Multi-omics signatures of the human early life exposome.

Environmental exposures during early life play a critical role in life-course health, yet the molecular phenotypes underlying environmental effects on health are poorly understood. In the Human Early Life Exposome (HELIX) project, a multi-centre cohort of 1301 mother-child pairs, we associate individual exposomes consisting of >100 chemical, outdoor, social and lifestyle exposures assessed in pregnancy and childhood, with multi-omics profiles (methylome, transcriptome, proteins and metabolites) in childhood. We identify 1170 associations, 249 in pregnancy and 921 in childhood, which reveal potential biological responses and sources of exposure. Pregnancy exposures, including maternal smoking, cadmium and molybdenum, are predominantly associated with child DNA methylation changes. In contrast, childhood exposures are associated with features across all omics layers, most frequently the serum metabolome, revealing signatures for diet, toxic chemical compounds, essential trace elements, and weather conditions, among others. Our comprehensive and unique resource of all associations ( https://helixomics.isglobal.org/ ) will serve to guide future investigation into the biological imprints of the early life exposome.

Study of the Combined Effect of Maternal Tobacco Smoking and Polygenic Risk Scores on Birth Weight and Body Mass Index in Childhood.

Background: Maternal smoking during pregnancy has adverse health effects on the offspring, including lower birth weight and increased risk for obesity. These outcomes are also influenced by common genetic polymorphisms. We aimed to investigate the combined effect of maternal smoking during pregnancy and genetic predisposition on birth weight and body mass index (BMI)-related traits in 1,086 children of the Human Early Life Exposome (HELIX) project. Methods: Maternal smoking during pregnancy was self-reported. Phenotypic traits were assessed at birth or at the age of 8 years. Ten polygenic risk scores (PRSs) per trait were calculated using the PRSice v2 program. For birth weight, we estimated two sets of PRSs based on two different base GWAS summary statistics: PRS-EGG, which includes HELIX children, and PRS-PanUK, which is completely independent. The best PRS per trait (highest R 2) was selected for downstream analyses, and it was treated in continuous or categorized into three groups. Multivariate linear regression models were applied to evaluate the association of the explanatory variables with the traits of interest. The combined effect was evaluated by including an interaction term in the regression models and then running models stratified by the PRS group. Results: BMI-related traits were correlated among them but not with birth weight. A similar pattern was observed for their PRSs. On average, the PRSs explained ∼4% of the phenotypic variation, with higher PRS values related to higher trait values (p-value <5.55E-08). Sustained maternal smoking was associated with lower birth weight and higher BMI and related traits (p-value <2.99E-02). We identified a gene by environment (GxE) interaction for birth weight between sustained maternal smoking and the PRS-EGG in three groups (p-value interaction = 0.01), which was not replicated with the PRS-PanUK (p-value interaction = 0.341). Finally, we did not find any statistically significant GxE interaction for BMI-related traits (p-value interaction >0.237). Conclusion: Sustained maternal smoking and the PRSs were independently associated with birth weight and childhood BMI-related traits. There was low evidence of GxE interactions.

Urban Environment and Health: A Cross-Sectional Study of the Influence of Environmental Quality and Physical Activity on Blood Pressure.

Few studies have examined the relation between urban built environment and the prevalence of hypertension. This cross-sectional study aimed at assessing the relationship between the environmental quality, physical activity, and stress on hypertension among citizens of Kaunas city, Lithuania. We conducted a survey of 1086 citizens residing in 11 districts to determine their perceptions of environmental quality, health behavior, and health indices. The independent variables included residential traffic flows, access to public transportation and green spaces. Dependent variables included physician-diagnosed hypertension, systolic and diastolic blood pressure, and stress level. We used multivariable logistic regression to assess the associations as odds ratios (OR). The environmental factors beneficially associated with meeting the physical activity recommendations were opportunities for walking to reach the city's green spaces and available relaxation areas. Residents of high noise level districts aged 45-64 years had a significantly higher OR of stress and a higher prevalence of hypertension when age, sex, education status, family status, and smoking were accounted for. However, meeting the physical activity recommendations had a beneficial effect on the risk of hypertension. This study provided evidence that improvement of the district-level built environment supporting citizens' physical activity might reduce the risk of hypertension.

