title: Breast milk protects against the development of necrotizing enterocolitis through inhibition of Toll-like receptor 4 in the intestinal epithelium via activation of the epidermal growth factor receptor.
abstract: Breast milk is the most effective strategy to protect infants against necrotizing enterocolitis (NEC), a devastating disease that is characterized by severe intestinal necrosis. Previous studies have demonstrated that the lipopolysaccharide receptor Toll-like receptor 4 (TLR4) plays a critical role in NEC development via deleterious effects on mucosal injury and repair. We now hypothesize that breast milk protects against NEC by inhibiting TLR4 within the intestinal epithelium, and sought to determine the mechanisms involved. Breast milk protected against NEC and reduced TLR4 signaling in wild-type neonatal mice, but not in mice lacking the epidermal growth factor receptor (EGFR), whereas selective removal of EGF from breast milk reduced its protective properties, indicating that breast milk inhibits NEC and attenuates TLR4 signaling via EGF/EGFR activation. Overexpression of TLR4 in the intestinal epithelium reversed the protective effects of breast milk. The protective effects of breast milk occurred via inhibition of enterocyte apoptosis and restoration of enterocyte proliferation. Importantly, in IEC-6 enterocytes, breast milk inhibited TLR4 signaling via inhibition of glycogen synthase kinase-3β (GSK3β). Taken together, these findings offer mechanistic insights into the protective role for breast milk in NEC, and support a link between growth factor and innate immune receptors in NEC pathogenesis.
authors.affiliation: 1] Division of Newborn Medicine, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: M
authors.initials: M
authors.lastname: Good
authors.affiliation: 1] Division of General Pediatric Surgery, Johns Hopkins University and Bloomberg Children's Center, Johns Hopkins Hospital, Baltimore, Maryland, USA [2] Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: C P
authors.initials: CP
authors.lastname: Sodhi
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Egan
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: A
authors.initials: A
authors.lastname: Afrazi
authors.affiliation: 1] Division of General Pediatric Surgery, Johns Hopkins University and Bloomberg Children's Center, Johns Hopkins Hospital, Baltimore, Maryland, USA [2] Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: H
authors.initials: H
authors.lastname: Jia
authors.affiliation: 1] Division of General Pediatric Surgery, Johns Hopkins University and Bloomberg Children's Center, Johns Hopkins Hospital, Baltimore, Maryland, USA [2] Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Yamaguchi
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: P
authors.initials: P
authors.lastname: Lu
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: M F
authors.initials: MF
authors.lastname: Branca
authors.affiliation: 1] Division of Newborn Medicine, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: C
authors.initials: C
authors.lastname: Ma
authors.affiliation: 1] Division of General Pediatric Surgery, Johns Hopkins University and Bloomberg Children's Center, Johns Hopkins Hospital, Baltimore, Maryland, USA [2] Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: T
authors.initials: T
authors.lastname: Prindle
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: S
authors.initials: S
authors.lastname: Mielo
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: A
authors.initials: A
authors.lastname: Pompa
authors.affiliation: 1] Division of Pediatric Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Hodzic
authors.affiliation: 1] Division of Pediatric Pathology, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, USA [2] Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Ozolek
authors.affiliation: 1] Division of General Pediatric Surgery, Johns Hopkins University and Bloomberg Children's Center, Johns Hopkins Hospital, Baltimore, Maryland, USA [2] Department of Surgery, Johns Hopkins University, Baltimore, Maryland, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Hackam
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D020345:Enterocolitis, Necrotizing / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000517:prevention & control*; D020895:Enterocytes / Q000276:immunology* / Q000473:pathology; D004815:Epidermal Growth Factor / Q000235:genetics / Q000276:immunology; D066246:ErbB Receptors / Q000235:genetics / Q000276:immunology; D005260:Female; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000276:immunology; D000071679:Glycogen Synthase Kinase 3 beta; D051379:Mice; D008892:Milk / Q000276:immunology*; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000276:immunology*
chemical_list: C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D004815:Epidermal Growth Factor; C541485:EGFR protein, mouse; D066246:ErbB Receptors; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D038362:Glycogen Synthase Kinase 3