GPCR). It is an immune checkpoint in the tumor micro-environment and has become an emerging target for cancer treatment. In this study, we aimed to explore the effects of cancer-patient-derived A2AAR mutations on ligand binding and receptor functions. The wild-type A2AAR and 15 mutants identified by Genomic Data Commons (GDC) in human cancers were expressed in HEK293T cells. Firstly, we found that the binding affinity for agonist NECA was decreased in six mutants but increased for the V275A mutant. Mutations A165V and A265V decreased the binding affinity for antagonist ZM241385. Secondly, we found that the potency of NECA (EC50) in an impedance-based cell-morphology assay was mostly correlated with the binding affinity for the different mutants. Moreover, S132L and H278N were found to shift the A2AAR towards the inactive state. Importantly, we found that ZM241385 could not inhibit the activation of V275A and P285L stimulated by NECA. Taken together, the cancer-associated mutations of A2AAR modulated ligand binding and receptor functions. This study provides fundamental insights into the structure–activity relationship of the A2AAR and provides insights for A2AAR-related personalized treatment in cancer.

Excuse me, there is a mutant in my bioactivity soup! A comprehensive analysis of the genetic variability landscape of bioactivity databases and its effect on activity modelling

“…Variant annotation in particular is one of the key aspects that should be considered when analysing bioactivity data 23 . The same compound can have a very different bioactivity on different genetic variants of the same protein [24][25][26][27] . In fact, some compounds are explicitly designed to have differential bioactivity across variants to, for example, reduce side effects by avoiding targeting the wild-type (WT) protein in anti-cancer therapies, or to target escape variants in antibiotics or antivirals 28,29 .…”

3DDPDs: describing protein dynamics for proteochemometric bioactivity prediction. A case for (mutant) G protein-coupled receptors

“…In the last decades, the scientific community has seen an increasing interest in the dynamic aspects of GPCRs, resulting in community efforts such as the GPCRmd database, where curated GPCR MD simulations are publicly available [23]. Simultaneously, GPCR research in the context of oncological therapies is gaining momentum [24], with several in vitro studies showing how cancer-related somatic mutations affect receptor function and/or pharmacological intervention [25][26][27]. Some of the physiological effects observed in mutants have been associated with changes in receptor dynamics thanks to MD simulations [28].…”

3DDPDs: Describing protein dynamics for proteochemometric bioactivity prediction. A case for (mutant) G protein-coupled receptors

“…In the last decades, the scientific community has seen an increasing interest in the dynamic aspects of GPCRs, resulting in community efforts such as the GPCRmd database, where curated GPCR MD simulations are publicly available [22]. Simultaneously, GPCR research in the context of oncological therapies is gaining momentum [23], with a number of in vitro studies showing how cancer-related somatic mutations affect receptor function and/or pharmacological intervention [24][25][26]. Some of the physiological effects observed in mutants have been associated to changes in receptor dynamics thanks to MD simulations [27].…”