Alterations in human milk leptin and insulin are associated with early changes in the infant intestinal microbiome.

BACKGROUND Increased maternal body mass index (BMI) is a robust risk factor for later pediatric obesity. Accumulating evidence suggests that human milk (HM) may attenuate the transfer of obesity from mother to offspring, potentially through its effects on early development of the infant microbiome. OBJECTIVES Our objective was to identify early differences in intestinal microbiota in a cohort of breastfeeding infants born to obese compared with normal-weight (NW) mothers. We also investigated relations between HM hormones (leptin and insulin) and both the taxonomic and functional potentials of the infant microbiome. DESIGN Clinical data and infant stool and fasting HM samples were collected from 18 NW [prepregnancy BMI (in kg/m(2)) <24.0] and 12 obese (prepregnancy BMI >30.0) mothers and their exclusively breastfed infants at 2 wk postpartum. Infant body composition at 2 wk was determined by air-displacement plethysmography. Infant gastrointestinal microbes were estimated by using 16S amplicon and whole-genome sequencing. HM insulin and leptin were determined by ELISA; short-chain fatty acids (SCFAs) were measured in stool samples by using gas chromatography. Power was set at 80%. RESULTS Infants born to obese mothers were exposed to 2-fold higher HM insulin and leptin concentrations (P < 0.01) and showed a significant reduction in the early pioneering bacteria Gammaproteobacteria (P = 0.03) and exhibited a trend for elevated total SCFA content (P < 0.06). Independent of maternal prepregnancy BMI, HM insulin was positively associated with both microbial taxonomic diversity (P = 0.03) and Gammaproteobacteria (e.g., Enterobacteriaceae; P = 0.04) and was negatively associated with Lactobacillales (e.g., Streptococcaceae; P = 0.05). Metagenomic analysis showed that HM leptin and insulin were associated with decreased bacterial proteases, which are implicated in intestinal permeability, and reduced concentrations of pyruvate kinase, a biomarker of pediatric gastrointestinal inflammation. CONCLUSION Our results indicate that, although maternal obesity may adversely affect the early infant intestinal microbiome, HM insulin and leptin are independently associated with beneficial microbial metabolic pathways predicted to increase intestinal barrier function and reduce intestinal inflammation. This trial was registered at clinicaltrials.gov as NCT01693406.

[1]  S. Abramson.  Quantity and Diversity of Environmental Microbial Exposure and Development of Asthma: A Birth Cohort Study , 2015, Pediatrics.

[2]  Ellen E. Blaak,et al.  Short-chain fatty acids in control of body weight and insulin sensitivity , 2015, Nature Reviews Endocrinology.

[3]  P. Hartmann,et al.  The Effects of Leptin on Breastfeeding Behaviour , 2015, International journal of environmental research and public health.

[4]  Lisa J. Martin,et al.  Does Breastfeeding Protect Against Childhood Obesity? Moving Beyond Observational Evidence , 2015, Current Obesity Reports.

[5]  Tommi Vatanen,et al.  The dynamics of the human infant gut microbiome in development and in progression toward type 1 diabetes. , 2015, Cell host & microbe.

[6]  Guowei Huang,et al.  The association between breastfeeding and childhood obesity: a meta-analysis , 2014, BMC Public Health.

[7]  J. Moczko,et al.  Comparison of fecal pyruvate kinase isoform M2 and calprotectin in acute diarrhea in hospitalized children , 2014, Scientific Reports.

[8]  A. Petrova,et al.  Is variation in total antioxidant capacity of human milk associated with levels of bio-active proteins? , 2014, Journal of Perinatology.

[9]  K. Flegal,et al.  Prevalence of childhood and adult obesity in the United States, 2011-2012. , 2014, JAMA.

[10]  D. Sandoval.  Old dog, new trick: a direct role for leptin in regulating microbiota composition. , 2014, Endocrinology.

[11]  Cara L. Wilhelm,et al.  Proteobacteria-specific IgA regulates maturation of the intestinal microbiota , 2014, Gut microbes.

[12]  C. Y. Ooi,et al.  Fecal Biomarkers of Intestinal Health and Disease in Children , 2013, Front. Pediatr..

[13]  E. Zoetendal,et al.  Human intestinal microbiota composition is associated with local and systemic inflammation in obesity , 2013, Obesity.

[14]  I. Carroll,et al.  Enteric bacterial proteases in inflammatory bowel disease- pathophysiology and clinical implications. , 2013, World journal of gastroenterology.

[15]  Anders F. Andersson,et al.  Decreased gut microbiota diversity, delayed Bacteroidetes colonisation and reduced Th1 responses in infants delivered by Caesarean section , 2013, Gut.

[16]  J. Garssen,et al.  Human milk: a source of more life than we imagine. , 2013, Beneficial microbes.

