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Necrotizing enterocolitis: specific human milk oligosaccharides prevent enteric glia loss and hypomotility

Pediatric Research (2025)Cite this article

Abstract

Background

Necrotizing enterocolitis (NEC) is mediated by toll-like receptor 4 (TLR4)-induced inflammation and is preceded by reduced intestinal motility. Human milk oligosaccharides (HMOs) are non-digestible components of breast milk that prevent NEC in preclinical models. We now hypothesize that HMOs can reduce the risk of NEC through restoration of intestinal motility and reduced TLR4-mediated inflammation.

Methods

NEC was induced in C57-BL/6 mice through the combination of formula gavage, hypoxia, and oral administration of NEC stool. Mice were administered either 2’-FL (5 g/L), 6’-SL (5 g/L), or a blend of 5 specific HMOs (5 g/L) containing 2’-FL (2.606 g/L), 3’-FL (0.652 g/L), LNT (1.304 g/L), 3’-SL (0.174 g/L), and 6’-SL (0.260 g/L). Gastrointestinal motility was assessed by 70 Kd FITC-dextran transit time. Enteric glia were quantified by immunohistochemistry and qRT-PCR expression.

Results

Administration of either 2’-FL, 6’-SL, or HMO blend significantly attenuated NEC severity and reversed intestinal hypomotility. HMOs prevented enteric glia loss and regulated key genes critical for enteric glia maintenance, attenuated pro-apoptotic genes, and increased anti-apoptotic genes in vitro, resulting in a reduction in apoptosis. Strikingly, HMOs reduced LPS-TLR4-induced NFκB signaling and ROS generation in enteric glia.

Conclusions

HMOs protect against NEC at least in part through protective effects on inflammation and the enteric nervous system.

Impact

  • This study sheds light on the role of certain human milk oligosaccharides in a clinically relevant mouse model of NEC and adds additional insights into their underlying mechanism of action by revealing a protective effect on the enteric nervous system.

  • These results reveal that HMOs prevent the loss of enteric glia in NEC and influence the expression of genes that regulate enteric glia maintenance.

  • HMOs also limit TLR4-NFkB signaling, providing an additional mechanism of enteric glia maintenance.

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Fig. 1: Administration of human milk oligosaccharides (HMOs) protects against NEC development in neonatal mice.
Fig. 2: Administration of human milk oligosaccharides (HMOs) rescues NEC-induced intestinal hypomotility in neonatal mice.
Fig. 3: Administration of human milk oligosaccharides (HMOs) prevents NEC-induced enteric glia and Bdnf loss in neonatal mice.
Fig. 4: Administration of human milk oligosaccharides (HMOs) prevents NEC-induced loss of prominent enteric neuronal signaling genes in neonatal mice.
Fig. 5: Administration of human milk oligosaccharides (HMOs) inhibits LPS-induced NF-κB translocation and reactive oxygen species (ROS) generation in cultured enteric glial cells (EGCs).
Fig. 6: Administration of human milk oligosaccharides (HMOs) prevents LPS-induced apoptosis and rescues LPS-induced cell proliferation loss in cultured enteric glial cells (EGCs).

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Data availability

All materials described in the manuscript, including all relevant raw data, are freely available to any researcher wishing to use them for non-commercial purposes after appropriate material transfer agreements.

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Funding

This study was supported in part by a sponsored research grant from Abbott Nutrition. D.J.H. is supported by R35 GM141956, and C.T., and D.S. are supported by T32DK007713.

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Authors and Affiliations

  1. Division of General Pediatric Surgery and Department of Surgery, Johns Hopkins University and Johns Hopkins Children’s Center, Baltimore, MD, USA

    Chhinder P. Sodhi, Daniel J. Scheese, Cody Tragesser, William B. Fulton, Johannes W. Duess, Koichi Tsuboi, Maame Efua S. Sampah, Thomas Prindle, Sanxia Wang, Menghan Wang & David J. Hackam

  2. Nutrition Division, Abbott, Columbus, OH, USA

    Rachael H. Buck, David R. Hill & Anice Sabag-Daigle

Authors
  1. Chhinder P. Sodhi
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  2. Daniel J. Scheese
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Contributions

D.J.H., C.P.S., D.J.S., C.T., W.B.F., J.W.D., K.T., M.E.S., R.H.B., D.R.H., A.S., T.P., S.W., and M.W. made substantial contributions to conception and design, acquisition of data, or analysis and interpretation of data. D.J.H., C.P.S., R.H.B., D.R.H., and A.S. drafted the article or revised it critically for important intellectual content, and all authors gave their final approval of the version to be published.

Corresponding authors

Correspondence to Chhinder P. Sodhi or David J. Hackam.

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Competing interests

R.H.B., D.R.H., and A.S.-D. are employees of Abbott, Nutrition Division, which supplied the products used in the current study and which provided funding for the current study.

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Sodhi, C.P., Scheese, D.J., Tragesser, C. et al. Necrotizing enterocolitis: specific human milk oligosaccharides prevent enteric glia loss and hypomotility. Pediatr Res (2025). https://doi.org/10.1038/s41390-025-04077-y

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