Elucidation of Breast Milk Composition and Structure Over the First Year of Lactation: UC Davis Lactation Study

Study Overview
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Brief Summary
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This observational study is designed to determine how sugars, lipids and proteins in breast milk vary among mothers; is affected by maternal diet, health and microbiota and vary over the course of lactation. Additionally, the study is designed to determine how the structure and composition of complex milk sugars, maternal health status and diet influence the development and maintenance of infant gut microflora.

Detailed Description
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This study's objectives include: 1) develop a better understanding of the composition and structures of breast milk over the early stages of lactation with specific emphasis on the glycans and lipids; 2) how glycan diversity relates to maternal and infant gut microbiota with an emphasis on bifidobacteria species; 3)how milk composition and structure relate to maternal health and diet. These samples will be analyzed using comprehensive gas and liquid chromatographic methods, nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS); and next generation sequencing, terminal restriction fragment length polymorphism and quantitative polymerase chain reaction. There are several cohorts to the main study. Gestational Diabetes Mellitus Cohort. The aim of this study is to investigate the differences between milk composition, infant and maternal gut microbiota from women with and without gestational diabetes mellitus or type 2 diabetes vs. controls. Fresh Milk Study. The aim of this study is to elucidate the relationship between the expression of glycan metabolizing genes of mammary epithelial cells and levels of milk glycans. RNA Study. The aim of this study is to compare the RNA of milk fat globule membrane crescents in human milk against epithelial expression profiles in non-human primate milk. Skin Study. The aim of this study is to provide skin samples from healthy term infants to act as the control group for a different study with preterm infants. Breast Milk, Gut Microbiome, and Immunity (BMMI) Project. The aim of this study is to provide healthy control samples for the BMMI project. The BMMI project is a multi-investigative project designed to elucidate how maternal nutrient status, and milk composition and structure influence infant gut microbiome from participants in developing countries.

Official Title
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Functional Deconstruction of Human Milk Oligosaccharides and Lipids

Conditions
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Healthy, Gestational Diabetes Mellitus, Type 2 Diabetes Mellitus

Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Pregnant women 18 years and older who plan to breastfeed for at least 6 months Women who are in good health Women with or without diagnosis of gestational diabetes mellitus, or type 2 diabetes mellitus or impaired glucose tolerance Exclusion Criteria: Women who do not plan to breastfeed for at least 6 months Women who give birth to pre-term infants Women who smoke, either currently or 1 year before becoming pregnant Women who have chronic medical conditions or communicable diseases Women who take metabolic altering drugs such as corticosteroids or thyroid medication

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 45 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Main Study<br>Women enrolled in this cohort of the study will collect their breast milk, infant urine and stool, and their urine and stool samples. Blood and saliva will be collected from these participants by study personnel. | |
| Gestational Diabetes Mellitus Cohort<br>Women who have been diagnosed with gestational diabetes mellitus, type 2 diabetes mellitus or impaired glucose tolerance will be enrolled in this cohort. Participants will be asked to report their fasting and postprandial blood glucose levels if they are monitoring these outcomes at home with a glucometer. | |
| Fresh Milk Cohort<br>Women enrolled in this cohort will provide fresh milk samples (stored in the refrigerator and picked up by study personnel within 1 hour of collection) for analysis of glycan composition and gene expression of glycan metabolizing enzymes of somatic cells in milk. | |
| RNA Milk Fat Cohort<br>Women enrolled in this cohort must have given birth to sons and will collect a fresh milk sample for transcriptomic analysis compared against non-human primate milk. | |
| Skin Study<br>This cohort includes mothers and their infants who will provide milk and infant stratum corneum cells to act as the control group for a different study designed to investigate skin function in premature infants. | |
| BMMI Project<br>Subjects enrolled in this cohort will be part of the control group for the BMMI Project. | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Human milk glycans | Human milk glycans: human milk oligosaccharides and glycoconjugates to lipids, proteins and peptides will be measured by MS methods. | Change from day 2 to day 366 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Microbiota | Maternal and infant gut microbiota, infant skin microbiota will be measured using next generation sequencing, terminal restriction fragment length polymorphism and quantitative polymerase chain reaction. | 3rd trimester pregnancy and Day 2-Day 366 postpartum | 
| Proteome | Milk and infant stratum corneum will be analyzed by MS methods. | Day 2-Day 366 postpartum | 
| Lipidome | Milk and skin will be analyzed by gas chromatography and MS methods. | Day 2-Day 366 postpartum | 
| Maternal plasma lipids | The plasma lipid profile will be analyzed by enzyme assay by UC Davis Pathology Lab. | 3rd trimester pregnancy and 2 months postpartum | 
| Peptidome | Milk and infant and maternal stool will be analyzed by MS methods. | 3rd trimester pregnancy and Day 2-Day 366 | 
| Metabolome by Nuclear Magnetic Spectroscopy (NMR) | Milk, maternal and infant urine and stool will be analyzed by NMR spectroscopy. | 3rd trimester of pregnancy and D2-Day 366 | 
| Metabolome by MS | Milk, maternal and infant urine and stool will be analyzed by MS. | Day 2-Day 366 postpartum | 
| Secretor status genotyping | Maternal secretor status genotyping will be determined by quantitative polymerase chain reaction for the functional fucosyltransferase 2 gene from maternal saliva. | 3rd trimester pregnancy and Day 60 postpartum | 
| Maternal fasting and postprandial blood sugar | Maternal blood sugar will be checked by finger stick using a glucometer in women diagnosed with gestational diabetes mellitus. | 3rd trimester of pregnancy and Day 3-7 | 
| Maternal plasma lipoprotein size distribution | Plasma lipoprotein size distribution and concentration will be analyzed by NMR spectroscopy by LipoScience. | 3rd trimester of pregnancy and Day 60 postpartum | 
| Human Milk Transcriptomics | Milk samples will be preserved with RNA-later, RNA extracted and analyzed by next generation sequencing techniques. | 3 and 6 months postpartum |