Background: DNA methylation (DNAm) may contribute to processes that underlie associations between air pollution and poor health. Therefore, our objective was to evaluate associations between DNAm and ambient concentrations of particulate matter (PM) ≤2.5, ≤10, and 2.5–10 μm in diameter (PM2.5; PM10; PM2.5–10). Methods: We conducted a methylome-wide association study among twelve cohort- and race/ethnicity-stratified subpopulations from the Women's Health Initiative and the Atherosclerosis Risk in Communities study (n = 8397; mean age: 61.5 years; 83% female; 45% African American; 9% Hispanic/Latino American). We averaged geocoded address-specific estimates of daily and monthly mean PM concentrations over 2, 7, 28, and 365 days and 1 and 12 months before exams at which we measured leukocyte DNAm in whole blood. We estimated subpopulation-specific, DNAm-PM associations at approximately 485,000 Cytosine-phosphate-Guanine (CpG) sites in multi-level, linear, mixed-effects models. We combined subpopulation- and site-specific estimates in fixed-effects, inverse variance-weighted meta-analyses, then for associations that exceeded methylome-wide significance and were not heterogeneous across subpopulations (P < 1.0 × 10−7; PCochran's Q > 0.10), we characterized associations using publicly accessible genomic databases and attempted replication in the Cooperative Health Research in the Region of Augsburg (KORA) study. Results: Analyses identified significant DNAm-PM associations at three CpG sites. Twenty-eight-day mean PM10 was positively associated with DNAm at cg19004594 (chromosome 20; MATN4; P = 3.33 × 10−8). One-month mean PM10 and PM2.5–10 were positively associated with DNAm at cg24102420 (chromosome 10; ARPP21; P = 5.84 × 10−8) and inversely associated with DNAm at cg12124767 (chromosome 7; CFTR; P = 9.86 × 10−8). The PM-sensitive CpG sites mapped to neurological, pulmonary, endocrine, and cardiovascular disease-related genes, but DNAm at those sites was not associated with gene expression in blood cells and did not replicate in KORA. Conclusions: Ambient PM concentrations were associated with DNAm at genomic regions potentially related to poor health among racially, ethnically and environmentally diverse populations of U.S. women and men. Further investigation is warranted to uncover mechanisms through which PM-induced epigenomic changes may cause disease.

Objective: Physical activity (PA) during pregnancy is associated with lower neonatal fat mass, but associations with child body composition are mixed. The purpose of this study was to examine associations between trimester-specific pregnancy PA and child body composition at 4 years. Methods: Participants of the Minnesota Infant Nutrition, Neurodevelopment, and Obesity Study were asked to recall participation in any moderate or vigorous PA in the first (T1), second (T2) and third (T3) trimesters at about 5 years postpartum. Child fat mass and fat-free mass were measured via air displacement plethysmography at 2 weeks, 3 months and 4 years of age. Multivariate linear regression was used for analyses. Results: Of 51 possible participants, 37 recalled pregnancy PA. Any vigorous PA in T3 was associated with lower child fat mass at 4 years (adjβ = −1.077, p < 0.05). Conclusion: Late pregnancy PA may have lasting benefits for child body composition. Replication of these findings is needed in a larger sample with prospective measures.

Genome-wide association studies (GWAS) have identified >250 loci for body mass index (BMI), implicating pathways related to neuronal biology. Most GWAS loci represent clusters of common, noncoding variants from which pinpointing causal genes remains challenging. Here we combined data from 718,734 individuals to discover rare and low-frequency (minor allele frequency (MAF) < 5%) coding variants associated with BMI. We identified 14 coding variants in 13 genes, of which 8 variants were in genes (ZBTB7B, ACHE, RAPGEF3, RAB21, ZFHX3, ENTPD6, ZFR2 and ZNF169) newly implicated in human obesity, 2 variants were in genes (MC4R and KSR2) previously observed to be mutated in extreme obesity and 2 variants were in GIPR. The effect sizes of rare variants are ~10 times larger than those of common variants, with the largest effect observed in carriers of an MC4R mutation introducing a stop codon (p.Tyr35Ter, MAF = 0.01%), who weighed ~7 kg more than non-carriers. Pathway analyses based on the variants associated with BMI confirm enrichment of neuronal genes and provide new evidence for adipocyte and energy expenditure biology, widening the potential of genetically supported therapeutic targets in obesity.

