The prospective 5-month follow-up study will involve 150 mother-infant dyads grouped based on maternal baseline BMI category (normal weight, overweight, obesity). The investigators plan 3 study visits (1, 3, and 6 months postpartum) with analysis the following parameters and outcomes: 1. maternal anthropometry and body composition; 2. maternal dietary intake and dietary patterns (with results of biomarker-based validation); 3. maternal status of lipophilic antioxidants, selected vitamins, and fatty acids profile that may be related to adipose tissue metabolism, inflammation, and cardiometabolic status; 4. maternal metabolomics, adipokines, insulin, selected biomarkers of inflammation, altered glucose metabolism, and oxidation; 5. breastmilk composition assessed by systemic approach (macronutrients, fatty acids profile, lipophilic antioxidants, adipokines, hormones, immunological profile, and preliminary lipidomic analysis in part of the study group); 6. infant growth trajectory, body composition, urine metabolomics, and biomarkers of oxidative stress. Additionally, maternal and infant stool, buccal swabs, and human milk samples will be banked for further microbiome analysis studies.
The global obesity pandemic continues to escalate, with projections showing 1 in 5 women and 1 in 7 men obese by 2030. Maternal obesity raises the risk of childhood obesity by 264%, and overweight/obese children are more prone to become obese adults. Obesity-associated metabolic dysfunctions (inflammation, insulin resistance, dyslipidemia) heighten morbidity and mortality risks. Breastfeeding mitigates overweight/obesity risks, yet mothers with overweight/obesity often encounter challenges in initiation, continuation, and altered milk composition, showing elevated levels of leptin, insulin, and n-6/n-3 fatty acids. However, findings on other components, like total fat, fatty acids, insulin, adipokines, carotenoids, and immune factors, remain inconsistent due to methodological limitations and lack of comprehensive analysis. Nonetheless, the potential pro-inflammatory and obesogenic properties of milk produced by mothers with overweight/obesity have been discussed. Recent systematic reviews link breastmilk protein, fat, leptin, and IL-6 to infant growth and adiposity. Findings for other components remain unclear due to small samples, methodological limitations, and lack of a systemic approach to analyze breastmilk as a complex biological system. Alterations in breastmilk composition may stem from disrupted de novo synthesis, transport across the blood-breastmilk barrier, or altered circulating levels. Recent studies showed impaired de novo synthesis of C15:0 and C16:1 fatty acids, probably caused by insulin resistance and reduced long-chain fatty acid transfer linked to inflammation (suppress lipoprotein lipase activity essential for transporting lipid compounds). Hence, poor cardiometabolic status may mediate altered breastfeeding outcomes and milk composition, but it requires further studies. Studies among non-lactating individuals suggested lifestyle and diet play crucial roles in mitigating overweight/obesity metabolic implications (e.g., metabolically healthy obese (MHO) and metabolically obese with normal weight (MONW) phenotypes). Maternal diet can influence breastmilk concentration of certain diet-dependent nutrients and bioactives (e.g., fatty acids, vitamins A, D, and carotenoids). Similarly, prohealthy/unhealthy dietary patterns seem to have beneficial/unfavorable effects on breastmilk composition and breastfeeding outcomes, but results remain inconclusive. Nonetheless, some suggest that diet may mitigate the effects of maternal overweight/obesity on breastmilk composition in mothers with overweight/obesity. The mechanisms linking obesity-related changes in breastmilk to infant development remain unclear. This study addresses this gap by studying the mother-breastmilk-infant triad, focusing on maternal cardiometabolic status, diet, breastmilk composition as a complex biological system, and anthropometric and metabolic outcomes. This study aims to elucidate the intricate interplay between maternal adiposity, cardiometabolic status, diet, and breastmilk composition and to understand the role of breastmilk in the transmission of obesity and the shaping of infant metabolic health. Our research questions will aim to determine/explore the following questions: Q1. How does the maternal BMI category differentiate breastmilk composition from a systemic perspective? Q2. How does maternal cardiometabolic status mediate OW/OB-related alterations in breastmilk composition? Q3. Whether (and how) maternal diet moderates OW/OB-related alterations in breastmilk composition? Q4. Whether (and how) maternal OW/OB differentiates infant metabolomic profile? Q5. Whether (and how) OW/OB-related alterations in breastmilk composition influence infant outcomes and obesity risk? Q6. Whether (and how) maternal dietary patterns moderate adverse health outcomes of maternal OW/OB in infants? The research hypotheses assume that: H1. Maternal OW and OB are associated with pro-inflammatory properties and altered breastmilk metabolome via obesity-related low-grade inflammation, metabolic alterations, and oxidative stress, not dietary differences. H2. Metabolically healthy OW and OB mothers produce less altered breastmilk than metabolically unhealthy mothers, as cardiometabolic status mediates between adiposity and breastmilk composition. H3. A pro-healthy dietary pattern attenuates, and a westernized dietary pattern exacerbates obesity-related alterations in breastmilk composition due to differences in concentrations of pro-, anti-inflammatory, and antioxidant dietary compounds. H4. Infants of OW and OB mothers have altered growth trajectories and metabolic pathways, higher fat mass, and oxidative stress compared to infants of NW mothers. H5. Obesity-related changes in breastmilk composition determine alterations in infant outcomes. H6. Maternal pro-health/westernized dietary patterns and metabolic health mitigate/exacerbate the adverse effects of overweight and obesity on infant outcomes. The investigators will verify these hypotheses based on results collected using gold standards and novel body composition assessment and analytical methods (DXA scans, HPLC, GC-MS, ELISA, and LC-MS), nutrient-based and dietary pattern-based dietary assessment with biomarkers validation, in-depth evaluation of maternal cardiometabolic status and infant outcomes, and a systemic approach to breastmilk composition analysis. The findings will lay the foundation for tailored, evidence-based interventions to disrupt the intergenerational transmission of obesity. Furthermore. contribute to advancing the health sciences discipline and decreasing the global health burden of overweight and obesity, supporting the realization of WHO's Global Nutrition Goals.
