The Human G Protein-Coupled ATP Receptor P2Y11 Is Associated With IL-10 Driven Macrophage Differentiation

Abstract

The G protein-coupled P2Y 11 receptor is known to sense extracellular ATP during inflammatory and immune responses. The dinucleotide NAD + has also been proposed to be a P2Y 11 receptor ligand but its role is less clear. Here, we have examined for the first time human P2Y 11 receptor protein levels and show that the receptor was upregulated during polarization of M2 macrophages. IL-10 reinforced P2Y 11 receptor expression during differentiation of M2c macrophages expressing CD163, CD16, and CD274 (PD-L1). Nutlin-3a mediated p53 stabilization further increased P2Y 11 receptor, CD16, and PD-L1 expression. AMP-activated kinase (AMPK), which mediates anti-inflammatory effects of IL-10, and nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme of the NAD + salvage pathway, which is under the control of AMPK, were also required for P2Y 11 receptor expression. The P2Y 11 receptor agonist ATPγS and NAD + could independently stimulate the production of IL-8 in M2 macrophages, however, only the ATPγS-induced response was mediated by P2Y 11 receptor. Both in a recombinant system and in macrophages, P2Y 11 receptor-driven IL-8 production predominantly depended on IkB kinase (IKK), and extracellular signal–regulated kinase (ERK). In conclusion, our data indicate that an AMPK-NAMPT-NAD + signaling axis promotes P2Y 11 receptor expression during M2 polarization of human macrophages in response to IL-10. PD-L1 expressing M2c macrophages that secrete the cancer-promoting chemokine IL-8 in response to P2Y 11 receptor stimulation may represent an important target in checkpoint blockade immunotherapy.

The human G protein‐coupled ATP receptor P2Y11 is a target for anti‐inflammatory strategies

“…While IL-1 receptor signalling was enhanced, TLR4 responses appeared to be suppressed. We have recently shown that P2Y 11 receptor-driven IL-8 secretion requires IKK and ERK as critical signalling components, both in the ectopic expression system and in primary human M2 macrophages (Gruenbacher et al, 2019). Our current observation that P2Y 11 receptors engage the IL-1 receptor signalling is in accordance with our previous findings, because IL-1 receptor responses likewise depend on IKK and ERK ( Figure S1) (Weber et al, 2010).…”

“…We have previously shown that P2Y 11 receptor expression is upregulated during human M2 macrophage differentiation (Gruenbacher et al, 2019), suggesting that expression of these receptors contributes to the anti-inflammatory function of this macrophage subset. To extend these studies, we have now examined ectopic P2Y 11 receptors in a recombinant cell system and native P2Y 11 receptors in human monocyte-derived M2 macrophages.…”

“…ATPγS (Sigma Aldrich, St. Louis, MO, USA) and NF546 (Tocris, Bristol, UK) were used as P2Y 11 receptor agonists (10 to 30 μM). NF340 (Santa Cruz, Dallas, TX, USA) served as P2Y 11 receptor antagonist (10 μM)(Gruenbacher et al, 2019). Additional reagents used in this study are as follows: P2X1 receptor antagonist NF449 (10 μM)(Tocris), adenylyl cyclase inhibitor SQ22536 (5 to 20 μM) (Tocris), Gq/11 inhibitor YM-254890 (0.1 to 0.5 μM) (Adipogen Life Sciences, Liestal, Switzerland), recombinant IL-1α and IL-1ß (50 to 1,000 pgÁml −1 ) (R&D, Bio-Techne), recombinant IL-1 receptor antagonist (IRAP; 0.05 to 0.2 μgÁml −1 ) (R&D, Bio-techne); nonselective phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) (20 to 200 μM) (Sigma-Aldrich); PDE4-selective inhibitor rolipram…”

Enhancement of Mast Cell Degranulation Mediated by Purinergic Receptor Activation and PI3K(δ)

“…The present data indicate that P2Y11R is linked to degranulation responses in LAD2 cells. This conclusion is based on (i) the expression of P2Y11 (Supplemental Figure 1), (ii) the lack of enhancing effect of purines and pyrimidines that target the other P2YR expressed by LAD2 cells, (iii) the strong enhancing effects of ATPγS, an agonist at the P2Y11R, [25][26][27] and (iv) the marked inhibition of ATPγS' effects by NF157, a selective P2Y11R antagonist. 28,29 Mechanistically, the P2Y11R effects involve the PI3K/Akt pathway but not the induction of intracellular Ca 2+ mobilization following WAS triggering.…”

Metabolic reprogramming of macrophages and its involvement in inflammatory diseases

“…Additionally, AMPK regulates nicotinamide phosphoribosyl transferase (NAMPT). NAMPT is the rate-limiting enzyme in the NAD+ salvage pathway and is associated with the activation of anti-inflammatory signaling (Audrito et al, 2015[ 2 ]; Gruenbacher et al, 2019[ 22 ]). IL-10 is an essential effector molecule of this process.…”