The early-life exposome and epigenetic age acceleration in children.

The early-life exposome influences future health and accelerated biological aging has been proposed as one of the underlying biological mechanisms. We investigated the association between more than 100 exposures assessed during pregnancy and in childhood (including indoor and outdoor air pollutants, built environment, green environments, tobacco smoking, lifestyle exposures, and biomarkers of chemical pollutants), and epigenetic age acceleration in 1,173 children aged 7 years old from the Human Early-Life Exposome project. Age acceleration was calculated based on Horvath's Skin and Blood clock using child blood DNA methylation measured by Infinium HumanMethylation450 BeadChips. We performed an exposure-wide association study between prenatal and childhood exposome and age acceleration. Maternal tobacco smoking during pregnancy was nominally associated with increased age acceleration. For childhood exposures, indoor particulate matter absorbance (PMabs) and parental smoking were nominally associated with an increase in age acceleration. Exposure to the organic pesticide dimethyl dithiophosphate and the persistent pollutant polychlorinated biphenyl-138 (inversely associated with child body mass index) were protective for age acceleration. None of the associations remained significant after multiple-testing correction. Pregnancy and childhood exposure to tobacco smoke and childhood exposure to indoor PMabs may accelerate epigenetic aging from an early age.

Early life multiple exposures and child cognitive function: A multi-centric birth cohort study in six European countries.

Epidemiological studies mostly focus on single environmental exposures. This study aims to systematically assess associations between a wide range of prenatal and childhood environmental exposures and cognition. The study sample included data of 1298 mother-child pairs, children were 6-11 years-old, from six European birth cohorts. We measured 87 exposures during pregnancy and 122 cross-sectionally during childhood, including air pollution, built environment, meteorology, natural spaces, traffic, noise, chemicals and life styles. The measured cognitive domains were fluid intelligence (Raven's Coloured Progressive Matrices test, CPM), attention (Attention Network Test, ANT) and working memory (N-Back task). We used two statistical approaches to assess associations between exposure and child cognition: the exposome-wide association study (ExWAS) considering each exposure independently, and the deletion-substitution-addition algorithm (DSA) considering all exposures simultaneously to build a final multiexposure model. Based on this multiexposure model that included the exposure variables selected by ExWAS and DSA models, child organic food intake was associated with higher fluid intelligence (CPM) scores (beta = 1.18; 95% CI = 0.50, 1.87) and higher working memory (N-Back) scores (0.23; 0.05, 0.41), and child fast food intake (-1.25; -2.10, -0.40), house crowding (-0.39; -0.62, -0.16), and child environmental tobacco smoke (ETS) (-0.89; -1.42, -0.35), were all associated with lower CPM scores. Indoor PM2.5 exposure was associated with lower N-Back scores (-0.09; -0.16, -0.02). Additional associations in the unexpected direction were found: Higher prenatal mercury levels, maternal alcohol consumption and child higher perfluorooctane sulfonic acid (PFOS) levels were associated with better cognitive performance; and higher green exposure during pregnancy with lower cognitive performance. This first comprehensive and systematic study of many prenatal and childhood environmental risk factors suggests that unfavourable child nutrition, family crowdedness and child indoor air pollution and ETS exposures adversely and cross-sectionally associate with cognitive function. Unexpected associations were also observed and maybe due to confounding and reverse causality.

Early-life environmental exposure determinants of child behavior in Europe: A longitudinal, population-based study.