Study Type
OBSERVATIONAL
Enrollment
165
Food, Nutrition and Health, Uviversity of British Columbia; Affiliated Investigator, Healthy Starts, BC Children's Hospital Research Institute & Women's Health Research Institute
Vancouver, British Columbia, Canada
Wroclaw Medical University
Wroclaw, Lower Silesian Voivodeship, Poland
National Institute of Public Health NIH - National Research Institute
Warsaw, Masovian Voivodeship, Poland
Department of Human Nutrition, Institute of Human Nutrition Sciences, Warsaw University of Life Sciences (SGGW-WULS)
Warsaw, Masovian Voivodeship, Poland
Difference between groups and change from 1 to 6 months postpartum in maternal body fat percentage
Assessment of changes in total body fat content, assessed by a bioelectrical impedance analysis (BIA) and a dual-energy X-ray absorptiometer (DXA). Results will be reported as a percentage (%).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal fat-free mass percentage
Assessment of changes in fat-free mass, assessed by a bioelectrical impedance analysis (BIA) and a dual-energy X-ray absorptiometer (DXA). Results will be reported as a percentage (%).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal Visceral Fat Area
Assessment of changes in visceral fat area, assessed by DXA. Results will be reported as square centimeters (cm\^2).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal Subcutaneous Fat Area
Assessment of changes in subcutaneous fat area, assessed by DXA. Results will be reported as square centimeters (cm\^2).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal Body Mass Index (BMI)
Weight and height will be combined to report BMI in kg/m\^2.
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal Waist-Hip Ratio (WHR)
Assessment of fat distribution calculated as the waist measurement divided by the hip measurement (dimensionless ratio).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal Waist-to-Height Ratio (WHtR)
Assessment of fat distribution calculated as the waist measurement divided by height (dimensionless ratio).
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in metabolic syndrome (MetS) incidence
Assessment of MetS incidences diagnosed based on the occurence of three from five metabolic abnormalities diagnosed based on International Diabetes Federation-established cut-off points for women for waist circumference (≥80 cm), fasting blood glucose (≥100 mg/dL), triglicerides (≥150 mg/dL), HDL cholesterol (\<50 mg/dL), blood pressure (systolic SBP ≥ 130 or diastolic ≥ 85 mmHg) as a marker of cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal glycated haemoglobin (HbA1c)
Assessment of changes in HbA1c (reported in %) as a marker of maternal cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in Homeostatic Model Assessment - Insulin Resistance (HOMA-IR)
Assessment of HOMA-IR calculated as the product of fasting insulin \[µIU/mL\] and fasting glucose \[µIU/mL\] divided by 22.5 (dimensionless ratio) as an indicator of maternal cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum leptin
Assessment of maternal serum leptin assessed by the ELISA method as an indicator of maternal cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum adiponectin
Assessment of maternal serum adiponectin assessed by the ELISA method as an indicator of maternal cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum and infant urine Retinol-Binding Protein-4 (RBP4)
Assessment of maternal serum and infant urine RBP-4 assessed by the ELISA method as an indicator of maternal and infant cardiometabolic health.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk macronutrients
Assessment of human milk macronutrients (fat, carbohydrate, dry material, protein, true protein) concentration assessed by mid-infrared (mid-IR) transmission spectroscopy method. Results will be reported in grams per 100 mL.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk energy value
Assessment of human milk energy value assessed by mid-infrared (mid-IR) transmission spectroscopy method. Results will be reported in kcal per 100 mL.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk secretory immunoglobulin A (S-IgA)
Assessment of human milk S-IgA assessed by Enzyme-Linked Immunosorbent Assay (ELISA) method. Results will be reported in grams per liter \[g/L\].
