title: Altered goblet cell differentiation and surface mucus properties in Hirschsprung disease.
abstract: Hirschsprung disease-associated enterocolitis (HAEC) leads to significant mortality and morbidity, but its pathogenesis remains unknown. Changes in the colonic epithelium related to goblet cells and the luminal mucus layer have been postulated to play a key role. Here we show that the colonic epithelium of both aganglionic and ganglionic segments are altered in patients and in mice with Hirschsprung disease (HSCR). Structurally, goblet cells were altered with increased goblet cell number and reduced intracellular mucins in the distal colon of biopsies from patients with HSCR. Endothelin receptor B (Ednrb) mutant mice showed increased goblet cell number and size and increased cell proliferation compared to wild-type mice in aganglionic segments, and reduced goblet cell size and number in ganglionic segments. Functionally, compared to littermates, Ednrb-/- mice showed increased transepithelial resistance, reduced stool water content and similar chloride secretion in the distal colon. Transcript levels of goblet cell differentiation factors SPDEF and Math1 were increased in the distal colon of Ednrb-/- mice. Both distal colon from Ednrb mice and biopsies from HSCR patients showed reduced Muc4 expression as compared to controls, but similar expression of Muc2. Particle tracking studies showed that mucus from Ednrb-/- mice provided a more significant barrier to diffusion of 200 nm nanoparticles as compared to wild-type mice. These results suggest that aganglionosis is associated with increased goblet cell proliferation and differentiation and subsequent altered surface mucus properties, prior to the development of inflammation in the distal colon epithelium. Restoration of normal goblet cell function and mucus layer properties in the colonic epithelium may represent a therapeutic strategy for prevention of HAEC.
authors.affiliation: Department of Gastroenterology and Nutrition, Boston Children's Hospital, Boston, Massachusetts, United States of America.
authors.forename: Jay R
authors.initials: JR
authors.lastname: Thiagarajah
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts, United States of America.
authors.forename: Hasan
authors.initials: H
authors.lastname: Yildiz
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts, United States of America.
authors.forename: Taylor
authors.initials: T
authors.lastname: Carlson
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Alyssa R
authors.initials: AR
authors.lastname: Thomas
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Casey
authors.initials: C
authors.lastname: Steiger
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Alberto
authors.initials: A
authors.lastname: Pieretti
authors.affiliation: Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Lawrence R
authors.initials: LR
authors.lastname: Zukerberg
authors.affiliation: Department of Chemical Engineering, Northeastern University, Boston, Massachusetts, United States of America.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Carrier
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
authors.forename: Allan M
authors.initials: AM
authors.lastname: Goldstein
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D001692:Biological Transport; D002452:Cell Count; D002454:Cell Differentiation; D048429:Cell Size; D003106:Colon / Q000378:metabolism / Q000473:pathology*; D005243:Feces / Q000737:chemistry; D005726:Ganglia, Parasympathetic / Q000473:pathology*; D017353:Gene Deletion; D005786:Gene Expression Regulation; D020397:Goblet Cells / Q000378:metabolism / Q000473:pathology*; D006627:Hirschsprung Disease / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D055262:Mucin-2 / Q000235:genetics / Q000378:metabolism; D055264:Mucin-4 / Q000235:genetics / Q000378:metabolism; D009093:Mucus / Q000378:metabolism*; D053758:Nanoparticles / Q000378:metabolism; D018408:Patch-Clamp Techniques; D050783:Proto-Oncogene Proteins c-ets / Q000235:genetics / Q000378:metabolism; D044023:Receptor, Endothelin B / Q000172:deficiency / Q000235:genetics*; D017466:Receptors, Endothelin / Q000235:genetics* / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D055262:Mucin-2; D055264:Mucin-4; D050783:Proto-Oncogene Proteins c-ets; D044023:Receptor, Endothelin B; D017466:Receptors, Endothelin; C500942:ATOH1 protein, human; C000588596:EDNRB protein, human; C524625:MUC2 protein, human; C524658:MUC4 protein, human; C403974:SPDEF protein, human