Objective: To better understand the relationship between cardiovascular disease risk and age-at-natural menopause using genetic data. Methods: Early menopause is associated with cardiovascular disease risk. We constructed a genetic risk score comprising 56 age-at-natural menopause decreasing alleles in men and women from the Framingham Heart Study, the Atherosclerosis Risk in Communities Study, and the Rotterdam Study. If the genetic predisposition to earlier age-at-natural menopause is associated with increased cardiovascular disease risk, it is reasonable to ask whether the risk is shared by men carrying the alleles, despite not experiencing menopause. We estimated the hazard ratio for the score for time to first cardiovascular event. To investigate the possible genetic pleiotropy between age-at-natural menopause and cardiovascular disease, we performed cross-trait linkage disequilibrium score regressions between age-at-natural menopause and cardiovascular disease and risk factors using genome-wide association studies. Results: Twenty-two thousand five hundred and sixty-eight cardiovascular disease-free participants at baseline were analyzed (9,808 men, 12,760 women). Each additional unit of the genetic propensity to earlier age-at-natural menopause increased the hazard of both cardiovascular disease and cardiac death in women (cardiovascular disease: hazard ratio 1.10 [1.04-1.16], P = 9.7 × 10 -4 ; cardiac death: 1.12 [1.02-1.24], P = 0.03), whereas no effect was observed for either outcome in men (hazard ratio 0.99 [0.95-1.04], P = 0.71; 1.05 [0.94-1.16], P = 0.34). We found significant negative genetic correlations in women, but not men, between age-at-natural menopause and cardiovascular disease and risk factors. Conclusion: Genetic variants associated with earlier age-at-natural menopause are associated with increased cardiovascular disease risk in women, but not men, suggesting sex-specific genetic effects on cardiovascular disease risk.

Background/Objectives:There is increasing evidence of a relationship between blood DNA methylation and body mass index (BMI). We aimed to assess associations of BMI with individual methylation measures (CpGs) through a cross-sectional genome-wide DNA methylation association study and a longitudinal analysis of repeated measurements over time.Subjects/Methods:Using the Illumina Infinium HumanMethylation450 BeadChip, DNA methylation measures were determined in baseline peripheral blood samples from 5361 adults recruited to the Melbourne Collaborative Cohort Study (MCCS) and selected for nested case-control studies, 2586 because they were subsequently diagnosed with cancer (cases) and 2775 as controls. For a subset of 1088 controls, these measures were repeated using blood samples collected at wave 2 follow-up, a median of 11 years later; weight was measured at both time points. Associations between BMI and blood DNA methylation were assessed using linear mixed-effects regression models adjusted for batch effects and potential confounders. These were applied to cases and controls separately, with results combined through fixed-effects meta-Analysis.Results:Cross-sectional analysis identified 310 CpGs associated with BMI with P<1.0 × 10 â '7, 225 of which had not been reported previously. Of these 225 novel associations, 172 were replicated (P<0.05) using the Atherosclerosis Risk in Communities (ARIC) study. We also replicated using MCCS data (P<0.05) 335 of 392 associations previously reported with P<1.0 × 10 â '7, including 60 that had not been replicated before. Associations between change in BMI and change in methylation were observed for 34 of the 310 strongest signals in our cross-sectional analysis, including 7 that had not been replicated using the ARIC study.Conclusions:Together, these findings suggest that BMI is associated with blood DNA methylation at a large number of CpGs across the genome, several of which are located in or near genes involved in ATP-binding cassette transportation, tumour necrosis factor signalling, insulin resistance and lipid metabolism.

