Gαi2 Signaling Regulates Inflammasome Priming and Cytokine Production by Biasing Macrophage Phenotype Determination

Abstract

Macrophages exist as innate immune subsets that exhibit phenotypic heterogeneity and functional plasticity. Their phenotypes are dictated by inputs from the tissue microenvironment. G-protein-coupled receptors are essential in transducing signals from the microenvironment, and heterotrimeric Gα signaling links these receptors to downstream effectors. Several Gα_i-coupled G-protein-coupled receptors have been implicated in macrophage polarization. In this study, we use genetically modified mice to investigate the role of Gα_i2 on inflammasome activity and macrophage polarization. We report that Gα_i2 in murine bone marrow-derived macrophages (BMDMs) regulates IL-1β release after activation of the NLRP3, AIM2, and NLRC4 inflammasomes. We show this regulation stems from the biased polarity of Gα_i2 deficient (<i>Gnai2</i> ^-/-) and RGS-insensitive Gα_i2 (<i>Gnai2</i> ^G184S/G184S) BMDMs. We determined that although <i>Gnai2</i> ^G184S/G184S BMDMs (excess Gα_i2 signaling) have a tendency toward classically activated proinflammatory (M1) phenotype, <i>Gnai2^-/-</i> BMDMs (Gα_i2 deficient) are biased toward alternatively activated anti-inflammatory (M2) phenotype. Finally, we find that Gα_i2-deficient macrophages have increased Akt activation and IFN-β production but defects in ERK1/2 and STAT3 activation after LPS stimulation. Gα_i2-deficient macrophages also exhibit increased STAT6 activation after IL-4 stimulation. In summary, our data indicates that excess Gα_i2 signaling promotes an M1 macrophage phenotype, whereas Gα_i2 signaling deficiency promotes an M2 phenotype. Understanding Gα_i2-mediated effects on macrophage polarization may bring to light insights regarding disease pathogenesis and the reprogramming of macrophages for the development of novel therapeutics.