The human intestine is home to a continuous balancing act between the host immune response, the large population of resident bacteria, and the thin epithelial layer that separates them. Disruptions in this fine balance lead to intestinal inflammation, a significant cause of morbidity and mortality worldwide. The idiopathic inflammatory bowel diseases (IBD) impose a significant health and monetary burden in the developed world, with roughly 1 in 200 people in the US affected. In the developing world, infection with microbial pathogens leads to about two billion cases of diarrheal disease annually and 1.5 million deaths, primarily among children. Diarrheal pathogens such as Salmonella Typhimurium cause inflammatory diarrhea that mimics several clinical hallmarks of IBD including massive neutrophil infiltration into the intestine. <p/>Understanding the shared mechanisms that drive neutrophil infiltration during disease is therefore of critical importance inpublic health. Hepoxilin A3 (HXA3) is a lipid secreted from the apical surface of Salmonella- infected epithelial cells that has been identified as a crucial and specific mediator of neutrophiltransepithelial migration in the intestine. HXA3 also drives neutrophil infiltration during intestial inflammation triggered by a variety of stimuli as well as during lung inflammation. In addition, wehave recently identified an activity secreted by uninfected epithelial cells that inhibits HXA3-mediated migration, which we refer to as AMEND. <p/>We hypothesize that the balance between AMEND and HXA3 activity in the intestine regulates the homeostatic set point that must be overcome for the induction of intestinal inflammation. In this application we seek to further understand the interaction between HXA3 and neutrophils, and to investigate the mechanisms by which AMEND regulates HXA3 activity during homeostasis and disease. We will integrate findings in vitro and in vivo to define precise mechanisms of HXA3 and AMEND function during Salmonella infection, and will broaden the applicability of these findings by investigating the regulation of HXA3 activity by AMEND in DSS colitis and T cell transfer, two established mouse models of non-infectious colitis. Identification of the HXA3 receptor will provide a major conceptual advance in the field and identify a potential therapeutic target in inflammatory disease. Similarly, identifying the mechanism(s) by which AMEND regulates HXA3-mediated inflammation will highlight pathways to target for therapeutic intervention. Current therapies for IBD suffer from damaging sequelae and an inability to prevent relapses, with surgical intervention eventually required in 35-40% of ulcerative colitis patients and 70-80% of Crohn's disease patients. <p/> Targeting local inflammatory responses rather than systemic therapies may help to reduce damaging sequelae, and directing therapies to block the early steps in initiation of inflammation may have better outcomes in preventing relapse. Furthermore, the studies proposed here will contribute greatly to understanding the basic biology of the epithelium and its ability to control neutrophil recruitment, opening up further research avenues in intestinal biology.
PUBLIC HEALTH RELEVANCE: It is estimated that as many as one million Americans have inflammatory bowel disease (IBD) with that number evenly split between Crohn's disease and ulcerative colitis. Males and females appear to be affected equally. IBDs including Crohn's disease and ulcerative colitis are chronic recurrent inflammatory disorders of the gastrointestinal tract that are of unkown etiology. Although the exact pathogenesis of IBDs is poorly understood, evidence indicates that it involves interaction between the immune system, genetic susceptibility, and the environment. These diseases are characterized by a pronounced infiltration of neutrophils (PMNs) into colon mucosal lesions, accompanied by epithelial cell necrosis and ulceration. Moreover, disease activity and patient symptoms correlate with the histologic finding of PMN infiltrates within the mucosa and surface epithelium. Curiously, this same histological event of massive transepithelial migration of PMNs occurs in the acute phase of gastroenteritis induced by Salmonella enteritis serovar Typhimurium (salmonellosis). Thus, while the evolution of these responses most likely initially targeted pathogens such as S. typhimurium, perhaps aberrant activation of such pathways also occurs, leading to inappropriate states of active intestinal inflammation. <p/>The goal of this proposal is to combine models of S. typhimurium induced neutrophil migration with in vivo models of idiopathic IBD to elucidate mechanisms that regulate neutrophil infiltration into the intestine in health and disease so that novel targets for therapeutic intervention in such mucosal inflammatory diseases can be identified.