Background: We conducted an epigenome-wide association study (EWAS) on obesity in healthy youth and young adults and further examined to what extent identified signals influenced gene expression and were independent of cell type composition and obesity-related cardio-metabolic risk factors. Genome-wide DNA methylation data from leukocytes were obtained from 700 African Americans aged 14-36. We also measured genome-wide DNA methylation data from neutrophils as well as genome-wide gene expression data from leukocytes in a subset of samples (n = 188). Results: The EWAS identified 76 obesity-related CpG sites in leukocytes with p < 1 × 10-7. In silico replication in the ARIC study of 2097 African Americans aged 47-70 validated 54 CpG sites. Out of the 54 CpG sites, 29 associations with obesity were novel and 37 were replicated in neutrophils. Fifty one CpG sites were associated with at least one cardio-metabolic risk factor; however, the number reduced to 9 after adjustment for obesity. Sixteen CpG sites were associated with expression of 17 genes in cis, of which 5 genes displayed differential expression between obese cases and lean controls. We also replicated 71.5% of obesity-related CpG sites previously reported. Conclusion: In this study of youth and young adults, we identified 29 novel CpG sites associated with obesity and replicated the majority of the CpG sites previously identified. We further demonstrated that the majority of the obesity-related CpG sites in leukocytes were not driven by cell composition or obesity-related cardio-metabolic risk factors. We also provided the direct link between DNA methylation-gene expression-obesity for 5 genes.

This study determined the effects of consuming a high-fructose corn syrup (HFCS)-sweetened beverage on breast milk fructose, glucose, and lactose concentrations in lactating women. At six weeks postpartum, lactating mothers (n = 41) were randomized to a crossover study to consume a commercially available HFCS-sweetened beverage or artificially sweetened control beverage. At each session, mothers pumped a complete breast milk expression every hour for six consecutive hours. The baseline fasting concentrations of breast milk fructose, glucose, and lactose were 5.0 ± 1.3 µg/mL, 0.6 ± 0.3 mg/mL, and 6.8 ± 1.6 g/dL, respectively. The changes over time in breast milk sugars were significant only for fructose (treatment × time, p < 0.01). Post hoc comparisons showed the HFCS-sweetened beverage vs. control beverage increased breast milk fructose at 120 min (8.8 ± 2.1 vs. 5.3 ± 1.9 µg/mL), 180 min (9.4 ± 1.9 vs. 5.2 ± 2.2 µg/mL), 240 min (7.8 ± 1.7 vs. 5.1 ± 1.9 µg/mL), and 300 min (6.9 ± 1.4 vs. 4.9 ± 1.9 µg/mL) (all p < 0.05). The mean incremental area under the curve for breast milk fructose was also different between treatments (14.7 ± 1.2 vs. −2.60 ± 1.2 µg/mL × 360 min, p < 0.01). There was no treatment × time interaction for breast milk glucose or lactose. Our data suggest that the consumption of an HFCS-sweetened beverage increased breast milk fructose concentrations, which remained elevated up to five hours post-consumption.

Background: Infants born prematurely are at risk for neurodevelopmental complications. Early growth is associated with improved later cognition. The relationship of early proportionality and body composition with later cognition is not well established. Objectives: To assess differences in fat-free mass and adiposity (fat mass, percent body fat) changes in preterm and full-term infants through preschool age and examine associations with early childhood cognition. Methods: This is a prospective, observational study in an appropriate for gestational age cohort of 71 patients (20 preterm and 51 full-term) from infancy through preschool age. Anthropometric and body composition measurements via air displacement plethysmography were obtained during infancy at term and 3-4 months (preterm corrected ages), and at 4 years. Cognitive testing occurred at 4 years. Associations of body composition changes between visits with cognitive function were tested using linear regression. Results: In the preterm group, higher term to 4-month corrected age percent body fat gains were associated with lower working memory performance (p = 0.01), and higher 4-month corrected age to 4-year fat-free mass gains were associated with higher full-scale IQ (p = 0.03) and speed of processing performance (p ≤ 0.02). In the full-term group, higher 4-month to 4-year fat mass gains were associated with lower full-scale IQ (p = 0.03). Conclusions: Body composition gains during different time periods are associated with varying areas of cognitive function. These findings may inform interventions aimed at optimal growth.

