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The human milk oligosaccharide 2′-fucosyllactose attenuates the severity of experimental necrotising enterocolitis by enhancing mesenteric perfusion in the neonatal intestine

  • Misty Good (a1) (a2), Chhinder P. Sodhi (a3) (a4), Yukihiro Yamaguchi (a3) (a4), Hongpeng Jia (a3) (a4), Peng Lu (a3) (a4), William B. Fulton (a3) (a4), Laura Y. Martin (a3) (a4), Thomas Prindle (a3) (a4), Diego F. Nino (a3) (a4), Qinjie Zhou (a3) (a4), Congrong Ma (a1) (a2), John A. Ozolek (a5) (a6), Rachael H. Buck (a7), Karen C. Goehring (a7) and David J. Hackam (a3) (a4)...

Abstract

Necrotising enterocolitis (NEC) is a common disease in premature infants characterised by intestinal ischaemia and necrosis. The only effective preventative strategy against NEC is the administration of breast milk, although the protective mechanisms remain unknown. We hypothesise that an abundant human milk oligosaccharide (HMO) in breast milk, 2′-fucosyllactose (2′FL), protects against NEC by enhancing intestinal mucosal blood flow, and we sought to determine the mechanisms underlying this protection. Administration of HMO-2′FL protected against NEC in neonatal wild-type mice, resulted in a decrease in pro-inflammatory markers and preserved the small intestinal mucosal architecture. These protective effects occurred via restoration of intestinal perfusion through up-regulation of the vasodilatory molecule endothelial nitric oxide synthase (eNOS), as administration of HMO-2′FL to eNOS-deficient mice or to mice that received eNOS inhibitors did not protect against NEC, and by 16S analysis HMO-2′FL affected the microbiota of the neonatal mouse gut, although these changes do not seem to be the primary mechanism of protection. Induction of eNOS by HMO-2′FL was also observed in cultured endothelial cells, providing a link between eNOS and HMO in the endothelium. These data demonstrate that HMO-2′FL protects against NEC in part through maintaining mesenteric perfusion via increased eNOS expression, and suggest that the 2′FL found in human milk may be mediating some of the protective benefits of breast milk in the clinical setting against NEC.

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Corresponding author

* Corresponding author: D. J. Hackam, fax +1 410 502 5314, email dhackam1@jhmi.edu

