Inflammatory bowel diseases (IBDs) including Crohn's disease (CD) and ulcerative colitis (UC) are chronic diseases characterized by a pronounced infiltration of neutrophils (PMNs) into colonic mucosal lesions, accompanied by epithelial cell necrosis and ulceration. Disease activity and patient symptoms also correlate with the histologic finding of PMN infiltrates within the mucosa and surface epithelium. This same histological event of massive transepithelial migration of PMNs occurs in the acute phase of gastroenteritis induced by Salmonella enterica serovar Typhimurium (salmonellosis). Thus, insight into pathogen elicited active inflammation of the intestine will serve as an important model to provide valuable information relating to active inflammation in IBD. Using the enteric pathogen S. typhimurium as a tool to investigate mechanisms underlying PMN recruitment across the intestinal epithelia, we have demonstrated a novel and central role for the eicosanoid hepoxilin A3 (HXA3). HXA3 is a derivative of arachidonic acid formed from the enzymatic action of the 12-lipoxygenase (12-LO) pathway. This observation has unlocked a previous unexplored phenomenon that mechanistically explains a primary histopathologic feature of active states of intestinal inflammation. Moreover, we have recently shown that HXA3 is secreted through an efflux transport system located at the apical surface of the intestinal epithelia that involves the ABC transporter MRP2. We also revealed that induction of intestinal inflammation corresponds to a profound up-regulation of the apical expression of MRP2. Such up-regulation of MRP2 was further demonstrated in vivo in chronic murine models of IBD and human intestinal biopsy tissues from patients with active CD or UC. Thus, the primary objectives of this project are to understand the molecular mechanisms underlying HXA3 synthesis (Specific Aim 1), as well as to determine how HXA3 production is regulated (Specific Aim 2). To accomplish these aims we plan to use a combination of complementary in vitro and in vivo models coupled with state of the art cellular, molecular, and biochemical techniques. We hypothesize that understanding HXA3/MRP2 based signaling, which directs the movement of PMNs across the intestinal epithelium, will lead to the development of important therapeutics designed to help control this inflammatory event in patients with infectious, allergic, and idiopathic (IBD) colitis.

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. The goal of this proposal is to understand the molecular mechanisms by which S. typhimurium triggers and controls the directed movement of PMN across the epithelial surface so that novel targets for therapeutic intervention in such inflammatory bowel diseases can be identified.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK056754-14
Application #
8534088
Study Section
Gastrointestinal Mucosal Pathobiology Study Section (GMPB)
Program Officer
Grey, Michael J
Project Start
1999-12-01
Project End
2015-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
14
Fiscal Year
2013
Total Cost
$292,843
Indirect Cost
$114,823
Name
University of Massachusetts Medical School Worcester
Department
Genetics
Type
Schools of Medicine
DUNS #
603847393
City
Worcester
State
MA
Country
United States
Zip Code
01655
Liu, Zongcai; Meng, Fanwei; Li, Chan et al. (2014) Human umbilical cord mesenchymal stromal cells rescue mice from acetaminophen-induced acute liver failure. Cytotherapy 16:1207-19
Mallick, Emily M; Garber, John J; Vanguri, Vijay K et al. (2014) The ability of an attaching and effacing pathogen to trigger localized actin assembly contributes to virulence by promoting mucosal attachment. Cell Microbiol 16:1405-24
Agbor, Terence A; Demma, Zachary; Mrsny, Randall J et al. (2014) The oxido-reductase enzyme glutathione peroxidase 4 (GPX4) governs Salmonella Typhimurium-induced neutrophil transepithelial migration. Cell Microbiol 16:1339-53
Bhowmick, Rudra; Tin Maung, Nang H; Hurley, Bryan P et al. (2013) Systemic disease during Streptococcus pneumoniae acute lung infection requires 12-lipoxygenase-dependent inflammation. J Immunol 191:5115-23
Jandhyala, Dakshina M; Vanguri, Vijay; Boll, Erik J et al. (2013) Shiga toxin-producing Escherichia coli O104:H4: an emerging pathogen with enhanced virulence. Infect Dis Clin North Am 27:631-49
Boll, Erik J; Struve, Carsten; Sander, Anja et al. (2012) Enteroaggregative Escherichia coli promotes transepithelial migration of neutrophils through a conserved 12-lipoxygenase pathway. Cell Microbiol 14:120-32
Maldonado-Contreras, A L; McCormick, Beth A (2011) Intestinal epithelial cells and their role in innate mucosal immunity. Cell Tissue Res 343:5-12
Hurley, Bryan P; Pirzai, Waheed; Mumy, Karen L et al. (2011) Selective eicosanoid-generating capacity of cytoplasmic phospholipase A2 in Pseudomonas aeruginosa-infected epithelial cells. Am J Physiol Lung Cell Mol Physiol 300:L286-94
Srikanth, C V; Mercado-Lubo, Regino; Hallstrom, Kelly et al. (2011) Salmonella effector proteins and host-cell responses. Cell Mol Life Sci 68:3687-97
Agbor, Terence A; Demma, Zachary C; Mumy, Karen L et al. (2011) The ERM protein, ezrin, regulates neutrophil transmigration by modulating the apical localization of MRP2 in response to the SipA effector protein during Salmonella Typhimurium infection. Cell Microbiol 13:2007-21

Showing the most recent 10 out of 40 publications