I am an environmental epidemiologist and serve as an Assistant Professor in the Department of Epidemiology and Population Health at Stanford University. I joined the faculty at Stanford School of Medicine in 2022.

My research focuses on characterizing molecular and epigenetic biomarkers and the extent to which these alterations contribute to disease risk throughout the life course. My group utilizes computational approaches to investigate environmental chemical mixtures, biological aging markers and fetal epigenetic programming. We have several studies looking at chemical and non-chemical stressors in early-life and subsequent health including; neurodevelopment, obesity and immune function.

My research examines the intersection of chemical and social environments in shaping health and disease.

Academic Appointments

Honors & Awards

  • Outstanding New Environmental Scientist (ONES) award, NIEHS/NIH (2020-2025)
  • Young Alumni Award, College of Science, Oregon State University (2020)

Stanford Advisees

All Publications

  • Carcinogenicity of talc and acrylonitrile. The Lancet. Oncology Stayner, L. T., Carreón-Valencia, T., Demers, P. A., Fritz, J. M., Sim, M. R., Stewart, P., Tsuda, H., Cardenas, A., Consonni, D., Davies, L., De Matteis, S., Felley-Bosco, E., Ghio, A. J., Göen, T., Grosse, Y., Gualtieri, A. F., Josephy, P. D., Koutros, S., Linhart, I., Louro, H., O'Brien, K. M., Panzacchi, S., Peña, L., Rössner, P., Schildkraut, J. M., Stefaniak, A. B., Wentzensen, N., Wild, P., Xu, Y., de Conti, A., Facchin, C., Wedekind, R., Ahmadi, A., Blanco, J., Chittiboyina, S., Kulasingam, S., MacLehose, R., Motlhale, M., Shah, S., Suonio, E., Mattock, H., Kunzmann, A., Madia, F., Pasqual, E., Benbrahim-Tallaa, L., Schubauer-Berigan, M. K. 2024

    View details for DOI 10.1016/S1470-2045(24)00384-X

    View details for PubMedID 38976996

  • Prenatal blood metals, per- and polyfluoroalkyl substances and antigen- or mitogen-stimulated cord blood lymphocyte proliferation and cytokine secretion. Environmental research Smith, A., Lin, P. D., Rifas-Shiman, S. L., Fleisch, A. F., Wright, R. O., Coull, B., Finn, P. W., Oken, E., Gold, D. R., Cardenas, A. 2024: 119555


    Evidence suggests that prenatal per- and polyfluoroalkyl substances (PFAS) and metals, two classes of chemicals found ubiquitously in human populations, influence immune system development and response.We evaluated whether first trimester blood PFAS and metals were associated with antigen- or mitogen-stimulated cord blood lymphocyte proliferation and cytokine secretion.We measured six PFAS, as well as six nonessential and four essential metals, in first trimester blood from participants in the longitudinal pre-birth Project Viva cohort, recruited between 1999-2000 in eastern Massachusetts. We measured antigen- or mitogen-stimulated cord blood mononuclear cell proliferation responses (n=269-314) and cytokine secretion (n=217-302). We used covariate-adjusted least absolute shrinkage and selection operator (LASSO) for variable selection and multivariable regression to estimate associations with the immune markers.Each ng/mL of MeFOSAA was associated with a 3.6% (1.4, 5.8) higher lymphocyte proliferation response after stimulation with egg antigen, as well as 0.8 (0.7, 1.0) reduced odds of having IFN-γ detected in response to dust mite. Each ng/g increment of cesium was associated with 27.8% (-45.1, -4.9) lower IL-10 levels in response to dust mite. Each ng/g increment of mercury was associated with 12.0% (1.3, 23.8) higher IL-13 levels in response to mitogen PHA. Each ng/g increment of selenium and zinc was associated with 0.2% (0.01, 0.4) and 0.01% (0.002, 0.02) higher TNF-α in response to mitogen PHA, respectively.Prenatal metals and PFAS influence cord blood lymphocyte proliferation and cytokine secretion in ways that may increase risk for atopic disease in childhood.

    View details for DOI 10.1016/j.envres.2024.119555

    View details for PubMedID 38964580

  • Prenatal exposure to environmental phenols and phthalates and altered patterns of DNA methylation in childhood. Environment international Khodasevich, D., Holland, N., Harley, K. G., Eskenazi, B., Barcellos, L. F., Cardenas, A. 2024; 190: 108862


    Epigenetic marks are key biomarkers linking the prenatal environment to health and development. However, DNA methylation associations and persistence of marks for prenatal exposure to multiple Endocrine Disrupting Chemicals (EDCs) in human populations have not been examined in great detail.We measured Bisphenol-A (BPA), triclosan, benzophenone-3 (BP3), methyl-paraben, propyl-paraben, and butyl-paraben, as well as 11 phthalate metabolites, in two pregnancy urine samples, at approximately 13 and 26 weeks of gestation in participants of the Center for the Health Assessment of Mothers and Children of Salinas (CHAMACOS) study (N = 309). DNA methylation of cord blood at birth and child peripheral blood at ages 9 and 14 years was measured with 450K and EPIC arrays. Robust linear regression was used to identify differentially methylated probes (DMPs), and comb-p was used to identify differentially methylated regions (DMRs) in association with pregnancy-averaged EDC concentrations. Quantile g-computation was used to assess associations of the whole phenol/phthalate mixture with DMPs and DMRs.Prenatal BPA exposure was associated with 1 CpG among males and Parabens were associated with 10 CpGs among females at Bonferroni-level significance in cord blood. Other suggestive DMPs (unadjusted p-value < 1 × 10-6) and several DMRs associated with the individual phenols and whole mixture were also identified. A total of 10 CpG sites at least suggestively associated with BPA, Triclosan, BP3, Parabens, and the whole mixture in cord blood were found to persist into adolescence in peripheral blood.We found sex-specific associations between prenatal phenol exposure and DNA methylation, particularly with BPA in males and Parabens in females. Additionally, we found several DMPs that maintained significant associations with prenatal EDC exposures at age 9 and age 14 years.

    View details for DOI 10.1016/j.envint.2024.108862

    View details for PubMedID 38972116

  • Precision Medicine for Asthma Treatment: Unlocking the Potential of the Epigenome and Microbiome. The Journal of allergy and clinical immunology Perez-Garcia, J., Cardenas, A., Lorenzo-Diaz, F., Pino-Yanes, M. 2024


    Asthma is a leading worldwide biomedical concern. Patients can experience life-threatening worsening episodes (exacerbations) usually controlled by anti-inflammatory and bronchodilator drugs. However, substantial heterogeneity in treatment response exists and a subset of patients with unresolved asthma carry the major burden of this disease. The study of the epigenome and microbiome might bridge the gap between human genetics and environmental exposures to partially explain the heterogeneity in drug response. This review aims to provide a critical examination of the existing literature on the microbiome and epigenetic studies examining associations with asthma treatments and drug response, highlight convergent pathways, address current challenges, and offer future perspectives. Current epigenetic and microbiome studies have shown the bilateral relationship between asthma pharmacological interventions and the human epigenome and microbiome. These studies, focusing on corticosteroids and to a lesser extent on bronchodilators, azithromycin, immunotherapy, and mepolizumab, have improved the understanding of the molecular basis of treatment response and identified promising biomarkers for drug response prediction. Immune and inflammatory pathways (i.e., IL-2, TNF-α, NF-κB, and CEBPs) underlie microbiome-epigenetic associations with asthma treatment, representing potential therapeutic pathways to be targeted. A comprehensive evaluation of these omic biomarkers could significantly contribute to precision medicine and new therapeutic target discovery.

    View details for DOI 10.1016/j.jaci.2024.06.010

    View details for PubMedID 38906272

  • Ambient air pollution and urological cancer risk: A systematic review and meta-analysis of epidemiological evidence. Nature communications Li, J., Deng, Z., Soerensen, S. J., Kachuri, L., Cardenas, A., Graff, R. E., Leppert, J. T., Langston, M. E., Chung, B. I. 2024; 15 (1): 5116


    Exposure to ambient air pollution has significant adverse health effects; however, whether air pollution is associated with urological cancer is largely unknown. We conduct a systematic review and meta-analysis with epidemiological studies, showing that a 5 μg/m3 increase in PM2.5 exposure is associated with a 6%, 7%, and 9%, increased risk of overall urological, bladder, and kidney cancer, respectively; and a 10 μg/m3 increase in NO2 is linked to a 3%, 4%, and 4% higher risk of overall urological, bladder, and prostate cancer, respectively. Were these associations to reflect causal relationships, lowering PM2.5 levels to 5.8 μg/m3 could reduce the age-standardized rate of urological cancer by 1.5 ~ 27/100,000 across the 15 countries with the highest PM2.5 level from the top 30 countries with the highest urological cancer burden. Implementing global health policies that can improve air quality could potentially reduce the risk of urologic cancer and alleviate its burden.

    View details for DOI 10.1038/s41467-024-48857-2

    View details for PubMedID 38879581

    View details for PubMedCentralID PMC11180144

  • Maternal age is related to offspring DNA methylation: A meta-analysis of results from the PACE consortium. Aging cell Yeung, E., Biedrzycki, R. J., Gómez Herrera, L. C., Issarapu, P., Dou, J., Marques, I. F., Mansuri, S. R., Page, C. M., Harbs, J., Khodasevich, D., Poisel, E., Niu, Z., Allard, C., Casey, E., Berstein, F. M., Mancano, G., Elliott, H. R., Richmond, R., He, Y., Ronkainen, J., Sebert, S., Bell, E. M., Sharp, G., Mumford, S. L., Schisterman, E. F., Chandak, G. R., Fall, C. H., Sahariah, S. A., Silver, M. J., Prentice, A. M., Bouchard, L., Domellof, M., West, C., Holland, N., Cardenas, A., Eskenazi, B., Zillich, L., Witt, S. H., Send, T., Breton, C., Bakulski, K. M., Fallin, M. D., Schmidt, R. J., Stein, D. J., Zar, H. J., Jaddoe, V. W., Wright, J., Grazuleviciene, R., Gutzkow, K. B., Sunyer, J., Huels, A., Vrijheid, M., Harlid, S., London, S., Hivert, M. F., Felix, J., Bustamante, M., Guan, W. 2024: e14194


    Worldwide trends to delay childbearing have increased parental ages at birth. Older parental age may harm offspring health, but mechanisms remain unclear. Alterations in offspring DNA methylation (DNAm) patterns could play a role as aging has been associated with methylation changes in gametes of older individuals. We meta-analyzed epigenome-wide associations of parental age with offspring blood DNAm of over 9500 newborns and 2000 children (5-10 years old) from the Pregnancy and Childhood Epigenetics consortium. In newborns, we identified 33 CpG sites in 13 loci with DNAm associated with maternal age (PFDR < 0.05). Eight of these CpGs were located near/in the MTNR1B gene, coding for a melatonin receptor. Regional analysis identified them together as a differentially methylated region consisting of 9 CpGs in/near MTNR1B, at which higher DNAm was associated with greater maternal age (PFDR = 6.92 × 10-8) in newborns. In childhood blood samples, these differences in blood DNAm of MTNR1B CpGs were nominally significant (p < 0.05) and retained the same positive direction, suggesting persistence of associations. Maternal age was also positively associated with higher DNA methylation at three CpGs in RTEL1-TNFRSF6B at birth (PFDR < 0.05) and nominally in childhood (p < 0.0001). Of the remaining 10 CpGs also persistent in childhood, methylation at cg26709300 in YPEL3/BOLA2B in external data was associated with expression of ITGAL, an immune regulator. While further study is needed to establish causality, particularly due to the small effect sizes observed, our results potentially support offspring DNAm as a mechanism underlying associations of maternal age with child health.

    View details for DOI 10.1111/acel.14194

    View details for PubMedID 38808605

  • Protocol for CARES-HAPIN: an ambidirectional cohort study on exposure to environmental tobacco smoke and risk of early childhood caries. BMJ open Patil, S. S., Puttaswamy, N., Cardenas, A., Barr, D. B., Ghosh, S., Balakrishnan, K. 2024; 14 (5): e083874


    Prenatal and postnatal exposure to environmental tobacco smoke (ETS) has been linked with early childhood caries (ECC), but the specific molecular mechanisms and pathways remain largely unknown. The Caries Risk from exposure to Environmental tobacco Smoke (CARES) within the Household Air Pollution Intervention Network (HAPIN) study aims to establish the association between ETS and ECC by employing epidemiological and novel biomarker-based approaches. Here, we outline the overall design and rationale of the project.We will leverage the infrastructure and data from the HAPIN trial (India) to mount the CARES study. In this ambidirectional cohort study, children (n=735, aged: 3-5 years) will undergo ECC examination by a trained dentist using standard criteria and calibrated methods. Structured questionnaires will be used to gather information on sociodemographic variables, dietary habits, oral hygiene, oral health-related quality of life and current exposure to ETS. We will collect non-invasive or minimally invasive biospecimens (i.e., saliva, buccal cells, dried blood spots and urine) from a subset of HAPIN children (n=120) to assess a battery of biomarkers indicative of exposure to ETS, early biological effect and epigenetic modifications. Both self-reported and objective measures of ETS exposure collected longitudinally during in utero and early postnatal periods will be accessed from the HAPIN database. We will apply current science data techniques to assess the association and interrelationships between ETS, ECC, and multiple biomarkers.Information gathered in this research will be published in peer-reviewed journals and summaries will be shared with the key stakeholders as well as patients and their parents/guardians involved in this study. Sri Ramachandra Institute of Higher Education and Research Ethics Board has approved the study protocol (IEC-NI22/JUL/83/82).NCT02944682.

    View details for DOI 10.1136/bmjopen-2024-083874

    View details for PubMedID 38749682

    View details for PubMedCentralID PMC11097839

  • Epigenome-wide association study of lung cancer among never smokers in two prospective cohorts in Shanghai, China. Thorax Rahman, M. L., Breeze, C. E., Shu, X. O., Wong, J. Y., Blechter, B., Cardenas, A., Wang, X., Ji, B. T., Hu, W., Cai, Q., Hosgood, H. D., Yang, G., Shi, J., Long, J., Gao, Y. T., Bell, D. A., Zheng, W., Rothman, N., Lan, Q. 2024


    The aetiology of lung cancer among individuals who never smoked remains elusive, despite 15% of lung cancer cases in men and 53% in women worldwide being unrelated to smoking. Epigenetic alterations, particularly DNA methylation (DNAm) changes, have emerged as potential drivers. Yet, few prospective epigenome-wide association studies (EWAS), primarily focusing on peripheral blood DNAm with limited representation of never smokers, have been conducted.We conducted a nested case-control study of 80 never-smoking incident lung cancer cases and 83 never-smoking controls within the Shanghai Women's Health Study and Shanghai Men's Health Study. DNAm was measured in prediagnostic oral rinse samples using Illumina MethylationEPIC array. Initially, we conducted an EWAS to identify differentially methylated positions (DMPs) associated with lung cancer in the discovery sample of 101 subjects. The top 50 DMPs were further evaluated in a replication sample of 62 subjects, and results were pooled using fixed-effect meta-analysis.Our study identified three DMPs significantly associated with lung cancer at the epigenome-wide significance level of p<8.22×10-8. These DMPs were identified as cg09198866 (MYH9; TXN2), cg01411366 (SLC9A10) and cg12787323. Furthermore, examination of the top 1000 DMPs indicated significant enrichment in epithelial regulatory regions and their involvement in small GTPase-mediated signal transduction pathways. Additionally, GrimAge acceleration was identified as a risk factor for lung cancer (OR=1.19 per year; 95% CI 1.06 to 1.34).While replication in a larger sample size is necessary, our findings suggest that DNAm patterns in prediagnostic oral rinse samples could provide novel insights into the underlying mechanisms of lung cancer in never smokers.

    View details for DOI 10.1136/thorax-2023-220352

    View details for PubMedID 38702190

  • Air pollution mixture associated with oxidative stress exacerbation and symptoms deterioration in allergic rhinitis patients: Evidence from a panel study. The Science of the total environment Li, J., Wu, H., Xing, W., Li, X., Han, Z., Ji, R., Deng, Z., Jung, M., Sun, S., Chung, B. I., Cardenas, A., Langston, M. E. 2024: 172688


    With allergic rhinitis (AR) on the rise globally, there has been a growing focus on the role of environmental pollutants in the onset of AR. However, the potential mechanisms by how and which these pollutants exacerbate AR conditions remain unknown. This panel study of 49 patients diagnosed with AR over one year aimed to assess the individual and combined effects of short-term exposure to multiple ambient pollutants on oxidative stress, symptoms, and quality of life among patients with AR. All participants underwent four repeated assessments of health conditions and personal environmental exposures (PM2.5, O3, SO2, and NO2) over warm and cold seasons during 2017-2018. We evaluated two oxidative stress biomarkers (malondialdehyde [MDA], and superoxide dismutase [SOD]) via nasal lavage. We collected information on self-reported symptoms and quality of life using the Rhinitis Symptom Scale (SRS), the Visual Analog Scale (VAS), and the Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) through in-person interviews. Bayesian kernel machine regression (BKMR) was used to evaluate the joint effects of pollutant mixture and identify key contributors. The results revealed a significant association of the pollutant mixture when all four pollutants were at or above their median levels, with increased oxidative stress. This was evidenced by elevated MDA and reduced SOD. We found a joint detrimental effect of the pollutant mixture on AR symptoms with a strong association with increased SRS scores, but a non-significant positive association with VAS and RQLQ scores. PM2.5, O3, and SO2 presented as the potentially primary contributors to the adverse health effects associated with the pollutant mixture in Taiyuan city. Patients with AR exposed to short-term air pollutant mixture are more likely to have greater nasal symptoms and worse quality of life from increased oxidative stress and reduced antioxidant capacity. Further research is warranted to better elucidate the underlying mechanisms.

    View details for DOI 10.1016/j.scitotenv.2024.172688

    View details for PubMedID 38663627

  • Associations of prenatal one-carbon metabolism nutrients and metals with epigenetic aging biomarkers at birth and in childhood in a US cohort. Aging Bozack, A. K., Rifas-Shiman, S. L., Baccarelli, A. A., Wright, R. O., Gold, D. R., Oken, E., Hivert, M. F., Cardenas, A. 2024; 16


    Epigenetic gestational age acceleration (EGAA) at birth and epigenetic age acceleration (EAA) in childhood may be biomarkers of the intrauterine environment. We investigated the extent to which first-trimester folate, B12, 5 essential, and 7 non-essential metals in maternal circulation are associated with EGAA and EAA in early life. Bohlin EGAA and Horvath pan-tissue and skin and blood EAA were calculated using DNA methylation measured in cord blood (N=351) and mid-childhood blood (N=326; median age = 7.7 years) in the Project Viva pre-birth cohort. A one standard deviation increase in individual essential metals (copper, manganese, and zinc) was associated with 0.94-1.2 weeks lower Horvath EAA at birth, and patterns of exposures identified by exploratory factor analysis suggested that a common source of essential metals was associated with Horvath EAA. We also observed evidence nonlinear associations of zinc with Bohlin EGAA, magnesium and lead with Horvath EAA, and cesium with skin and blood EAA at birth. Overall, associations at birth did not persist in mid-childhood; however, arsenic was associated with greater EAA at birth and in childhood. Prenatal metals, including essential metals and arsenic, are associated with epigenetic aging in early life, which might be associated with future health.

    View details for DOI 10.18632/aging.205602

    View details for PubMedID 38412256

  • DNA Methylation-Based Biomarkers of Protein Levels and Cardiovascular Disease Risk: Opportunities and Challenges for Precision Cardiology. Circulation. Genomic and precision medicine Bozack, A. K., Navas-Acien, A., Cardenas, A. 2024: e004571

    View details for DOI 10.1161/CIRCGEN.124.004571

    View details for PubMedID 38348680

  • Cross-sectional and prospective associations of early childhood circulating metals with early and mid-childhood cognition in the project viva cohort. Environmental research Thilakaratne, R., Lin, P. D., Rifas-Shiman, S. L., Landero, J., Wright, R. O., Bellinger, D., Oken, E., Cardenas, A. 2023: 118068


    BACKGROUND: Relatively little is known about the immediate and prospective neurodevelopmental impacts of joint exposure to multiple metals (i.e., metal mixtures) in early childhood.OBJECTIVES: To estimate associations of early childhood (3 years of age) blood metal concentrations with cognitive test scores at early and mid-childhood (8 years of age).METHODS: We studied children from the Project Viva cohort. We measured erythrocyte concentrations of seven essential (Co, Cu, Mg, Mn, Mo, Se, and Zn) and eight non-essential metals (As, Ba, Cd, Cs, Hg, Pb, Sn, and Sr) in early childhood blood samples. Trained research assistants administered cognitive tests assessing vocabulary, visual-motor ability, memory, and general intelligence (standard deviations: 10 points), in early and mid-childhood. We employed multivariable linear regression to examine associations of individual metals with test scores adjusting for confounders, other concurrently measured metals, and first-trimester maternal blood metals. We also estimated joint associations and explored interaction between metals in mixture analyses.RESULTS: We analyzed 349 children (median whole blood Pb 1 mug/dL). In cross-sectional analyses, each doubling of Pb was associated with lower visual-motor function (mean difference: -2.43 points, 95% confidence interval (CI): -4.01, -0.86) and receptive vocabulary, i.e., words understood (-1.45 points, 95% CI: -3.26, 0.36). Associations of Pb with mid-childhood cognition were weaker and less precise by comparison. Mg was positively associated with cognition in cross-sectional but not prospective analyses, and cross-sectional associations were attenuated in a sensitivity analysis removing adjustment for concurrent metals. We did not observe joint associations nor interactions.DISCUSSION: In this cohort with low blood Pb levels, increased blood Pb was robustly associated with lower cognitive ability in cross-sectional analyses, even after adjustment for prenatal Pb exposure, and regardless of adjustment for metal co-exposures. However, associations with mid-childhood cognition were attenuated and imprecise, suggesting some buffering of Pb neurotoxicity in early life.WHAT THIS STUDY ADDS: Relatively few studies have comprehensively separated the effects of neurotoxic metals such as lead (Pb) from pre- and postnatal co-occurring metals, nor examined persistence of associations across childhood. In a cohort of middle-class children, we found higher early childhood (3 y) blood Pb was associated with lower scores on cognitive tests, independent of other metals and prenatal blood Pb. However, early childhood Pb was only weakly associated with cognition in mid-childhood (8 y). Our results suggest the effects of low-level Pb exposure may attenuate over time in some populations, implying the presence of factors that may buffer Pb neurotoxicity in early life.

    View details for DOI 10.1016/j.envres.2023.118068

    View details for PubMedID 38157961

  • Infant Feeding Practices and Metal Concentrations in Children's Blood. JAMA network open Smith, A. R., Thilakaratne, R., Lin, P. D., Rifas-Shiman, S. L., Hivert, M., Oken, E., Cardenas, A. 2023; 6 (12): e2348230


    This cohort study assesses the association between 4 infant feeding practices and concentrations of 8 nonessential and 7 essential metals in red blood cells.

    View details for DOI 10.1001/jamanetworkopen.2023.48230

    View details for PubMedID 38109115

  • Associations between prenatal blood metals and vitamins and cord blood peptide hormone concentrations ENVIRONMENTAL EPIDEMIOLOGY Smith, A. R., Lin, P. D., Rifas-Shiman, S. L., Switkowski, K. M., Fleisch, A. F., Wright, R. O., Coull, B., Oken, E., Hivert, M., Cardenas, A. 2023; 7 (6)
  • Associations Between Prenatal Blood Metals and Vitamins and Cord Blood Peptide Hormone Concentrations. Global reproductive health Smith, A. R., Lin, P. D., Rifas-Shiman, S. L., Switkowski, K. M., Fleisch, A. F., Wright, R. O., Coull, B., Oken, E., Hivert, M. F., Cardenas, A. 2023; 7 (6)


    Nonessential metals have endocrine disrupting properties, interfere with cellular processes, generate reactive oxygen and deplete antioxidants, while essential metals and vitamins act as antioxidants. The extent to which prenatal metals and vitamins are associated with cord blood hormones involved in maternal and fetal metabolic and growth processes is unknown.We measured six nonessential (arsenic, barium, cadmium, cesium, lead, mercury) and four essential (magnesium, manganese, selenium, zinc) metals and trace elements, and two vitamins (B12 and folate) in first trimester blood from participants in the longitudinal pre-birth Project Viva cohort, who were recruited between 1999-2002 in eastern Massachusetts. We measured adiponectin, C-peptide, IGF-1, IGF-2, IGFBP-3, insulin, and leptin concentrations in cord blood (~n=695). We used covariate-adjusted quantile g-computation for mixtures and linear regression for individual exposures to estimate associations with cord blood peptide hormones.The essential metal mixture (magnesium, manganese, selenium, zinc) was associated with higher IGF-1 (β=3.20 ng/ml per quartile, 95% CI: 0.39, 6.01), IGF-2 (β=10.93 ng/ml, 95% CI: 0.08, 21.79), and leptin (β=1.03 ng/ml, 95% CI: 0.25, 1.80). Magnesium was associated with higher leptin (β=2.90 ng/ml, 95% CI: 0.89, 4.91), while B12 was associated with lower adiponectin, IGF-2, and leptin, but higher C-peptide. Other individual nonessential metals were associated with cord blood hormones.Our findings suggest that some prenatal metals and vitamins are associated with cord blood hormones, which may influence growth and development.

    View details for DOI 10.1097/EE9.0000000000000275

    View details for PubMedID 38645676

    View details for PubMedCentralID PMC11031200

  • Environmental tobacco smoke and children's health: a bibliometric and altmetric analysis of 100 most cited articles. BMC public health Patil, S. S., Puttaswamy, N., Sarode, S. C., Sarode, G. S., Patil, S. S., Cardenas, A., Gandhirajan, R. K., Balakrishnan, K. 2023; 23 (1): 2208


    BACKGROUND: Exposure to environmental tobacco smoke (ETS) is arguably the most ubiquitous and hazardous, even at very low levels, starting in early life. The objective of this study was to describe the state of research and future trends on ETS exposure and Children's Health (CH) topics with bibliometrics and altmetrics.METHODS: An electronic search was performed in Scopus database on January 31, 2023. Consensus was arrived on 100 most-cited articles by two reviewers. These papers were then cross matched with citations harvested from Web of Science (WoS) and Google Scholar. Altmetric Attention Score (AAS) and Dimension counts were also collected. Analysis and network visualization of authors, countries, and keywords were generated using VOSviewer software.RESULTS: Among a total of 1107 articles published on ETS and CH, the 100 top-cited articles appeared in 54 journals, with Pediatrics (n=12) contributing a maximum number of articles. The time period between 2000 and 2009 accounted for 44% of all publications. With respect to the research design employed across these studies, cross-sectional design took precedence over others accounting for approximately 40%. Predominantly,articles focused on childhood asthma; however, current research trends have shifted towards emerging fields such as children's oral health and DNA methylation. Twitter, policy documents, and news outlets were the main platforms where outputs were discussed. The AAS was not associated with journal impact factor or access type. Weak correlations were observed between AAS and citation count in Scopus, WoS, and Google Scholar (r=0.17 to 0.27) while a positive association existed between dimension count and the number of citations across all three databases (r=0.84 to 0.98).CONCLUSION: This study demonstrates the evolution, digital dissemination and research hotspots in the field of ETS and CH, predicting the possible future research directions. High-quality studies with more specific exposure classification are warranted to better understand the relationship between ETS and CH.