References

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1. Neu, J & Walker, WA (2011) Necrotizing enterocolitis. N Engl J Med 364, 255264.
2. Fitzgibbons, SC, Ching, Y, Yu, D, et al. (2009) Mortality of necrotizing enterocolitis expressed by birth weight categories. J Pediatr Surgery 44, 10721075 ; discussion 1075–1076.
3. Lin, PW & Stoll, BJ (2006) Necrotising enterocolitis. Lancet 368, 12711283.
4. Obladen, M (2009) Necrotizing enterocolitis – 150 years of fruitless search for the cause. Neonatology 96, 203210.
5. Sullivan, S, Schanler, RJ, Kim, JH, et al. (2010) An exclusively human milk-based diet is associated with a lower rate of necrotizing enterocolitis than a diet of human milk and bovine milk-based products. J Pediatr 156, 562-567.e1.
6. Lucas, A & Cole, TJ (1990) Breast milk and neonatal necrotising enterocolitis. Lancet 336, 15191523.
7. Newburg, DS & Walker, WA (2007) Protection of the neonate by the innate immune system of developing gut and of human milk. Pediatr Res 61, 28.
8. Meinzen-Derr, J, Poindexter, B, Wrage, L, et al. (2009) Role of human milk in extremely low birth weight infants’ risk of necrotizing enterocolitis or death. J Perinatol 29, 5762.
9. Leaphart, CL, Cavallo, J, Gribar, SC, et al. (2007) A critical role for TLR4 in the pathogenesis of necrotizing enterocolitis by modulating intestinal injury and repair. J Immunol 179, 48084820.
10. Sodhi, CP, Neal, MD, Siggers, R, et al. (2012) Intestinal epithelial toll-like receptor 4 regulates goblet cell development and is required for necrotizing enterocolitis in mice. Gastroenterology 143, 708-718.e1–e5.
11. Good, M, Siggers, RH, Sodhi, CP, et al. (2012) Amniotic fluid inhibits toll-like receptor 4 signaling in the fetal and neonatal intestinal epithelium. Proc Natl Acad Sci 109, 1133011335.
12. Good, M, Sodhi, CP, Egan, CE, et al. (2015) 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. Mucosal Immunol 8, 11661179.
13. Jilling, T, Simon, D, Lu, J, et al. (2006) The roles of bacteria and TLR4 in rat and murine models of necrotizing enterocolitis. J Immunol 177, 32733282.
14. Nino, DF, Sodhi, CP & Hackam, DJ (2016) Necrotizing enterocolitis: mechanisms and management. Nat Rev Gastroenterol Hepatol (In the press).
15. Afrazi, A, Branca, MF, Sodhi, CP, et al. (2014) Toll like receptor 4-mediated endoplasmic reticulum stress in intestinal crypts induces necrotizing enterocolitis. J Biol Chem 289, 95849599.
16. Egan, CE, Sodhi, CP, Good, M, et al. (2015) Toll-like receptor 4-mediated lymphocyte influx induces neonatal necrotizing enterocolitis. J Clin Invest 126, 495508.
17. Neal, MD, Sodhi, CP, Jia, H, et al. (2012) Toll-like receptor 4 is expressed on intestinal stem cells and regulates their proliferation and apoptosis via the p53 up-regulated modulator of apoptosis. J Biol Chem 287, 3729637308.
18. Yazji, I, Sodhi, CP, Lee, EK, et al. (2013) Endothelial TLR4 activation impairs intestinal microcirculatory perfusion in necrotizing enterocolitis via eNOS-NO-nitrite signaling. Proc Natl Acad Sci U S A 110, 94519456.
19. Jantscher-Krenn, E, Zherebtsov, M, Nissan, C, et al. (2012) The human milk oligosaccharide disialyllacto-N-tetraose prevents necrotising enterocolitis in neonatal rats. Gut 61, 14171425.
20. Manzoni, P, Rinaldi, M, Cattani, S, et al. (2009) Bovine lactoferrin supplementation for prevention of late-onset sepsis in very low-birth-weight neonates: a randomized trial. JAMA 302, 14211428.
21. Dvorak, B, Halpern, MD, Holubec, H, et al. (2002) Epidermal growth factor reduces the development of necrotizing enterocolitis in a neonatal rat model. Am J Physiol Gastrointest Liver Physiol 282, G156G164.
22. Radulescu, A, Zorko, NA, Yu, X, et al. (2009) Preclinical neonatal rat studies of heparin-binding EGF-like growth factor in protection of the intestines from necrotizing enterocolitis. Pediatr Res 65, 437442.
23. Newburg, DS (2009) Neonatal protection by an innate immune system of human milk consisting of oligosaccharides and glycans. J Anim Sci 87, 2634.
24. Newburg, DS, Ruiz-Palacios, GM & Morrow, AL (2005) Human milk glycans protect infants against enteric pathogens. Annu Rev Nutr 25, 3758.
25. Bode, L (2009) Human milk oligosaccharides: prebiotics and beyond. Nutr Rev 67, Suppl. 2, S183S191.
26. Bode, L (2012) Human milk oligosaccharides: every baby needs a sugar mama. Glycobiology 22, 11471162.
27. Bode, L, Kunz, C, Muhly-Reinholz, M, et al. (2004) Inhibition of monocyte, lymphocyte, and neutrophil adhesion to endothelial cells by human milk oligosaccharides. Thromb Haemost 92, 14021410.
28. Wang, M, Li, M, Wu, S, et al. (2015) Fecal microbiota composition of breast-fed infants is correlated with human milk oligosaccharides consumed. J Pediatr Gastroenterol Nutr 60, 825833.
29. Zivkovic, AM, German, JB, Lebrilla, CB, et al. (2011) Human milk glycobiome and its impact on the infant gastrointestinal microbiota. Proc Natl Acad Sci U S A 108, Suppl. 1, 46534658.
30. Marcobal, A, Barboza, M, Froehlich, JW, et al. (2010) Consumption of human milk oligosaccharides by gut-related microbes. J Agric Food Chem 58, 53345340.
31. Ruiz-Palacios, GM, Cervantes, LE, Ramos, P, et al. (2003) Campylobacter jejuni binds intestinal H(O) antigen (Fuc alpha 1, 2Gal beta 1, 4GlcNAc), and fucosyloligosaccharides of human milk inhibit its binding and infection. J Biol Chem 278, 1411214120.
32. Goehring, KC, Kennedy, AD, Prieto, PA, et al. (2014) Direct evidence for the presence of human milk oligosaccharides in the circulation of breastfed infants. PLOS ONE 9, e101692.
