2. Academic Validation
  3. The human milk oligosaccharides 2'-fucosyllactose and 6'-sialyllactose protect against the development of necrotizing enterocolitis by inhibiting toll-like receptor 4 signaling

The human milk oligosaccharides 2'-fucosyllactose and 6'-sialyllactose protect against the development of necrotizing enterocolitis by inhibiting toll-like receptor 4 signaling

  • Pediatr Res. 2021 Jan;89(1):91-101. doi: 10.1038/s41390-020-0852-3.
Chhinder P Sodhi 1 2,  Peter Wipf 3,  Yukihiro Yamaguchi 1 2,  William B Fulton 1 2,  Mark Kovler 1 2,  Diego F Niño 1 2,  Qinjie Zhou 1 2,  Emilyn Banfield 1,  Adam D Werts 3,  Mitchell R Ladd 1 2,  Rachael H Buck 4,  Karen C Goehring 4,  Thomas Prindle Jr 1 2,  Sanxia Wang 1 2,  Hongpeng Jia 1 2,  Peng Lu 1 2,  David J Hackam 5 6
Affiliations

Affiliations

  • 1 Division of General Pediatric Surgery, Johns Hopkins University and Johns Hopkins Children's Center, Baltimore, MD, 21287, USA.
  • 2 Department of Surgery, Johns Hopkins University and Johns Hopkins Children's Center, Baltimore, MD, 21287, USA.
  • 3 Department of Chemistry, University of Pittsburgh, Pittsburgh, PA, USA.
  • 4 Abbott Nutrition, a Division of Abbott Laboratories, Columbus, OH, USA.
  • 5 Division of General Pediatric Surgery, Johns Hopkins University and Johns Hopkins Children's Center, Baltimore, MD, 21287, USA. [email protected].
  • 6 Department of Surgery, Johns Hopkins University and Johns Hopkins Children's Center, Baltimore, MD, 21287, USA. [email protected].
PMID: 32221473 DOI: 10.1038/s41390-020-0852-3
Abstract

Background: Necrotizing enterocolitis (NEC) develops through exaggerated Toll-like Receptor 4 (TLR4) signaling in the intestinal epithelium. Breast milk is rich in non-digestible oligosaccharides and prevents NEC through unclear mechanisms. We now hypothesize that the human milk oligosaccharides 2'-fucosyllactose (2'-FL) and 6'-sialyllactose (6'-SL) can reduce NEC through inhibition of TLR4 signaling.

Methods: NEC was induced in newborn mice and premature piglets and infant formula was supplemented with 2'-FL, 6'-SL, or lactose. Intestinal tissue was obtained at surgical resection. HMO inhibition of TLR4 was assessed in IEC-6 enterocytes, mice, and human tissue explants and via in silico modeling.

Results: Supplementation of infant formula with either 2'-FL and/or 6'-SL, but not the parent sugar lactose, reduced NEC in mice and piglets via reduced Apoptosis, inflammation, weight loss, and histological appearance. Mechanistically, both 2'-FL and 6'-SL, but not lactose, reduced TLR4-mediated nuclear factor kappa light-chain enhancer of activated B cells (NF-kB) inflammatory signaling in the mouse and human intestine. Strikingly, in silico modeling revealed 2'-FL and 6'-SL, but not lactose, to DOCK into the binding pocket of the TLR4-MD2 complex, explaining their ability to inhibit TLR4 signaling.

Conclusions: 2'-FL and 6'-SL, but not lactose, prevent NEC in mice and piglet models and attenuate NEC inflammation in the human ileum, in part through TLR4 inhibition.

Impact: Necrotizing enterocolitis (NEC) is a major cause of morbidity and mortality in premature infants that occurs in the setting of Bacterial colonization of the gut and administration of formula feeds and activation by the innate immune receptor Toll-like Receptor 4 (TLR4). Breast milk prevents NEC through unclear mechanisms. We now show that breast milk-enriched human milk oligosaccharides (HMOs) that are derived from lactose prevent NEC through inhibition of TLR4. The human milk oligosaccharides 2'-FL and 6'-SL, but not the backbone sugar lactose, prevent NEC in mice and piglets. 2'-FL and 6'-SL but not lactose inhibited TLR4 signaling in cultured enterocytes, in enteroids derived from mouse intestine, and in human intestinal explants obtained at the time of surgical resection for patients with NEC. In seeking the mechanisms involved, 2'-FL and 6'-SL but not lactose were found to directly bind to TLR4, explaining the inhibition and protection against NEC. These findings may impact clinical practice by suggesting that administration of HMOs could serve as a preventive strategy for premature infants at risk for NEC development.

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