BACKGROUND: Environmental exposures in early life influence the development of behavioral outcomes in children, but research has not considered multiple exposures. We therefore aimed to investigate the impact of a broad spectrum of pre- and postnatal environmental exposures on child behavior. METHODS AND FINDINGS: We used data from the HELIX (Human Early Life Exposome) project, which was based on six longitudinal population-based birth cohorts in Europe. At 6-11 years, children underwent a follow-up to characterize their exposures and assess behavioral problems. We measured 88 prenatal and 123 childhood environmental factors, including outdoor, indoor, chemical, lifestyle and social exposures. Parent-reported behavioral problems included (1) internalizing, (2) externalizing scores, using the child behavior checklist (CBCL), and (3) the Conner's Attention Deficit Hyperactivity Disorder (ADHD) index, all outcomes being discrete raw counts. We applied LASSO penalized negative binomial regression models to identify which exposures were associated with the outcomes, while adjusting for co-exposures. In the 1287 children (mean age 8.0 years), 7.3% had a neuropsychiatric medical diagnosis according to parent's reports. During pregnancy, smoking and car traffic showing the strongest associations (e.g. smoking with ADHD index, aMR:1.31 [1.09; 1.59]) among the 13 exposures selected by LASSO, for at least one of the outcomes. During childhood, longer sleep duration, healthy diet and higher family social capital were associated with reduced scores whereas higher exposure to lead, copper, indoor air pollution, unhealthy diet were associated with increased scores. Unexpected decreases in behavioral scores were found with polychlorinated biphenyls (PCBs) and organophosphate (OP) pesticides. CONCLUSIONS: Our systematic exposome approach identified several environmental contaminants and healthy lifestyle habits that may influence behavioral problems in children. Modifying environmental exposures early in life may limit lifetime mental health risk.

In Utero Exposure to Mercury Is Associated With Increased Susceptibility to Liver Injury and Inflammation in Childhood.

BACKGROUND AND AIMS: Nonalcoholic fatty liver disease (NAFLD) is the most prevalent cause of liver disease in children. Mercury (Hg), a ubiquitous toxic metal, has been proposed as an environmental factor contributing to toxicant-associated fatty liver disease. APPROACH AND RESULTS: We investigated the effect of prenatal exposure to Hg on childhood liver injury by combining epidemiological results from a multicenter mother-child cohort with complementary in vitro experiments on monocyte cells that are known to play a key role in liver immune homeostasis and NAFLD. We used data from 872 mothers and their children (median age, 8.1 years; interquartile range [IQR], 6.5-8.7) from the European Human Early-Life Exposome cohort. We measured Hg concentration in maternal blood during pregnancy (median, 2.0 µg/L; IQR, 1.1-3.6). We also assessed serum levels of alanine aminotransferase (ALT), a common screening tool for pediatric NAFLD, and plasma concentrations of inflammation-related cytokines in children. We found that prenatal Hg exposure was associated with a phenotype in children that was characterized by elevated ALT (≥22.1 U/L for females and ≥25.8 U/L for males) and increased concentrations of circulating IL-1ß, IL-6, IL-8, and TNF-α. Consistently, inflammatory monocytes exposed in vitro to a physiologically relevant dose of Hg demonstrated significant up-regulation of genes encoding these four cytokines and increased concentrations of IL-8 and TNF-α in the supernatants. CONCLUSIONS: These findings suggest that developmental exposure to Hg can contribute to inflammation and increased NAFLD risk in early life.

Neighborhood Social and Built Environment and Disparities in the Risk of Hypertension: A Cross-Sectional Study.

