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Human Milk Oligosaccharides Protect against Necrotizing Enterocolitis by Activating Intestinal Cell Differentiation
Bo Li1,2, Richard You Wu1,3,4, Rachael G Horne3
1Translational Medicine Program, The Hospital for Sick Children, Toronto, M5G 1X8, Canada.
Insights
Human milk oligosaccharides (HMOs) protect preterm infants from necrotizing enterocolitis (NEC) by enhancing intestinal cell differentiation. This study reveals molecular mechanisms, showing HMOs alter gene expression related to gut health.
Area of Science:
- Neonatal research
- Gastroenterology
- Molecular biology
Background:
- Necrotizing enterocolitis (NEC) is a critical gastrointestinal condition and a primary cause of death in preterm infants.
- Human milk oligosaccharides (HMOs) show promise in reducing NEC incidence, but their protective mechanisms remain unclear.
Purpose of the Study:
- To investigate the molecular mechanisms by which HMOs protect against NEC.
- To explore the impact of HMOs on the intestinal epithelial transcriptome.
Main Methods:
- Genome-wide transcriptome profiling using RNA-sequencing in response to HMOs.
- Validation of cell differentiation markers (HMGCS2, MUC2) in Caco-2 intestinal cells.
- In vivo NEC model using HMO-gavaged neonatal pups.
Main Results:
- HMOs modulated 225 host genes involved in cell proliferation and differentiation, including upregulation of HMGCS2.
- Alcian Blue staining and TER recordings confirmed enhanced differentiation in Caco-2 cells.
- HMO-fed pups exhibited increased MUC2 differentiation and HMGCS2 expression.
Conclusions:
- HMOs offer protection against NEC by influencing the differentiation of the intestinal crypt-villus axis.
- Pooled HMOs initiate biological processes that provide mechanistic insights into intestinal protection.
Scope:
Necrotizing enterocolitis (NEC) is a devastating gastrointestinal emergency and currently the leading cause of mortality in preterm infants. Recent studies show that human milk oligosaccharides (HMOs) reduce the frequency and incidence of NEC; however, the molecular mechanisms for their protection are largely unexplored.
Methods And Results:
To address this gap, a genome-wide profiling of the intestinal epithelial transcriptome in response to HMOs using RNA-sequencing is performed. It is found that HMOs alter the host transcriptome in 225 unique target genes pertaining to cell proliferation and differentiation, including upregulation of stem cell differentiation marker HMGCS2. To validate these results, differentiation in Caco-2Bbe1 (Caco-2) intestinal cells is verified by Alcian Blue staining and transepithelial electrical resistance (TER) recordings. Furthermore, an in vivo model of NEC is also employed whereby neonatal pups are gavage fed HMOs. Interestingly, HMOs-fed pups show enhanced cell MUC2 differentiation and HMGCS2 expression.
Conclusions:
These findings demonstrate HMOs protect against NEC in part by altering the differentiation of the crypt-villus axis. In addition, this study suggests that pooled HMOs directly induce a series of biological processes, which provide mechanistic insights to how HMOs protect the host intestine.
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