33. Ruhaak, LR, Stroble, C, Underwood, MA, et al. (2014) Detection of milk oligosaccharides in plasma of infants. Anal Bioanal Chem 406, 57755784.
34. Afrazi, A, Sodhi, CP, Good, M, et al. (2012) Intracellular heat shock protein-70 negatively regulates TLR4 signaling in the newborn intestinal epithelium. J Immunol 188, 45434557.
35. Good, M, Sodhi, CP, Ozolek, JA, et al. (2014) Lactobacillus rhamnosus HN001 decreases the severity of necrotizing enterocolitis in neonatal mice and preterm piglets: evidence in mice for a role of TLR9. Am J Physiol Gastrointest Liver Physiol 306, G1021G1032.
36. Anand, RJ, Leaphart, CL, Mollen, KP, et al. (2007) The role of the intestinal barrier in the pathogenesis of necrotizing enterocolitis. Shock 27, 124133.
37. Magoc, T & Salzberg, SL (2011) FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics 27, 29572963.
38. Bolger, AM, Lohse, M & Usadel, B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 21142120.
39. Kuczynski, J, Stombaugh, J, Walters, WA, et al. (2011) Using QIIME to analyze 16S rRNA gene sequences from microbial communities. Curr Protoc Bioinformatics 36, 10.7.110.7.20. Chapter 10, Unit 10.17.
40. Caporaso, JG, Bittinger, K, Bushman, FD, et al. (2010) PyNAST: a flexible tool for aligning sequences to a template alignment. Bioinformatics 26, 266267.
41. Edgar, RC, Haas, BJ, Clemente, JC, et al. (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27, 21942200.
42. Langmead, B & Salzberg, SL (2012) Fast gapped-read alignment with Bowtie 2. Nature Methods 9, 357359.
43. McDonald, D, Price, MN, Goodrich, J, et al. (2012) An improved Greengenes taxonomy with explicit ranks for ecological and evolutionary analyses of bacteria and archaea. ISME J 6, 610618.
44. Wang, Q, Garrity, GM, Tiedje, JM, et al. (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73, 52615267.
45. Salter, SJ, Cox, MJ, Turek, EM, et al. (2014) Reagent and laboratory contamination can critically impact sequence-based microbiome analyses. BMC Biol 12, 87.
46. Benjamini, Y, Drai, D, Elmer, G, et al. (2001) Controlling the false discovery rate in behavior genetics research. Behav Brain Res 125, 279284.
47. McMurdie, PJ & Holmes, S (2013) phyloseq: An R package for reproducible interactive analysis and graphics of microbiome census data. PLOS ONE 8, e61217.
48. Jain, SK, Baggerman, EW, Mohankumar, K, et al. (2014) Amniotic fluid-borne hepatocyte growth factor protects rat pups against experimental necrotizing enterocolitis. Am J Physiol Gastrointest Liver Physiol 306, G361G369.
49. Shesely, EG, Maeda, N, Kim, HS, et al. (1996) Elevated blood pressures in mice lacking endothelial nitric oxide synthase. Proc Natl Acad Sci U S A 93, 1317613181.
50. Yu, X, Radulescu, A, Zorko, N, et al. (2009) Heparin-binding EGF-like growth factor increases intestinal microvascular blood flow in necrotizing enterocolitis. Gastroenterology 137, 221230.
51. Demmert, M, Schaper, A, Pagel, J, et al. (2015) FUT 2 polymorphism and outcome in very-low-birth-weight infants. Pediatr Res 77, 586590.
52. Wacklin, P, Makivuokko, H, Alakulppi, N, et al. (2011) Secretor genotype (FUT2 gene) is strongly associated with the composition of Bifidobacteria in the human intestine. PLoS ONE 6, e20113.
53. Garrido, D, Barile, D & Mills, DA (2012) A molecular basis for bifidobacterial enrichment in the infant gastrointestinal tract. Adv Nutr 3, 415S421S.
54. Marcobal, A & Sonnenburg, JL (2012) Human milk oligosaccharide consumption by intestinal microbiota. Clin Microbiol Infect 18, Suppl. 4, 1215.
55. Kosloske, AM & Ulrich, JA (1980) A bacteriologic basis for the clinical presentations of necrotizing enterocolitis. J Pediatr Surg 15, 558564.
56. Morrow, AL, Ruiz-Palacios, GM, Jiang, X, et al. (2005) Human-milk glycans that inhibit pathogen binding protect breast-feeding infants against infectious diarrhea. J Nutr 135, 13041307.
57. Gribar, SC, Sodhi, CP, Richardson, WM, et al. (2009) Reciprocal expression and signaling of TLR4 and TLR9 in the pathogenesis and treatment of necrotizing enterocolitis. J Immunol 182, 636646.
58. La Rosa, PS, Warner, BB, Zhou, Y, et al. (2014) Patterned progression of bacterial populations in the premature infant gut. Proc Natl Acad Sci U S A 111, 1252212527.
59. Underwood, MA, Gaerlan, S, De Leoz, ML, et al. (2015) Human milk oligosaccharides in premature infants: absorption, excretion, and influence on the intestinal microbiota. Pediatr Res 78, 670677.
60. Santos-Fandila, A, Zafra-Gomez, A, Vazquez, E, et al. (2014) Ultra high performance liquid chromatography-tandem mass spectrometry method for the determination of soluble milk glycans in rat serum. Talanta 118, 137146.
61. Prieto, PA, Mukerji, P, Kelder, B, et al. (1995) Remodeling of mouse milk glycoconjugates by transgenic expression of a human glycosyltransferase. J Biol Chem 270, 2951529519.
62. Neal, MD, Jia, H, Eyer, B, et al. (2013) Discovery and validation of a new class of small molecule Toll-like receptor 4 (TLR4) inhibitors. PLOS ONE 8, e65779.
63. He, Y, Liu, S, Kling, DE, et al. (2016) The human milk oligosaccharide 2’-fucosyllactose modulates CD14 expression in human enterocytes, thereby attenuating LPS-induced inflammation. Gut 65, 3346.
64. Eidelman, AI, Schanler, RJ, Johnston, M, et al. (2012) Breastfeeding and the use of human milk. Pediatrics 129, e827e841.
65. Nutrition, ECo, Arslanoglu, S, Corpeleijn, W, et al. (2013) Donor human milk for preterm infants: current evidence and research directions. J Pediatr Gastroenterol Nutr 57, 535542.

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