    View details for DOI 10.1186/s12889-023-16242-1

    View details for PubMedID 37946187

  • Associations Between Epigenetic Age Acceleration and microRNA Expression Among U.S. Firefighters. Epigenetics insights Jung, A. M., Furlong, M. A., Goodrich, J. M., Cardenas, A., Beitel, S. C., Littau, S. R., Caban-Martinez, A. J., Gulotta, J. J., Wallentine, D. D., Urwin, D., Gabriel, J., Hughes, J., Graber, J. M., Grant, C., Burgess, J. L. 2023; 16: 25168657231206301


    Epigenetic changes may be biomarkers of health. Epigenetic age acceleration (EAA), the discrepancy between epigenetic age measured via epigenetic clocks and chronological age, is associated with morbidity and mortality. However, the intersection of epigenetic clocks with microRNAs (miRNAs) and corresponding miRNA-based health implications have not been evaluated. We analyzed DNA methylation and miRNA profiles from blood sampled among 332 individuals enrolled across 2 U.S.-based firefighter occupational studies (2015-2018 and 2018-2020). We considered 7 measures of EAA in leukocytes (PhenoAge, GrimAge, Horvath, skin-blood, and Hannum epigenetic clocks, and extrinsic and intrinsic epigenetic age acceleration). We identified miRNAs associated with EAA using individual linear regression models, adjusted for sex, race/ethnicity, chronological age, and cell type estimates, and investigated downstream effects of associated miRNAs with miRNA enrichment analyses and genomic annotations. On average, participants were 38 years old, 88% male, and 75% non-Hispanic white. We identified 183 of 798 miRNAs associated with EAA (FDR q < 0.05); 126 with PhenoAge, 59 with GrimAge, 1 with Horvath, and 1 with the skin-blood clock. Among miRNAs associated with Horvath and GrimAge, there were 61 significantly enriched disease annotations including age-related metabolic and cardiovascular conditions and several cancers. Enriched pathways included those related to proteins and protein modification. We identified miRNAs associated with EAA of multiple epigenetic clocks. PhenoAge had more associations with individual miRNAs, but GrimAge and Horvath had greater implications for miRNA-associated pathways. Understanding the relationship between these epigenetic markers could contribute to our understanding of the molecular underpinnings of aging and aging-related diseases.

    View details for DOI 10.1177/25168657231206301

    View details for PubMedID 37953967

    View details for PubMedCentralID PMC10634256

  • Maternal adverse childhood experiences (ACEs) and DNA methylation of newborns in cord blood. Clinical epigenetics Collender, P., Bozack, A. K., Veazie, S., Nwanaji-Enwerem, J. C., Van Der Laan, L., Kogut, K., Riddell, C., Eskenazi, B., Holland, N., Deardorff, J., Cardenas, A. 2023; 15 (1): 162


    Adverse childhood experiences (ACEs) increase the risk of poor health outcomes later in life. Psychosocial stressors may also have intergenerational health effects by which parental ACEs are associated with mental and physical health of children. Epigenetic programming may be one mechanism linking parental ACEs to child health. This study aimed to investigate epigenome-wide associations of maternal preconception ACEs with DNA methylation patterns of children. In the Center for the Health Assessment of Mothers and Children of Salinas study, cord blood DNA methylation was measured using the Illumina HumanMethylation450 BeadChip. Preconception ACEs, which occurred during the mothers' childhoods, were collected using a standard ACE questionnaire including 10 ACE indicators. Maternal ACE exposures were defined in this study as (1) the total number of ACEs; (2) the total number of ACEs categorized as 0, 1-3, and > 4; and (3) individual ACEs. Associations of ACE exposures with differential methylated positions, regions, and CpG modules determined using weighted gene co-expression network analysis were evaluated adjusting for covariates.Data on maternal ACEs and cord blood DNA methylation were available for 196 mother/newborn pairs. One differential methylated position was associated with maternal experience of emotional abuse (cg05486260/FAM135B gene; q value < 0.05). Five differential methylated regions were significantly associated with the total number of ACEs, and 36 unique differential methylated regions were associated with individual ACEs (Šidák p value < 0.05). Fifteen CpG modules were significantly correlated with the total number of ACEs or individual ACEs, of which 8 remained significant in fully adjusted models (p value < 0.05). Significant modules were enriched for pathways related to neurological and immune development and function.Maternal ACEs prior to conception were associated with cord blood DNA methylation of offspring at birth. Although there was limited overlap between differential methylated regions and CpGs in modules associated with ACE exposures, statistically significant regions and networks were related to genes involved in neurological and immune function. Findings may provide insights to pathways linking psychosocial stressors to health. Further research is needed to understand the relationship between changes in DNA methylation and child health.

    View details for DOI 10.1186/s13148-023-01581-y

    View details for PubMedID 37845746

    View details for PubMedCentralID PMC10577922

  • Maternal prenatal social experiences and offspring epigenetic aging from birth to mid-childhood. Annals of epidemiology Laubach, Z. M., Bozack, A., Aris, I. M., Slopen, N., Tiemeier, H., Hivert, M. F., Cardenas, A., Perng, W. 2023


    Investigate associations of maternal social experiences with offspring epigenetic age acceleration (EAA) from birth through mid-childhood among 205 mother-offspring dyads of minoritized racial and ethnic groups.We used linear regression to examine associations of maternal experiences of racial bias or discrimination (0=none, 1-2=intermediate, or 3+=high), social support (tertile 1=low, 2=intermediate, 3=high), and socioeconomic status index (tertile 1=low, 2=intermediate, 3=high) during the prenatal period with offspring EAA according to Horvath's Pan-Tissue, Horvath's Skin & Blood, and Intrinsic EAA clocks at birth, 3 years, and 7 years.In comparison to children of women who did not experience any racial bias or discrimination, those whose mothers reported highest levels of racial bias or discrimination had lower Pan-Tissue clock EAA in early (-0.50 years; 90% CI: -0.91, -0.09) and mid-childhood (-0.75 years; -1.41, -0.08). We observed similar associations for the Skin & Blood clock and Intrinsic EAA. Maternal experiences of discrimination were not associated with Pan-Tissue EAA at birth. Neither maternal social support nor socioeconomic status predicted offspring EAA.Children whose mothers experienced racial bias or discrimination exhibited slower EAA. Future studies are warranted to confirm these findings and establish associations of early-life EAA with long-term health outcomes.

    View details for DOI 10.1016/j.annepidem.2023.10.003

    View details for PubMedID 37839726

  • Parenting Practices May Buffer the Impact of Adversity on Epigenetic Age Acceleration Among Young Children With Developmental Delays. Psychological science Sullivan, A. D., Bozack, A. K., Cardenas, A., Comer, J. S., Bagner, D. M., Forehand, R., Parent, J. 2023: 9567976231194221


    This study examined whether children exposed to adversity would exhibit lower epigenetic age acceleration in the context of improved parenting. Children with developmental delays and externalizing behavior problems (N = 62; Mage = 36.26 months; 70.97% boys, 29.03% girls; 71% Latinx, 22.6% Black) were drawn from a larger randomized controlled trial (RCT), which randomized them to receive Internet-delivered parent-child interaction therapy (iPCIT; n = 30) or community referrals as usual (RAU; n = 32). Epigenetic age acceleration was estimated with the pediatric buccal epigenetic clock, using saliva. Adversity was assessed using parent, family, and neighborhood-level cumulative-risk indicators. Adversity interacted with Time 2 (T2) observations of positive and negative-parenting practices to predict epigenetic age acceleration 1.5 years later, regardless of treatment assignment. Children exposed to more adversity displayed lower epigenetic age acceleration when parents evidenced increased positive (b = -0.15, p = .001) and decreased negative (b = -0.12, p = .01) parenting practices.

    View details for DOI 10.1177/09567976231194221

    View details for PubMedID 37733001

  • Climate change and epigenetic biomarkers in allergic and airway diseases. The Journal of allergy and clinical immunology Cardenas, A., Fadadu, R., Bunyavanich, S. 2023


    Human epigenetic variation is associated with both environmental exposures and allergic diseases and can potentially serve as a biomarker connecting climate change with allergy and airway diseases. In this narrative review, we summarize recent human epigenetic studies examining exposure to temperature, precipitation, extreme weather events, and malnutrition to discuss findings as they relate to allergic and airway diseases. Temperature has been the most widely studied exposure, with the studies implicating both short-term and long-term exposure with epigenetic alterations and epigenetic aging. Few studies have examined natural disasters or extreme weather events. The studies available have reported differential DNA methylation of multiple genes and pathways, some of which were previously associated with asthma or allergy. Few studies have integrated climate-related events, epigenetic biomarkers, and allergic disease together. Prospective longitudinal studies are needed along with the collection of target tissues beyond blood samples, such as nasal and skin cells. Finally, global collaboration to increase diverse representation of study participants, particularly those most affected by climate injustice, and strengthen replication, validation, and harmonization of measurements will be needed to elucidate the impact of climate change on the human epigenome.

    View details for DOI 10.1016/j.jaci.2023.09.011

    View details for PubMedID 37741554

  • Evaluation of pediatric epigenetic clocks across multiple tissues. Clinical epigenetics Fang, F., Zhou, L., Perng, W., Marsit, C. J., Knight, A. K., Cardenas, A., Aung, M. T., Hivert, M., Aris, I. M., Goodrich, J. M., Smith, A. K., Gaylord, A., Fry, R. C., Oken, E., O'Connor, G., Ruden, D. M., Trasande, L., Herbstman, J. B., Camargo, C. A., Bush, N. R., Dunlop, A. L., Dabelea, D. M., Karagas, M. R., Breton, C. V., Ober, C., Everson, T. M., Page, G. P., Ladd-Acosta, C., program collaborators for Environmental influences on Child Health Outcomes, Smith, P. B., Newby, L. K., Jacobson, L. P., Catellier, D. J., Gershon, R., Cella, D., Laham, F. R., Mansbach, J. M., Wu, S., Spergel, J. M., Celedon, J. C., Puls, H. T., Teach, S. J., Porter, S. C., Waynik, I. Y., Iyer, S. S., Samuels-Kalow, M. E., D Thompson, A., Stevenson, M. D., Bauer, C. S., Inhofe, N. R., Boos, M., Macias, C. G., Gern, J., Jackson, D., Bacharier, L., Kattan, M., Wood, R., Rivera-Spoljaric, K., Bacharier, L., Bastain, T., Farzan, S., Habre, R., Karr, C., Tylavsky, F., Mason, A., Zhao, Q., Sathyanarayana, S., Bush, N., LeWinn, K. Z., Lester, B., Carter, B., Pastyrnak, S., Neal, C., Smith, L., Helderman, J., McEvoy, C., Tepper, R., Lyall, K., Volk, H., Schmidt, R., Croen, L., O'Shea, M., Vaidya, R., Obeid, R., Rollins, C., Bear, K., Pastyrnak, S., Lenski, M., Singh, R., Msall, M., Frazier, J., Gogcu, S., Montgomery, A., Kuban, K., Douglass, L., Jara, H., Joseph, R., Kerver, J. M., Perera, F. 2023; 15 (1): 142


    BACKGROUND: Epigenetic clocks are promising tools for assessing biological age. We assessed the accuracy of pediatric epigenetic clocks in gestational and chronological age determination.RESULTS: Our study used data from seven tissue types on three DNA methylation profiling microarrays and found that the Knight and Bohlin clocks performed similarly for blood cells, while the Lee clock was superior for placental samples. The pediatric-buccal-epigenetic clock performed the best for pediatric buccal samples, while the Horvath clock is recommended for children's blood cell samples. The NeoAge clock stands out for its unique ability to predict post-menstrual age with high correlation with the observed age in infant buccal cell samples.CONCLUSIONS: Our findings provide valuable guidance for future research and development of epigenetic clocks in pediatric samples, enabling more accurate assessments of biological age.

    View details for DOI 10.1186/s13148-023-01552-3

    View details for PubMedID 37660147

  • Mixtures of Metals and Micronutrients in Early Pregnancy and Cognition in Early and Mid-Childhood: Findings from the Project Viva Cohort. Environmental health perspectives Thilakaratne, R., Lin, P. D., Rifas-Shiman, S. L., Wright, R. O., Hubbard, A., Hivert, M. F., Bellinger, D., Oken, E., Cardenas, A. 2023; 131 (8): 87008


    The developing fetal brain is sensitive to many environmental exposures. However, the independent and joint effects of prenatal exposure to metals and micronutrients on child cognition are not well understood.Our aim was to evaluate associations of first-trimester (∼10 wk) maternal erythrocyte concentrations of mixtures of nonessential and essential metals and micronutrients with early (∼3 y) and mid-childhood (∼8 y) cognitive test scores in Project Viva, a prebirth cohort in Boston, Massachusetts, USA.We measured concentrations of five essential metals (Cu, Mg, Mn, Se, Zn) and two micronutrients (vitamin B12 and folate), together termed the "nutrient mixture," as well as six nonessential metals (As, Ba, Cd, Cs, Hg, Pb), together termed the "neurotoxic mixture," in first-trimester (∼10 wk) maternal erythrocytes (metals) or plasma (micronutrients). We assessed visual-motor function and receptive vocabulary in early childhood (∼3 y), and visual-motor function, visual memory, and fluid and crystallized intelligence in mid-childhood (∼8 y). We employed adjusted quantile g-computation and linear regression to estimate mixture and individual component associations, respectively.Analyses included 900 mother-child pairs (74% college graduates; 52% male children). In mixture analyses, a quartile increase in the nutrient mixture was associated with a mean difference in early childhood receptive vocabulary score of 1.58 points [95% confidence interval (CI): 0.06, 3.10], driven by Zn and Se. A quartile increase in the neurotoxic mixture was associated with a mean difference in mid-childhood visual-motor score of -3.01 points (95% CI: -5.55, -0.47), driven by Ba and Cs. Linear regressions supported quantile g-computation findings for mixture component contributions.Maternal circulating concentrations of several essential (Zn and Se) and nonessential (Ba and Cs) metals were associated with some domains of child cognition. In this folate-replete cohort, first-trimester circulating concentrations of known neurotoxic metals, such as Pb, were not associated with child cognition.

    View details for DOI 10.1289/EHP12016

    View details for PubMedID 37585348

  • Household air pollution and epigenetic aging in Xuanwei, China. Environment international Blechter, B., Cardenas, A., Shi, J., Wong, J. Y., Hu, W., Rahman, M. L., Breeze, C., Downward, G. S., Portengen, L., Zhang, Y., Ning, B., Ji, B. T., Cawthon, R., Li, J., Yang, K., Bozack, A., Dean Hosgood, H., Silverman, D. T., Huang, Y., Rothman, N., Vermeulen, R., Lan, Q. 2023; 178: 108041


    Household air pollution (HAP) from indoor combustion of solid fuel is a global health burden linked to lung cancer. In Xuanwei, China, lung cancer rate for nonsmoking women is among the highest in the world and largely attributed to high levels of polycyclic aromatic hydrocarbons (PAHs) that are produced from combustion of smoky (bituminous) coal used for cooking and heating. Epigenetic age acceleration (EAA), a DNA methylation-based biomarker of aging, has been shown to be highly correlated with biological processes underlying the susceptibility of age-related diseases. We aim to assess the association between HAP exposure and EAA.We analyzed data from 106 never-smoking women from Xuanwei, China. Information on fuel type was collected using a questionnaire, and validated exposure models were used to predict levels of 43 HAP constituents. Exposure clusters were identified using hierarchical clustering. EAA was derived for five epigenetic clocks defined as the residuals resulting from regressing each clock on chronological age. We used generalized estimating equations to test associations between exposure clusters derived from predicted levels of HAP exposure, ambient 5-methylchrysene (5-MC), a PAH previously found to be associated with risk of lung cancer, and EAA, while accounting for repeated-measurements and confounders.We observed an increase in GrimAge EAA for clusters with 31 and 33 PAHs reflecting current (β = 0.77 y per standard deviation (SD) increase, 95 % CI:0.36,1.19) and childhood (β = 0.92 y per SD, 95 % CI:0.40,1.45) exposure, respectively. 5-MC (ng/m3-year) was found to be associated with GrimAge EAA for current (β = 0.15 y, 95 % CI:0.05,0.25) and childhood (β = 0.30 y, 95 % CI:0.13,0.47) exposure.Our findings suggest that exposure to PAHs from indoor smoky coal combustion, particularly 5-MC, is associated with GrimAge EAA, a biomarker of mortality.

    View details for DOI 10.1016/j.envint.2023.108041

    View details for PubMedID 37354880

  • Epigenome-Wide Association Studies of Allergic Disease and the Environment. The Journal of allergy and clinical immunology Cardenas, A., Fadadu, R. P., Koppelman, G. H. 2023


    The epigenome is at the intersection of the environment, genotype, and cellular response. DNA methylation of cytosine nucleotides, the most studied epigenetic modification, has been systematically evaluated in human studies using untargeted epigenome-wide association studies (EWAS) and shown to be both sensitive to environmental exposures and associated with allergic diseases. In this narrative review, we summarize findings from key EWAS previously conducted on this topic, interpret results from recent studies, and discuss the strengths, challenges, and opportunities regarding epigenetics research on the environment-allergy relationship. The majority of EWAS have systematically investigated select environmental exposures, during the prenatal and early childhood period, and allergic diseases mostly in leukocyte-isolated DNA and more recently in nasal cells. Overall, many studies found consistent DNA methylation associations across cohorts for certain exposures, such as smoking (e.g., AHRR gene), and allergic diseases (e.g., EPX gene). We recommend the integration of both environmental exposures and allergy or asthma within long-term prospective designs to strengthen causality as well as biomarker development. Future studies should collect paired target tissues to examine compartment-specific epigenetic response, incorporate genetic influences in DNA methylation (meQTL), replicate findings across diverse populations, and carefully interpret epigenetic signatures from bulk, target tissue or isolated cells.

    View details for DOI 10.1016/j.jaci.2023.05.020

    View details for PubMedID 37295475

  • Long-term exposure to ambient fine particulate matter and carotid intima media thickness at bilateral, left and right in adults from Mexico City: Results from GEA study. Chemosphere Torrico-Lavayen, R., Vargas-Alarcón, G., Riojas-Rodriguez, H., Sánchez Guerra, M. A., Texcalac-Sangrador, J. L., Ortiz-Panozo, E., Gutiérrez-Avila, I., De Vizcaya-Ruiz, A., Cardenas, A., Posadas-Sánchez, R., Osorio-Yáñez C, C. 2023: 139009


    PM2.5 exposure has been associated with intima-media thickness (cIMT) increase. However, very few studies distinguished between left and right cIMT in relation to PM2.5 exposure.To evaluate associations between chronic exposure to PM2.5 and cIMT at bilateral, left, and right in adults from Mexico City.This study comprised 913 participants from the control group, participants without personal or family history of cardiovascular disease, of the Genetics of Atherosclerosis Disease Mexican study (GEA acronym in Spanish), recruited at the Instituto Nacional de Cardiología Ignacio Chávez from June 2008 to January 2013. To assess the associations between chronic exposure to PM2.5 (per 5 μg/m3 increase) at different lag years (1-4 years) and cIMT (bilateral, left, and right) we applied distributed lag non-linear models (DLNMs).The median and interquartile range for cIMT at bilateral, left, and right, were 630 (555, 735), 640 (550, 750), and 620 (530, 720) μm, respectively. Annual average PM2.5 exposure was 26.64 μg/m3, with median and IQR, of 24.46 (23.5-25.46) μg/m3. Results from DLNMs adjusted for age, sex, body mass index, low-density lipoproteins, and glucose, showed that PM2.5 exposure for year 1 and 2, were positively and significantly associated with right-cIMT [6.99% (95% CI: 3.67; 10.42) and 2.98% (0.03; 6.01), respectively]. Negative associations were observed for PM2.5 at year 3 and 4 and right-cIMT; however only year 3 was statistically significant [-2.83% (95% CI: 5.12; -0.50)]. Left-cIMT was not associated with PM2.5 exposure at any lag year. The increase in bilateral cIMT followed a similar pattern as that observed for right-cIMT, but with lower estimates.Our results suggest different susceptibility between left and right cIMT associated with PM2.5 exposure highlighting the need of measuring both, left and right cIMT, regarding ambient air pollution in epidemiological studies.

    View details for DOI 10.1016/j.chemosphere.2023.139009

    View details for PubMedID 37245594

  • Associations between prenatal phthalate exposure and childhood epigenetic age acceleration. Environmental research Khodasevich, D., Holland, N., Hubbard, A., Harley, K., Deardorff, J., Eskenazi, B., Cardenas, A. 2023: 116067


    BACKGROUND: Phthalates, a group of pervasive endocrine-disrupting chemicals found in plastics and personal care products, have been associated with a wide range of developmental and health outcomes. However, their impact on biomarkers of aging has not been characterized. We tested associations between prenatal exposure to 11 phthalate metabolites on epigenetic aging in children at birth, 7, 9, and 14 years of age. We hypothesized that prenatal phthalate exposure will be associated with epigenetic age acceleration measures at birth and in early childhood, with patterns dependent on sex and timing of DNAm measurement.METHODS: Among 385 mother-child pairs from the CHAMACOS cohort, we measured DNAm at birth, 7, 9, and 14 years of age, and utilized adjusted linear regression to assess the association between prenatal phthalate exposure and Bohlin's Gestational Age Acceleration (GAA) at birth and Intrinsic Epigenetic Age Acceleration (IEAA) throughout childhood. Additionally, quantile g-computation was utilized to assess the effect of the phthalate mixture on GAA at birth and IEAA throughout childhood.RESULTS: We found a negative association between prenatal di (2-ethylhexyl) phthalate (DEHP) exposure and IEAA among males at age 7 (-0.58 years; 95% CI: 1.02 to -0.13), and a marginal negative association between the whole phthalate mixture and GAA among males at birth (-1.54 days, 95% CI: 2.79 to -0.28), while most other associations were nonsignificant.CONCLUSIONS: Our results suggest that prenatal exposure to certain phthalates is associated with epigenetic aging in children. Additionally, our findings suggest that the influence of prenatal exposures on epigenetic age may only manifest during specific periods of child development, and studies relying on DNAm measurements solely from cord blood or single time points may overlook potential relationships.

    View details for DOI 10.1016/j.envres.2023.116067

    View details for PubMedID 37149020

  • Association of Early Pregnancy Perfluoroalkyl and Polyfluoroalkyl Substance Exposure With Birth Outcomes. JAMA network open Zhang, Y., Mustieles, V., Sun, Q., Coull, B., McElrath, T., Rifas-Shiman, S. L., Martin, L., Sun, Y., Wang, Y. X., Oken, E., Cardenas, A., Messerlian, C. 2023; 6 (5): e2314934


    Prenatal perfluoroalkyl and polyfluoroalkyl substances (PFAS) have been linked to adverse birth outcomes. Previous research showed that higher folate concentrations are associated with lower blood PFAS concentrations in adolescents and adults. Further studies are needed to explore whether prenatal folate status mitigates PFAS-related adverse birth outcomes.To examine whether prenatal folate status modifies the negative associations between pregnancy PFAS concentrations, birth weight, and gestational age previously observed in a US cohort.In a prospective design, a prebirth cohort of mothers or pregnant women was recruited between April 1999 and November 2002, in Project Viva, a study conducted in eastern Massachusetts. Statistical analyses were performed from May 24 and October 25, 2022.Plasma concentrations of 6 PFAS compounds were measured in early pregnancy (median gestational week, 9.6). Folate status was assessed through a food frequency questionnaire and measured in plasma samples collected in early pregnancy.Birth weight and gestational age, abstracted from delivery records; birth weight z score, standardized by gestational age and infant sex; low birth weight, defined as birth weight less than 2500 g; and preterm birth, defined as birth at less than 37 completed gestational weeks.The cohort included a total of 1400 mother-singleton pairs. The mean (SD) age of the mothers was 32.21 (4.89) years. Most of the mothers were White (73.2%) and had a college degree or higher (69.1%). Early pregnancy plasma perfluorooctanoic acid concentration was associated with lower birth weight and birth weight z score only among mothers whose dietary folate intake (birth weight: β, -89.13 g; 95% CI, -166.84 to -11.42 g; birth weight z score: -0.13; 95% CI, -0.26 to -0.003) or plasma folate concentration (birth weight: -87.03 g; 95% CI, -180.11 to 6.05 g; birth weight z score: -0.14; 95% CI, -0.30 to 0.02) were below the 25th percentile (dietary: 660 μg/d, plasma: 14 ng/mL). No associations were found among mothers in the higher folate level groups, although the tests for heterogeneity did not reject the null. Associations between plasma perfluorooctane sulfonic acid and perfluorononanoate (PFNA) concentrations and lower birth weight, and between PFNA and earlier gestational age were noted only among mothers whose prenatal dietary folate intake or plasma folate concentration was in the lowest quartile range. No associations were found among mothers in higher folate status quartile groups.In this large, US prebirth cohort, early pregnancy exposure to select PFAS compounds was associated with adverse birth outcomes only among mothers below the 25th percentile of prenatal dietary or plasma folate levels.

