title: Intestinal dysbiosis and bacterial enteroinvasion in a murine model of Hirschsprung's disease.
abstract: OBJECTIVE
  Hirschsprung's disease (HSCR), characterized by the absence of ganglia in the distal colon, results in functional obstruction. Despite surgical resection of the aganglionic segment, around 40% of patients suffer recurrent life threatening Hirschsprung's-associated enterocolitis (HAEC). The aim of this study was to investigate whether gut microbiota and intestinal immunity changes contribute to the HAEC risk in an HSCR model.
  
  
  METHODS
  Mice with neural crest conditional deletion of Endothelin receptor B (EdnrB) and their littermate controls were used (EdnrB-null and EdnrB-het). Bacterial DNA was prepared from cecal contents of P16-18 and P21-24 animals and pyrosequencing employed for microbiome analysis. Ileal tissue was isolated and secretory phospholipase A2 (sPLA2) expression and activity determined. Enteroinvasion of Escherichia coli into ileal explants was measured using an ex vivo organ culture system.
  
  
  RESULTS
  EdnrB-het and EdnrB-nulls displayed similar flora, sPLA2 expression and activity at P16-18. However, by P21-24, EdnrB-hets demonstrated increased Lactobacillus and decreased Bacteroides and Clostridium, while EdnrB-nulls exhibited reciprocal changes. EdnrB-nulls also showed reduced sPLA2 expression and luminal activity at this stage. Functionally, EdnrB-nulls were more susceptible to enteroinvasion with E. coli ex vivo and released less sPLA2 than EdnrB-hets.
  
  
  CONCLUSIONS
  Initially, EdnrB-het and EdnrB-nulls contain similar cecal flora but then undergo reciprocal changes. EdnrB-nulls display dysbiosis, demonstrate impaired mucosal defense, decreased luminal sPLA2 and increased enteroinvasion of E. coli just prior to robust colonic inflammation and death. These findings suggest a role for the intestinal microbiome in the development of HAEC.
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Joseph F
authors.initials: JF
authors.lastname: Pierre
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Barlow-Anacker
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Erickson
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Aaron F
authors.initials: AF
authors.lastname: Heneghan
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Glen E
authors.initials: GE
authors.lastname: Leverson
authors.affiliation: Research and Testing Laboratory, Lubbock, TX, USA.
authors.forename: Scot E
authors.initials: SE
authors.lastname: Dowd
authors.affiliation: Department of Neuroscience, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
authors.forename: Miles L
authors.initials: ML
authors.lastname: Epstein
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA; Veteran Administration Surgical Service, William S. Middleton Memorial Veterans Hospital, Madison, WI, USA.
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Kudsk
authors.affiliation: Department of Surgery, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA; Department of Neuroscience, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA. Electronic address: gosain@surgery.wisc.edu.
authors.forename: Ankush
authors.initials: A
authors.lastname: Gosain
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis; D004195:Disease Models, Animal; D064806:Dysbiosis / Q000175:diagnosis / Q000209:etiology* / Q000276:immunology; D004760:Enterocolitis / Q000175:diagnosis / Q000209:etiology* / Q000276:immunology; D006627:Hirschsprung Disease / Q000150:complications* / Q000175:diagnosis / Q000276:immunology; D007111:Immunity, Cellular; D007413:Intestinal Mucosa / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D007422:Intestines / Q000382:microbiology*; D051379:Mice; D008822:Mice, Transgenic; D012307:Risk Factors
chemical_list: D004269:DNA, Bacterial
keywords: Dysbiosis; Hirschsprung’s disease; Hirschsprung’s-associated enterocolitis; Microbiome; Mucosal immunity; Secretory phospholipase A2
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