Diet-Dependent Changes in The Intestinal DNA Methylome After Introduction of Enteral Feeding in Preterm Pigs

Abstract

Aim: To examine how enteral feeding affects the intestinal epigenome and gene expression just after preterm birth. Materials & methods: Intestinal tissue from preterm pigs, modeling preterm infants, was collected at birth and 5 days after gradual introduction of infant formula or bovine colostrum. The intestinal tissue was analyzed by reduced representation bisulfite sequencing and real-time qPCR. Results: Relative to colostrum, formula increased bacterial epithelial adherence and lipopolysaccharide binding protein (LBP) expression, which was regulated by promoter methylation. Diet-dependent changes in DNA methylation and/or mRNA expression were related to innate immune response, hypoxia, angiogenesis and epithelial–mesenchymal transition pathways (e.g., TTC38, IL8, C3, HIF1A and VEGFR1). Conclusion: Epigenetic changes may mediate important effects of the first feeding on intestinal development in preterm neonates.

Keywords::

• DNA methylation
• epigenetic gene regulation
• immunity
• microbiota
• nutrition
• preterm birth

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/suppl/10.2217/epi-2017-0122

Data availability

All RRBS sequencing and processed data will be deposited in the Gene Expression Omnibus (GEO).

Author’s contributions

X Pan performed the experimental work. X Pan analyzed and interpreted the RRBS and qPCR data with help from D Gong. X Pan was the major contributor in writing the manuscript. PT Sangild and F Gao took part in the main study design, and critically reviewed the manuscript. All authors read and approved the final manuscript.

Acknowledgements

The authors thank T Thymann, R Liang, E Skytte, K M⊘ller, J Povlsen and A Brunse for their technical support with animal procedures and laboratory analyses.

Financial & competing interests disclosure

This work was supported by the Danish Strategic Research Council under Grant 12–132401, the Agricultural Science and Technology Innovation Program (ASTIP) of China, the Science and Technology Project of Shenzhen (JSGG20160427104724699) and the China Scholarship Council under Grant 201406150073. University of Copenhagen holds a patent on use of bovine colostrum for human preterm infants. PT Sangild is listed as sole inventor but has declined any share of potential revenue arising from commercial exploitation of such a patent. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Additional information

Funding

This work was supported by the Danish Strategic Research Council under Grant 12–132401, the Agricultural Science and Technology Innovation Program (ASTIP) of China, the Science and Technology Project of Shenzhen (JSGG20160427104724699) and the China Scholarship Council under Grant 201406150073. University of Copenhagen holds a patent on use of bovine colostrum for human preterm infants. PT Sangild is listed as sole inventor but has declined any share of potential revenue arising from commercial exploitation of such a patent. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.