    View details for DOI 10.1001/jamanetworkopen.2023.14934

    View details for PubMedID 37256622

  • DNA methylation age at birth and childhood: performance of epigenetic clocks and characteristics associated with epigenetic age acceleration in the Project Viva cohort. Clinical epigenetics Bozack, A. K., Rifas-Shiman, S. L., Gold, D. R., Laubach, Z. M., Perng, W., Hivert, M. F., Cardenas, A. 2023; 15 (1): 62


    Epigenetic age acceleration (EAA) and epigenetic gestational age acceleration (EGAA) are biomarkers of physiological development and may be affected by the perinatal environment. The aim of this study was to evaluate performance of epigenetic clocks and to identify biological and sociodemographic correlates of EGAA and EAA at birth and in childhood. In the Project Viva pre-birth cohort, DNA methylation was measured in nucleated cells in cord blood (leukocytes and nucleated red blood cells, N = 485) and leukocytes in early (N = 120, median age = 3.2 years) and mid-childhood (N = 460, median age = 7.7 years). We calculated epigenetic gestational age (EGA; Bohlin and Knight clocks) and epigenetic age (EA; Horvath and skin & blood clocks), and respective measures of EGAA and EAA. We evaluated the performance of clocks relative to chronological age using correlations and median absolute error. We tested for associations of maternal-child characteristics with EGAA and EAA using mutually adjusted linear models controlling for estimated cell type proportions. We also tested associations of Horvath EA at birth with childhood EAA.Bohlin EGA was strongly correlated with chronological gestational age (Bohlin EGA r = 0.82, p < 0.001). Horvath and skin & blood EA were weakly correlated with gestational age, but moderately correlated with chronological age in childhood (r = 0.45-0.65). Maternal smoking during pregnancy was associated with higher skin & blood EAA at birth [B (95% CI) = 1.17 weeks (- 0.09, 2.42)] and in early childhood [0.34 years (0.03, 0.64)]. Female newborns and children had lower Bohlin EGAA [- 0.17 weeks (- 0.30, - 0.04)] and Horvath EAA at birth [B (95% CI) = - 2.88 weeks (- 4.41, - 1.35)] and in childhood [early childhood: - 0.3 years (- 0.60, 0.01); mid-childhood: - 0.48 years (- 0.77, - 0.18)] than males. When comparing self-reported Asian, Black, Hispanic, and more than one race or other racial/ethnic groups to White, we identified significant differences in EGAA and EAA at birth and in mid-childhood, but associations varied across clocks. Horvath EA at birth was positively associated with childhood Horvath and skin & blood EAA.Maternal smoking during pregnancy and child sex were associated with EGAA and EAA at multiple timepoints. Further research may provide insight into the relationship between perinatal factors, pediatric epigenetic aging, and health and development across the lifespan.

    View details for DOI 10.1186/s13148-023-01480-2

    View details for PubMedID 37046280

    View details for PubMedCentralID 8901047

  • The prenatal environment and its influence on maternal and child mitochondrial DNA copy number and methylation: A review of the literature. Environmental research Smith, A. R., Hinojosa Briseno, A., Picard, M., Cardenas, A. 2023; 227: 115798


    Mitochondrial DNA (mtDNA) is sensitive to environmental stressors and associated with human health. We reviewed epidemiological literature examining associations between prenatal environmental, dietary, and social exposures and alterations in maternal/child mtDNA copy number (mtDNAcn) and mtDNA methylation. Evidence exists that prenatal maternal exposures are associated with alterations in mtDNAcn for air pollution, chemicals (e.g. metals), cigarette smoke, human immunodeficiency virus (HIV) infection and treatment. Evidence for their associations with mtDNA methylation was limited. Given its potential implications as a disease pathway biomarker, studies with sufficient biological specificity should examine the long-term implications of prenatal and early-life mtDNA alterations in response to prenatal exposures.

    View details for DOI 10.1016/j.envres.2023.115798

    View details for PubMedID 37001851

  • Occupational Years of Service and Leukocyte Epigenetic Aging: Relationships in United States Firefighters. Journal of occupational and environmental medicine Nwanaji-Enwerem, J. C., Cardenas, A., Goodrich, J. M., Furlong, M. A., Jung, A. M., Collender, P., Caban-Martinez, A. J., Grant, C., Beitel, S., Littau, S., Urwin, D., Gabriel, J., Hughes, J., Gulotta, J., Wallentine, D., Burgess, J. L. 2023


    OBJECTIVE: To examine associations between years of firefighting service and eight chronological age-adjusted measures of blood leukocyte epigenetic age acceleration: Horvath, Hannum, SkinBloodClock, Intrinsic, Extrinsic, PhenoAge, GrimAge, and DNAm telomere length.METHODS: A repeated measures analysis of data from 379 incumbent firefighters from eight career departments and 100 recruit firefighters from two of the departments, across the United States.RESULTS: Incumbent firefighters had on average greater epigenetic age acceleration compared to recruit firefighters, potentially due to the cumulative effect of occupational exposures. However, among incumbent firefighters, additional years of service were associated with epigenetic age deceleration, particularly for GrimAge, a strong predictor of mortality.CONCLUSIONS: Long-term studies with more specific occupational exposure classification are needed to better understand the relationship between years of service and aging biomarkers.

    View details for DOI 10.1097/JOM.0000000000002817

    View details for PubMedID 36787539

  • Analysis of Pregnancy Complications and Epigenetic Gestational Age of Newborns. JAMA network open Ladd-Acosta, C., Vang, E., Barrett, E. S., Bulka, C. M., Bush, N. R., Cardenas, A., Dabelea, D., Dunlop, A. L., Fry, R. C., Gao, X., Goodrich, J. M., Herbstman, J., Hivert, M. F., Kahn, L. G., Karagas, M. R., Kennedy, E. M., Knight, A. K., Mohazzab-Hosseinian, S., Morin, A., Niu, Z., O'Shea, T. M., Palmore, M., Ruden, D., Schmidt, R. J., Smith, A. K., Song, A., Spindel, E. R., Trasande, L., Volk, H., Weisenberger, D. J., Breton, C. V. 2023; 6 (2): e230672


    Preeclampsia, gestational hypertension, and gestational diabetes, the most common pregnancy complications, are associated with substantial morbidity and mortality in mothers and children. Little is known about the biological processes that link the occurrence of these pregnancy complications with adverse child outcomes; altered biological aging of the growing fetus up to birth is one molecular pathway of increasing interest.To evaluate whether exposure to each of these 3 pregnancy complications (gestational diabetes, gestational hypertension, and preeclampsia) is associated with accelerated or decelerated gestational biological age in children at birth.Children included in these analyses were born between 1998 and 2018 and spanned multiple geographic areas of the US. Pregnancy complication information was obtained from maternal self-report and/or medical record data. DNA methylation measures were obtained from blood biospecimens collected from offspring at birth. The study used data from the national Environmental Influences on Child Health Outcomes (ECHO) multisite cohort study collected and recorded as of the August 31, 2021, data lock date. Data analysis was performed from September 2021 to December 2022.Three pregnancy conditions were examined: gestational hypertension, preeclampsia, and gestational diabetes.Accelerated or decelerated biological gestational age at birth, estimated using existing epigenetic gestational age clock algorithms.A total of 1801 child participants (880 male [48.9%]; median [range] chronological gestational age at birth, 39 [30-43] weeks) from 12 ECHO cohorts met the analytic inclusion criteria. Reported races included Asian (49 participants [2.7%]), Black (390 participants [21.7%]), White (1026 participants [57.0%]), and other races (92 participants [5.1%]) (ie, American Indian or Alaska Native, Native Hawaiian or other Pacific Islander, multiple races, and other race not specified). In total, 524 participants (29.0%) reported Hispanic ethnicity. Maternal ages ranged from 16 to 45 years of age with a median of 29 in the analytic sample. A range of maternal education levels, from less than high school (260 participants [14.4%]) to Bachelor's degree and above (629 participants [34.9%]), were reported. In adjusted regression models, prenatal exposure to maternal gestational diabetes (β, -0.423; 95% CI, -0.709 to -0.138) and preeclampsia (β, -0.513; 95% CI, -0.857 to -0.170), but not gestational hypertension (β, 0.003; 95% CI, -0.338 to 0.344), were associated with decelerated epigenetic aging among exposed neonates vs those who were unexposed. Modification of these associations, by sex, was observed with exposure to preeclampsia (β, -0.700; 95% CI, -1.189 to -0.210) and gestational diabetes (β, -0.636; 95% CI, -1.070 to -0.200), with associations observed among female but not male participants.This US cohort study of neonate biological changes related to exposure to maternal pregnancy conditions found evidence that preeclampsia and gestational diabetes delay biological maturity, especially in female offspring.

    View details for DOI 10.1001/jamanetworkopen.2023.0672

    View details for PubMedID 36826815

  • OxPhos defects cause hypermetabolism and reduce lifespan in cells and in patients with mitochondrial diseases. Communications biology Sturm, G., Karan, K. R., Monzel, A. S., Santhanam, B., Taivassalo, T., Bris, C., Ware, S. A., Cross, M., Towheed, A., Higgins-Chen, A., McManus, M. J., Cardenas, A., Lin, J., Epel, E. S., Rahman, S., Vissing, J., Grassi, B., Levine, M., Horvath, S., Haller, R. G., Lenaers, G., Wallace, D. C., St-Onge, M. P., Tavazoie, S., Procaccio, V., Kaufman, B. A., Seifert, E. L., Hirano, M., Picard, M. 2023; 6 (1): 22


    Patients with primary mitochondrial oxidative phosphorylation (OxPhos) defects present with fatigue and multi-system disorders, are often lean, and die prematurely, but the mechanistic basis for this clinical picture remains unclear. By integrating data from 17 cohorts of patients with mitochondrial diseases (n = 690) we find evidence that these disorders increase resting energy expenditure, a state termed hypermetabolism. We examine this phenomenon longitudinally in patient-derived fibroblasts from multiple donors. Genetically or pharmacologically disrupting OxPhos approximately doubles cellular energy expenditure. This cell-autonomous state of hypermetabolism occurs despite near-normal OxPhos coupling efficiency, excluding uncoupling as a general mechanism. Instead, hypermetabolism is associated with mitochondrial DNA instability, activation of the integrated stress response (ISR), and increased extracellular secretion of age-related cytokines and metabokines including GDF15. In parallel, OxPhos defects accelerate telomere erosion and epigenetic aging per cell division, consistent with evidence that excess energy expenditure accelerates biological aging. To explore potential mechanisms for these effects, we generate a longitudinal RNASeq and DNA methylation resource dataset, which reveals conserved, energetically demanding, genome-wide recalibrations. Taken together, these findings highlight the need to understand how OxPhos defects influence the energetic cost of living, and the link between hypermetabolism and aging in cells and patients with mitochondrial diseases.

    View details for DOI 10.1038/s42003-022-04303-x

    View details for PubMedID 36635485

  • Associations of Prenatal First Trimester Essential and Nonessential Metal Mixtures with Body Size and Adiposity in Childhood. Epidemiology (Cambridge, Mass.) Smith, A. R., Lin, P. D., Rifas-Shiman, S. L., Wright, R. O., Coull, B., Hivert, M. F., Hubbard, A., Oken, E., Cardenas, A. 2023; 34 (1): 80-89


    Prenatal nonessential metals may contribute to postnatal adiposity, whereas essential metals may have metabolic benefits. We evaluated joint and individual associations between prenatal metals and childhood adiposity.We measured concentrations of six nonessential (arsenic, barium, cadmium, cesium, lead, and mercury) and four essential (magnesium, manganese, selenium, and zinc) metals in first trimester maternal blood from a prebirth cohort. We collected anthropometric measures in early childhood, mid-childhood, and early adolescence including subscapular+tricep skinfold thickness (mm) (N = 715-859), waist circumference (cm) (N = 717-882), and body mass index (BMI) (z-score) (N = 716-875). We measured adiposity in mid-childhood and early adolescence using bone densitometry total- and trunk- fat mass index (kg/m2) (N = 511-599). We estimated associations using adjusted quantile g-computation and linear regression.The nonessential metal mixture was associated with higher total (β = 0.07, 95% CI = 0.01, 0.12) and trunk fat mass index (β = 0.12, CI = 0.02, 0.22), waist circumference (β = 0.01, CI = 0.00, 0.01), and BMI (β = 0.24, CI = 0.07, 0.41) in mid-childhood, and total fat mass index (β = 0.07, CI = 0.01, 0.14), and BMI (β = 0.19, CI = 0.02, 0.37) in early adolescence. The essential metal mixture was associated with lower early adolescence total-(β = -0.11, CI = -0.17, -0.04) and trunk- fat mass index (β = -0.13, CI = -0.21, -0.05), subscapular+tricep skinfold thickness (β = -0.02, CI = -0.03, -0.00), waist circumference (β = -0.003, CI = -0.01, -0.00), and BMI (β = -0.16, CI = -0.28, -0.04). Cadmium and cesium were individually associated with childhood adiposity at different timepoints.Prenatal first-trimester essential metals were associated with lower childhood adiposity, whereas nonessential metals were associated with higher adiposity into adolescence.

    View details for DOI 10.1097/EDE.0000000000001560

    View details for PubMedID 36455248

  • Psychological Stress and Epigenetic Aging in Older Men: The VA Normative Aging Study. Translational medicine of aging Nwanaji-Enwerem, J. C., Cardenas, A., Gao, X., Wang, C., Vokonas, P., Spiro, A., Osborne, A. D., Kosheleva, A., Hou, L., Baccarelli, A. A., Schwartz, J. 2023; 7: 66-74


    Psychological stress remains an important risk factor for morbidity and mortality throughout the life course. However, there have been counterintuitive findings reported in previous studies of older persons that examine the relationships of perceived psychological stress with DNA methylation-based markers of aging, which also serve as predictors of morbidity and mortality (epigenetic age/clocks). We aimed to replicate and expand findings from existing work by examining relationships of self-reported stress with nine epigenetic clocks: Hannum, Horvath, Intrinsic, Extrinsic, SkinBloodClock, PhenoAge, GrimAge, DNAm Telomere Length, and Pace of Aging. We analyzed data from 607 male participants (mean age 73.2 years) of the VA Normative Aging Study with one to two study visits from 1999 to 2007 (observations = 956). Stress was assessed via the 14-item Perceived Stress Scale (PSS). Epigenetic age was calculated from DNA methylation measured in leukocytes with the HumanMethylation450 BeadChip. In linear mixed effects models adjusted for demographic/lifestyle/health factors, a standard deviation (sd) increase in PSS was associated with Horvath (beta = -0.35-years, 95%CI: -0.61, -0.09, P=0.008) and Intrinsic (beta = -0.40-years, 95%CI: -0.67, -0.13, P=0.004) epigenetic age deceleration. However, in models limited to participants with the highest levels of stress (≥ 75th-percentile), Horvath (beta = 2.29-years, 95%CI: 0.16, 4.41, P=0.04) and Intrinsic (beta = 2.06-years, 95%CI: -0.17, 4.28, P=0.07) age acceleration associations were observed. Our results reinforce the complexity of psychological stress and epigenetic aging relationships and lay a foundation for future studies that explore longitudinal relationships with other adult stress metrics and factors that can influence stress such as resilience measures.

    View details for DOI 10.1016/j.tma.2023.06.003

    View details for PubMedID 37576443

  • Prenatal trimester-specific intake of micronutrients: global DNA methylation and hydroxymethylation at birth and persistence in childhood. Journal of developmental origins of health and disease Parsons, E., Rifas-Shiman, S. L., Bozack, A. K., Baccarelli, A. A., DeMeo, D. L., Hivert, M., Godderis, L., Duca, R., Oken, E., Cardenas, A. 2022: 1-8


    The prenatal environment may program health and disease susceptibility via epigenetic mechanisms. We evaluated associations of maternal trimester-specific intake of micronutrients with global DNA methylation (%5mC) and 5-hydroxymethylation (%5hmC) at birth in cord blood and tested for persistence into childhood. We quantified global %5mC and %5hmC in cord blood cells (n = 434) and in leukocytes collected in early (n = 108) and mid-childhood (n = 390) from children in Project Viva, a pre-birth cohort from Boston, MA. Validated food frequency questionnaires estimated maternal first- and second-trimester intakes of vitamin B2, vitamin B6, vitamin B12, folate, betaine, choline, methionine, iron, and zinc. Mean (SD) cord blood %5mC and %5hmC was 5.62% (2.04) and 0.25% (0.15), respectively. Each mug increase in first-trimester B12 intake was associated with 0.002 lower %5hmC in cord blood (95% CI: -0.005, -0.0003), and this association persisted in early childhood (beta = -0.007; 95% CI: -0.01, -0.001) but not mid-childhood. Second-trimester iron (mg) was associated with 0.01 lower %5mC (95% CI: -0.02, -0.002) and 0.001 lower %5hmC (95% CI: -0.01, -0.00001) in cord blood only. Increased second-trimester zinc (mg) intake was associated with 0.003 greater %5hmC in early childhood (beta = 0.003; 95% CI: 0.0004, 0.006). Second-trimester folate was positively associated with %5hmC in early childhood only (beta = 0.08, 95% CI: 0.003, 0.16). Associations did not survive multiple testing adjustment; future replication is needed. Trimester-specific nutrients may impact various sensitive windows of epigenetic programming some with lasting effects in childhood. Further research is needed to understand the role of gene-specific epigenetic changes and how global DNA methylation measures relate to child health.

    View details for DOI 10.1017/S2040174422000642

    View details for PubMedID 36515010

  • Early pregnancy essential and non-essential metal mixtures and maternal antepartum and postpartum depressive symptoms. Neurotoxicology Rokoff, L. B., Cardenas, A., Lin, P. D., Rifas-Shiman, S. L., Wright, R. O., Enlow, M. B., Coull, B. A., Oken, E., Korrick, S. A. 2022


    Mood disorders are common during and after pregnancy, and environmental metals may contribute to increased risk. Antepartum metal exposures have not been well characterized in relation to maternal depression. We evaluated the extent to which early pregnancy erythrocyte concentrations of essential and non-essential metals were prospectively associated with antepartum and postpartum depressive symptoms.Participants were 1226 women in Project Viva, a longitudinal cohort recruited during pregnancy (1999-2002). We measured concentrations of 11 metals in maternal first trimester erythrocytes (arsenic, barium, cadmium, cesium, copper, mercury, magnesium, manganese, lead, selenium, zinc). Using the Edinburgh Postnatal Depression Scale (EPDS), we assessed elevated depressive symptoms (≥13; 0-30 scale) at mid-pregnancy and at 6 and 12 months postpartum. We applied latent class mixed modeling to identify symptom trajectories. Adjusting for maternal sociodemographics and co-exposures, we examined associations between the metal mixture and depressive symptoms using logistic (for EPDS≥13)/multinomial (for symptom trajectories) regression and quantile g-computation.In this cohort of moderately high socioeconomic status participants (e.g., 72% college graduate), low-level metal concentrations were weakly to moderately correlated (Spearman: -0.24 to 0.59); the prevalence of depressive symptoms ranged from 9% (mid-pregnancy) to 6% (12 months postpartum); and three trajectories (stable low; elevated mid-pregnancy, then decreasing; moderate mid-pregnancy, then increasing) best fit the EPDS data. The early pregnancy erythrocyte metal mixture was not associated with maternal depressive symptoms in logistic, multinomial, or mixture models. For individual metals, most confidence intervals (CI) included the null. There was weak evidence that arsenic, lead, and selenium were moderately associated with elevated odds of depressive symptoms and/or trajectories. However, the odds ratios (95% CI) per doubling of these three metals were imprecise [e.g., arsenic: 1.13 (0.94, 1.40) for EPDS≥13 at six months postpartum; lead: 1.19 (0.80, 1.77) for EPDS≥13 at mid-pregnancy; selenium: 2.35 (0.84, 6.57) for elevated mid-pregnancy, then decreasing versus stable low trajectory].We did not observe strong, consistent evidence of associations between early pregnancy erythrocyte metal concentrations and subsequent maternal antepartum and postpartum depressive symptoms.

    View details for DOI 10.1016/j.neuro.2022.12.005

    View details for PubMedID 36526156

  • A multi-omics longitudinal aging dataset in primary human fibroblasts with mitochondrial perturbations. Scientific data Sturm, G., Monzel, A. S., Karan, K. R., Michelson, J., Ware, S. A., Cardenas, A., Lin, J., Bris, C., Santhanam, B., Murphy, M. P., Levine, M. E., Horvath, S., Belsky, D. W., Wang, S., Procaccio, V., Kaufman, B. A., Hirano, M., Picard, M. 2022; 9 (1): 751


    Aging is a process of progressive change. To develop biological models of aging, longitudinal datasets with high temporal resolution are needed. Here we report a multi-omics longitudinal dataset for cultured primary human fibroblasts measured across their replicative lifespans. Fibroblasts were sourced from both healthy donors (n=6) and individuals with lifespan-shortening mitochondrial disease (n=3). The dataset includes cytological, bioenergetic, DNA methylation, gene expression, secreted proteins, mitochondrial DNA copy number and mutations, cell-free DNA, telomere length, and whole-genome sequencing data. This dataset enables the bridging of mechanistic processes of aging as outlined by the "hallmarks of aging", with the descriptive characterization of aging such as epigenetic age clocks. Here we focus on bridging the gap for the hallmark mitochondrial metabolism. Our dataset includes measurement of healthy cells, and cells subjected to over a dozen experimental manipulations targeting oxidative phosphorylation (OxPhos), glycolysis, and glucocorticoid signaling, among others. These experiments provide opportunities to test how cellular energetics affect the biology of cellular aging. All data are publicly available at our webtool:

    View details for DOI 10.1038/s41597-022-01852-y

    View details for PubMedID 36463290

  • Non-essential and essential trace element mixtures and kidney function in early pregnancy - A cross-sectional analysis in project viva. Environmental research Lin, P. D., Cardenas, A., Rifas-Shiman, S. L., Zota, A. R., Hivert, M. F., Aris, I. M., Sanders, A. P. 2022; 216 (Pt 4): 114846


    Some trace elements are established nephrotoxicants, yet their associations with kidney function remain understudied in the context of pregnancy, a time of substantial change in kidney physiology and function. We aimed to estimate the individual and joint associations of trace element mixtures with maternal kidney function during the 1st trimester of pregnancy (mean 9.7 gestational weeks). 1040 women from Project Viva contributed blood samples which were assessed for erythrocyte non-essential [arsenic (As), cadmium (Cd), cesium (Cs), mercury (Hg), lead (Pb)] and essential [barium (Ba), magnesium (Mg), manganese (Mn), selenium (Se), and Zinc (Zn)] trace elements, and plasma creatinine for kidney function. We estimated glomerular filtration rate using the Chronic Kidney Disease Epidemiology Collaboration (eGFRCKD-EPI) equation without race-adjustment factors. We examined associations of eGFRCKD-EPI with individual trace elements using multivariable linear regression and their mixtures using quantile-based g-computation, adjusting for sociodemographics, pregnancy characteristics, and diet. Participants in our study were predominantly White (75%), college graduates (72%), and had household income >$70,000/year (63%). After adjusting for covariates, higher Pb (β -3.51 ml/min/1.73 m2; 95% CI -5.83, -1.18) concentrations were associated with lower eGFRCKD-EPI, while higher Mg (β 10.53 ml/min/1.73 m2; 95% CI 5.35, 15.71), Se (β 5.56 ml/min/1.73 m2; 95% CI 0.82, 10.31), and Zn (β 5.88 ml/min/1.73 m2; 95% CI 0.51, 11.26) concentrations were associated with higher eGFRCKD-EPI. In mixture analyses, higher non-essential trace elements mixture concentration was associated with reduced eGFRCKD-EPI (Ψ -1.03 ml/min/1.73 m2; 95% CI: 1.92, -0.14). Conversely, higher essential trace elements mixture concentration was associated with higher eGFR (Ψ 1.42; 95% CI: 0.48, 2.37). Exposure to trace elements in early pregnancy may influence women's kidney function although reverse causation cannot be eliminated in this cross-sectional analysis. These findings have important implications for long-term cardiovascular and postpartum kidney health that warrant additional studies.

    View details for DOI 10.1016/j.envres.2022.114846

    View details for PubMedID 36402181

  • Increased epigenetic age acceleration in the hidradenitis suppurativa skin. Archives of dermatological research Lukac, D., Pagani, K., Collender, P. A., Fadadu, R. P., Cardenas, A., McGee, J. S. 2022


    Epigenetic (or DNA methylation) age is calculated based on methylation of certain cytosine-guanine (CpG) repeats, and it can accurately estimate one's chronologic age. Importantly, epigenetic age acceleration (EAA) is highly predictive of age-associated morbidity and all-cause mortality. Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease with significant systemic disease burden. Here, we performed a pilot study to calculate EAA from formalin-fixed paraffin-embedded skin samples using Illumina Infinium MethylationEpic BeadChip arrays. Our results demonstrated no significant difference in intrinsic EAA among HS compared to controls (-1.00years, p-value=0.52), significant increases in both extrinsic EAA (13.72years, p-value<0.001) and PhenoAge acceleration (7.72years, p-value=0.003), and asignificant decrease in GrimAge acceleration (-5.14years, p-value<0.001). Our findings suggest that the acceleration of epigenetic age in the HS skin may be associated with extrinsic immune-related changes and can potentially serve as a biomarker of the present and/or future disease burden in HS patients.

    View details for DOI 10.1007/s00403-022-02432-1

    View details for PubMedID 36308559

  • Short-Term Exposure of PM2.5 and Epigenetic Aging: A Quasi-Experimental Study. Environmental science & technology Gao, X., Huang, J., Cardenas, A., Zhao, Y., Sun, Y., Wang, J., Xue, L., Baccarelli, A. A., Guo, X., Zhang, L., Wu, S. 2022


    Epigenetic age (EA) is an emerging DNA methylation-based biomarker of biological aging, but whether EA is causally associated with short-term PM2.5 exposure remains unknown. We conducted a quasi-experimental study of 26 healthy adults to test whether short-term PM2.5 exposure accelerates seven EAs with three health examinations performed before, during, and after multiple PM2.5 pollution waves. Seven EAs were derived from the DNA methylation profiles of the Illumina HumanMethylationEPIC BeadChip from CD4+ T-helper cells. We found that an increase of 10 mug/m3 in the 0-24 h personal PM2.5 exposure prior to health examinations was associated with a 0.035, 0.035, 0.050, 0.055, 0.052, and 0.037-unit increase in the changes of z-scored DNA methylation age acceleration (AA,Horvath), AA (Hannum), AA (GrimAge), DunedinPoAm, mortality risk score (MS), and epiTOC, respectively (p-values < 0.05). The same increase in the 24-48 h average personal PM2.5 exposure yielded smaller effects but was still robustly associated with the changes in AA (GrimAge), DunedinPoAm, and MS. Such acute aging effects of PM2.5 were mediated by the changes in several circulating biomarkers, including EC-SOD and sCD40L, with up to 28% mediated proportions. Our findings demonstrated that short-term PM2.5 exposure could accelerate aging reflected by DNA methylation profiles via blood coagulation, oxidative stress, and systematic inflammation.