Citizens' participation in urban environmental quality assessment is important when identifying local problems in the sustainable development and environmental planning policy. The principal aim of this study was to analyze whether any social differences exist between the joint effect of built neighborhood quality and exposure to urban green spaces and the risk of hypertension. The study sample consisted of 580 participants residing in 11 districts in Kaunas city, Lithuania. Using geographic information systems (GIS), individual data on the socioeconomic status (SES) and health were linked to the participants' perceptions of the environmental quality and exposure to green spaces (NDVI). We used multivariate logistic regression to estimate associations as odds ratios (OR). Those study participants with lower education and those study participants with higher education on low incomes rated their health significantly worse. Low SES persons residing in areas with low exposure to green spaces had a significantly higher risk of hypertension when sex, age, family status, smoking, and income were accounted for (OR 1.83, 95% CI 1.01-3.36). This citizen science study provided evidence that the social environment and the quality of the built environment had a complex effect on disparities in the risk of hypertension.

In utero and childhood exposure to tobacco smoke and multi-layer molecular signatures in children.

BACKGROUND: The adverse health effects of early life exposure to tobacco smoking have been widely reported. In spite of this, the underlying molecular mechanisms of in utero and postnatal exposure to tobacco smoke are only partially understood. Here, we aimed to identify multi-layer molecular signatures associated with exposure to tobacco smoke in these two exposure windows. METHODS: We investigated the associations of maternal smoking during pregnancy and childhood secondhand smoke (SHS) exposure with molecular features measured in 1203 European children (mean age 8.1 years) from the Human Early Life Exposome (HELIX) project. Molecular features, covering 4 layers, included blood DNA methylation and gene and miRNA transcription, plasma proteins, and sera and urinary metabolites. RESULTS: Maternal smoking during pregnancy was associated with DNA methylation changes at 18 loci in child blood. DNA methylation at 5 of these loci was related to expression of the nearby genes. However, the expression of these genes themselves was only weakly associated with maternal smoking. Conversely, childhood SHS was not associated with blood DNA methylation or transcription patterns, but with reduced levels of several serum metabolites and with increased plasma PAI1 (plasminogen activator inhibitor-1), a protein that inhibits fibrinolysis. Some of the in utero and childhood smoking-related molecular marks showed dose-response trends, with stronger effects with higher dose or longer duration of the exposure. CONCLUSION: In this first study covering multi-layer molecular features, pregnancy and childhood exposure to tobacco smoke were associated with distinct molecular phenotypes in children. The persistent and dose-dependent changes in the methylome make CpGs good candidates to develop biomarkers of past exposure. Moreover, compared to methylation, the weak association of maternal smoking in pregnancy with gene expression suggests different reversal rates and a methylation-based memory to past exposures. Finally, certain metabolites and protein markers evidenced potential early biological effects of postnatal SHS, such as fibrinolysis.

Early-Life Environmental Exposures and Childhood Obesity: An Exposome-Wide Approach.

BACKGROUND: Chemical and nonchemical environmental exposures are increasingly suspected to influence the development of obesity, especially during early life, but studies mostly consider single exposure groups. OBJECTIVES: Our study aimed to systematically assess the association between a wide array of early-life environmental exposures and childhood obesity, using an exposome-wide approach. METHODS: The HELIX (Human Early Life Exposome) study measured child body mass index (BMI), waist circumference, skinfold thickness, and body fat mass in 1,301 children from six European birth cohorts age 6-11 y. We estimated 77 prenatal exposures and 96 childhood exposures (cross-sectionally), including indoor and outdoor air pollutants, built environment, green spaces, tobacco smoking, and biomarkers of chemical pollutants (persistent organic pollutants, metals, phthalates, phenols, and pesticides). We used an exposure-wide association study (ExWAS) to screen all exposure-outcome associations independently and used the deletion-substitution-addition (DSA) variable selection algorithm to build a final multiexposure model. RESULTS: The prevalence of overweight and obesity combined was 28.8%. Maternal smoking was the only prenatal exposure variable associated with higher child BMI (z-score increase of 0.28, 95% confidence interval: 0.09, 0.48, for active vs. no smoking). For childhood exposures, the multiexposure model identified particulate and nitrogen dioxide air pollution inside the home, urine cotinine levels indicative of secondhand smoke exposure, and residence in more densely populated areas and in areas with fewer facilities to be associated with increased child BMI. Child blood levels of copper and cesium were associated with higher BMI, and levels of organochlorine pollutants, cobalt, and molybdenum were associated with lower BMI. Similar results were found for the other adiposity outcomes. DISCUSSION: This first comprehensive and systematic analysis of many suspected environmental obesogens strengthens evidence for an association of smoking, air pollution exposure, and characteristics of the built environment with childhood obesity risk. Cross-sectional biomarker results may suffer from reverse causality bias, whereby obesity status influenced the biomarker concentration. https://doi.org/10.1289/EHP5975.

