Intestinal dysbiosis and bacterial enteroinvasion in a murine model of Hirschsprung's disease

Joseph Pierre, Amanda J. Barlow-Anacker, Christopher S. Erickson, Aaron F. Heneghan, Glen E. Leverson, Scot E. Dowd, Miles L. Epstein, Kenneth A. Kudsk, Ankush Gosain

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Background/purpose 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.

Original languageEnglish (US)
Pages (from-to)1242-1251
Number of pages10
JournalJournal of pediatric surgery
Volume49
Issue number8
DOIs
StatePublished - Jan 1 2014

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Dysbiosis
Endothelin B Receptors
Hirschsprung Disease
Secretory Phospholipase A2
Enterocolitis
Escherichia coli
Phospholipase A2 Receptors
Endothelin Receptors
Bacterial DNA
Bacteroides
Clostridium
Neural Crest
Organ Culture Techniques
Microbiota
Lactobacillus
Ganglia

All Science Journal Classification (ASJC) codes

  • Surgery
  • Pediatrics, Perinatology, and Child Health

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Intestinal dysbiosis and bacterial enteroinvasion in a murine model of Hirschsprung's disease. / Pierre, Joseph; Barlow-Anacker, Amanda J.; Erickson, Christopher S.; Heneghan, Aaron F.; Leverson, Glen E.; Dowd, Scot E.; Epstein, Miles L.; Kudsk, Kenneth A.; Gosain, Ankush.

In: Journal of pediatric surgery, Vol. 49, No. 8, 01.01.2014, p. 1242-1251.

Research output: Contribution to journalArticle

Pierre, J, Barlow-Anacker, AJ, Erickson, CS, Heneghan, AF, Leverson, GE, Dowd, SE, Epstein, ML, Kudsk, KA & Gosain, A 2014, 'Intestinal dysbiosis and bacterial enteroinvasion in a murine model of Hirschsprung's disease', Journal of pediatric surgery, vol. 49, no. 8, pp. 1242-1251. https://doi.org/10.1016/j.jpedsurg.2014.01.060
Pierre, Joseph ; Barlow-Anacker, Amanda J. ; Erickson, Christopher S. ; Heneghan, Aaron F. ; Leverson, Glen E. ; Dowd, Scot E. ; Epstein, Miles L. ; Kudsk, Kenneth A. ; Gosain, Ankush. / Intestinal dysbiosis and bacterial enteroinvasion in a murine model of Hirschsprung's disease. In: Journal of pediatric surgery. 2014 ; Vol. 49, No. 8. pp. 1242-1251.
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abstract = "Background/purpose 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.",
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AU - Pierre, Joseph

AU - Barlow-Anacker, Amanda J.

AU - Erickson, Christopher S.

AU - Heneghan, Aaron F.

AU - Leverson, Glen E.

AU - Dowd, Scot E.

AU - Epstein, Miles L.

AU - Kudsk, Kenneth A.

AU - Gosain, Ankush

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N2 - Background/purpose 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.

AB - Background/purpose 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.

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