    View details for DOI 10.1021/acs.est.2c05534

    View details for PubMedID 36197060

  • Epigenome-wide analysis of DNA methylation and optimism in women and men. Psychosomatic medicine Wang, C., DeMeo, D. L., Kim, E. S., Cardenas, A., Fong, K. C., Lee, L. O., Spiro, A. 3., Whitsel, E. A., Horvath, S., Hou, L., Baccarelli, A. A., Li, Y., Stewart, J. D., Manson, J. E., Grodstein, F., Kubzansky, L. D., Schwartz, J. D. 2022


    OBJECTIVE: Higher optimism is associated with reduced mortality and a lower risk of age-related chronic diseases. DNA methylation (DNAm) may provide insight into mechanisms underlying these relationships. We hypothesized DNAm would differ among older individuals who are more versus less optimistic.METHODS: Using cross-sectional data from two population-based cohorts of women with diverse races/ethnicities (N = 3,816) and men (only white, N = 667), we investigated the associations of optimism with epigenome-wide leukocyte DNAm. Random-effects meta-analyses were subsequently used to pool the inabldividual results. Significantly differentially methylated cytosine-phosphate-guanines (CpGs) were identified by the number of independent degrees of freedom approach: effective degrees of freedom correction using the number of principal components (PCs), explaining>95% of the variation of the DNAm data (PC-correction). We performed regional analyses using comb-p and pathways analyses using the Ingenuity Pathway Analysis software.RESULTS: We found essentially all CpGs (total probe N = 359,862) were homogeneous across sex and race/ethnicity in the DNAm~optimism association. In the single CpG site analyses based on homogeneous CpGs, we identified 13 significantly differentially methylated probes using PC-correction. We found four significantly differentially methylated regions and two significantly differentially methylated pathways. The annotated genes from the single CpG site and regional analyses are involved in psychiatric disorders, cardiovascular disease, cognitive impairment, and cancer. Identified pathways were related to cancer, neurodevelopmental and neurodegenerative disorders.CONCLUSION: Our findings provide new insights into possible mechanisms underlying optimism and health.

    View details for DOI 10.1097/PSY.0000000000001147

    View details for PubMedID 36201768

  • Consensus on the Key Characteristics of Immunotoxic Agents as a Basis for Hazard Identification. Environmental health perspectives Germolec, D. R., Lebrec, H., Anderson, S. E., Burleson, G. R., Cardenas, A., Corsini, E., Elmore, S. E., Kaplan, B. L., Lawrence, B. P., Lehmann, G. M., Maier, C. C., McHale, C. M., Myers, L. P., Pallardy, M., Rooney, A. A., Zeise, L., Zhang, L., Smith, M. T. 2022; 130 (10): 105001


    BACKGROUND: Key characteristics (KCs), properties of agents or exposures that confer potential hazard, have been developed for carcinogens and other toxicant classes. KCs have been used in the systematic assessment of hazards and to identify assay and data gaps that limit screening and risk assessment. Many of the mechanisms through which pharmaceuticals and occupational or environmental agents modulate immune function are well recognized. Thus KCs could be identified for immunoactive substances and applied to improve hazard assessment of immunodulatory agents.OBJECTIVES: The goal was to generate a consensus-based synthesis of scientific evidence describing the KCs of agents known to cause immunotoxicity and potential applications, such as assays to measure the KCs.METHODS: A committee of 18 experts with diverse specialties identified 10 KCs of immunotoxic agents, namely, 1) covalently binds to proteins to form novel antigens, 2) affects antigen processing and presentation, 3) alters immune cell signaling, 4) alters immune cell proliferation, 5) modifies cellular differentiation, 6) alters immune cell-cell communication, 7) alters effector function of specific cell types, 8) alters immune cell trafficking, 9) alters cell death processes, and 10) breaks down immune tolerance. The group considered how these KCs could influence immune processes and contribute to hypersensitivity, inappropriate enhancement, immunosuppression, or autoimmunity.DISCUSSION: KCs can be used to improve efforts to identify agents that cause immunotoxicity via one or more mechanisms, to develop better testing and biomarker approaches to evaluate immunotoxicity, and to enable a more comprehensive and mechanistic understanding of adverse effects of exposures on the immune system.

    View details for DOI 10.1289/EHP10800

    View details for PubMedID 36201310

  • Associations of cord blood leukocyte telomere length with adiposity growth from infancy to adolescence. Pediatric obesity Qureshi, F., Aris, I. M., Rifas-Shiman, S. L., Perng, W., Oken, E., Rich-Edwards, J., Cardenas, A., Baccarelli, A. A., Enlow, M. B., Belfort, M. B., Tiemeier, H. 2022: e12977


    OBJECTIVE: Leukocyte telomere length (LTL) may be a biomarker for chronic disease susceptibility, but no work has tested this hypothesis directly. Our study investigated associations of LTL at birth with markers of adiposity growth that are linked with cardiometabolic health later in life.METHODS: Participants were 375 children in Project Viva (48% female, 71% White). Body mass index (BMI) trajectories from birth to 18years were tracked using repeated measures of BMI collected in physical examinations and via medical records, then used to predict age (months) and magnitude (kg/m2 ) of BMI peak and rebound. LTL was measured from cord blood via duplex quantitative PCR. A binary variable indicating LTL shorter than the reference population average was the primary exposure.RESULTS: LTL was unrelated to BMI at peak or rebound, but associations were apparent with the timing of BMI growth milestones. Short LTL was related to a later age of peak for females (beta=0.99, 95% CI=0.16, 1.82; psex interaction =0.015) and an earlier age of rebound for both males and females (betacombined =-5.26, 95% CI=-9.44, -1.08).CONCLUSION: LTL at birth may be an early biomarker of altered adiposity growth. Newborn telomere biology may shed new insight into the developmental origins of health and disease.

    View details for DOI 10.1111/ijpo.12977

    View details for PubMedID 36085441

  • Health Effects of Pesticide Exposure in Latin American and the Caribbean Populations: A Scoping Review. Environmental health perspectives Zuniga-Venegas, L. A., Hyland, C., Munoz-Quezada, M. T., Quiros-Alcala, L., Butinof, M., Buralli, R., Cardenas, A., Fernandez, R. A., Foerster, C., Gouveia, N., Gutierrez Jara, J. P., Lucero, B. A., Munoz, M. P., Ramirez-Santana, M., Smith, A. R., Tirado, N., van Wendel de Joode, B., Calaf, G. M., Handal, A. J., Soares da Silva, A., Cortes, S., Mora, A. M. 2022; 130 (9): 96002


    BACKGROUND: Multiple epidemiological studies have shown that exposure to pesticides is associated with adverse health outcomes. However, the literature on pesticide-related health effects in the Latin American and the Caribbean (LAC) region, an area of intensive agricultural and residential pesticide use, is sparse. We conducted a scoping review to describe the current state of research on the health effects of pesticide exposure in LAC populations with the goal of identifying knowledge gaps and research capacity building needs.METHODS: We searched PubMed and SciELO for epidemiological studies on pesticide exposure and human health in LAC populations published between January 2007 and December 2021. We identified 233 publications from 16 countries that met our inclusion criteria and grouped them by health outcome (genotoxicity, neurobehavioral outcomes, placental outcomes and teratogenicity, cancer, thyroid function, reproductive outcomes, birth outcomes and child growth, and others).RESULTS: Most published studies were conducted in Brazil (37%, n=88) and Mexico (20%, n=46), were cross-sectional in design (72%, n=167), and focused on farmworkers (45%, n=105) or children (21%, n=48). The most frequently studied health effects included genotoxicity (24%, n=62) and neurobehavioral outcomes (21%, n=54), and organophosphate (OP) pesticides were the most frequently examined (26%, n=81). Forty-seven percent (n=112) of the studies relied only on indirect pesticide exposure assessment methods. Exposure to OP pesticides, carbamates, or to multiple pesticide classes was consistently associated with markers of genotoxicity and adverse neurobehavioral outcomes, particularly among children and farmworkers.DISCUSSION: Our scoping review provides some evidence that exposure to pesticides may adversely impact the health of LAC populations, but methodological limitations and inconsistencies undermine the strength of the conclusions. It is critical to increase capacity building, integrate research initiatives, and conduct more rigorous epidemiological studies in the region to address these limitations, better inform public health surveillance systems, and maximize the impact of research on public policies.

    View details for DOI 10.1289/EHP9934

    View details for PubMedID 36173136

  • Corrigendum to "Association of DNA methylation in circulating CD4+ T cells with short-term PM2.5 pollution waves: A quasi-experimental study of healthy young adults" [Ecotoxicol. Environ. Saf. 239 (2022) 1-8/113634]. Ecotoxicology and environmental safety Jiang, M., Wang, X., Gao, X., Cardenas, A., Baccarelli, A. A., Guo, X., Huang, J., Wu, S. 2022; 243: 113970

    View details for DOI 10.1016/j.ecoenv.2022.113970

    View details for PubMedID 35964396

  • Prenatal and birth associations of epigenetic gestational age acceleration in the Center for the Health Assessment of Mothers and Children of Salinas (CHAMACOS) cohort EPIGENETICS Daredia, S., Huen, K., van der Laan, L., Collender, P. A., Nwanaji-Enwerem, J. C., Harley, K., Deardorff, J., Eskenazi, B., Holland, N., Cardenas, A. 2022: 1-16


    Gestational age (GA) is an important determinant of child health and disease risk. Two epigenetic GA clocks have been developed using DNA methylation (DNAm) patterns in cord blood. We investigate the accuracy of GA clocks and determinants of epigenetic GA acceleration (GAA), a biomarker of biological ageing. We hypothesize that prenatal and birth characteristics are associated with altered GAA, thereby disrupting foetal biological ageing. We examined 372 mother-child pairs from the Center for the Health Assessment of Mothers and Children of Salinas study of primarily Latino farmworkers in California. Chronological GA was robustly correlated with epigenetic GA (DNAm GA) estimated by the Knight (r = 0.48, p < 2.2x10-16) and Bohlin clocks (r = 0.67, p < 2.2x10-16) using the Illumina 450K array in cord blood samples collected at birth. GA clock performance was robust, though slightly lower, using DNAm profiles from the Illumina EPIC array in a smaller subsample (Knight: r = 0.39, p < 3.5x10-5; Bohlin: r = 0.60, p < 7.7x10-12). After adjusting for confounders, high maternal serum triglyceride levels (Bohlin: β = -0.01 days per mg/dL, p = 0.03), high maternal serum lipid levels (Bohlin: β = -4.31x10-3 days per mg/dL, p = 0.04), preterm delivery (Bohlin: β = -4.03 days, p = 9.64x10-4), greater maternal parity (Knight: β = -4.07 days, p = 0.01; Bohlin: β = -2.43 days, p = 0.01), and male infant sex (Knight: β = -3.15 days, p = 3.10x10-3) were associated with decreased GAA.Prenatal and birth characteristics affect GAA in newborns. Understanding factors that accelerate or delay biological ageing at birth may identify early-life targets for disease prevention and improve ageing across the life-course. Future research should test the impact of altered GAA on the long-term burden of age-related diseases.

    View details for DOI 10.1080/15592294.2022.2102846

    View details for Web of Science ID 000834893500001

    View details for PubMedID 35912433

  • A mass-balance approach to evaluate arsenic intake and excretion in different populations ENVIRONMENT INTERNATIONAL Beene, D., Collender, P., Cardenas, A., Harvey, C., Huhmann, L., Lin, Y., Lewis, J., LoIacono, N., Navas-Acien, A., Nigra, A., Steinmaus, C., van Geen, A. 2022; 166: 107371


    Unless a toxicant builds up in a deep compartment, intake by the human body must on average balance the amount that is lost. We apply this idea to assess arsenic (As) exposure misclassification in three previously studied populations in rural Bangladesh (n = 11,224), Navajo Nation in the Southwestern United States (n = 619), and northern Chile (n = 630), under varying assumptions about As sources. Relationships between As intake and excretion were simulated by taking into account additional sources, as well as variability in urine dilution inferred from urinary creatinine. The simulations bring As intake closer to As excretion but also indicate that some exposure misclassification remains. In rural Bangladesh, accounting for intake from more than one well and rice improved the alignment of intake and excretion, especially at low exposure. In Navajo Nation, comparing intake and excretion revealed home dust as an important source. Finally, in northern Chile, while food-frequency questionnaires and urinary As speciation indicate fish and shellfish sources, persistent imbalance of intake and excretion suggests imprecise measures of drinking water arsenic as a major cause of exposure misclassification. The mass-balance approach could prove to be useful for evaluating sources of exposure to toxicants in other settings.

    View details for DOI 10.1016/j.envint.2022.107371

    View details for Web of Science ID 000828016800003

    View details for PubMedID 35809487

  • Long-Term Exposure to Ozone and Fine Particulate Matter and Risk of Premature Coronary Artery Disease: Results from Genetics of Atherosclerotic Disease Mexican Study. Biology Posadas-Sanchez, R., Vargas-Alarcon, G., Cardenas, A., Texcalac-Sangrador, J. L., Osorio-Yanez, C., Sanchez-Guerra, M. 2022; 11 (8)


    (1) Background: Epidemiological studies have identified associations between fine particulate matter (PM2.5) and ozone exposure with cardiovascular disease; however, studies linking ambient air pollution and premature coronary artery disease (pCAD) in Latin America are non-existing. (2) Methods: Our study was a case-control analysis nested in the Genetics of Atherosclerotic Disease (GEA) Mexican study. We included 1615 participants (869 controls and 746 patients with pCAD), recruited at the Instituto Nacional de Cardiologia Ignacio Chavez from June 2008 to January 2013. We defined pCAD as history of myocardial infarction, angioplasty, revascularization surgery or coronary stenosis > 50% diagnosed before age 55 in men and age 65 in women. Controls were healthy individuals without personal or family history of pCAD and with coronary artery calcification equal to zero. Hourly measurements of ozone and PM2.5 from the Atmospheric Monitoring System in Mexico City (SIMAT in Spanish; Sistema de Monitero Atmosferico de la Ciudad de Mexico) were used to calculate annual exposure to ozone and PM2.5 in the study participants. (3) Results: Each ppb increase in ozone at 1-year, 2-year, 3-year and 5-year averages was significantly associated with increased odds (OR = 1.10; 95% CI: 1.03-1.18; OR = 1.17; 95% CI: 1.05-1.30; OR = 1.18; 95% CI: 1.05-1.33, and OR = 1.13; 95% CI: 1.04-1.23, respectively) of pCAD. We observed higher risk of pCAD for each 5 g/m3 increase only for the 5-year average of PM2.5 exposure (OR = 2.75; 95% CI: 1.47-5.16), compared to controls. (4) Conclusions: Ozone exposure at different time points and PM2.5 exposure at 5 years were associated with increased odds of pCAD. Our results highlight the importance of reducing long-term exposure to ambient air pollution levels to reduce the burden of cardiovascular disease in Mexico City and other metropolitan areas.

    View details for DOI 10.3390/biology11081122

    View details for PubMedID 35892978

  • Comparison of DNA methylation measurements from EPIC BeadChip and SeqCap targeted bisulphite sequencing in PON1 and nine additional candidate genes EPIGENETICS Khodasevich, D., Smith, A. R., Huen, K., Eskenazi, B., Cardenas, A., Holland, N. 2022: 1-12


    Epigenome-wide association studies (EWAS) are widely implemented in epidemiology, and the Illumina HumanMethylationEPIC BeadChip (EPIC) DNA microarray is the most-used technology. Recently, next-generation sequencing (NGS)-based methods, which assess DNA methylation at single-base resolution, have become more affordable and technically feasible. While the content of microarray technology is fixed, NGS-based approaches, such as the Roche Nimblegen, SeqCap Epi Enrichment System (SeqCap), offer the flexibility of targeting most CpGs in a gene. With the current usage of microarrays and emerging NGS-based technologies, it is important to establish whether data generated from the two platforms are comparable. We harnessed 112 samples from the Center for the Health Assessment of Mothers and Children of Salinas (CHAMACOS) birth cohort study and compared DNA methylation between the EPIC microarray and SeqCap for PON1 and nine additional candidate genes, by evaluating epigenomic coverage and correlations. We conducted multivariable linear regression and principal component analyses to assess the ability of the EPIC array and SeqCap to detect biological differences in gene methylation by the PON1-108 single nucleotide polymorphism. We found an overall high concordance (r = 0.84) between SeqCap and EPIC DNA methylation, among highly methylated and minimally methylated regions. However, substantial disagreement was present between the two methods in moderately methylated regions, with SeqCap measurements exhibiting greater within-site variation. Additionally, SeqCap did not capture PON1 SNP associated differences in DNA methylation that were evident with the EPIC array. Our findings indicate that microarrays perform well for analysing DNA methylation in large cohort studies but with limited coverage.

    View details for DOI 10.1080/15592294.2022.2091818

    View details for Web of Science ID 000820075200001

    View details for PubMedID 35786310

  • Association of DNA methylation in circulating CD4+T cells with short-term PM2.5 pollution waves: A quasi-experimental study of healthy young adults ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY Jiang, M., Wang, X., Gao, X., Cardenas, A., Baccarelli, A. A., Guo, X., Huang, J., Wu, S. 2022; 239: 113634


    Fine particulate matter (PM2.5) is a modifiable environmental risk factor with established adverse effects on human health. However, associations between acute PM2.5 fluctuation and DNA methylation remain unknown.A quasi-experimental study utilizing naturally occurring PM2.5 pollution waves (PPWs) was conducted on 32 healthy young adults. Repeated follow-up measurements were performed and participants served as their own controls before, during, and after PPWs. Exposure measurements including indoor and ambient PM2.5 levels, and equivalent personal PM2.5 exposure were further estimated based on the time-location information. DNA methylation profiles of circulating CD4+T cells were obtained using Illumina HumanMethylationEPIC BeadChip. Linear mixed-effect models were applied to estimate the associations between two scenarios (during-PPWs vs. pre-PPWs periods and during-PPWs vs. post-PPWs periods) and methylation level of each CpG site. We further validated their associations with the personal PM2.5 exposure, and GO and KEGG analyses and mediation analysis were conducted accordingly.Data from 26 participants were included in final analysis after quality control. Short-term high PM2.5 exposure was associated with DNA methylation changes of participants. Nine differently methylated CpG sites were not only significantly associated with PPWs periods but also with personal PM2.5 exposure in 24-h prior to the health examinations (p < 0.01). Gene ontology analysis found that five sites were associated with two pathways relating to membrane protein synthesis. PM2.5-related changes in CpG sites were mediated by sP-selectin, 8-isoPGF2α, EGF, GRO, IL-15, and IFN-α2, with mediated proportions ranging from 9.65% to 23.40%.This is the first quasi-experimental study showing that short-term high PM2.5 exposure could alter the DNA methylation of CD4+T cells, which provided valuable information for further exploring underlying biological mechanisms and epigenetic biomarkers for PM2.5-related acute health effects.

    View details for DOI 10.1016/j.ecoenv.2022.113634

    View details for Web of Science ID 000817576600005

    View details for PubMedID 35617899

  • The impact of prenatal and early-life arsenic exposure on epigenetic age acceleration among adults in Northern Chile ENVIRONMENTAL EPIGENETICS Bozack, A. K., Boileau, P., Hubbard, A. E., Sille, F. M., Ferreccio, C., Steinmaus, C. M., Smith, M. T., Cardenas, A. 2022; 8 (1): dvac014


    Exposure to arsenic affects millions of people globally. Changes in the epigenome may be involved in pathways linking arsenic to health or serve as biomarkers of exposure. This study investigated associations between prenatal and early-life arsenic exposure and epigenetic age acceleration (EAA) in adults, a biomarker of morbidity and mortality. DNA methylation was measured in peripheral blood mononuclear cells (PBMCs) and buccal cells from 40 adults (median age = 49 years) in Chile with and without high prenatal and early-life arsenic exposure. EAA was calculated using the Horvath, Hannum, PhenoAge, skin and blood, GrimAge, and DNA methylation telomere length clocks. We evaluated associations between arsenic exposure and EAA using robust linear models. Participants classified as with and without arsenic exposure had a median drinking water arsenic concentration at birth of 555 and 2 μg/l, respectively. In PBMCs, adjusting for sex and smoking, exposure was associated with a 6-year PhenoAge acceleration [B (95% CI) = 6.01 (2.60, 9.42)]. After adjusting for cell-type composition, we found positive associations with Hannum EAA [B (95% CI) = 3.11 (0.13, 6.10)], skin and blood EAA [B (95% CI) = 1.77 (0.51, 3.03)], and extrinsic EAA [B (95% CI) = 4.90 (1.22, 8.57)]. The association with PhenoAge acceleration in buccal cells was positive but not statistically significant [B (95% CI) = 4.88 (-1.60, 11.36)]. Arsenic exposure limited to early-life stages may be associated with biological aging in adulthood. Future research may provide information on how EAA programmed in early life is related to health.

    View details for DOI 10.1093/eep/dvac014

    View details for Web of Science ID 000816745100001

    View details for PubMedID 35769198

    View details for PubMedCentralID PMC9235373

  • In vitro relationships of galactic cosmic radiation and epigenetic clocks in human bronchial epithelial cells ENVIRONMENTAL AND MOLECULAR MUTAGENESIS Nwanaji-Enwerem, J. C., Boileau, P., Galazka, J. M., Cardenas, A. 2022; 63 (4): 184-189


    Ionizing radiation is a well-appreciated health risk, precipitant of DNA damage, and contributor to DNA methylation variability. Nevertheless, relationships of ionizing radiation with DNA methylation-based markers of biological age (i.e. epigenetic clocks) remain poorly understood. Using existing data from human bronchial epithelial cells, we examined in vitro relationships of three epigenetic clock measures (Horvath DNAmAge, MiAge, and epiTOC2) with galactic cosmic radiation (GCR), which is particularly hazardous due to its high linear energy transfer (LET) heavy-ion components. High-LET 56Fe was significantly associated with accelerations in epiTOC2 (β = 192 cell divisions, 95% CI: 71, 313, p-value = .003). We also observed a significant, positive interaction of 56Fe ions and time-in-culture with epiTOC2 (95% CI: 42, 441, p-value = .019). However, only the direct 56Fe ion association remained statistically significant after adjusting for multiple hypothesis testing. Epigenetic clocks were not significantly associated with high-LET 28Si and low-LET X-rays. Our results demonstrate sensitivities of specific epigenetic clock measures to certain forms of GCR. These findings suggest that epigenetic clocks may have some utility for monitoring and better understanding the health impacts of GCR.

    View details for DOI 10.1002/em.22483

    View details for Web of Science ID 000793690300001

    View details for PubMedID 35470505

    View details for PubMedCentralID PMC9233067

  • Epigenome-wide association study and epigenetic age acceleration associated with cigarette smoking among Costa Rican adults. Scientific reports Cardenas, A., Ecker, S., Fadadu, R. P., Huen, K., Orozco, A., McEwen, L. M., Engelbrecht, H., Gladish, N., Kobor, M. S., Rosero-Bixby, L., Dow, W. H., Rehkopf, D. H. 2022; 12 (1): 4277


    Smoking-associated DNA methylation (DNAm) signatures are reproducible among studies of mostly European descent, with mixed evidence if smoking accelerates epigenetic aging and its relationship to longevity. We evaluated smoking-associated DNAm signatures in the Costa Rican Study on Longevity and Healthy Aging (CRELES), including participants from the high longevity region of Nicoya.We measured genome-wide DNAm in leukocytes, tested Epigenetic Age Acceleration (EAA) from five clocks and estimates of telomere length (DNAmTL), and examined effect modification by the high longevity region.489 participants had a mean (SD) age of 79.4 (10.8) years, and 18% were from Nicoya. Overall, 7.6% reported currently smoking, 35% were former smokers, and 57.4% never smoked. 46 CpGs and five regions (e.g. AHRR, SCARNA6/SNORD39, SNORA20, and F2RL3) were differentially methylated for current smokers. Former smokers had increased Horvath's EAA (1.69-years; 95% CI 0.72, 2.67), Hannum's EAA (0.77-years; 95% CI 0.01, 1.52), GrimAge (2.34-years; 95% CI1.66, 3.02), extrinsic EAA (1.27-years; 95% CI 0.34, 2.21), intrinsic EAA (1.03-years; 95% CI 0.12, 1.94) and shorter DNAmTL (-0.04-kb; 95% CI -0.08, -0.01) relative to non-smokers. There was no evidence of effect modification among residents of Nicoya. Our findings recapitulate previously reported and novel smoking-associated DNAm changes in a Latino cohort.

    View details for DOI 10.1038/s41598-022-08160-w

    View details for PubMedID 35277542

  • Meta-analysis of epigenome-wide associations between DNA methylation at birth and childhood cognitive skills MOLECULAR PSYCHIATRY Caramaschi, D., Neumann, A., Cardenas, A., Tindula, G., Alemany, S., Zillich, L., Pesce, G., Lahti, J. T., Havdahl, A., Mulder, R., Felix, J. F., Tiemeier, H., Sirignano, L., Frank, J., Witt, S. H., Rietschel, M., Deuschle, M., Huen, K., Eskenazi, B., Send, T., Ferrer, M., Gilles, M., de Agostini, M., Baiz, N., Rifas-Shiman, S. L., Kvist, T., Czamara, D., Tuominen, S. T., Relton, C. L., Rai, D., London, S. J., Raikkonen, K., Holland, N., Annesi-Maesano, I., Streit, F., Hivert, M., Oken, E., Sunyer, J., Cecil, C. M., Sharp, G. 2022; 27 (4): 2126-2135


    Cognitive skills are a strong predictor of a wide range of later life outcomes. Genetic and epigenetic associations across the genome explain some of the variation in general cognitive abilities in the general population and it is plausible that epigenetic associations might arise from prenatal environmental exposures and/or genetic variation early in life. We investigated the association between cord blood DNA methylation at birth and cognitive skills assessed in children from eight pregnancy cohorts within the Pregnancy And Childhood Epigenetics (PACE) Consortium across overall (total N = 2196), verbal (total N = 2206) and non-verbal cognitive scores (total N = 3300). The associations at single CpG sites were weak for all of the cognitive domains investigated. One region near DUSP22 on chromosome 6 was associated with non-verbal cognition in a model adjusted for maternal IQ. We conclude that there is little evidence to support the idea that variation in cord blood DNA methylation at single CpG sites is associated with cognitive skills and further studies are needed to confirm the association at DUSP22.

