NTPDase8 protects mice from intestinal inflammation by limiting P2Y6 receptor activation: identification of a new pathway of inflammation for the potential treatment of IBD

• Published in: Gut Vol. 71; no. 1; pp. 43 - 54
• Main Authors: Salem, Mabrouka, Lecka, Joanna, Pelletier, Julie, Gomes Marconato, Danielle, Dumas, Aline, Vallières, Luc, Brochu, Gaetan, Robaye, Bernard, Jobin, Christian, Sévigny, Jean
• Format: Journal Article
• Language: English
• Published: London BMJ Publishing Group Ltd and British Society of Gastroenterology 01.01.2022; BMJ Publishing Group LTD
• Subjects: Antibodies; Apoptosis; Bone marrow; Bone marrow transplantation; Colitis; Colon; Cytokines; Dextran; Drinking water; Ectonucleotidase; Enzymes; Epithelial cells; Epithelium; gastrointestinal immune response; Immunohistochemistry; Inflammation; Inflammatory bowel disease; Inflammatory bowel diseases; intestinal epithelium; Intestine; Nucleotides; Pathogens; Purine P2 receptors; Purine P2Y receptors; Receptor mechanisms; RNA-directed DNA polymerase; Sodium sulfate; Canada; California; United States > US; Massachusetts; Quebec Canada
• ISSN: 0017-5749; 1468-3288
• DOI: 10.1136/gutjnl-2020-320937

ObjectiveNucleotides are danger signals that activate inflammatory responses via binding P2 receptors. The nucleoside triphosphate diphosphohydrolase-8 (NTPDase8) is an ectonucleotidase that hydrolyses P2 receptor ligands. We investigated the role of NTPDase8 in intestinal inflammation.DesignWe generated NTPDase8-deficient (Entpd8 –/–) mice to define the role of NTPDase8 in the dextran sodium sulfate (DSS) colitis model. To assess inflammation, colons were collected and analysed by histopathology, reverse transcriptase-quantitative real-time PCR (RT-qPCR) and immunohistochemistry. P2 receptor expression was analysed by RT-qPCR on primary intestinal epithelium and NTPDase8 activity by histochemistry. The role of intestinal P2Y6 receptors was assessed by bone marrow transplantation experiments and with a P2Y6 receptor antagonist.ResultsNTPDase8 is the dominant enzyme responsible for the hydrolysis of nucleotides in the lumen of the colon. Compared with wild-type (WT) control mice, the colon of Entpd8 –/– mice treated with DSS displayed significantly more histological damage, immune cell infiltration, apoptosis and increased expression of several proinflammatory cytokines. P2Y6 was the dominant P2Y receptor expressed at the mRNA level by the colonic epithelia. Irradiated P2ry6 –/– mice transplanted with WT bone marrow were fully protected from DSS-induced intestinal inflammation. In agreement, the daily intrarectal injection of a P2Y6 antagonist protected mice from DSS-induced intestinal inflammation in a dose-dependent manner. Finally, human intestinal epithelial cells express NTPDase8 and P2Y6 similarly as in mice.ConclusionNTPDase8 protects the intestine from inflammation most probably by limiting the activation of P2Y6 receptors in colonic epithelial cells. This may provide a novel therapeutic strategy for the treatment of inflammatory bowel disease.