Human Early Life Exposome (HELIX) study: a European population-based exposome cohort.

PURPOSE: Essential to exposome research is the collection of data on many environmental exposures from different domains in the same subjects. The aim of the Human Early Life Exposome (HELIX) study was to measure and describe multiple environmental exposures during early life (pregnancy and childhood) in a prospective cohort and associate these exposures with molecular omics signatures and child health outcomes. Here, we describe recruitment, measurements available and baseline data of the HELIX study populations. PARTICIPANTS: The HELIX study represents a collaborative project across six established and ongoing longitudinal population-based birth cohort studies in six European countries (France, Greece, Lithuania, Norway, Spain and the UK). HELIX used a multilevel study design with the entire study population totalling 31 472 mother-child pairs, recruited during pregnancy, in the six existing cohorts (first level); a subcohort of 1301 mother-child pairs where biomarkers, omics signatures and child health outcomes were measured at age 6-11 years (second level) and repeat-sampling panel studies with around 150 children and 150 pregnant women aimed at collecting personal exposure data (third level). FINDINGS TO DATE: Cohort data include urban environment, hazardous substances and lifestyle-related exposures for women during pregnancy and their offspring from birth until 6-11 years. Common, standardised protocols were used to collect biological samples, measure exposure biomarkers and omics signatures and assess child health across the six cohorts. Baseline data of the cohort show substantial variation in health outcomes and determinants between the six countries, for example, in family affluence levels, tobacco smoking, physical activity, dietary habits and prevalence of childhood obesity, asthma, allergies and attention deficit hyperactivity disorder. FUTURE PLANS: HELIX study results will inform on the early life exposome and its association with molecular omics signatures and child health outcomes. Cohort data are accessible for future research involving researchers external to the project.

Impact of the Social and Natural Environment on Preschool-Age Children Weight.

Background: The complex impact of environmental and social factors on preschool children being overweight/obese is unclear. We examined the associations between the levels of green space exposure and the risk of being overweight/obese for 4-6 year-old children and assessed the impact of maternal education on these associations. Methods: This cross-sectional study included 1489 mother-child pairs living in Kaunas, Lithuania, in 2012-2013. We assessed children overweight/obesity by standardized questionnaires using international body mass index cut-off points, and the level of greenness exposures by satellite-derived normalized difference vegetation index (NDVI) of each child's home and by the distance to a nearest city park. The maternal education was used as the SES indicator. We used logistic regression models to investigate the strength of the associations. Results: Children from families with poorer maternal education, pathological mother-child relations and smoking mothers, and living in areas with less greenness exposure (NDVI-100 m), had significantly higher odds ratios of being overweight/obese. Lower maternal education and distance to a city park modified the effect of greenness cover level exposure on the risk of children being overweight/obese. Conclusions: Higher greenness exposure in the residential settings has beneficial effects on children's physical development. The green spaces exposures for psychosocial stress management is recommended as a measure to prevent overweight/obesity among children.

Effect of diet and maternal education on allergies among preschool children: A case-control study.