    View details for DOI 10.1038/s41380-022-01441-w

    View details for Web of Science ID 000753754100004

    View details for PubMedID 35145228

    View details for PubMedCentralID PMC9126809

  • Environment-induced epigenetic modifications as therapeutic targets for prevention and treatment Environment-induced epigenetic modifications as therapeutic targets for prevention and treatment Ward-Caviness, C., Kraus, W. E., Dhingra, R., Martin, C. L., Cardenas, A., Nwanaji-Enwerem, J., Cascio, W. E. Elsevier. 2022
  • Maternal adverse childhood experiences before pregnancy are associated with epigenetic aging changes in their children AGING-US Nwanaji-Enwerem, J. C., van der Laan, L., Kogut, K., Eskenazi, B., Holland, N., Deardorff, J., Cardenas, A. 2021; 13 (24): 25653-25669


    Emerging research suggests associations of physical and psychosocial stressors with epigenetic aging. Although this work has included early-life exposures, data on maternal exposures and epigenetic aging of their children remain sparse. Using longitudinally collected data from the California, Salinas Valley CHAMACOS study, we examined relationships between maternal Adverse Childhood Experiences (ACEs) reported up to 18 years of life, prior to pregnancy, with eight measures (Horvath, Hannum, SkinBloodClock, Intrinsic, Extrinsic, PhenoAge, GrimAge, and DNAm telomere length) of blood leukocyte epigenetic age acceleration (EAA) in their children at ages 7, 9, and 14 years (N = 238 participants with 483 observations). After adjusting for maternal chronological age at delivery, pregnancy smoking/alcohol use, parity, child gestational age, and estimated leukocyte proportions, higher maternal ACEs were significantly associated with at least a 0.76-year increase in child Horvath and Intrinsic EAA. Higher maternal ACEs were also associated with a 0.04 kb greater DNAm estimate of telomere length of children. Overall, our data suggests that maternal preconception ACEs are associated with biological aging in their offspring in childhood and that preconception ACEs have differential relationships with EAA measures, suggesting different physiologic utilities of EEA measures. Studies are necessary to confirm these findings and to elucidate potential pathways to explain these relationships, which may include intergenerational epigenetic inheritance and persistent physical and social exposomes.

    View details for Web of Science ID 000740024100004

    View details for PubMedID 34923483

    View details for PubMedCentralID PMC8751604

  • An epigenetic aging analysis of randomized metformin and weight loss interventions in overweight postmenopausal breast cancer survivors CLINICAL EPIGENETICS Nwanaji-Enwerem, J. C., Chung, F., Van der Laan, L., Novoloaca, A., Cuenin, C., Johansson, H., Bonanni, B., Hubbard, A. E., Smith, M. T., Hartman, S. J., Cardenas, A., Sears, D. D., Herceg, Z. 2021; 13 (1): 224


    Metformin and weight loss relationships with epigenetic age measures-biological aging biomarkers-remain understudied. We performed a post-hoc analysis of a randomized controlled trial among overweight/obese breast cancer survivors (N = 192) assigned to metformin, placebo, weight loss with metformin, or weight loss with placebo interventions for 6 months. Epigenetic age was correlated with chronological age (r = 0.20-0.86; P < 0.005). However, no significant epigenetic aging associations were observed by intervention arms. Consistent with published reports in non-cancer patients, 6 months of metformin therapy may be inadequate to observe expected epigenetic age deceleration. Longer duration studies are needed to better characterize these relationships.Trial Registration: Registry Name: ClincialTrials.Gov.Registration Number: NCT01302379.Date of Registration: February 2011.URL:

    View details for DOI 10.1186/s13148-021-01218-y

    View details for Web of Science ID 000731259800001

    View details for PubMedID 34920739

    View details for PubMedCentralID PMC8684118

  • Detecting cord blood cell type-specific epigenetic associations with gestational diabetes mellitus and early childhood growth CLINICAL EPIGENETICS Lu, T., Cardenas, A., Perron, P., Hivert, M., Bouchard, L., Greenwood, C. T. 2021; 13 (1): 131


    Epigenome-wide association studies (EWAS) have provided opportunities to understand the role of epigenetic mechanisms in development and pathophysiology of many chronic diseases. However, an important limitation of conventional EWAS is that profiles of epigenetic variability are often obtained in samples of mixed cell types. Here, we aim to assess whether changes in cord blood DNA methylation (DNAm) associated with gestational diabetes mellitus (GDM) exposure and early childhood growth markers occur in a cell type-specific manner.We analyzed 275 cord blood samples collected at delivery from a prospective pre-birth cohort with genome-wide DNAm profiled by the Illumina MethylationEPIC array. We estimated proportions of seven common cell types in each sample using a cord blood-specific DNAm reference panel. Leveraging a recently developed approach named CellDMC, we performed cell type-specific EWAS to identify CpG loci significantly associated with GDM, or 3-year-old body mass index (BMI) z-score. A total of 1410 CpG loci displayed significant cell type-specific differences in methylation level between 23 GDM cases and 252 controls with a false discovery rate < 0.05. Gene Ontology enrichment analysis indicated that LDL transportation emerged from CpG specifically identified from B-cells DNAm analyses and the mitogen-activated protein kinase pathway emerged from CpG specifically identified from natural killer cells DNAm analyses. In addition, we identified four and six loci associated with 3-year-old BMI z-score that were specific to CD8+ T-cells and monocytes, respectively. By performing genome-wide permutation tests, we validated that most of our detected signals had low false positive rates.Compared to conventional EWAS adjusting for the effects of cell type heterogeneity, the proposed approach based on cell type-specific EWAS could provide additional biologically meaningful associations between CpG methylation, prenatal maternal GDM or 3-year-old BMI. With careful validation, these findings may provide new insights into the pathogenesis, programming, and consequences of related childhood metabolic dysregulation. Therefore, we propose that cell type-specific analyses are worth cautious explorations.

    View details for DOI 10.1186/s13148-021-01114-5

    View details for Web of Science ID 000666861700001

    View details for PubMedID 34174944

    View details for PubMedCentralID PMC8236204

  • Prenatal metal exposure, cord blood DNA methylation and persistence in childhood: an epigenome-wide association study of 12 metals CLINICAL EPIGENETICS Bozack, A. K., Rifas-Shiman, S. L., Coull, B. A., Baccarelli, A. A., Wright, R. O., Amarasiriwardena, C., Gold, D. R., Oken, E., Hivert, M., Cardenas, A. 2021; 13 (1): 208


    Prenatal exposure to essential and non-essential metals impacts birth and child health, including fetal growth and neurodevelopment. DNA methylation (DNAm) may be involved in pathways linking prenatal metal exposure and health. In the Project Viva cohort, we analyzed the extent to which metals (As, Ba, Cd, Cr, Cs, Cu, Hg, Mg, Mn, Pb, Se, and Zn) measured in maternal erythrocytes were associated with differentially methylated positions (DMPs) and regions (DMRs) in cord blood and tested if associations persisted in blood collected in mid-childhood. We measured metal concentrations in first-trimester maternal erythrocytes, and DNAm in cord blood (N = 361) and mid-childhood blood (N = 333, 6-10 years) with the Illumina HumanMethylation450 BeadChip. For each metal individually, we tested for DMPs using linear models (considered significant at FDR < 0.05), and for DMRs using comb-p (Sidak p < 0.05). Covariates included biologically relevant variables and estimated cell-type composition. We also performed sex-stratified analyses.Pb was associated with decreased methylation of cg20608990 (CASP8) (FDR = 0.04), and Mn was associated with increased methylation of cg02042823 (A2BP1) in cord blood (FDR = 9.73 × 10-6). Both associations remained significant but attenuated in blood DNAm collected at mid-childhood (p < 0.01). Two and nine Mn-associated DMPs were identified in male and female infants, respectively (FDR < 0.05), with two and six persisting in mid-childhood (p < 0.05). All metals except Ba and Pb were associated with ≥ 1 DMR among all infants (Sidak p < 0.05). Overlapping DMRs annotated to genes in the human leukocyte antigen (HLA) region were identified for Cr, Cs, Cu, Hg, Mg, and Mn.Prenatal metal exposure is associated with DNAm, including DMRs annotated to genes involved in neurodevelopment. Future research is needed to determine if DNAm partially explains the relationship between prenatal metal exposures and health outcomes.

    View details for DOI 10.1186/s13148-021-01198-z

    View details for Web of Science ID 000720641900001

    View details for PubMedID 34798907

    View details for PubMedCentralID PMC8605513

  • Prospective Associations of Early Pregnancy Metal Mixtures with Mitochondria DNA Copy Number and Telomere Length in Maternal and Cord Blood ENVIRONMENTAL HEALTH PERSPECTIVES Smith, A. R., Lin, P. D., Rifas-Shiman, S. L., Rahman, M. L., Gold, D. R., Baccarelli, A. A., Henn, B., Amarasiriwardena, C., Wright, R. O., Coull, B., Hivert, M., Oken, E., Cardenas, A. 2021; 129 (11): 117007


    Metal exposure during pregnancy influences maternal and child health. Oxidative stress and inflammation may mediate adverse effects of heavy metals, whereas essential metals may act as antioxidants. Mitochondrial DNA is a prime target for metal-induced oxidative damage. Telomere dysfunction is attributed to imbalances between reactive oxidant species and antioxidants.We evaluated individual and joint associations of prenatal metals with mitochondrial DNA copy number (mtDNAcn) and telomere length (TL) in maternal and cord blood as biomarkers of inflammation and oxidative stress.We measured six nonessential metals (arsenic, barium, cadmium, cesium, lead, mercury) and four essential metals (magnesium, manganese, selenium, zinc) in first-trimester maternal red blood cells in Project Viva, a U.S. prebirth cohort. We measured relative mtDNAcn (n=898) and TL (n=893) in second-trimester maternal blood and mtDNAcn (n=419) and TL (n=408) in cord blood. We used multivariable linear regression and quantile g-computation to estimate associations between prenatal metals and the biomarkers. We used generalized additive models and Bayesian kernel machine regression to examine nonlinearity and interactions.A 2-fold increase in maternal magnesium was associated with lower maternal [β=-0.07, 95% confidence interval (CI): -0.10, -0.01] and cord blood (β=-0.08, 95% CI: -0.20, -0.01) mtDNAcn. Lead was associated with higher maternal mtDNAcn (β=0.04, 95% CI: 0.01, 0.06). Selenium was associated with longer cord blood TL (β=0.30, 95% CI: 0.01 0.50). An association was observed between the nonessential metal mixture and higher maternal mtDNAcn (β=0.04, 95% CI: 0.01, 0.07). There was a nonlinear relationship between cord blood mtDNAcn and magnesium; maternal mtDNAcn and barium, lead, and mercury; and maternal TL and barium.Maternal exposure to metals such as lead, magnesium, and selenium was associated with mtDNAcn and TL in maternal second trimester and cord blood. Future work will evaluate whether these biomarkers are associated with child health.

    View details for DOI 10.1289/EHP9294

    View details for Web of Science ID 000726754800012

    View details for PubMedID 34797165

    View details for PubMedCentralID PMC8604047

  • Early pregnancy exposure to metal mixture and birth outcomes - A prospective study in Project Viva. Environment international Rahman, M. L., Oken, E., Hivert, M. F., Rifas-Shiman, S., Lin, P. D., Colicino, E., Wright, R. O., Amarasiriwardena, C., Claus Henn, B. G., Gold, D. R., Coull, B. A., Cardenas, A. 2021; 156: 106714


    Prenatal exposure to metals has been individually associated with birth outcomes. However, little is known about the effect of metal mixture, particularly at low exposure levels.To estimate individual and joint effects of metal mixture components on birth outcomes.We used data from 1,391 mother-infant pairs in Project Viva (1999-2002). We measured 11 metals in maternal 1st trimester erythrocyte; abstracted birth weight from medical records; calculated gestational age from last menstrual period or ultrasound; and obtained birth length (n = 729) and head circumference (n = 791) from research measurements. We estimated individual and joint effects of metals using multivariable linear and Bayesian kernel machine regressions.In both single metal and metal mixture analyses, exposure to higher concentrations of arsenic was associated with lower birth weight in males, zinc with higher head circumference in females, and manganese with higher birth length in sex-combined analysis. We also observed sex-specific metal interactions with birth outcomes. Arsenic and manganese showed a synergistic association with birth weight in males, in whom an interquartile range (IQR) increase in arsenic was associated with 25.3 g (95% CI: -79.9, 29.3), 47.9 g (95% CI: -98.0, 2.1), and 72.2 g (95% CI: -129.8, -14.7) lower birth weight when manganese concentrations were at 25th, 50th, and 75th percentiles, respectively. Lead and zinc showed an antagonistic association with head circumference in males, where an IQR increase in lead was associated with 0.18 cm (95% CI: -0.35, -0.02), 0.10 cm (95% CI: -0.25, 0.04), 0.03 cm (95% CI: -0.2, 0.14) smaller head circumference when zinc concentrations were at 25th, 50th, and 75th percentiles, respectively. Exposure to higher concentrations of arsenic was also associated with lower gestational age in males when concentrations of manganese and lead were higher.Maternal erythrocyte concentrations of arsenic, manganese, lead, and zinc were individually and interactively associated with birth outcomes. The associations varied by infant sex and exposure level of other mixture components.

    View details for DOI 10.1016/j.envint.2021.106714

    View details for PubMedID 34147999

    View details for PubMedCentralID PMC8842844

  • Short- and intermediate-term exposure to ambient fine particulate elements and leukocyte epigenome-wide DNA methylation in older men: the Normative Aging Study ENVIRONMENT INTERNATIONAL Wang, C., Cardenas, A., Hutchinson, J. N., Just, A., Heiss, J., Hou, L., Zheng, Y., Coull, B. A., Kosheleva, A., Koutrakis, P., Baccarelli, A. A., Schwartz, J. D. 2022; 158: 106955


    Several epigenome-wide association studies (EWAS) of ambient particulate matter with aerodynamic diameter ≤ 2.5 µm (PM2.5) have been reported. However, EWAS of PM2.5 elements (PEs), reflecting different emission sources, are very limited.We performed EWAS of short- and intermediate-term exposure to PM2.5 and 13 PEs. We hypothesized that significant changes in DNAm may vary by PM2.5 mass and its elements.We repeatedly collected blood samples in the Normative Aging Study and measured leukocyte DNA methylation (DNAm) with the Illumina HumanMethylation450K BeadChip. We collected daily PM2.5 and 13 PEs at a fixed central site. To estimate the associations between each PE and DNAm at individual cytosine-phosphate-guanine (CpG) sites, we incorporated a distributed-lag (0-27 d) term in the setting of median regression with subject-specific intercept and examined cumulative lag associations. We also accounted for selection bias due to loss to follow-up and mortality prior to enrollment. Significantly differentially methylated probes (DMPs) were identified using Bonferroni correction for multiple testing. We further conducted regional and pathway analyses to identify significantly differentially methylated regions (DMRs) and pathways.We included 695 men with 1,266 visits between 1999 and 2013. The subjects had a mean age of 75 years. The significant DMPs, DMRs, and pathways varied by to PM2.5 total mass and PEs. For example, PM2.5 total mass was associated with 2,717 DMPs and 10,470 DMRs whereas Pb was associated with 3,173 DMPs and 637 DMRs. The identified pathways by PM2.5 mass were mostly involved in mood disorders, neuroplasticity, immunity, and inflammation, whereas the pathways associated with motor vehicles (BC, Cu, Pb, and Zn) were related with cardiovascular disease and cancer (e.g., "PPARs signaling").PM2.5 and PE were associated with methylation changes at multiple probes and along multiple pathways, in ways that varied by particle components.

    View details for DOI 10.1016/j.envint.2021.106955

    View details for Web of Science ID 000717953600019

    View details for PubMedID 34717175

    View details for PubMedCentralID PMC8710082

  • Associations between an integrated component of maternal glycemic regulation in pregnancy and cord blood DNA methylation EPIGENOMICS Juvinao-Quintero, D. L., Cardenas, A., Perron, P., Bouchard, L., Lutz, S. M., Hivert, M. 2021; 13 (18): 1459-1472


    Background: Previous studies suggest that fetal programming to hyperglycemia in pregnancy is due to modulation of DNA methylation (DNAm), but they have been limited in their maternal glycemic characterization. Methods: In the Gen3G study, we used a principal component analysis to integrate multiple glucose and insulin values measured during the second trimester oral glucose tolerance test. We investigated associations between principal components and cord blood DNAm levels in an epigenome-wide analysis among 430 mother-child pairs. Results: The first principal component was robustly associated with lower DNAm at cg26974062 (TXNIP; p = 9.9 × 10-9) in cord blood. TXNIP is a well-known DNAm marker for type 2 diabetes in adults. Conclusion: We hypothesize that abnormal glucose metabolism in pregnancy may program dysregulation of TXNIP across the life course.

    View details for DOI 10.2217/epi-2021-0220

    View details for Web of Science ID 000702421300001

    View details for PubMedID 34596421

    View details for PubMedCentralID PMC8503802

  • Nasal epigenetic age and systemic steroid response in pediatric emergency department asthma patients ALLERGY Nwanaji-Enwerem, J. C., Osborne, A., Cardenas, A. 2022; 77 (1): 307-309

    View details for DOI 10.1111/all.15102

    View details for Web of Science ID 000700083200001

    View details for PubMedID 34542184

    View details for PubMedCentralID PMC8716423

  • Epigenetic aging biomarkers and occupational exposure to benzene, trichloroethylene and formaldehyde ENVIRONMENT INTERNATIONAL Van der Laan, L., Cardenas, A., Vermeulen, R., Fadadu, R. P., Hubbard, A. E., Phillips, R., Zhang, L., Breeze, C., Hu, W., Wen, C., Huang, Y., Tang, X., Smith, M. T., Rothman, N., Lan, Q. 2022; 158: 106871


    Epigenetic aging biomarkers are associated with increased morbidity and mortality. We evaluated if occupational exposure to three established chemical carcinogens is associated with acceleration of epigenetic aging. We studied workers in China occupationally exposed to benzene, trichloroethylene (TCE) or formaldehyde by measuring personal air exposures prior to blood collection. Unexposed controls matched by age and sex were selected from nearby factories. We measured leukocyte DNA methylation (DNAm) in peripheral white blood cells using the Infinium HumanMethylation450 BeadChip to calculate five epigenetic aging clocks and DNAmTL, a biomarker associated with leukocyte telomere length and cell replication. We tested associations between exposure intensity and epigenetic age acceleration (EAA), defined as the residuals of regressing the DNAm aging biomarker on chronological age, matching factors and potential confounders. Median differences in EAA between exposure groups were tested using a permutation test with exact p-values. Epigenetic clocks were strongly correlated with age (Spearman r > 0.8) in all three occupational studies. There was a positive exposure-response relationship between benzene and the Skin-Blood Clock EAA biomarker: median EAA was -0.91 years in controls (n = 44), 0.78 years in workers exposed to <10 ppm (n = 41; mean benzene = 1.35 ppm; p = 0.034 vs. controls), and 2.10 years in workers exposed to ≥10 ppm (n = 9; mean benzene = 27.3 ppm; p = 0.019 vs. controls; ptrend = 0.0021). In the TCE study, control workers had a median Skin-Blood Clock EAA of -0.54 years (n = 71) compared to 1.63 years among workers exposed to <10 ppm of TCE (n = 27; mean TCE = 4.22 ppm; p = 0.035). We observed no evidence of EAA associations with formaldehyde exposure (39 controls, 31 exposed). Occupational benzene and TCE exposure were associated with increased epigenetic age acceleration measured by the Skin-Blood Clock. For TCE, there was some evidence of epigenetic age acceleration for lower exposures compared to controls. Our results suggest that some chemical carcinogens may accelerate epigenetic aging.

    View details for DOI 10.1016/j.envint.2021.106871

    View details for Web of Science ID 000705001600005

    View details for PubMedID 34560324

    View details for PubMedCentralID PMC9084243

  • Impact of paternal education on epigenetic ageing in adolescence and mid-adulthood: a multi-cohort study in the USA and Mexico INTERNATIONAL JOURNAL OF EPIDEMIOLOGY Joyce, B. T., Gao, T., Koss, K., Zheng, Y., Cardenas, A., Heiss, J., Just, A., Zhang, K., van Horn, L., Allen, N., Greenland, P., Cohen, S., Gordon-Larsen, P., Mitchell, C., McLanahan, S., Schneper, L., Notterman, D., Rifas-Shiman, S. L., Oken, E., Hivert, M., Wright, R., Baccarelli, A., Lloyd-Jones, D., Hou, L. 2022; 51 (3): 870-884


    Both parental and neighbourhood socio-economic status (SES) are linked to poorer health independently of personal SES measures, but the biological mechanisms are unclear. Our objective was to examine these influences via epigenetic age acceleration (EAA)-the discrepancy between chronological and epigenetic ages.We examined three USA-based [Coronary Artery Risk Disease in Adults (CARDIA) study, Fragile Families and Child Wellbeing Study (FFCWS) and Programming Research in Obesity, Growth, Environment and Social Stressors (PROGRESS)] and one Mexico-based (Project Viva) cohort. DNA methylation was measured using Illumina arrays, personal/parental SES by questionnaire and neighbourhood disadvantage from geocoded address. In CARDIA, we examined the most strongly associated personal, parental and neighbourhood SES measures with EAA (Hannum's method) at study years 15 and 20 separately and combined using a generalized estimating equation (GEE) and compared with other EAA measures (Horvath's EAA, PhenoAge and GrimAge calculators, and DunedinPoAm).EAA was associated with paternal education in CARDIA [GEEs: βsome college = -1.01 years (-1.91, -0.11) and β

    View details for DOI 10.1093/ije/dyab196

    View details for Web of Science ID 000790162200001

    View details for PubMedID 34534313

    View details for PubMedCentralID PMC9189973

  • FUNCTION-ON-FUNCTION REGRESSION FOR THE IDENTIFICATION OF EPIGENETIC REGIONS EXHIBITING WINDOWS OF SUSCEPTIBILITY TO ENVIRONMENTAL EXPOSURES. The annals of applied statistics Zemplenyi, M., Meyer, M. J., Cardenas, A., Hivert, M. F., Rifas-Shiman, S. L., Gibson, H., Kloog, I., Schwartz, J., Oken, E., DeMeo, D. L., Gold, D. R., Coull, B. A. 2021; 15 (3): 1366-1385


    The ability to identify time periods when individuals are most susceptible to exposures as well as the biological mechanisms through which these exposures act is of great public health interest. Growing evidence supports an association between prenatal exposure to air pollution and epigenetic marks, such as DNA methylation, but the timing and gene-specific effects of these epigenetic changes are not well understood. Here, we present the first study that aims to identify prenatal windows of susceptibility to air pollution exposures in cord blood DNA methylation. In particular, we propose a function-on-function regression model that leverages data from nearby DNA methylation probes to identify epigenetic regions that exhibit windows of susceptibility to ambient particulate matter less than 2.5 microns (PM2.5). By incorporating the covariance structure among both the multivariate DNA methylation outcome and the time-varying exposure under study, this framework yields greater power to detect windows of susceptibility and greater control of false discoveries than methods that model probes independently. We compare our method to a distributed lag model approach that models DNA methylation in a probe-by-probe manner, both in simulation and by application to motivating data from the Project Viva birth cohort. We identify a window of susceptibility to PM2.5 exposure in the middle of the third trimester of pregnancy in an epigenetic region selected based on prior studies of air pollution effects on epigenome-wide methylation.

    View details for DOI 10.1214/20-aoas1425

    View details for PubMedID 36313278

    View details for PubMedCentralID PMC9615608

  • FUNCTION-ON-FUNCTION REGRESSION FOR THE IDENTIFICATION OF EPIGENETIC REGIONS EXHIBITING WINDOWS OF SUSCEPTIBILITY TO ENVIRONMENTAL EXPOSURES ANNALS OF APPLIED STATISTICS Zemplenyi, M., Meyer, M. J., Cardenas, A., Hivert, M., Rifas-Shiman, S. L., Gibson, H., Kloog, I., Schwartz, J., Oken, E., DeMeo, D. L., Gold, D. R., Coull, B. A. 2021; 15 (3): 1366-1385
  • Placental DNA methylation signatures of maternal smoking during pregnancy and potential impacts on fetal growth NATURE COMMUNICATIONS Everson, T. M., Vives-Usano, M., Seyve, E., Cardenas, A., Lacasana, M., Craig, J. M., Lesseur, C., Baker, E. R., Fernandez-Jimenez, N., Heude, B., Perron, P., Gonzalez-Alzaga, B., Halliday, J., Deyssenroth, M. A., Karagas, M. R., Iniguez, C., Bouchard, L., Carmona-Saez, P., Loke, Y. J., Hao, K., Belmonte, T., Charles, M. A., Martorell-Marugan, J., Muggli, E., Chen, J., Fernandez, M. F., Tost, J., Gomez-Martin, A., London, S. J., Sunyer, J., Marsit, C. J., Lepeule, J., Hivert, M., Bustamante, M. 2021; 12 (1): 5095


    Maternal smoking during pregnancy (MSDP) contributes to poor birth outcomes, in part through disrupted placental functions, which may be reflected in the placental epigenome. Here we present a meta-analysis of the associations between MSDP and placental DNA methylation (DNAm) and between DNAm and birth outcomes within the Pregnancy And Childhood Epigenetics (PACE) consortium (N = 1700, 344 with MSDP). We identify 443 CpGs that are associated with MSDP, of which 142 associated with birth outcomes, 40 associated with gene expression, and 13 CpGs are associated with all three. Only two CpGs have consistent associations from a prior meta-analysis of cord blood DNAm, demonstrating substantial tissue-specific responses to MSDP. The placental MSDP-associated CpGs are enriched for environmental response genes, growth-factor signaling, and inflammation, which play important roles in placental function. We demonstrate links between placental DNAm, MSDP and poor birth outcomes, which may better inform the mechanisms through which MSDP impacts placental function and fetal growth.