[17]  Leah M. Feazel,et al.  Sex Differences in the Gut Microbiome Drive Hormone-Dependent Regulation of Autoimmunity , 2013, Science.

[18]  Pelin Yilmaz,et al.  The SILVA ribosomal RNA gene database project: improved data processing and web-based tools , 2012, Nucleic Acids Res..

[19]  D. Frank,et al.  Prevention of Virus-Induced Type 1 Diabetes with Antibiotic Therapy , 2012, The Journal of Immunology.

[20]  H. Schellhorn,et al.  Regulators of oxidative stress response genes in Escherichia coli and their functional conservation in bacteria. , 2012, Archives of biochemistry and biophysics.

[21]  S. Salminen,et al.  The human milk microbiome changes over lactation and is shaped by maternal weight and mode of delivery. , 2012, The American journal of clinical nutrition.

[22]  Lars Bode,et al.  Human milk oligosaccharides: every baby needs a sugar mama. , 2012, Glycobiology.

[23]  D. Frank,et al.  Impact of the Gut Microbiota on the Development of Obesity: Current Concepts , 2012 .

[24]  Andreas Wilke,et al.  The M5nr: a novel non-redundant database containing protein sequences and annotations from multiple sources and associated tools , 2012, BMC Bioinformatics.

[25]  A. Macpherson,et al.  Interactions Between the Microbiota and the Immune System , 2012, Science.

[26]  A. Dattilo,et al.  Need for Early Interventions in the Prevention of Pediatric Overweight: A Review and Upcoming Directions , 2012, Journal of obesity.

[27]  John J. McNeil,et al.  Obesity and Trends in Life Expectancy , 2012, Journal of obesity.

[28]  Elmar Pruesse,et al.  SINA: Accurate high-throughput multiple sequence alignment of ribosomal RNA genes , 2012, Bioinform..

[29]  B. Zinman,et al.  Associations of prenatal metabolic abnormalities with insulin and adiponectin concentrations in human milk. , 2012, The American journal of clinical nutrition.

[30]  P. Hartmann,et al.  Analysis of Insulin in Human Breast Milk in Mothers with Type 1 and Type 2 Diabetes Mellitus , 2012, International journal of endocrinology.

[31]  Anders F. Andersson,et al.  Low diversity of the gut microbiota in infants with atopic eczema. , 2012, The Journal of allergy and clinical immunology.

[32]  A. Pontecorvi,et al.  Male Fertility and Reduction in Semen Parameters: A Single Tertiary-Care Center Experience , 2012, International journal of endocrinology.

[33]  Howard C. Tenenbaum,et al.  Bacterial biogeography of the human digestive tract , 2011, Scientific reports.

[34]  Susumu Goto,et al.  KEGG for integration and interpretation of large-scale molecular data sets , 2011, Nucleic Acids Res..

[35]  Sebastian Tims,et al.  Global and deep molecular analysis of microbiota signatures in fecal samples from patients with irritable bowel syndrome. , 2011, Gastroenterology.

[36]  C. Hsieh,et al.  Peripheral education of the immune system by colonic commensal microbiota , 2011, Nature.

[37]  J. Gordon,et al.  Human nutrition, the gut microbiome and the immune system , 2011, Nature.

[38]  T. Harder,et al.  Leptin determination in colostrum and early human milk from mothers of preterm and term infants. , 2011, Early human development.

[39]  M. Netea,et al.  Trained immunity: a memory for innate host defense. , 2011, Cell host & microbe.

[40]  L. Poston,et al.  Obesity in Pregnancy: Implications for the Mother and Lifelong Health of the Child. A Consensus Statement , 2011, Pediatric Research.

[41]  D. Baird,et al.  Development of gut microbiota in infants not exposed to medical interventions , 2011, APMIS : acta pathologica, microbiologica, et immunologica Scandinavica.

[42]  N. Pace,et al.  Disease phenotype and genotype are associated with shifts in intestinal‐associated microbiota in inflammatory bowel diseases , 2011, Inflammatory bowel diseases.

[43]  P. Harris,et al.  Research electronic data capture (REDCap) - A metadata-driven methodology and workflow process for providing translational research informatics support , 2009, J. Biomed. Informatics.

[44]  M. Crowell,et al.  Human gut microbiota in obesity and after gastric bypass , 2009, Proceedings of the National Academy of Sciences.

[45]  B. Roe,et al.  A core gut microbiome in obese and lean twins , 2008, Nature.

[46]  L. Grummer-Strawn,et al.  Infant Feeding Practices Study II: Study Methods , 2008, Pediatrics.

[47]  S. Salminen,et al.  Early differences in fecal microbiota composition in children may predict overweight. , 2008, The American journal of clinical nutrition.

