The ATP-hydrolyzing ectoenzyme E-NTPD8 attenuates colitis through modulation of P2X4 receptor–dependent metabolism in myeloid cells

Abstract

Extracellular adenosine triphosphate (ATP) released by mucosal immune cells and by microbiota in the intestinal lumen elicits diverse immune responses that mediate the intestinal homeostasis via P2 purinergic receptors, while overactivation of ATP signaling leads to mucosal immune system disruption, which leads to pathogenesis of intestinal inflammation. In the small intestine, hydrolysis of luminal ATP by ectonucleoside triphosphate diphosphohydrolase (E-NTPD)7 in epithelial cells is essential for control of the number of T helper 17 (Th17) cells. However, the molecular mechanism by which microbiota-derived ATP in the colon is regulated remains poorly understood. Here, we show that E-NTPD8 is highly expressed in large-intestinal epithelial cells and hydrolyzes microbiota-derived luminal ATP. Compared with wild-type mice, Entpd8−/− mice develop more severe dextran sodium sulfate–induced colitis, which can be ameliorated by either the depletion of neutrophils and monocytes by injecting with anti–Gr-1 antibody or the introduction of P2rx4 deficiency into hematopoietic cells. An increased level of luminal ATP in the colon of Entpd8−/− mice promotes glycolysis in neutrophils through P2x4 receptor–dependent Ca2+ influx, which is linked to prolonged survival and elevated reactive oxygen species production in these cells. Thus, E-NTPD8 limits intestinal inflammation by controlling metabolic alteration toward glycolysis via the P2X4 receptor in myeloid cells.

Fatecode: Cell fate regulator prediction using classification autoencoder perturbation

“…Our results are consistent with Tani et al who reported that Entpd8 -/- mice have a higher number of neutrophils, monocytes, and DCs. Furthermore, hydrolysis of luminal ATP by Entpd8 prevents innate intestinal pathology 44 Fatecode also reveals Nlrp6 as one of the master regulators in neutrophil and monocyte differentiation. Cai et al showed that the number of hematopoietic stem cells and granulocyte-monocyte progenitors is reduced in Kp-infected Nlrp6 -/- mice, while the survival of matured neutrophils in bone marrow is increased 45 .…”

“…Our results are consistent with Tani et al who reported that Entpd8 -/mice have a higher number of neutrophils, monocytes, and DCs. Furthermore, hydrolysis of luminal ATP by Entpd8 prevents innate intestinal pathology 44 . Fatecode also reveals Nlrp6 as one of the master regulators in neutrophil and monocyte differentiation.…”

Exploration of the link between gut microbiota and purinergic signalling

“…ENTPDase8 is highly expressed in large intestinal epithelial cells and can regulate eATP concentration in the large intestine [35]. In fecal microbiota of P2rx4 − / − mice, the relative abundance of Bacteroidetes was greater than that in WT mice, while the relative abundance of Proteobacteria was lower [35].…”

Ion channel regulation of gut immunity

“…Among them, P2X7 purinergic receptor (P2X7R), which mediates transmembrane K + efflux and Ca 2+ influx in immune cells, engages with extracellular ATP released by mucosal immune cells and microbiota in the gut and is critically involved in intestinal homeostasis through eliciting diverse immune responses ( Cheng et al, 2021 ). Besides P2X7R, genetic manipulation of neutrophil-expressing P2X4R showed that the ATP–P2X4R signaling axis in myeloid cells causes Ca 2+ influx that promotes glycolysis in response to an increase in microbiota-derived luminal ATP, thereby regulating intestinal inflammation ( Tani et al, 2021 ; Fig. 1 ).…”