    View details for DOI 10.1038/s41467-021-24558-y

    View details for Web of Science ID 000688006200005

    View details for PubMedID 34429407

    View details for PubMedCentralID PMC8384884

  • DNA methylation as a mediator of associations between the environment and chronic diseases: A scoping review on application of mediation analysis EPIGENETICS Fujii, R., Sato, S., Tsuboi, Y., Cardenas, A., Suzuki, K. 2022; 17 (7): 759-785


    DNA methylation (DNAm) is one of the most studied epigenetic modifications. DNAm has emerged as a key biological mechanism and biomarkers to test associations between environmental exposure and outcomes in epidemiological studies. Although previous studies have focused on associations between DNAm and either exposure/outcomes, it is useful to test for mediation of the association between exposure and outcome by DNAm. The purpose of this scoping review is to introduce the methodological essence of statistical mediation analysis and to examine emerging epidemiological research applying mediation analyses. We conducted this scoping review for published peer-reviewed journals on this topic using online databases (PubMed, Scopus, Cochrane, and CINAHL) ending in December 2020. We extracted a total of 219 articles by initial screening. After reviewing titles, abstracts, and full texts, a total of 69 articles were eligible for this review. The breakdown of studies assigned to each category was 13 for smoking (18.8%), 8 for dietary intake and famine (11.6%), 6 for other lifestyle factors (8.7%), 8 for clinical endpoints (11.6%), 22 for environmental chemical exposures (31.9%), 2 for socioeconomic status (SES) (2.9%), and 10 for genetic factors and race (14.5%). In this review, we provide an exposure-wide summary for the mediation analysis using DNAm levels. However, we found heterogenous methods and interpretations in mediation analysis with typical issues such as different cell compositions and tissue-specificity. Further accumulation of evidence with diverse exposures, populations and with rigorous methodology will be expected to provide further insight in the role of DNAm in disease susceptibility.

    View details for DOI 10.1080/15592294.2021.1959736

    View details for Web of Science ID 000684435300001

    View details for PubMedID 34384035

    View details for PubMedCentralID PMC9336467

  • Early-pregnancy maternal body mass index is associated with common DNA methylation markers in cord blood and placenta: a paired-tissue epigenome-wide association study EPIGENETICS Ghildayal, N., Fore, R., Lutz, S. M., Cardenas, A., Perron, P., Bouchard, L., Hivert, M. 2022; 17 (7): 808-818


    Women entering pregnancy with elevated body mass index (BMI) face greater risk of adverse outcomes during pregnancy, delivery, and for their offspring later in life, potentially via epigenetics. If epigenetic programming occurs early during in utero development, the differential marks should be detectable in multiple tissues despite the known unique epigenetic profile in each.We used early-pregnancy BMI as reflection of maternal metabolic milieu exposure in peri-conception and early-pregnancy period. We analysed DNA methylation in paired cord blood and placenta samples among 437 newborns from Gen3G, a pre-birth prospective cohort of primarily European descent. We measured DNA methylation in both tissues across the genome in >720,000 CpG sites using the Illumina MethylationEPIC array. At each site, we used linear mixed models (LMMs) with an unstructured variance-covariance matrix to test for an association between maternal early-pregnancy BMI and DNA methylation in both tissues (modelled as M-values). We adjusted for tissue-specific covariates, offspring sex, gestational age at delivery, and maternal smoking and age.Women had a mean (SD) BMI of 25.4 (5.7) kg/m2 measured at first trimester visit (mean=9.9 weeks). Early-pregnancy BMI was associated with differential DNA methylation levels in paired-tissue analyses at two sites: cg10593758 (β=0.0126, SE=0.0025; P=4.07e-7), annotated to CRHBP, and cg0762168 (β=-0.0094, SE=0.0018; P=2.78e-7), annotated to CCDC97.Application of LMMs in DNA methylation data from distinct fetal-origin tissues allowed us to identify CpG sites at which early-pregnancy BMI may have an epigenetic 'programming' effect on overall fetus development. One site (CRHBP) may play a role in hypothalamic-pituitary-adrenal axis regulation.

    View details for DOI 10.1080/15592294.2021.1959975

    View details for Web of Science ID 000684478500001

    View details for PubMedID 34384032

    View details for PubMedCentralID PMC9336499

  • Dietary fat intake during early pregnancy is associated with cord blood DNA methylation at IGF2 and H19 genes in newborns ENVIRONMENTAL AND MOLECULAR MUTAGENESIS Chiu, Y., Fadadu, R. P., Gaskins, A. J., Rifas-Shiman, S. L., Laue, H. E., Moley, K. H., Hivert, M., Baccarelli, A., Oken, E., Chavarro, J. E., Cardenas, A. 2021; 62 (7): 388-398


    Maternal fat intake during pregnancy affects fetal growth, but mechanisms underlying this relationship are unclear. We performed an exploratory study of the associations of fat consumption during pregnancy with cord blood DNA methylation of the insulin-like growth factor 2 (IGF2) and H19 genes. We used data from 96 uncomplicated full-term pregnancies of mothers of whom majority had normal body mass index (BMI) (66%) in Project Viva, a prospective pre-birth cohort. We assessed maternal diet with validated food frequency questionnaires during the first and second trimesters and measured DNA methylation in segments of the IGF2- and H19-differentially methylated regions (DMRs) by pyrosequencing DNA extracted from umbilical cord blood samples. Mean (SD) age was 32.8 (4.1) years and prepregnancy BMI was 24.0 (4.4) kg/m2 . Mean DNA methylation was 56.3% (3.9%) for IGF2-DMR and 44.6% (1.9%) for H19-DMR. Greater first trimester intake of omega-6 polyunsaturated fat (effect per 1% of calories at the expense of carbohydrates) was associated with lower DNA methylation of IGF2-DMR (-1.2%; 95% confidence interval [CI]: -2.2%, -0.2%) and higher DNA methylation at H19-DMR (0.8%; 95% CI: 0.3%, 1.3%). On the other hand, greater first trimester intake of omega-3 polyunsaturated fat was associated with lower DNA methylation of the H19-DMR (-4.3%; 95% CI: -7.9%, -0.8%). We did not find significant associations of IGF2 and H19 methylation with IGF2 cord blood levels. Our findings suggest that early prenatal fat intake (omega-3, omega-6, and saturated fatty acids) may influence DNA methylation at the IGF2 and H19 locus, which could impact fetal development and long-term health.

    View details for DOI 10.1002/em.22452

    View details for Web of Science ID 000679683500001

    View details for PubMedID 34288135

    View details for PubMedCentralID PMC8364885

  • Temporal trends of concentrations of per- and polyfluoroalkyl substances among adults with overweight and obesity in the United States: Results from the Diabetes Prevention Program and NHANES ENVIRONMENT INTERNATIONAL Lin, P. D., Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2021; 157: 106789


    Understanding the temporal trends and change of concentrations of per- and polyfluoroalkyl substances (PFAS) is important to evaluate the health impact of PFAS at both the individual- and population-level, however, limited information is available for pre-diabetic adults in the U.S.Determine trends and rate of change of plasma PFAS concentrations in overweight or obese U.S. adults and evaluate variation by sex, race/ethnicity, and age.We described temporal trends of plasma PFAS concentrations using samples collected in 1996-1998, 1999-2001, and 2011-2012 from 957 pre-diabetic adults enrolled in the Diabetes Prevention Program (DPP) trial and Outcomes Study (DPPOS) and compared to serum concentrations from the National Health and Nutrition Examination Survey (NHANES 1999-2000, 2003-2016, adults with BMI ≥ 24 kg/m2). We examined associations between participants' characteristics and PFAS concentrations and estimated the rate of change using repeated measures in DPP/DPPOS assuming a first-order elimination model.Longitudinal measures of PFAS concentrations in DPP/DPPOS individuals were comparable to NHANES cross-sectional populational means. Plasma concentrations of perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid, perfluorohexanesulfonic acid (PFHxS), N-ethyl-perfluorooctane sulfonamido acetic acid (EtFOSAA), and N-methylperfluorooctane sulfonamido acetic acid (MeFOSAA) started to decline after the year 2000 and concentrations of perfluorononanoic acid (PFNA) increased after 2000 and, for NHANES, decreased after 2012. We consistently observed higher PFOS, PFHxS and PFNA among male, compared to female, and higher PFOS and PFNA among Black, compared to white, participants. The estimated time for concentrations to decrease by half ranged from 3.39 years for EtFOSAA to 17.56 years for PFHxS.We observed a downward temporal trend in plasma PFOS concentrations that was consistent with the timing for U.S. manufacturers' phaseout. Male and Black participants consistently showed higher PFOS and PFNA than female and white participants, likely due to differences in exposure patterns, metabolism or elimination kinetics.

    View details for DOI 10.1016/j.envint.2021.106789

    View details for Web of Science ID 000704786000007

    View details for PubMedID 34333293

    View details for PubMedCentralID PMC8490287

  • Exposure to arsenic at different life-stages and DNA methylation meta-analysis in buccal cells and leukocytes ENVIRONMENTAL HEALTH Bozack, A. K., Boileau, P., Wei, L., Hubbard, A. E., Sille, F. M., Ferreccio, C., Acevedo, J., Hou, L., Ilievski, V., Steinmaus, C. M., Smith, M. T., Navas-Acien, A., Gamble, M., Cardenas, A. 2021; 20 (1): 79


    Arsenic (As) exposure through drinking water is a global public health concern. Epigenetic dysregulation including changes in DNA methylation (DNAm), may be involved in arsenic toxicity. Epigenome-wide association studies (EWAS) of arsenic exposure have been restricted to single populations and comparison across EWAS has been limited by methodological differences. Leveraging data from epidemiological studies conducted in Chile and Bangladesh, we use a harmonized data processing and analysis pipeline and meta-analysis to combine results from four EWAS.DNAm was measured among adults in Chile with and without prenatal and early-life As exposure in PBMCs and buccal cells (N = 40, 850K array) and among men in Bangladesh with high and low As exposure in PBMCs (N = 32, 850K array; N = 48, 450K array). Linear models were used to identify differentially methylated positions (DMPs) and differentially variable positions (DVPs) adjusting for age, smoking, cell type, and sex in the Chile cohort. Probes common across EWAS were meta-analyzed using METAL, and differentially methylated and variable regions (DMRs and DVRs, respectively) were identified using comb-p. KEGG pathway analysis was used to understand biological functions of DMPs and DVPs.In a meta-analysis restricted to PBMCs, we identified one DMP and 23 DVPs associated with arsenic exposure; including buccal cells, we identified 3 DMPs and 19 DVPs (FDR < 0.05). Using meta-analyzed results, we identified 11 DMRs and 11 DVRs in PBMC samples, and 16 DMRs and 19 DVRs in PBMC and buccal cell samples. One region annotated to LRRC27 was identified as a DMR and DVR. Arsenic-associated KEGG pathways included lysosome, autophagy, and mTOR signaling, AMPK signaling, and one carbon pool by folate.Using a two-step process of (1) harmonized data processing and analysis and (2) meta-analysis, we leverage four DNAm datasets from two continents of individuals exposed to high levels of As prenatally and during adulthood to identify DMPs and DVPs associated with arsenic exposure. Our approach suggests that standardizing analytical pipelines can aid in identifying biological meaningful signals.

    View details for DOI 10.1186/s12940-021-00754-7

    View details for Web of Science ID 000671560400001

    View details for PubMedID 34243768

    View details for PubMedCentralID PMC8272372

  • Prenatal exposure to a mixture of elements and neurobehavioral outcomes in mid-childhood: Results from Project Viva ENVIRONMENTAL RESEARCH Fruh, V., Rifas-Shiman, S. L., Coull, B. A., Devick, K. L., Amarasiriwardena, C., Cardenas, A., Bellinger, D. C., Wise, L. A., White, R. F., Wright, R. O., Oken, E., Henn, B. 2021; 201: 111540


    Lead (Pb), manganese (Mn), selenium (Se) and methylmercury (MeHg) can be neurotoxic individually, despite Mn and Se also being essential elements. Little is known about the joint effects of essential and non-essential elements on neurobehavior, particularly for prenatal exposures.To evaluate associations of prenatal exposure to multiple elements with executive function and neurobehavior in children.Participants included 1009 mother-child pairs from the Project Viva pre-birth cohort. We estimated maternal erythrocyte Pb, Mn, Se, and Hg concentrations prenatally. In 6-11-year old children (median 7.6 years), parents and teachers rated children's executive function-related behaviors using the Behavior Rating Inventory of Executive Function (BRIEF) Global Executive Composite score and behavioral difficulties using the Strengths and Difficulties Questionnaire (SDQ) total difficulties score. We evaluated associations of element mixtures with neurobehavior using Bayesian kernel machine regression (BKMR), multivariable linear regression, and quantile g-computation.Median erythrocyte Pb, Mn, Se, and Hg concentrations were 1.1 μg/dL, 33.1 μg/L, 204.5 ng/mL, and 3.1 ng/g, respectively. Findings from BKMR and quantile g-computation models both showed worse (higher) parent-rated BRIEF and SDQ z-scores with higher concentrations of the mixture, although estimates were imprecise. When remaining elements were set at their median within BKMR models, increases in Pb and Se from the 25th to 75th percentile of exposure distributions were associated with 0.08 (95% CI: 0.02, 0.19) and 0.07 (95% CI: 0.03, 0.16) standard deviation increases in parent-rated BRIEF scores, and 0.08 (95% CI: 0.02, 0.17) and 0.05 (95% CI: 0.03, 0.13) standard deviation increases in SDQ scores, respectively. There was no evidence of element interactions.Although associations were small in magnitude, we found a trend of worsening neurobehavioral ratings with increasing prenatal exposure to an element mixture. However, we may be observing a limited range of dose-dependent impacts given the levels of exposure within our population.

    View details for DOI 10.1016/j.envres.2021.111540

    View details for Web of Science ID 000703937500001

    View details for PubMedID 34166661

    View details for PubMedCentralID PMC8502495

  • Early pregnancy exposure to metal mixture and birth outcomes-A prospective study in Project Viva ENVIRONMENT INTERNATIONAL Rahman, M. L., Oken, E., Hivert, M., Rifas-Shiman, S., Lin, P. D., Colicino, E., Wright, R. O., Amarasiriwardena, C., Henn, B., Gold, D. R., Coull, B. A., Cardenas, A. 2021; 156
  • Early pregnancy essential and non-essential metal mixtures and gestational glucose concentrations in the 2nd trimester: Results from project viva ENVIRONMENT INTERNATIONAL Zheng, Y., Lin, P., Williams, P. L., Weisskopf, M. G., Cardenas, A., Rifas-Shiman, S. L., Wright, R. O., Amarasiriwardena, C., Henn, B., Hivert, M., Oken, E., James-Todd, T. 2021; 155: 106690


    Metals are involved in glucose metabolism, and some may alter glycemic regulation. However, joint effects of essential and non-essential metals on glucose concentrations during pregnancy are unclear. This study explored the joint associations of pregnancy exposures to essential (copper, magnesium, manganese, selenium, zinc) and non-essential (arsenic, barium, cadmium, cesium, lead, mercury) metals with gestational glucose concentrations using 1,311 women enrolled 1999-2002 in Project Viva, a Boston, MA-area pregnancy cohort. The study measured erythrocyte metal concentrations from 1st trimester blood samples and used glucose concentrations measured 1 h after non-fasting 50-gram glucose challenge tests (GCT) from clinical gestational diabetes screening at 26-28 weeks gestation. Bayesian Kernel Machine Regression (BKMR) and quantile-based g-computation were applied to model the associations of metal mixtures-including their interactions-with glucose concentrations post-GCT. We tested for reproducibility of BKMR results using generalized additive models. The BKMR model showed an inverse U-shaped association for barium and a linear inverse association for mercury. Specifically, estimated mean glucose concentrations were highest around 75th percentile of barium concentrations [2.1 (95% confidence interval: -0.2, 4.4) mg/dL higher comparing to the 25th percentile], and each interquartile range increase of erythrocyte mercury was associated with 1.9 mg/dL lower mean glucose concentrations (95% credible interval: -4.2, 0.4). Quantile g-computation showed joint associations of all metals, essential-metals, and non-essential metals on gestational glucose concentrations were all null, however, we observed evidences of interaction for barium and lead. Overall, we found early pregnancy barium and mercury erythrocytic concentrations were associated with altered post-load glucose concentrations in later pregnancy, with potential interactions between barium and lead.

    View details for DOI 10.1016/j.envint.2021.106690

    View details for Web of Science ID 000679283700002

    View details for PubMedID 34120006

  • Diet and erythrocyte metal concentrations in early pregnancy-cross-sectional analysis in Project Viva AMERICAN JOURNAL OF CLINICAL NUTRITION Lin, P. D., Cardenas, A., Rifas-Shiman, S. L., Hivert, M., James-Todd, T., Amarasiriwardena, C., Wright, R. O., Rahman, M. L., Oken, E. 2021; 114 (2): 540-549


    Dietary sources of metals are not well established among pregnant women in the United States.We aimed to perform a diet-wide association study (DWAS) of metals during the first trimester of pregnancy.In early pregnancy (11.3 ± 2.8 weeks of gestation), 1196 women from Project Viva (recruited 1999-2002 in eastern Massachusetts) completed a validated FFQ (135 food items) and underwent measurements of erythrocyte metals [arsenic (As), barium, cadmium, cesium (Cs), copper, mercury (Hg), magnesium, manganese, lead (Pb), selenium (Se), zinc]. The DWAS involved a systematic evaluation and visualization of all bivariate relations for each food-metal combination. For dietary items with strong associations with erythrocyte metals, we applied targeted maximum likelihood estimations and substitution models to evaluate how hypothetical dietary interventions would influence metals' concentrations.Participants' mean ± SD age was 32.5 ± 4.5 y and prepregnancy BMI was 24.8 ± 5.4 kg/m2; they were mostly white (75.9%), college graduates (72.4%), married or cohabitating (94.6%), had a household income >$70,000/y (63.5%), and had never smoked (67.1%). Compared with other US-based cohorts, the overall diet quality of participants was above average, and concentrations of erythrocyte metals were lower. The DWAS identified significant associations of several food items with As, Hg, Pb, Cs, and Se; for example, As was higher for each SD increment in fresh fruit (11.5%; 95% CI: 4.9%, 18.4%), white rice (17.9%; 95% CI: 9.4%, 26.9%), and seafood (50.9%; 95% CI: 42.8%, 59.3%). Following the guidelines for pregnant women to consume ≤3 servings/wk of seafood was associated with lower As (-0.55 ng/g; 95% CI: -0.82, -0.28 ng/g) and lower Hg (-2.67 ng/g; 95% CI: -3.55, -1.80 ng/g). Substituting white rice with bread, pasta, tortilla, and potato was also associated with lower As (35%-50%) and Hg (35%-70%).Our DWAS provides a systematic evaluation of diet-metals relations. Prenatal diet may be an important source of exposures to metals.

    View details for DOI 10.1093/ajcn/nqab088

    View details for Web of Science ID 000685072000021

    View details for PubMedID 34038956

    View details for PubMedCentralID PMC8326032

  • Residential PM2.5 exposure and the nasal methylome in children ENVIRONMENT INTERNATIONAL Sordillo, J. E., Cardenas, A., Qi, C., Rifas-Shiman, S. L., Coull, B., Luttmann-Gibson, H., Schwartz, J., Kloog, I., Hivert, M., DeMeo, D. L., Baccarelli, A. A., Xu, C., Gehring, U., Vonk, J. M., Koppelman, G., Oken, E., Gold, D. R. 2021; 153: 106505


    PM2.5-induced adverse effects on respiratory health may be driven by epigenetic modifications in airway cells. The potential impact of exposure duration on epigenetic alterations in the airways is not yet known.We aimed to study associations of fine particulate matter PM2.5 exposure with DNA methylation in nasal cells.We conducted nasal epigenome-wide association analyses within 503 children from Project Viva (mean age 12.9 y), and examined various exposure durations (1-day, 1-week, 1-month, 3-months and 1-year) prior to nasal sampling. We used residential addresses to estimate average daily PM2.5 at 1 km resolution. We collected nasal swabs from the anterior nares and measured DNA methylation (DNAm) using the Illumina MethylationEPIC BeadChip. We tested 719,075 high quality autosomal CpGs using CpG-by-CpG and regional DNAm analyses controlling for multiple comparisons, and adjusted for maternal education, household smokers, child sex, race/ethnicity, BMI z-score, age, season at sample collection and cell-type heterogeneity. We further corrected for bias and genomic inflation. We tested for replication in a cohort from the Netherlands (PIAMA).In adjusted analyses, we found 362 CpGs associated with 1-year PM2.5 (FDR < 0.05), 20 CpGs passing Bonferroni correction (P < 7.0x10-8) and 10 Differentially Methylated Regions (DMRs). In 445 PIAMA participants (mean age 16.3 years) 11 of 203 available CpGs replicated at P < 0.05. We observed differential DNAm at/near genes implicated in cell cycle, immune and inflammatory responses. There were no CpGs or regions associated with PM2.5 levels at 1-day, 1-week, or 1-month prior to sample collection, although 2 CpGs were associated with past 3-month PM2.5.We observed wide-spread DNAm variability associated with average past year PM2.5 exposure but we did not detect associations with shorter-term exposure. Our results suggest that nasal DNAm marks reflect chronic air pollution exposure.

    View details for DOI 10.1016/j.envint.2021.106505

    View details for Web of Science ID 000663330500004

    View details for PubMedID 33872926

    View details for PubMedCentralID PMC8823376

  • Controlled human exposures to diesel exhaust: a human epigenome-wide experiment of target bronchial epithelial cells ENVIRONMENTAL EPIGENETICS Cardenas, A., Fadadu, R. P., Van der Laan, L., Ward-Caviness, C., Granger, L., Diaz-Sanchez, D., Devlin, R. B., Bind, M. 2021; 7 (1): dvab003


    Diesel exhaust (DE) is a major contributor to ambient air pollution around the world. It is a known human carcinogen that targets the respiratory system and increases risk for many diseases, but there is limited research on the effects of DE exposure on the epigenome of human bronchial epithelial cells. Understanding the epigenetic impact of this environmental pollutant can elucidate biological mechanisms involved in the pathogenesis of harmful DE-related health effects. To estimate the causal effect of short-term DE exposure on the bronchial epithelial epigenome, we conducted a controlled single-blinded randomized crossover human experiment of exposure to DE and used bronchoscopy and Illumina 450K arrays for data collection and analysis, respectively. Of the 13 participants, 11 (85%) were male and 2 (15%) were female, and 12 (92%) were White and one (8%) was Hispanic; the mean age was 26 years (SD = 3.8 years). Eighty CpGs were differentially methylated, achieving the minimum possible exact P-value of P = 2.44 × 10-4 (i.e. 2/213). In regional analyses, we found two differentially methylated regions (DMRs) annotated to the chromosome 5 open reading frame 63 genes (C5orf63; 7-CpGs) and unc-45 myosin chaperone A gene (UNC45A; 5-CpGs). Both DMRs showed increased DNA methylation after DE exposure. The average causal effects for the DMRs ranged from 1.5% to 6.0% increases in DNA methylation at individual CpGs. In conclusion, we found that short-term DE alters DNA methylation of genes in target bronchial epithelial cells, demonstrating epigenetic level effects of exposure that could be implicated in pulmonary pathologies.

    View details for DOI 10.1093/eep/dvab003

    View details for Web of Science ID 000674770100001

    View details for PubMedID 33859829

    View details for PubMedCentralID PMC8035831

  • Adopting a "Compound" Exposome Approach in Environmental Aging Biomarker Research: A Call to Action for Advancing Racial Health Equity ENVIRONMENTAL HEALTH PERSPECTIVES Nwanaji-Enwerem, J. C., Jackson, C. L., Ottinger, M., Cardenas, A., James, K. A., Malecki, K. C., Chen, J., Geller, A. M., Mitchell, U. A. 2021; 129 (4): 45001


    In June 2020, the National Academies of Sciences, Engineering, and Medicine hosted a virtual workshop focused on integrating the science of aging and environmental health research. The concurrent COVID-19 pandemic and national attention on racism exposed shortcomings in the environmental research field's conceptualization and methodological use of race, which have subsequently hindered the ability of research to address racial health disparities. By the workshop's conclusion, the authors deduced that the utility of environmental aging biomarkers-aging biomarkers shown to be specifically influenced by environmental exposures-would be greatly diminished if these biomarkers are developed absent of considerations of broader societal factors-like structural racism-that impinge on racial health equity.The authors reached a post-workshop consensus recommendation: To advance racial health equity, a "compound" exposome approach should be widely adopted in environmental aging biomarker research. We present this recommendation here.The authors believe that without explicit considerations of racial health equity, people in most need of the benefits afforded by a better understanding of the relationships between exposures and aging will be the least likely to receive them because biomarkers may not encompass cumulative impacts from their unique social and environmental stressors. Employing an exposome approach that allows for more comprehensive exposure-disease pathway characterization across broad domains, including the social exposome and neighborhood factors, is the first step. Exposome-centered study designs must then be supported with efforts aimed at increasing the recruitment and retention of racially diverse study populations and researchers and further "compounded" with strategies directed at improving the use and interpretation of race throughout the publication and dissemination process. This compound exposome approach maximizes the ability of our science to identify environmental aging biomarkers that explicate racial disparities in health and best positions the environmental research community to contribute to the elimination of racial health disparities.

    View details for DOI 10.1289/EHP8392

    View details for Web of Science ID 000662049200008

    View details for PubMedID 33822649

    View details for PubMedCentralID PMC8043128

  • DNA methylation architecture of the ACE2 gene in nasal cells of children SCIENTIFIC REPORTS Cardenas, A., Rifas-Shiman, S. L., Sordillo, J. E., DeMeo, D. L., Baccarelli, A. A., Hivert, M., Gold, D. R., Oken, E. 2021; 11 (1): 7107


    Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has led to the global coronavirus disease 2019 (COVID-19) pandemic. SARS-CoV-2 enters cells via angiotensin-Converting Enzyme 2 (ACE2) receptors, highly expressed in nasal epithelium with parallel high infectivity.1,2 The nasal epigenome is in direct contact with the environment and could explain COVID-19 disparities by reflecting social and environmental influences on ACE2 regulation. We collected nasal swabs from anterior nares of 547 children, measured DNA methylation (DNAm), and tested differences at 15 ACE2 CpGs by sex, age, race/ethnicity and epigenetic age. ACE2 CpGs were differentially methylated by sex with 12 sites having lower DNAm (mean = 12.71%) and 3 sites greater DNAm (mean = 1.45%) among females relative to males. We observed differential DNAm at 5 CpGs for Hispanic females (mean absolute difference = 3.22%) and lower DNAm at 8 CpGs for Black males (mean absolute difference = 1.33%), relative to white participants. Longer DNAm telomere length was associated with greater ACE2 DNAm at 11 and 13 CpGs among males (mean absolute difference = 7.86%) and females (mean absolute difference = 8.21%), respectively. Nasal ACE2 DNAm differences could contribute to our understanding COVID-19 severity and disparities reflecting upstream environmental and social influences. Findings need to be confirmed among adults and patients with risk factors for COVID-19 severity.