[48]  H. Brenner,et al.  Adipokines in Human Milk and Risk of Overweight in Early Childhood: A Prospective Cohort Study , 2007, Epidemiology.

[49]  N. Pace,et al.  Molecular-phylogenetic characterization of microbial community imbalances in human inflammatory bowel diseases , 2007, Proceedings of the National Academy of Sciences.

[50]  Daniel B. DiGiulio,et al.  Development of the Human Infant Intestinal Microbiota , 2007, PLoS biology.

[51]  J. Doré,et al.  Bacterial Imprinting of the Neonatal Immune System: Lessons From Maternal Cells? , 2007, Pediatrics.

[52]  R. Shamir,et al.  Gastrointestinal Tract as a Target Organ for Orally Administered Insulin , 2006, Journal of pediatric gastroenterology and nutrition.

[53]  J. Bauer,et al.  Effects of Prenatal Lipopolysaccharide Exposure on Epithelial Development and Function in Newborn Rat Intestine , 2006, Journal of pediatric gastroenterology and nutrition.

[54]  C. Picó,et al.  A Physiological Role of Breast Milk Leptin in Body Weight Control in Developing Infants , 2006, Obesity.

[55]  P. A. van den Brandt,et al.  Factors Influencing the Composition of the Intestinal Microbiota in Early Infancy , 2006, Pediatrics.

[56]  C. Mccusker,et al.  Neonatal exposure with LPS and/or allergen prevents experimental allergic airways disease: development of tolerance using environmental antigens. , 2006, The Journal of allergy and clinical immunology.

[57]  F. Shi,et al.  Dietary Insulin Affects Leucine Aminopeptidase, Growth Hormone, Insulin-Like Growth Factor I and Insulin Receptors in the Intestinal Mucosa of Neonatal Pigs , 2006, Neonatology.

[58]  Tao Cai,et al.  Automated genome annotation and pathway identification using the KEGG Orthology (KO) as a controlled vocabulary , 2005, Bioinform..

[59]  Korbinian Strimmer,et al.  APE: Analyses of Phylogenetics and Evolution in R language , 2004, Bioinform..

[60]  M. Muñoz-Navas,et al.  Distribution of the long leptin receptor isoform in brush border, basolateral membrane, and cytoplasm of enterocytes , 2002, Gut.

[61]  R. Shulman,et al.  Effect of enteral administration of insulin on intestinal development and feeding tolerance in preterm infants: a pilot study , 2002, Archives of disease in childhood. Fetal and neonatal edition.

[62]  P. Clifton,et al.  Short-chain fatty acids and human colonic function: roles of resistant starch and nonstarch polysaccharides. , 2001, Physiological reviews.

[63]  Marti J. Anderson,et al.  A new method for non-parametric multivariate analysis of variance in ecology , 2001 .

[64]  F. Casanueva,et al.  Presence of leptin in colostrum and/or breast milk from lactating mothers: a potential role in the regulation of neonatal food intake. , 1997, The Journal of clinical endocrinology and metabolism.

[65]  E. Sokal,et al.  Expression of insulin receptors and of 60-kDa receptor substrate in rat mature and immature enterocytes. , 1997, The American journal of physiology.

[66]  P. Mortensen,et al.  Kinetic studies on colonocyte metabolism of short chain fatty acids and glucose in ulcerative colitis. , 1995, Gut.

[67]  N. Levasseur,et al.  Breastfeeding: A Guide for the Medical Profession , 1995 .

[68]  T. Spector,et al.  Refinement of the coomassie blue method of protein quantitation. A simple and linear spectrophotometric assay for less than or equal to 0.5 to 50 microgram of protein. , 1983, Analytical biochemistry.

[69]  Mitchell J. Mergenthaler.  Nonparametrics: Statistical Methods Based on Ranks , 1979 .

[70]  W. Walker,et al.  Breast milk, microbiota, and intestinal immune homeostasis , 2015, Pediatric Research.

[71]  L. Morelli,et al.  Human milk and infant intestinal mucosal glycans guide succession of the neonatal intestinal microbiota , 2015, Pediatric Research.

[72]  R Core Team,et al.  R: A language and environment for statistical computing. , 2014 .

[73]  R. Shamir,et al.  Insulin in human milk and the use of hormones in infant formulas. , 2013, Nestle Nutrition Institute workshop series.

[74]  P. Legendre,et al.  vegan : Community Ecology Package. R package version 1.8-5 , 2007 .

[75]  C. Hoyen.  Factors Influencing the Composition of the Intestinal Microbiota in Early Infancy , 2007 .

[76]  Ruth A.Lawrence Robert M.Lawrence.  Breast-feeding, a guide for the medical profession , 1980 .

[77]  phylogenetic and functional analysis of metagenomes , 2022 .