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk lactoferrin
Assessment of human milk lactoferrin assessed by ELISA method. Results will be reported in grams per liter \[g/L\].
Time frame: 1, 3, and 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk leptin
Assessment of human milk leptin assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk adiponectin
Assessment of human milk adiponectin assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk insulin
Assessment of human milk insulin assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk High-Sensitivity C-Reactive Protein (hs-CRP)
Assessment of human milk hsCRP assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk interleukin 1 (IL-1)
Assessment of human milk IL-1 assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk interleukin 6 (IL-6)
Assessment of human milk IL-6 assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk Tumor Necrosis Factor-alpha (TNF-alfa)
Assessment of human milk TNF-alfa assessed by ELISA method.
Time frame: 1, 3, 6 months of postpartum
Difference between groups and change from 1 to 6 months postpartum in human milk fatty acid profile
Assessment of human milk fatty acid profile analyzed as methyl ester (FAME) by gas chromatography. Results will be reported as concentration within a total fat \[% wt/wt\].
Time frame: 1, 3, 6 months of postpartum
Difference between groups and the change from 1 to 6 months of life in infant anthropometric development
Assessment of anthropometric development calculated as weight-, length-, BMI-, and head circumference z-scores for age and sex, analyzed based on WHO standards.
Time frame: 1, 3, 6 months of life
Difference between groups and the change from 1 to 6 months of life in infant body fat-free mass percentage
Assessment of changes in total body fat-free mass content, assessed by the whole-body DXA scans with a vacuum cushion to prevent movement. Results will be reported as a percentage (%).
Time frame: 1, 6 months of life
Difference between groups and the change from 1 to 6 months of life in infant body fat percentage
Assessment of changes in total body fat content, assessed by the whole-body DXA scans with a vacuum cushion to prevent movement. Results will be reported as a percentage (%).
Time frame: 1, 6 months of life
Difference between groups and the change from 1 to 6 months postpartum in the maternal resting metabolic rate (RMR)
Assessment of the rate of maternal energy metabolism assessed by indirect calorimetry. Results will be reported as kilocalories per day \[kcal/d\].
Time frame: 1, 3, 6 months postpartum
Difference between groups and the change from 1 to 6 months postpartum in maternal dietary intake
Assessment of maternal dietary intake using 3-day food records.
Time frame: 1, 3, 6 months postpartum
Difference between groups and the change from 1 to 6 months postpartum in maternal dietary habits
Assessment of maternal dietary intake using a food frequency questionnaire (FFQ).
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal short- and long-term fatty acid status
Assessment of serum blood and erythrocyte membranes fatty acids profile analyzed by gas chromatography. Results will be reported as concentration within a total fat \[% wt/wt\].
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal and human milk lipophilic antioxidants and vitamins
Assessment of plasma and human milk carotenoids, retinol, and vitamin E, as well as serum vitamin D (25(OH)D) analyzed by high-performance liquid chromatography (HPLC). Results will be reported as micromoles per liter \[µmol/L\].
Time frame: 1, 3, 6 months postpartum
Difference between groups and the change from 1 to 6 months of life in infant skin carotenoid status
Assessment of skin carotenoids assessed by reflectance spectroscopy with Veggie Meter (Longevity Link, Ltd.). Results will be reported as a skin carotenoid score (SCS; 0-800, higher values mean more carotenoids)
Time frame: 1, 3, 6 months of life
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Difference between groups and change from 1 to 6 months postpartum in maternal and infant urine 8-isoprostane
Assessment of maternal and infant urine 8-isoprostane analyzed by the ELISA as an indicator of oxidative stress. Results will be reported as picograms per milligram of creatinine \[pg per mg of creatinine\].
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum hsCRP
Assessment of maternal serum hsCRP assessed by the ELISA method as an indicator of systemic inflammation.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum IL-1
Assessment of maternal serum IL-1 assessed by the ELISA method as an indicator of systemic inflammation.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum IL-6
Assessment of maternal serum IL-6 assessed by the ELISA method as an indicator of systemic inflammation.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 6 months postpartum in maternal serum TNF-alfa
Assessment of maternal serum TNF-alfa assessed by the ELISA method as an indicator of systemic inflammation.
Time frame: 1, 3, 6 months postpartum
Difference between groups and change from 1 to 3 months postpartum in human milk lipidomics
Assessment of human milk triacylglycerols (TG), diacylglycerols (DG), phosphatidylcholine (PC), sphingomyelin (SM), and phosphatidylethanolamine (PE) assessed by targeted lipidomic analysis using the LC-MS. Results expressed as the percentage relative amount of many lipid species.
Time frame: 1, 3 months postpartum
Difference between groups and change from 1 to 6 months postpartum in metabolomic profile of maternal serum blood and infant urine
Assessment of the serum and urine non-targeted metabolomic profiles.
Time frame: 1, 3, 6 months postpartum