    View details for DOI 10.1038/s41598-021-86494-7

    View details for Web of Science ID 000636358400050

    View details for PubMedID 33782449

    View details for PubMedCentralID PMC8007733

  • Exposure to violence, chronic stress, nasal DNA methylation, and atopic asthma in children PEDIATRIC PULMONOLOGY Yan, Q., Forno, E., Cardenas, A., Qi, C., Han, Y., Acosta-Perez, E., Kim, S., Zhang, R., Boutaoui, N., Canino, G., Vonk, J. M., Xu, C., Chen, W., Marsland, A., Oken, E., Gold, D. R., Koppelman, G. H., Celedon, J. C. 2021; 56 (7): 1896-1905


    Exposure to violence (ETV) or chronic stress may influence asthma through unclear mechanisms.Epigenome-wide association study (EWAS) of ETV or chronic stress measures and DNA methylation in nasal epithelium from 487 Puerto Ricans aged 9-20 years who participated in the Epigenetic Variation and Childhood Asthma in Puerto Ricans study [EVA-PR]). We assessed four measures of ETV and chronic stress in children (ETV scale, gun violence, and perceived stress) and their mothers (perceived stress). Each EWAS was conducted using linear regression, with CpGs as dependent variables and the stress/violence measure as a predictor, adjusting for age, sex, the top five principal components, and SVA latent factors. We then selected the top 100 CpGs (by p value) associated with each stress/violence measure in EVA-PR and conducted a meta-analysis of the selected CpGs and atopic asthma using data from EVA-PR and two additional cohorts (Project Viva and PIAMA).Three CpGs (in SNN, PTPRN2, and LINC01164) were associated with maternal perceived stress or gun violence (p = 1.28-3.36 × 10-7 ), but not with atopic asthma, in EVA-PR. In a meta-analysis of three cohorts, which included the top CpGs associated with stress/violence measures in EVA-PR, 12 CpGs (in STARD3NL, SLC35F4, TSR3, CDC42SE2, KLHL25, PLCB1, BUD13, OR2B3, GALR1, TMEM196, TEAD4, and ANAPC13) were associated with atopic asthma at FDR-p < .05.Pending confirmation in longitudinal studies, our findings suggest that nasal epithelial methylation markers associated with measures of ETV and chronic stress may be linked to atopic asthma in children and adolescents.

    View details for DOI 10.1002/ppul.25372

    View details for Web of Science ID 000631206100001

    View details for PubMedID 33751861

    View details for PubMedCentralID PMC8217314

  • Comparative epigenome-wide analysis highlights placenta-specific differentially methylated regions EPIGENOMICS Groleau, M., White, F., Cardenas, A., Perron, P., Hivert, M., Bouchard, L., Jacques, P. 2021; 13 (05): 357-368


    Aim: The placenta undergoes DNA methylation (DNAm) programming that is unique compared with all other fetal tissues. We aim to decipher some of the physiologic roles of the placenta by comparing its DNAm profile with that of another fetal tissue. Materials & methods: We performed a comparative analysis of genome-wide DNAm of 444 placentas paired with cord blood samples collected at birth. Gene ontology term analyses were conducted on the resulting differentially methylated regions. Results: Genomic regions upstream of transcription start sites showing lower DNAm in the placenta were enriched with terms related to miRNA functions and genes encoding G-protein-coupled receptors. Conclusion: These results highlight genomic regions that are differentially methylated in the placenta in contrast to fetal blood.

    View details for DOI 10.2217/epi-2020-0271

    View details for Web of Science ID 000625227100001

    View details for PubMedID 33661023

  • Detecting differentially methylated regions with multiple distinct associations EPIGENOMICS Lent, S., Cardenas, A., Rifas-Shiman, S. L., Perron, P., Bouchard, L., Liu, C., Hivert, M., Dupuis, J. 2021; 13 (06): 451-464


    Aim: We evaluated five methods for detecting differentially methylated regions (DMRs): DMRcate, comb-p, seqlm, GlobalP and dmrff. Materials & methods: We used a simulation study and real data analysis to evaluate performance. Additionally, we evaluated the use of an ancestry-matched reference cohort to estimate correlations between CpG sites in cord blood. Results: Several methods had inflated Type I error, which increased at more stringent significant levels. In power simulations with 1-2 causal CpG sites with the same direction of effect, dmrff was consistently among the most powerful methods. Conclusion: This study illustrates the need for more thorough simulation studies when evaluating novel methods. More work must be done to develop methods with well-controlled Type I error that do not require individual-level data.

    View details for DOI 10.2217/epi-2020-0344

    View details for Web of Science ID 000623328200001

    View details for PubMedID 33641349

    View details for PubMedCentralID PMC8023344

  • Per- and polyfluoroalkyl substances and calcifications of the coronary and aortic arteries in adults with prediabetes: Results from the diabetes prevention program outcomes study ENVIRONMENT INTERNATIONAL Osorio-Yanez, C., Sanchez-Guerra, M., Cardenas, A., Lin, P. D., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Fleisch, A. F., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2021; 151: 106446


    Per- and polyfluoroalkyl substances (PFAS) are endocrine disrupting chemicals that have been associated with cardiovascular risk factors including elevated body weight and hypercholesterolemia. Therefore, PFAS may contribute to the development of atherosclerosis and cardiovascular disease (CVD). However, no previous study has evaluated associations between PFAS exposure and arterial calcification.This study used data from 666 prediabetic adults enrolled in the Diabetes Prevention Program trial who had six PFAS quantified in plasma at baseline and two years after randomization, as well as measurements of coronary artery calcium (CAC) and ascending (AsAC) and descending (DAC) thoracic aortic calcification 13-14 years after baseline. We performed multinomial regression to test associations between PFAS and CAC categorized according to Agatston score [low (<10), moderate (11-400) and severe (>400)]. We used logistic regression to assess associations between PFAS and presence of AsAC and DAC. We adjusted models for baseline sex, age, BMI, race/ethnicity, cigarette smoking, education, treatment assignment (placebo or lifestyle intervention), and statin use. PFAS concentrations were similar to national means; 53.9% of participants had CAC > 11, 7.7% had AsAC, and 42.6% had DAC. Each doubling of the mean sum of plasma concentrations of linear and branched isomers of perfluorooctane sulfonic acid (PFOS) was associated with 1.49-fold greater odds (95% CI: 1.01, 2.21) of severe versus low CAC. This association was driven mainly by the linear (n-PFOS) isomer [1.54 (95% CI: 1.05, 2.25) greater odds of severe versus low CAC]. Each doubling of mean plasma N-ethyl-perfluorooctane sulfonamido acetic acid concentration was associated with greater odds of CAC in a dose-dependent manner [OR = 1.26 (95% CI:1.08, 1.47) for moderate CAC and OR = 1.37 (95% CI:1.07, 1.74) for severe CAC, compared to low CAC)]. Mean plasma PFOS and n-PFOS were also associated with greater odds of AsAC [OR = 1.67 (95% CI:1.10, 2.54) and OR = 1.70 (95% CI:1.13, 2.56), respectively], but not DAC. Other PFAS were not associated with outcomes.Prediabetic adults with higher plasma concentrations of select PFAS had higher risk of coronary and thoracic aorta calcification. PFAS exposure may be a risk factor for adverse cardiovascular health among high-risk populations.

    View details for DOI 10.1016/j.envint.2021.106446

    View details for Web of Science ID 000632313500001

    View details for PubMedID 33631604

    View details for PubMedCentralID PMC8721596

  • Epigenome-wide association study of maternal hemoglobin A1c in pregnancy and cord blood DNA methylation EPIGENOMICS Juvinao-Quintero, D. L., Starling, A. P., Cardenas, A., Powe, C. E., Perron, P., Bouchard, L., Dabelea, D., Hivert, M. 2021; 13 (03): 203-218


    Background: Gestational hyperglycemia is associated with adverse perinatal outcomes and long-term offspring metabolic programming, likely through dysregulation of DNA methylation (DNAm). Materials & methods: We tested associations between maternal HbA1c and cord blood DNAm among 412 mother-child pairs in the genetics of glucose regulation in gestation and growth (Gen3G) and implemented Mendelian randomization to infer causality. We sought replication in an independent sample from Healthy Start. Results: Higher second trimester HbA1c levels were associated with lower DNAm at cg21645848 (p = 3.9 × 10-11) near URGCP. Mendelian randomization and replication analyses showed same direction of effect between HbA1c and DNAm at cg21645848, but did not reach statistical significance. Conclusion: We found that higher maternal glycemia reflected by HbA1c is associated with cord blood DNAm at URGCP, a gene related with inflammatory pathways.

    View details for DOI 10.2217/epi-2020-0279

    View details for Web of Science ID 000618389700004

    View details for PubMedID 33406918

    View details for PubMedCentralID PMC7907967

  • Blood DNA methylation biomarkers of cumulative lead exposure in adults JOURNAL OF EXPOSURE SCIENCE AND ENVIRONMENTAL EPIDEMIOLOGY Colicino, E., Just, A., Kioumourtzoglou, M., Vokonas, P., Cardenas, A., Sparrow, D., Weisskopf, M., Nie, L. H., Hu, H., Schwartz, J. D., Wright, R. O., Baccarelli, A. A. 2021; 31 (1): 108-116


    Lead is a ubiquitous toxicant following three compartment kinetics with the longest half-life found in bones. Patella and tibia lead levels-validated measures of cumulative exposure-require specialized X-ray-fluorescence-spectroscopy available only in a few centers worldwide. We developed minimally invasive biomarkers reflecting individual cumulative lead exposure using blood DNA methylation profiles-obtainable via Illumina 450K or IlluminaEPIC bead-chip assays.We developed and tested two methylation-based biomarkers from 348 Normative Aging Study (NAS) elderly men. We selected methylation sites with strong associations with bone lead levels via robust regressions analysis and constructed the biomarkers using elastic nets. Results were validated in a NAS subset, reporting specificity, and sensitivity.Participants were 73 years old on average (standard deviation, SD = 6), with moderate lead levels of (mean ± SD patella: 27 ± 18 µg/g; tibia:21 ± 13 µg/g). Methylation-based biomarkers for lead in patella and tibia included 59 and 138 DNA methylation sites, respectively. Estimated lead levels were significantly correlated with actual measured values, (r = 0.62 patella, r = 0.59 tibia) and had low mean square error (MSE) (MSE = 0.68 patella, MSE = 0.53 tibia). Means and distributions of the estimated and actual lead levels were not significantly different across patella and tibia bones (p > 0.05). Methylation-based biomarkers discriminated participants highly exposed (>median) to lead with a specificity of 74 and 73% for patella and tibia lead levels, respectively, with 70% sensitivity.DNA methylation-based lead biomarkers are novel tools that can be used to reconstruct decades' worth of individual cumulative lead exposure using only blood DNA methylation profiles and may help identify the consequences of cumulative exposure.

    View details for DOI 10.1038/s41370-019-0183-9

    View details for Web of Science ID 000607303900011

    View details for PubMedID 31636367

    View details for PubMedCentralID PMC7170756

  • Per- and polyfluoroalkyl substances and kidney function: Follow-up results from the Diabetes Prevention Program trial ENVIRONMENT INTERNATIONAL Lin, P. D., Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2021; 148: 106375


    Per- and polyfluoroalkyl substances (PFAS) are ubiquitously detected in populations worldwide and may hinder kidney function. The objective of the study was to determine longitudinal associations of plasma PFAS concentrations with estimated glomerular filtration rate (eGFR) and evaluate whether a lifestyle intervention modify the associations. We studied 875 participants initially randomized to the lifestyle or placebo arms in the Diabetes Prevention Program (DPP, 1996-2002) trial and Outcomes Study (DPPOS, 2002-2014). We ran generalized linear mixed models accounting a priori covariates to evaluate the associations between baseline PFAS concentrations and repeated measures of eGFR, separately, for six PFAS (PFOS, PFOA, PFHxS, EtFOSAA, MeFOSAA, PFNA); then used quantile-based g-computation to evaluate the effects of the six PFAS chemicals as a mixture. The cohort was 64.9% female; 73.4% 40-64 years-old; 29.4% with hypertension; 50.5% randomized to lifestyle intervention and 49.5% to placebo and had similar plasma PFAS concentrations as the general U.S. population in 1999-2000. Most participants had normal kidney function (eGFR > 90 mL/min/1.73 m2) over the approximately 14 years of follow-up. We found that plasma PFAS concentrations during DPP were inversely associated with eGFR during DPPOS follow-up. Each quartile increase in baseline plasma concentration of the 6 PFAS as a mixture was associated with 2.26 mL/min/1.73 m2 lower eGFR (95% CI: -4.12, -0.39) at DPPOS Year 5, approximately 9 years since DPP randomization and PFAS measurements. The lifestyle intervention did not modify associations, but inverse associations were stronger among participants with hypertension at baseline. Among prediabetic adults, we found inverse associations between baseline plasma PFAS concentrations and measures of eGFR throughout 14 years of follow-up. The lifestyle intervention of diet, exercise and behavioral changes did not modify the associations, but persons with hypertension may have heightened susceptibility.

    View details for DOI 10.1016/j.envint.2020.106375

    View details for Web of Science ID 000743579500007

    View details for PubMedID 33482440

    View details for PubMedCentralID PMC7929640

  • Added sugar intake during pregnancy: Fetal behavior, birth outcomes, and placental DNA methylation DEVELOPMENTAL PSYCHOBIOLOGY Trumpff, C., Sturm, G., Picard, M., Foss, S., Lee, S., Feng, T., Cardenas, A., McCormack, C., Champagne, F. A., Monk, C. 2021; 63 (5): 878-889


    Pregnancy is a critical time for the effects of environmental factors on children's development. The effect of added sugar intake on fetal development and pregnancy outcomes remains understudied despite increasing dietary intake in the United States. This study investigated the effect of added sugar on fetal programming by examining the association between maternal added sugar consumption, fetal movement, birth outcomes, and placental DNA methylation. Further, primary human fibroblasts were cultured under normal or high glucose conditions to assess the effect of high glucose exposure on cells' DNA methylation. We found that higher added sugar intake across pregnancy was associated with reduced 3rd-trimester fetal movement (p < .05) and shorter gestation (p < .01). Our sample size was not powered to detect the alteration of individual placental CpG with genome-wide significance. However, a secondary analysis suggested that added sugar consumption was associated with differential methylation of functionally related gene families across pregnancy. Consistent with this, high glucose exposure in primary cultured human fibroblasts altered the methylation of 17% of all CpGs, providing converging evidence for an effect of sugar on DNA methylation. Our results suggest that diets high in added sugar during pregnancy may have implications for offspring health via prenatal programming effects measurable before birth.

    View details for DOI 10.1002/dev.22088

    View details for Web of Science ID 000605560300001

    View details for PubMedID 33415750

  • Maternal anxiety during pregnancy and newborn epigenome-wide DNA methylation MOLECULAR PSYCHIATRY Sammallahti, S., Cortes Hidalgo, A. P., Tuominen, S., Malmberg, A., Mulder, R. H., Brunst, K. J., Alemany, S., McBride, N. S., Yousefi, P., Heiss, J. A., McRae, N., Page, C. M., Jin, J., Pesce, G., Caramaschi, D., Rifas-Shiman, S. L., Koen, N., Adams, C. D., Magnus, M. C., Baiz, N., Ratanatharathorn, A., Czamara, D., Haberg, S. E., Colicino, E., Baccarelli, A. A., Cardenas, A., DeMeo, D. L., Lawlor, D. A., Relton, C. L., Felix, J. F., van IJzendoorn, M. H., Bakermans-Kranenburg, M. J., Kajantie, E., Raikkonen, K., Sunyer, J., Sharp, G. C., Houtepen, L. C., Nohr, E. A., Sorensen, T. A., Tellez-Rojo, M. M., Wright, R. O., Annesi-Maesano, I., Wright, J., Hivert, M., Wright, R. J., Zar, H. J., Stein, D. J., London, S. J., Cecil, C. M., Tiemeier, H., Lahti, J. 2021; 26 (6): 1832-1845


    Maternal anxiety during pregnancy is associated with adverse foetal, neonatal, and child outcomes, but biological mechanisms remain unclear. Altered foetal DNA methylation (DNAm) has been proposed as a potential underlying mechanism. In the current study, we performed a meta-analysis to examine the associations between maternal anxiety, measured prospectively during pregnancy, and genome-wide DNAm from umbilical cord blood. Sixteen non-overlapping cohorts from 12 independent longitudinal studies of the Pregnancy And Childhood Epigenetics Consortium participated, resulting in a combined dataset of 7243 mother-child dyads. We examined prenatal anxiety in relation to genome-wide DNAm and differentially methylated regions. We observed no association between the general symptoms of anxiety during pregnancy or pregnancy-related anxiety, and DNAm at any of the CpG sites, after multiple-testing correction. Furthermore, we identify no differentially methylated regions associated with maternal anxiety. At the cohort-level, of the 21 associations observed in individual cohorts, none replicated consistently in the other cohorts. In conclusion, contrary to some previous studies proposing cord blood DNAm as a promising potential mechanism explaining the link between maternal anxiety during pregnancy and adverse outcomes in offspring, we found no consistent evidence for any robust associations between maternal anxiety and DNAm in cord blood. Larger studies and analysis of DNAm in other tissues may be needed to establish subtle or subgroup-specific associations between maternal anxiety and the foetal epigenome.

    View details for DOI 10.1038/s41380-020-00976-0

    View details for Web of Science ID 000605874700002

    View details for PubMedID 33414500

    View details for PubMedCentralID PMC8595870

  • Associations of DNA Methylation Mortality Risk Markers with Congenital Microcephaly from Zika Virus: A Study of Brazilian Children Less than 4 Years of Age. Journal of tropical pediatrics Nwanaji-Enwerem, J. C., Laan, L. V., Avakame, E. F., Scott, K. A., Burris, H. H., Cardenas, A. 2021; 67 (1)


    BACKGROUND: Zika virus (ZIKV)-associated congenital microcephaly is an important contributor to pediatric death, and more robust pediatric mortality risk metrics are needed to help guide life plans and clinical decision making for these patients. Although common etiologies of pediatric and adult mortality differ, early life health can impact adult outcomes-potentially through DNA methylation. Hence, in this pilot study, we take an early step in identifying pediatric mortality risk metrics by examining associations of ZIKV infection and associated congenital microcephaly with existing adult DNA methylation-based mortality biomarkers: GrimAge and Zhang's mortality score (ZMS).METHODS: Mortality measures were calculated from previously published HumanMethylationEPIC BeadChip data from 44 Brazilian children aged 5-40months (18 with ZIKV-associated microcephaly; 7 normocephalic, exposed to ZIKV in utero; and 19 unexposed controls). We used linear models adjusted for chronological age, sex, methylation batch and white blood cell proportions to evaluate ZIKV and mortality marker relationships.RESULTS: We observed significant decreases in GrimAge-component plasminogen activator inhibitor-1 [PAI-1; beta=-2453.06pg/ml, 95% confidence interval (CI) -3652.96, -1253.16, p=0.0002], and ZMS-site cg14975410 methylation (beta=-0.06, 95% CI -0.09, -0.03, p=0.0003) among children with microcephaly compared to controls. PAI-1 (beta=-2448.70pg/ml, 95% CI -4384.45, -512.95, p=0.01) and cg14975410 (beta=0.01, 95% CI -0.04, 0.06, p=0.64) results in comparisons of normocephalic, ZIKV-exposed children to controls were not statistically significant.CONCLUSION: Our results suggest that elements of previously-identified adult epigenetic markers of mortality risk are associated with ZIKV-associated microcephaly, a known contributor to pediatric mortality risk. These findings may provide insights for efforts aimed at developing pediatric mortality markers.

    View details for DOI 10.1093/tropej/fmab020

    View details for PubMedID 33822234

  • Prenatal depression and offspring DNA methylation The Neuroscience of Depression Faleschini, S., Cardenas, A. 2021: 537-545
  • Long-term Aspirin Use and Epigenetic Mitotic Clocks for Cancer Risk Prediction: Findings in Healthy Colon Mucosa and Recommendations for Future Epigenetic Aging Studies. Epigenetics communications Nwanaji-Enwerem, J. C., Nze, C., Cardenas, A. 2021; 1 (1)


    Background: Despite the known role of mitosis in colorectal cancer, previous associations of long-term aspirin use with suppressed cancer-related epigenetic aging did not involve epigenetic mitotic clocks. We investigated these relationships using three epigenetic mitotic clocks developed for cancer risk prediction: EpiTOC, EpiTOC2, and MiAge. We utilized publicly available HumanMethylationEPIC BeadChip data from 112 healthy colon (proximal and distal) mucosal samples taken at baseline (T1) and at 10-years follow-up (T2) from a screening cohort of 28 Polish women (11 non-users and 17 long-term [≥ 2 years] aspirin users). Mitotic clock values were divided by chronological age at each timepoint to obtain intrinsic rates (IRs). We evaluated differences in residuals of the mitotic clock IRs taken from linear mixed effects models adjusted for BMI, polyp status, and DNA methylation batch.Findings: EpiTOC, EpiTOC2 and MiAge were significantly correlated with chronological age (P < 0.05) with correlations ranging from 0.41 to 0.63. The EpiTOC, EpiTOC2 and MiAge clocks were strongly correlated with each other in proximal and distal samples (r > 0.79, P < 0.0001). We observed proximal within group median clock IR deceleration for EpiTOC (-0.0004 DNAm, P = 0.008), EpiTOC2 (-16 cell divisions, P = 0.009), and MiAge (-3 cell divisions, P = 0.002) for long-term aspirin users from T1 to T2 but not for non-users. In distal samples, only the long-term user MiAge IR was significantly deaccelerated (-3 cell divisions, P = 0.009).Conclusions: Our observed findings support previously reported longitudinal associations of aspirin use with deceleration of other epigenetic age measures in the proximal colon. Our mitotic clock results suggest that cell proliferation could play a role in some aspirin relationships with epigenetic aging. Furthermore, the findings provide added impetus for establishing gold standards for epigenetic aging and consensus guidelines for more comprehensive reporting in future epigenetic aging cancer studies.

    View details for DOI 10.1186/s43682-021-00004-4

    View details for PubMedID 35493198

  • A Longitudinal Epigenetic Aging and Leukocyte Analysis of Simulated Space Travel: The Mars-500 Mission CELL REPORTS Nwanaji-Enwerem, J. C., Nwanaji-Enwerem, U., Van der Laan, L., Galazka, J. M., Redeker, N. S., Cardenas, A. 2020; 33 (10): 108406


    Astronauts undertaking long-duration space missions may be vulnerable to unique stressors that can impact human aging. Nevertheless, few studies have examined the relationship of mission duration with DNA-methylation-based biomarkers of aging in astronauts. Using data from the six participants of the Mars-500 mission, a high-fidelity 520-day ground simulation experiment, we tested relationships of mission duration with five longitudinally measured blood DNA-methylation-based metrics: DNAmGrimAge, DNAmPhenoAge, DNA-methylation-based estimator of telomere length (DNAmTL), mitotic divisions (epigenetic mitotic clock [epiTOC2]), and pace of aging (PoA). We provide evidence that, relative to baseline, mission duration was associated with significant decreases in epigenetic aging. However, only decreases in DNAmPhenoAge remained significant 7 days post-mission. We also observed significant changes in estimated proportions of plasmablasts, CD4T, CD8 naive, and natural killer (NK) cells. Only decreases in NK cells remained significant post-mission. If confirmed more broadly, these findings contribute insights to improve the understanding of the biological aging implications for individuals experiencing long-duration space travel.

    View details for DOI 10.1016/j.celrep.2020.108406

    View details for Web of Science ID 000596872600001

    View details for PubMedID 33242403

    View details for PubMedCentralID PMC7786521

  • Exposure to violence, chronic stress, nasal DNA methylation, and atopic asthma in children. medRxiv : the preprint server for health sciences Yan, Q., Forno, E., Cardenas, A., Qi, C., Han, Y., Acosta-Perez, E., Kim, S., Zhang, R., Boutaoui, N., Canino, G., Vonk, J. M., Xu, C., Chen, W., Oken, E., Gold, D. R., Koppelman, G. H., Celedon, J. C. 2020


    BACKGROUND: Exposure to violence (ETV) or stress may cause asthma through unclear mechanisms.METHODS: Epigenome-wide association study (EWAS) of DNA methylation in nasal epithelium and four ETV or chronic stress measures in 487 Puerto Ricans aged 9-20 years who participated in the Epigenetic Variation and Childhood Asthma in Puerto Ricans study [EVA-PR]). We assessed measures of ETV or chronic stress in children (ETV scale, gun violence, and perceived stress) and their mothers (perceived stress). Each EWAS was conducted using linear regression, with CpGs as dependent variables and the stress/violence measure as a predictor, adjusting for age, sex, the top five principal components, and SVA latent factors. We then selected the top 100 CpGs (by P-value) associated with each stress/violence measure in EVA-PR and conducted a meta-analysis of the selected CpGs and atopic asthma using data from EVA-PR and two additional cohorts (Project Viva and PIAMA).RESULTS: In the EWAS of stress/violence in EVA-PR, gun violence was associated with methylation of cg18961589 in LINC01164 (beta=0.03, P =1.28*10 -7 ), and maternal stress was associated with methylation of cg03402351 in SNN (beta=0.04, P =1.69*10 -7 ) and cg19064846 in PTPRN2 (beta=0.03, P =3.36*10 -7 ). In a meta-analysis of three cohorts, which included the top CpGs associated with stress/violence in EVA-PR, CpGs in STARD3NL, SLC35F4, TSR3, CDC42SE2, KLHL25, PLCB1, BUD13, OR2B3, GALR1, TMEM196, TEAD4 and ANAPC13 were associated with atopic asthma at FDR- P < 0.05.CONCLUSIONS: ETV and chronic stress may increase the risk of atopic asthma through DNA methylation in airway epithelium, though this needs confirmation in future longitudinal studies.