INTRODUCTION: The prevalence of asthma and allergy has increased among children. This increase in prevalence might be related to dietary patterns. AIMS: The present epidemiological study investigated the relationship between the consumption of fruit, vegetables, nuts, meat and fish, and the prevalence of wheeze, asthma, and eczema among preschool children. METHODS: This nested case-control study included 1489 children aged 4-6 years and residing in Kaunas city, Lithuania. The subjects were recruited to the KANC newborn cohort study during 2007-2009. Parents' responses to questionnaires were used to collect information on allergic diseases, diet, and other variables. The association between dietary patterns and children's allergic diseases were tested by using logistic regressions, after adjustment for maternal education level, smoking during pregnancy, parental asthma, children's sex, parity, and antibiotic usage during the first year of life. RESULTS: In this study, 83.3% of all children consumed fresh fruit and/or vegetables at least three times per week. A significantly lower adjusted odds ratio (aOR) of wheeze was found among children who ate fruit than among those who did not (aOR: 0.48; 95% CI: 0.22-0.96). The consumption of nuts was also associated with a lower 61% risk of eczema among 4-6 years old children. CONCLUSION: The results indicated a beneficial effect of a frequent consumption of fresh fruit and nuts on the prevalence of allergies among children. These results might have important implications for children's health.

Psychosocial stress and obesity among children residing in Kaunas City.

INTRODUCTION: An unfavorable psychosocial environment has been associated with an increased prevalence of obesity among children. However, the available evidence on the association of low socioeconomic status and parent-child relationships with childhood obesity is scarce. The aim of our study was to conduct a simultaneous evaluation of the risks associated with pathological mother-child relationships, education level, and overweight/obesity among 4-6 year-old children. METHODS: This cross-sectional study included 1489 mother-child pairs living in Kaunas city, Lithuania. The Parenting Stress Index was measured using the Parent-Child Dysfunctional Interaction subscale. Children's overweight/obesity was defined as the body mass index ≥18kg/m2. Logistic regression models as well as crude and adjusted odds ratios (OR) and their 95% confidence intervals (CI) were used to indicate the strength of the associations between childhood overweight/obesity, maternal education level, and psychosocial stress. RESULTS: The percentage of children with overweight/obesity rose with an increasing Parenting Stress Index score. The percentage of children with overweight/obesity in the group of parents with better education and normal mother-child relations was 6.0%, while in the group of less educated parents and pathological mother-child relations, this percentage reached 13.9%. The stratified multivariable model showed that, with reference to the group of better educated parents and normal mother-child relations, lower education level and pathological mother-child relations were statistically significant risk factors for overweight/obesity in 4-6 year-old children, increasing the OR of overweight/obesity (aOR: 2.43; 95% CI: 1.31-4.51). Pathological mother-child relations and maternal smoking mediated the effect of low maternal education level on children's BMI z-scores. CONCLUSION: Pathological mother-child relations, lower parental education levels, and smoking may be predictors of children's overweight/obesity. Measures oriented towards health behavior and psychosocial stress management should be encouraged among parents in order to decrease the risk of overweight/obesity in their children.

The impact of tobacco smoke exposure on wheezing and overweight in 4-6-year-old children.

AIM: To investigate the association between maternal smoking during pregnancy, second-hand tobacco smoke (STS) exposure, education level, and preschool children's wheezing and overweight. METHODS: This cohort study used data of the KANC cohort--1,489 4-6-year-old children from Kaunas city, Lithuania. Multivariate logistic regression was employed to study the influence of prenatal and postnatal STS exposure on the prevalence of wheezing and overweight, controlling for potential confounders. RESULTS: Children exposed to maternal smoking during pregnancy had a slightly increased prevalence of wheezing and overweight. Postnatal exposure to STS was associated with a statistically significantly increased risk of wheezing and overweight in children born to mothers with lower education levels (OR 2.12; 95% CI 1.04-4.35 and 3.57; 95% CI 1.76-7.21, accordingly). CONCLUSIONS: The present study findings suggest that both maternal smoking during pregnancy and STS increase the risk of childhood wheezing and overweight, whereas lower maternal education might have a synergetic effect. Targeted interventions must to take this into account and address household smoking.