Current knowledge of the genetic architecture of key reproductive events across the female life course is largely based on association studies of European descent women. The relevance of known loci for age at menarche (AAM) and age at natural menopause (ANM) in diverse populations remains unclear. We investigated 32 AAM and 14 ANM previously-identified loci and sought to identify novel loci in a trans-ethnic array-wide study of 196,483 SNPs on the MetaboChip (Illumina, Inc.). A total of 45,364 women of diverse ancestries (African, Hispanic/Latina, Asian American and American Indian/Alaskan Native) in the Population Architecture using Genomics and Epidemiology (PAGE) Study were included in cross-sectional analyses of AAM and ANM. Within each study we conducted a linear regression of SNP associations with self-reported or medical record-derived AAM or ANM (in years), adjusting for birth year, population stratification, and center/region, as appropriate, and meta-analyzed results across studies using multiple meta-analytic techniques. For both AAM and ANM, we observed more directionally consistent associations with the previously reported risk alleles than expected by chance (p-valuesbinomial0.01). Eight densely genotyped reproductive loci generalized significantly to at least one non-European population. We identified one trans-ethnic array-wide SNP association with AAM and two significant associations with ANM, which have not been described previously. Additionally, we observed evidence of independent secondary signals at three of six AAM trans-ethnic loci. Our findings support the transferability of reproductive trait loci discovered in European women to women of other race/ethnicities and indicate the presence of additional trans-ethnic associations both at both novel and established loci. These findings suggest the benefit of including diverse populations in future studies of the genetic architecture of female growth and development.

Background: Very preterm (VPT) infants are at-risk for altered growth, slower speed of processing (SOP), and hypertension. This study assesses the relationship between postnatal body composition (BC), neurodevelopment (indexed by SOP), and blood pressure (BP) in VPT infants. Methods: Thirty-four VPT infants underwent weekly measurements and BC testing until discharge and post-discharge at 4 mos CGA and 4 yrs. At post-discharge visits, SOP was assessed using visual evoked potentials and the NIH Toolbox; BP was also measured. Results: In-hospital rate of weight, length and fat-free mass (FFM) gains were associated with faster SOP at 4 yrs. Higher rate of gains in weight and FFM from discharge to 4 mos CGA were associated with faster SOP at 4 mos CGA, while higher fat mass (FM) gains during the same time were positively associated with BP at 4 yrs. BC at 4 yrs nor gains beyond 4 mos CGA were associated with outcomes. Conclusions: In VPT infants, early FFM gains are associated with faster SOP, whereas post-discharge FM gains are associated with higher BPs at 4 yrs. This shows birth to 4 mos CGA is a sensitive period for growth and its relation to neurodevelopmental and metabolic outcomes. Close monitoring and early nutritional adjustments to optimize quality of gains may improve outcomes.

