Differential involvement of LUBAC‐mediated linear ubiquitination in intestinal epithelial cells and macrophages during intestinal inflammation

Abstract Disruption of the intestinal epithelial barrier and dysregulation of macrophages are major factors contributing to the pathogenesis of inflammatory bowel diseases (IBDs). Activation of NF‐κB and cell death are involved in maintaining intestinal homeostasis in a cell type‐dependent manner. Although both are regulated by linear ubiquitin chain assembly complex (LUBAC)‐mediated linear ubiquitination, the physiological relevance of linear ubiquitination to intestinal inflammation remains unexplored. Here, we used two experimental mouse models of IBD (intraperitoneal LPS and oral dextran sodium sulfate [DSS] administration) to examine the role of linear ubiquitination in intestinal epithelial cells (IECs) and macrophages during intestinal inflammation. We did this by deleting the linear ubiquitination activity of LUBAC specifically from IECs or macrophages. Upon LPS administration, loss of ligase activity in IECs induced mucosal inflammation and augmented IEC death. LPS‐mediated death of LUBAC‐defective IECs was triggered by TNF. IEC death was rescued by an anti‐TNF antibody, and TNF (but not LPS) induced apoptosis of organoids derived from LUBAC‐defective IECs. However, augmented TNF‐mediated IEC death did not overtly affect the severity of colitis after DSS administration. By contrast, defective LUBAC ligase activity in macrophages ameliorated DSS‐induced colitis by attenuating both infiltration of macrophages and expression of inflammatory cytokines. Decreased production of macrophage chemoattractant MCP‐1/CCL2, as well as pro‐inflammatory IL‐6 and TNF, occurred through impaired activation of NF‐κB and ERK via loss of ligase activity in macrophages. Taken together, these results indicate that both intraperitoneal LPS and oral DSS administrations are beneficial for evaluating epithelial integrity under inflammatory conditions, as well as macrophage functions in the event of an epithelial barrier breach. The data clarify the cell‐specific roles of linear ubiquitination as a critical regulator of TNF‐mediated epithelial integrity and macrophage pro‐inflammatory responses during intestinal inflammation. © 2022 The Pathological Society of Great Britain and Ireland.