Elucidating Interactome Dynamics of the A2A Adenosine Receptor in the Presence of Polydeoxyribonucleotide

Polydeoxyribonucleotide (PDRN) is a tissue regeneration substance that resembles human DNA and is present in mammalian cells. It stimulates physiological regeneration and metabolic activity, and the effects of PDRN are attributed to the activation of Adenosine A2A receptors (ADORA2A). Several previous studies have demonstrated that PDRN activates ADORA2A, which has been shown with various models including rat fibroblasts, human umbilical vein endothelial cells (HUVECs), and the experimental autoimmune uveitis (EAU) model, resulting in the upregulation of angiogenesis and growth factor. In this work, we primarily used HUVECs to study the overall changes and dynamics of proteins related to their ADORA2A activation. We conducted selected reaction monitoring-mass spectrometry (SRM-MS), and we founded 374 proteins and 193 peptides demonstrating both upregulation and downregulation in expression. We conducted gene ontology (GO) analysis and physical network analysis and discovered novel networks and indirect interactions of proteins related to neuronal differentiation among the ADORA2A-associated protein. Finally, using the AlphaFold multimer, we were able to predict interaction sites among ADORA2A-associated proteins in the network associated with neuronal cell differentiation. Specifically, we predicted five interaction sites between ADORA2A and Neurotrophic Receptor Tyrosine Kinase 1 (NTRK1), forming the edge. Thus, we provided indicators for further research using ADORA2A present in a robust network and highlighted the potential of PDRN to impact neuronal differentiation.