Obesity (Silver Spring) 2017; 25:2092-2099. doi:10.1002/oby.22025 The authors of this article recently discovered that the C-reactive protein values used in the analysis were approximately threefold higher than the true concentration. The manufacturers of the ELISA kit recently informed the authors that the kit instruction manual gave the wrong volume of buffer needed to reconstitute the standard. This laboratory error affected the data presented in Tables and and Figure of the manuscript. After rerunning all of the statistical analyses with the corrected C-reactive protein values, there are no changes in either the statistical significance of the associations or the conclusions and interpretation of the findings. The authors apologize for this error. Summary statistics for breast milk inflammatory markers at 1 and 3 months post partum (Table presented.) Data presented as median (interquartile range) or mean ± SD. aP value testing for differences between 1- and 3-month summary statistics using Wilcoxon signed rank sum test for nontransformed data and paired t tests for log-transformed data. Bolded values are statistically significant (P<0.05). bThe value of 1 was added to the raw interleukin-6 values before log transformations were performed to avoid negative values on the log scale. Associations of maternal anthropometrics with log-transformed breast milk C-reactive protein at 1 and 3 months post partuma (Table presented.) aAll continuous independent variables (pre-pregnancy BMI, gestational weight gain, and postpartum weight loss) were standardized to a mean of 0 and a standard deviation of 1. Bolded values are statistically significant (P<0.05). bModel 1 is the crude model. cModel 2 adjusts for maternal education and gestational age at delivery. dModel 3 adjusts for covariates in Model 2, as well as pre-pregnancy BMI. eCalculated using the 2009 Institute of Medicine gestational weight gain guidelines, based on pre-pregnancy BMI. fPostpartum weight loss was calculated as maternal weight at delivery minus maternal weight at 1 and 3 months post partum and was entered into the mixed effects models as a time-varying independent variable. (Figure presented.) Adjusted means of back-transformed C-reactive protein (CRP) at 1 and 3 months post partum by pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) category. Differences between the mean values for each pre-pregnancy BMI and GWG category are denoted by different superscript letters (a, b, c, or d). In the top panel, at 1 month post partum, women with normal weight who gained weight below/within the guidelines (a) had significantly lower breast milk CRP than all other groups (b), except overweight women who gained weight below/within the guidelines. Women with obesity who gained below/within GWG guidelines (c) had significantly higher CRP than women with overweight, regardless of GWG category, and women with normal weight who gained below/within GWG guidelines (d). In the bottom panel, at 3 months post partum, women with normal weight who gained weight below/within guidelines (a) had significantly lower CRP than women with normal weight who exceeded GWG guidelines and women with obesity, regardless of GWG category (b). Women with obesity who experienced excessive weight gain (c) had significantly higher CRP than all normal weight and overweight groups (d; P<0.05 for all).

Background: Even before the onset of age-related diseases, obesity might be a contributing factor to the cumulative burden of oxidative stress and chronic inflammation throughout the life course. Obesity may therefore contribute to accelerated shortening of telomeres. Consequently, obese persons are more likely to have shorter telomeres, but the association between body mass index (BMI) and leukocyte telomere length (TL) might differ across the life span and between ethnicities and sexes. Objective: A collaborative cross-sectionalmeta-analysis of observational studies was conducted to investigate the associations between BMI and TL across the life span. Design: Eighty-seven distinct study samples were included in the meta-analysis capturing data from 146,114 individuals. Studyspecific age- and sex-adjusted regression coefficients were combined by using a random-effects model in which absolute [base pairs (bp)] and relative telomere to single-copy gene ratio (T/S ratio) TLs were regressed against BMI. Stratified analysis was performed by 3 age categories ("young": 18-60 y; "middle": 61-75 y; and "old": >75 y), sex, and ethnicity. Results: Each unit increase in BMI corresponded to a-3.99 bp (95% CI: -5.17, -2.81 bp) difference in TL in the total pooled sample; among young adults, each unit increase in BMI corresponded to a -7.67 bp (95% CI:-10.03,-5.31 bp) difference. Each unit increase in BMI corresponded to a -1.58 × 10-3 unit T/S ratio (0.16% decrease; 95% CI: -2.14 × 10-3, -1.01 × 10-3) difference in ageand sex-adjusted relative TL in the total pooled sample; among young adults, each unit increase in BMI corresponded to a -2.58 × 10-3 unit T/S ratio (0.26% decrease; 95% CI: -3.92 × 10-3, -1.25 × 10-3). The associations were predominantly for the white pooled population. No sex differences were observed. Conclusions: A higher BMI is associated with shorter telomeres, especially in younger individuals. The presently observed difference is not negligible. Meta-analyses of longitudinal studies evaluating change in body weight alongside change in TL arewarranted.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC Osteosarcoma (OS) is the most common form of bone cancer in children and displays a peak incidence that coincides with the pubertal growth spurt. Consequently a number of studies have investigated whether cases are taller at diagnosis than the general pediatric population; a recent analysis which pooled seven such studies found that cases were disproportionately in the 51st-89th and especially the >90th percentiles of height-for-age (HFA). However, no study to our knowledge has examined whether OS patients exhibit greater HFA at earlier ages. During 2008-2010 we enrolled 290 OS cases <20 years of age diagnosed at Children's Oncology Group institutions in the United States and Canada in a genetic epidemiologic study. In addition to collecting buccal cell samples for genetic analysis we requested permission to obtain length/height data from medical records from birth to diagnosis. Records were obtained for 153 male and 111 female participants. Anthropometrics and date of measurement were doubly entered into a custom database, and inconsistencies were resolved by a third abstraction. Between 1 and 46 measurements (median = 11) were recorded up to the earlier of diagnosis or 12 years of age. Mixed effects cubic spline models were applied to length/height to produce individual growth curves using Stata. Sex was included as a main effect and as interactions with the model slope terms to account for gender differences in growth. Individual estimates of length/height at every three months of age between birth and age two years, and at each year of age thereafter, were computed and transformed to Z-scores according to the CDC 2000 reference. OS cases of both sexes were consistently longer or taller than reference data at all ages, as at no age did the 95% confidence interval around mean Z-score include 0 (i.e. the population mean and median). Under 2 years of age length was examined; mean Z-score at birth was 0.56 (71st %ile), rose to 0.72 (76th %ile) at 0.5 years, and fell to 0.31 (62nd %ile) at 2 years. Over 2 years of age height was examined; mean Z-score was 0.48 (68th %ile) at 4 years, dipped to 0.33 (63rd %ile) at 8-9 years, and rose to 0.49 (69th %ile) at 12 years. No differences in mean Z-score at each age were observed by age at diagnosis (sex specific inter-quartile range of age at diagnosis vs. other), location of tumor (long bone of the lower limb vs. other), or race (White non-Hispanic vs. other). Our data clearly indicate that pediatric OS patients are substantially longer/taller than the United States national norm at all ages before puberty. Citation Format: Logan G. Spector, Kathryn Ritter, Ellen W. Demerath, Charles Sklar, Julie A. Ross, Mark Krailo, Rajaram Nagarajan, David Malkin, Tracy L. Bergemann, Sharon A. Savage, William Johnson. Pediatric osteosarcoma patients are taller than average from birth to age twelve: a report from the Children's Oncology Group. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2532. doi:10.1158/1538-7445.AM2013-2532

34 Human milk is a complex mix of nutritional and bioactive components that provide complete nutrition for 35 the infant. However, we lack a systematic knowledge of the factors shaping milk composition and how 36 milk variation influences infant health. Here, we used multi-omic profiling to characterize interactions 37 between maternal genetics, milk gene expression, milk composition, and the infant fecal microbiome in 38 242 exclusively breastfeeding mother-infant pairs. We identified 487 genetic loci associated with milk 39 gene expression unique to the lactating mammary gland, including loci that impacted breast cancer risk 40 and human milk oligosaccharide concentration. Integrative analyses uncovered connections between 41 milk gene expression and infant gut microbiome, including an association between the expression of 42 inflammation-related genes with IL-6 concentration in milk and the abundance of Bifidobacteria in the 43 infant gut. Our results show how an improved understanding of the genetics and genomics of human 44 milk connects lactation biology with maternal and infant health. 45 46. CC-BY 4.0 International license available under a (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made The copyright holder for this preprint this version posted January 25, 2023. ; https://doi.