    View details for DOI 10.1101/2020.11.03.20225250

    View details for PubMedID 33173928

  • Placental Epigenome-Wide Association Study Identified Loci Associated with Childhood Adiposity at 3 Years of Age INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Gagne-Ouellet, V., Breton, E., Thibeault, K., Fortin, C., Desgagne, V., Tremblay, E., Cardenas, A., Guerin, R., Perron, P., Hivert, M., Bouchard, L. 2020; 21 (19)


    The aim of this study was to identify placental DNA methylation (DNAm) variations associated with adiposity at 3 years of age. We quantified placental DNAm using the Infinium MethylationEPIC BeadChips. We assessed associations between DNAm at single-CpGs and skinfold thickness using robust linear regression models adjusted for gestational age, child's sex, age at follow-up and cellular heterogeneity. We sought replication of DNAm association with child adiposity in an independent cohort. We quantified placental mRNA levels for annotated gene using qRT-PCR and tested for correlation with DNAm. Lower DNAm at cg22593959 and cg22436429 was associated with higher adiposity (β = -1.18, q = 0.002 and β = -0.82, q = 0.04). The cg22593959 is located in an intergenic region (chr7q31.3), whereas cg22436429 is within the TFAP2E gene (1p34.3). DNAm at cg22593959 and cg22436429 was correlated with mRNA levels at FAM3C (rs = -0.279, p = 0.005) and TFAP2E (rs = 0.216, p = 0.03). In an independent cohort, the association between placental DNAm at cg22593959 and childhood adiposity was of similar strength and direction (β = -3.8 ± 4.1, p = 0.36), yet non-significant. Four genomic regions were also associated with skinfold thickness within FMN1, MAGI2, SKAP2 and BMPR1B genes. We identified placental epigenetic variations associated with adiposity at 3 years of age suggesting that childhood fat accretion patterns might be established during fetal life.

    View details for DOI 10.3390/ijms21197201

    View details for Web of Science ID 000586432300001

    View details for PubMedID 33003475

    View details for PubMedCentralID PMC7582906

  • Heterogeneous ozone effects on the DNA methylome of bronchial cells observed in a crossover study SCIENTIFIC REPORTS Bind, M. C., Rubin, D. B., Cardenas, A., Dhingra, R., Ward-Caviness, C., Liu, Z., Mirowsky, J., Schwartz, J. D., Diaz-Sanchez, D., Devlin, R. B. 2020; 10 (1): 15739


    We used a randomized crossover experiment to estimate the effects of ozone (vs. clean air) exposure on genome-wide DNA methylation of target bronchial epithelial cells, using 17 volunteers, each randomly exposed on two separated occasions to clean air or 0.3-ppm ozone for two hours. Twenty-four hours after exposure, participants underwent bronchoscopy to collect epithelial cells whose DNA methylation was measured using the Illumina 450 K platform. We performed global and regional tests examining the ozone versus clean air effect on the DNA methylome and calculated Fisher-exact p-values for a series of univariate tests. We found little evidence of an overall effect of ozone on the DNA methylome but some suggestive changes in PLSCR1, HCAR1, and LINC00336 DNA methylation after ozone exposure relative to clean air. We observed some participant-to-participant heterogeneity in ozone responses.

    View details for DOI 10.1038/s41598-020-72068-6

    View details for Web of Science ID 000577248100014

    View details for PubMedID 32978449

    View details for PubMedCentralID PMC7519112

  • DNA Methylation Architecture of the ACE2 gene in Nasal Cells. medRxiv : the preprint server for health sciences Cardenas, A., Rifas-Shiman, S. L., Sordillo, J. E., DeMeo, D. L., Baccarelli, A. A., Hivert, M., Gold, D. R., Oken, E. 2020


    Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has led to the global coronavirus disease 2019 (COVID-19) pandemic. SARS-CoV-2 enters cells via angiotensin-Converting Enzyme 2 (ACE2) receptors, highly expressed in nasal epithelium with parallel high infectivity.1,2 The nasal epigenome is in direct contact with the environment and could explain COVID-19 disparities by reflecting social and environmental influences on ACE2 regulation. We collected nasal swabs from anterior nares of 547 children, measured DNA methylation (DNAm), and tested differences at 15 ACE2 CpGs by sex, age, race/ethnicity and epigenetic age. ACE2 CpGs were differentially methylated by sex with 12 sites having lower DNAm (mean=12.71%) and 3 sites greater DNAm (mean=1.45%) among females relative to males. We observed differential DNAm at 5 CpGs for Hispanic females (mean absolute difference=3.22%) and lower DNAm at 8 CpGs for Black males (mean absolute difference=1.33%), relative to white participants. Longer DNAm telomere length was associated with greater ACE2 DNAm at 11 and 13 CpGs among males (mean absolute difference=7.86%) and females (mean absolute difference=8.21%), respectively. Nasal ACE2 DNAm differences could contribute to our understanding COVID-19 severity and disparities reflecting upstream environmental and social influences.

    View details for DOI 10.1101/2020.08.25.20182105

    View details for PubMedID 32995808

  • Prospective Association Between Manganese in Early Pregnancy and the Risk of Preeclampsia EPIDEMIOLOGY Liu, T., Hivert, M., Rifas-Shiman, S. L., Rahman, M. L., Oken, E., Cardenas, A., Mueller, N. T. 2020; 31 (5): 677-680


    Manganese, an essential micronutrient, has been found in lower concentrations among women with preeclampsia in cross-sectional and case-control studies without establishment of a temporal relationship.We evaluated the prospective association of manganese (in red blood cells) in first trimester of pregnancy with incidence of preeclampsia (ascertained by reviewing medical records) among 1,312 women in eastern Massachusetts (Project Viva, 1999-2002). We used log-binomial regression to examine the manganese-preeclampsia relationship, adjusting for maternal age, race/ethnicity, parity, prepregnancy body mass index, blood pressure, and hematocrit.The median (25th, 75th percentile) manganese concentrationin red blood cells was 16.2 ng/g (13.1, 20.4) and 48 (4%) women developed preeclampsia. We observed an inverse dose-response relationship between manganese and preeclampsia. Compared with women in the lowest tertile, women in the middle manganese tertile had 0.81 times the risk of preeclampsia (95% CI: 0.43, 1.5) and those in the highest tertile had 0.50 (95% CI: 0.25, 0.99) times the risk.Our results provide insight into a potentially modifiable way to prevent preeclampsia.

    View details for DOI 10.1097/EDE.0000000000001227

    View details for Web of Science ID 000559080200010

    View details for PubMedID 32618710

    View details for PubMedCentralID PMC7398820

  • Serum dioxin levels and sperm DNA methylation age: Findings in Vietnam war veterans exposed to Agent Orange REPRODUCTIVE TOXICOLOGY Nwanaji-Enwerem, J. C., Jenkins, T. G., Colicino, E., Cardenas, A., Baccarelli, A. A., Boyer, E. W. 2020; 96: 27-35


    Exposure to dioxin, a known endocrine disruptor and carcinogen, is associated with poor reproductive outcomes. Yet, few studies have explored the role of DNA methylation in these relationships. Utilizing a publicly available dataset from 37 male Air Force Health Study participants exposed to dioxin-contaminated Agent Orange during the Vietnam war, we cross-sectionally examined the relationship of serum dioxin levels with a novel DNA methylation-based measure of sperm age (DNAm-agesperm). DNAm-agesperm was calculated using CpG sites on the Illumina HumanMethylation450 BeadChip. We estimated associations of dioxin levels with DNAm-agesperm using linear regression models adjusted for chronological age, body mass index, and smoking status. Chronological age was highly correlated with DNAmagesperm (r = 0.80). In fully-adjusted linear models, a one percent increase in serum dioxin levels was significantly associated with a 0.0126-year (i.e. 4.6-day) increase in DNAm-agesperm (95%CI: 0.003, 0.022, p = 0.01). Further analyses demonstrated significant negative associations of dioxin levels (β = -0.0005, 95%CI: -0.0010, 0.00004, P = 0.03) and DNAm-agesperm (β = -0.02, 95%CI: -0.04, -0.001, P = 0.03) with methylation levels of FOXK2 - a gene previously reported to be hypomethylated in infertile men. In sum, we demonstrate associations of dioxin with increased methylation aging of sperm. DNAm-agesperm may provide utility for understanding how dioxin levels impact sperm health and potentially male reproductive capacity in human population studies. Moreover, our pilot study contributes further evidence that some environmental toxicants are associated with methylation aging. Additional studies are necessary to confirm these findings, and better characterize dioxin and sperm methylation relationships with male reproductive health.

    View details for DOI 10.1016/j.reprotox.2020.06.004

    View details for Web of Science ID 000582635800004

    View details for PubMedID 32522586

  • Locus -Specific Differential DNA Methylation and Urinary Arsenic: An Epigenome-Wide Association Study in Blood among Adults with Low -to -Moderate Arsenic Exposure ENVIRONMENTAL HEALTH PERSPECTIVES Bozack, A. K., Domingo-Relloso, A., Haack, K., Gamble, M. V., Tellez-Plaza, M., Umans, J. G., Best, L. G., Yracheta, J., Gribble, M. O., Cardenas, A., Francesconi, K. A., Goessler, W., Tang, W., Fallin, M., Cole, S. A., Navas-Acien, A. 2020; 128 (6): 67015


    Chronic exposure to arsenic (As), a human toxicant and carcinogen, remains a global public health problem. Health risks persist after As exposure has ended, suggesting epigenetic dysregulation as a mechanistic link between exposure and health outcomes.We investigated the association between total urinary As and locus-specific DNA methylation in the Strong Heart Study, a cohort of American Indian adults with low-to-moderate As exposure [total urinary As, mean (±SD) μg/g creatinine: 11.7 (10.6)].DNA methylation was measured in 2,325 participants using the Illumina MethylationEPIC array. We implemented linear models to test differentially methylated positions (DMPs) and the DMRcate method to identify regions (DMRs) and conducted gene ontology enrichment analysis. Models were adjusted for estimated cell type proportions, age, sex, body mass index, smoking, education, estimated glomerular filtration rate, and study center. Arsenic was measured in urine as the sum of inorganic and methylated species.In adjusted models, methylation at 20 CpGs was associated with urinary As after false discovery rate (FDR) correction (FDR< 0.05). After Bonferroni correction, 5 CpGs remained associated with total urinary As (pBonferroni<0.05), located in SLC7A11, ANKS3, LINGO3, CSNK1D, ADAMTSL4. We identified one DMR on chromosome 11 (chr11:2,322,050-2,323,247), annotated to C11orf2; TSPAN32 genes.This is one of the first epigenome-wide association studies to investigate As exposure and locus-specific DNA methylation using the Illumina MethylationEPIC array and the largest epigenome-wide study of As exposure. The top DMP was located in SLC7A11A, a gene involved in cystine/glutamate transport and the biosynthesis of glutathione, an antioxidant that may protect against As-induced oxidative stress. Additional DMPs were located in genes associated with tumor development and glucose metabolism. Further research is needed, including research in more diverse populations, to investigate whether As-related DNA methylation signatures are associated with gene expression or may serve as biomarkers of disease development.

    View details for DOI 10.1289/EHP6263

    View details for Web of Science ID 000548216800007

    View details for PubMedID 32603190

    View details for PubMedCentralID PMC7534587

  • Cannabis Exposure During Critical Windows of Development: Epigenetic and Molecular Pathways Implicated in Neuropsychiatric Disease CURRENT ENVIRONMENTAL HEALTH REPORTS Smith, A., Kaufman, F., Sandy, M. S., Cardenas, A. 2020; 7 (3): 325-342


    Cannabis exposure during critical windows of development may have intergenerational physiological consequences disrupting epigenetic programming and marks. This review examines the literature relating to pre-gestational and prenatal cannabinoid exposure and its effect on genes and molecular pathways related to the development of psychiatric disease.Developmental cannabis exposure alters epigenetic processes with functional gene consequences. These include potentially heritable alterations in genes and molecular pathways critical for brain development and associated with autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD), schizophrenia, addiction, and other psychiatric diseases. Cannabis consumption and mental health illness in adolescents and young adults are increasing in the United States (U.S.), and recent studies suggest that cannabis consumption during critical periods of brain development could contribute to mental health illness through epigenetic mechanisms. These findings warrant future studies and consideration by regulators and health communicators.

    View details for DOI 10.1007/s40572-020-00275-4

    View details for Web of Science ID 000538169400001

    View details for PubMedID 32441004

    View details for PubMedCentralID PMC7458902

  • Per- and polyfluoroalkyl substances and blood pressure in pre-diabetic adults-cross-sectional and longitudinal analyses of the diabetes prevention program outcomes study ENVIRONMENT INTERNATIONAL Lin, P. D., Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2020; 137: 105573


    The relationship of plasma concentration of per- and polyfluoroalkyl substances (PFAS) with blood pressure (BP) is uncertain. This study examined cross-sectional and prospective associations of PFAS with BP and hypertension. We quantified plasma PFAS concentrations from 957 participants enrolled in the lifestyle and placebo arms of the Diabetes Prevention Program (DPP), a randomized controlled trial with approximately 15 years of follow-up. We used multivariable linear and logistic regressions to test cross-sectional associations of six PFAS, including perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), N-ethyl-perfluorooctane sulfonamido acetic acid (EtFOSAA), N-methyl-perfluorooctane sulfonamido acetic acid (MeFOSAA), and perfluorononanoic acid (PFNA), with BP and hypertension prevalence, respectively, at baseline. We used generalized linear mixed models to estimate longitudinal associations between baseline PFAS and the rate of BP changes, and Cox-Proportional hazard models to estimate risk of developing hypertension relative to baseline PFAS. Models were adjusted for baseline age, sex, race/ethnicity, treatment arm, educational attainment, income, marital status, smoking habit, alcohol drinking, and diet. We tested for effect modification by the treatment arm and sex, and accounted for multiple comparisons using the False-Discovery Rate (FDR). PFAS concentrations and hypertension prevalence within the study population (65.3% female, 57.7% White, 65.3% aged 40-59 years) were comparable to the general U.S. population. Cross-sectionally, we found small but statistically significant associations of baseline plasma concentrations of PFOA with systolic BP (β per doubling: 1.49 mmHg, 95% CI: 0.29, 2.70); and MeFOSAA with hypertension (RR = 1.09 per doubling, 95% CI: 1.01, 1.19). Estimates were not statistically significant after FDR adjustment. Longitudinally, we observed null associations in the placebo arm, but some inverse associations of baseline PFOS and MeFOSAA with systolic BP in the lifestyle arm, perhaps due to regression toward the mean. Baseline PFAS concentrations also were not prospectively associated with hypertension risk. Overall, there were modest and mostly null associations of plasma PFAS concentrations with BP and hypertension.

    View details for DOI 10.1016/j.envint.2020.105573

    View details for Web of Science ID 000517970800061

    View details for PubMedID 32088543

    View details for PubMedCentralID PMC7094005

  • Cord blood DNA methylation of DNMT3A mediates the association between in utero arsenic exposure and birth outcomes: Results from a prospective birth cohort in Bangladesh ENVIRONMENTAL RESEARCH Bozack, A. K., Cardenas, A., Geldhof, J., Quamruzzaman, Q., Rahman, M., Mostofa, G., Christiani, D. C., Kile, M. L. 2020; 183: 109134


    Fetal epigenetic programming plays a critical role in development. DNA methyltransferase 3 alpha (DNMT3A), which is involved in de novo DNA methylation (DNAm), is a prime candidate gene as a mediator between prenatal exposures and birth outcomes. We evaluated the relationships between in utero arsenic (As) exposure, birth outcomes, and DNMT3A DNAm.In a prospective Bangladeshi birth cohort, cord blood DNAm of three DNMT3A CpGs was measured using bisulfite pyrosequencing. Maternal toenail As concentrations at birth were measured to estimate in utero exposure. Among vaginal births (N = 413), structural equation models (SEMs) were used to evaluate relationships between DNMT3A methylation, log2 (toenail As), birth weight, and gestational age.In an adjusted SEM including birth weight and gestational age, maternal toenail As levels were associated with DNMT3A DNAm (B = 0.40; 95% CI: 0.15, 0.66) and gestational age (B = -0.19 weeks; 95% CI: 0.36, -0.03). DNMT3A DNAm was associated with gestational age (B = -0.10 weeks; 95% CI: 0.16, -0.04) and birth weight (B = -11.0 g; 95% CI: 21.5, 0.4). There was an indirect effect of As on gestational age mediated through DNMT3A DNAm (B = -0.04; 95% CI: 0.08, -0.01), and there were indirect effects of maternal toenail As levels on birth weight through pathways including gestational age (B = -14.4 g; 95% CI: 29.2, -1.9), DNMT3A DNAm and gestational age (B = -3.1 g; 95% CI: 6.6, -0.8), and maternal weight gain and gestational age (B = -5.1 g; 95% CI: 9.6, -1.5). The total effect of a doubling in maternal toenail As concentration is a decrease in gestational age of 2.1 days (95% CI: 0.9, 3.3) and a decrease in birth weight of 29 g (95% CI: 14, 46).DNMT3A plays a critical role in fetal epigenetic programming. In utero arsenic exposure was associated with greater methylation of CpGs in DNMT3A which partially mediated associations between prenatal As exposure and birth outcomes. Additional studies are needed to verify this finding.

    View details for DOI 10.1016/j.envres.2020.109134

    View details for Web of Science ID 000526177500124

    View details for PubMedID 32018205

    View details for PubMedCentralID PMC7167334

  • Dietary characteristics associated with plasma concentrations of per- and polyfluoroalkyl substances among adults with pre-diabetes: Cross-sectional results from the Diabetes Prevention Program Trial ENVIRONMENT INTERNATIONAL Lin, P. D., Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Fleisch, A. F., Calafat, A. M., Sanchez-Guerra, M., Osorio-Yanez, C., Webster, T. F., Horton, E. S., Oken, E. 2020; 137: 105217


    Diet is assumed to be the main source of exposure to per- and polyfluoroalkyl substances (PFAS) in non-occupationally exposed populations, but studies on the diet-PFAS relationship in the United States are scarce. We extracted multiple dietary variables, including daily intakes of food group, diet scores, and dietary patterns, from self-reported dietary data collected at baseline (1996-1999) from adults with pre-diabetes enrolled in the Diabetes Prevention Program, and used linear regression models to evaluate relationships of each dietary variable with plasma concentrations of six PFAS (perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorohexane sulfonic acid (PFHxS), 2-(N-ethyl-perfluorooctane sulfonamido) acetic acid (EtFOSAA), 2-(N-methyl-perfluorooctane sulfonamido) acetic acid (MeFOSAA), perfluorononanoic acid (PFNA) adjusting for covariates. Participants (N = 941, 65% female, 58% Caucasian, 68% married, 75% with higher education, 95% nonsmoker) had similar PFAS concentrations compared to the general U.S. population during 1999-2000. Using a single food group approach, fried fish, other fish/shellfish, meat and poultry had positive associations with most PFAS plasma concentrations. The strongest effect estimate detected was between fried fish and PFNA [13.6% (95% CI: 7.7, 19.9) increase in median concentration per SD increase]. Low-carbohydrate and high protein diet score had positive association with plasma PFHxS. Some food groups, mostly vegetables and fruits, and the Dietary Approaches to Stop Hypertension diet score had inverse associations with PFOS and MeFOSAA. A vegetable diet pattern was associated with lower plasma concentrations of MeFOSAA, while high-fat meat and low-fiber and high-fat grains diet patterns were associated with higher plasma concentrations of PFOS, PFHxS, MeFOSAA and PFNA. We summarized four major dietary characteristics associated with variations in PFAS plasma concentrations in this population. Specifically, consuming more meat/fish/shellfish (especially fried fish, and excluding Omega3-rich fish), low-fiber and high-fat bread/cereal/rice/pasta, and coffee/tea was associated with higher plasma concentrations while dietary patterns of vegetables, fruits and Omega-3 rich fish were associated with lower plasma concentrations of some PFAS.

    View details for DOI 10.1016/j.envint.2019.105217

    View details for Web of Science ID 000517970800022

    View details for PubMedID 32086073

    View details for PubMedCentralID PMC7517661

  • The Key Characteristics of Carcinogens: Relationship to the Hallmarks of Cancer, Relevant Biomarkers, and Assays to Measure Them. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology Smith, M. T., Guyton, K. Z., Kleinstreuer, N., Borrel, A., Cardenas, A., Chiu, W. A., Felsher, D. W., Gibbons, C. F., Goodson, W. H., Houck, K. A., Kane, A., La Merrill, M. A., Lebrec, H., Lowe, L., McHale, C. M., Minocherhomji, S., Rieswijk, L., Sandy, M. S., Sone, H., Wang, A., Zhang, L., Zeise, L., Fielden, M. 2020


    BACKGROUND: The key characteristics (KCs) of human carcinogens provide a uniform approach to evaluating mechanistic evidence in cancer hazard identification. Refinements to the approach were requested by organizations and individuals applying the KCs.METHODS: We assembled an expert committee with knowledge of carcinogenesis and experience in applying the KCs in cancer hazard identification. We leveraged this expertise and an examination of the literature to more clearly describe each KC; identify current and emerging assays and in vivo biomarkers that can be used to measure them; and, make recommendations for future assay development.RESULTS: We found that the KCs are clearly distinct from the Hallmarks of Cancer, that interrelationships among the KCs can be leveraged to strengthen the KC approach (and an understanding of environmental carcinogenesis), and that the KC approach is applicable to the systematic evaluation of a broad range of potential cancer hazards in vivo and in vitro. We identified gaps in coverage of the KCs by current assays.CONCLUSION: Future efforts should expand the breadth, specificity and sensitivity of validated assays and biomarkers that can measure the 10 KCs.IMPACT: Refinement of the KC approach will enhance and accelerate carcinogen identification, a first step in cancer prevention.

    View details for DOI 10.1158/1055-9965.EPI-19-1346

    View details for PubMedID 32152214

  • Epigenome-wide meta-analysis of blood DNA methylation in newborns and children identifies numerous loci related to gestational age GENOME MEDICINE Merid, S., Novoloaca, A., Sharp, G. C., Kupers, L. K., Kho, A. T., Roy, R., Gao, L., Annesi-Maesano, I., Jain, P., Plusquin, M., Kogevinas, M., Allard, C., Vehmeijer, F. O., Kazmi, N., Salas, L. A., Rezwan, F. I., Zhang, H., Sebert, S., Czamara, D., Rifas-Shiman, S. L., Melton, P. E., Lawlor, D. A., Pershagen, G., Breton, C. V., Huen, K., Baiz, N., Gagliardi, L., Nawrot, T. S., Corpeleijn, E., Perron, P., Duijts, L., Nohr, E., Bustamante, M., Ewart, S. L., Karmaus, W., Zhao, S., Page, C. M., Herceg, Z., Jarvelin, M., Lahti, J., Baccarelli, A. A., Anderson, D., Kachroo, P., Relton, C. L., Bergstrom, A., Eskenazi, B., Soomro, M., Vineis, P., Snieder, H., Bouchard, L., Jaddoe, V. W., Sorensen, T. A., Vrijheid, M., Arshad, S., Holloway, J. W., Haberg, S. E., Magnus, P., Dwyer, T., Binder, E. B., DeMeo, D. L., Vonk, J. M., Newnham, J., Tantisira, K. G., Kull, I., Wiemels, J. L., Heude, B., Sunyer, J., Nystad, W., Munthe-Kaas, M. C., Raikkonen, K., Oken, E., Huang, R., Weiss, S. T., Anto, J., Bousquet, J., Kumar, A., Soderhall, C., Almqvist, C., Cardenas, A., Gruzieva, O., Xu, C., Reese, S. E., Kere, J., Brodin, P., Solomon, O., Wielscher, M., Holland, N., Ghantous, A., Hivert, M., Felix, J. F., Koppelman, G. H., London, S. J., Melen, E. 2020; 12 (1): 25


    Preterm birth and shorter duration of pregnancy are associated with increased morbidity in neonatal and later life. As the epigenome is known to have an important role during fetal development, we investigated associations between gestational age and blood DNA methylation in children.We performed meta-analysis of Illumina's HumanMethylation450-array associations between gestational age and cord blood DNA methylation in 3648 newborns from 17 cohorts without common pregnancy complications, induced delivery or caesarean section. We also explored associations of gestational age with DNA methylation measured at 4-18 years in additional pediatric cohorts. Follow-up analyses of DNA methylation and gene expression correlations were performed in cord blood. DNA methylation profiles were also explored in tissues relevant for gestational age health effects: fetal brain and lung.We identified 8899 CpGs in cord blood that were associated with gestational age (range 27-42 weeks), at Bonferroni significance, P < 1.06 × 10- 7, of which 3343 were novel. These were annotated to 4966 genes. After restricting findings to at least three significant adjacent CpGs, we identified 1276 CpGs annotated to 325 genes. Results were generally consistent when analyses were restricted to term births. Cord blood findings tended not to persist into childhood and adolescence. Pathway analyses identified enrichment for biological processes critical to embryonic development. Follow-up of identified genes showed correlations between gestational age and DNA methylation levels in fetal brain and lung tissue, as well as correlation with expression levels.We identified numerous CpGs differentially methylated in relation to gestational age at birth that appear to reflect fetal developmental processes across tissues. These findings may contribute to understanding mechanisms linking gestational age to health effects.

    View details for DOI 10.1186/s13073-020-0716-9

    View details for Web of Science ID 000517604800001

    View details for PubMedID 32114984

    View details for PubMedCentralID PMC7050134

  • Interplay of Placental DNA Methylation and Maternal Insulin Sensitivity in Pregnancy DIABETES Hivert, M., Cardenas, A., Allard, C., Doyon, M., Powe, C. E., Catalano, P. M., Perron, P., Bouchard, L. 2020; 69 (3): 484-492


    The placenta participates in maternal insulin sensitivity changes during pregnancy; however, mechanisms remain unclear. We investigated associations between maternal insulin sensitivity and placental DNA methylation markers across the genome. We analyzed data from 430 mother-offspring dyads in the Gen3G cohort. All women underwent 75-g oral glucose tolerance tests at ∼26 weeks of gestation; we used glucose and insulin measures to estimate insulin sensitivity (Matsuda index). At delivery, we collected samples from placenta (fetal side) and measured DNA methylation using Illumina EPIC arrays. Using linear regression models to quantify associations at 720,077 cytosine-guanine dinucleotides (CpGs), with adjustment for maternal age, gravidity, smoking, BMI, child sex, and gestational age at delivery, we identified 188 CpG sites where placental DNA methylation was associated with Matsuda index (P < 6.94 × 10-8). Among genes annotated to these 188 CpGs, we found enrichment in targets for miRNAs, in histone modifications, and in parent-of-origin DNA methylation including the H19/MIR675 locus (paternally imprinted). We identified 12 known placenta imprinted genes, including KCNQ1 Mendelian randomization analyses revealed five loci where placenta DNA methylation may causally influence maternal insulin sensitivity, including the maternally imprinted gene DLGAP2. Our results suggest that placental DNA methylation is fundamentally linked to the regulation of maternal insulin sensitivity in pregnancy.

    View details for DOI 10.2337/db19-0798

    View details for Web of Science ID 000515719900021

    View details for PubMedID 31882564

    View details for PubMedCentralID PMC7213861

  • Mediation Analysis Supports a Causal Relationship between Maternal Hyperglycemia and Placental DNA Methylation Variations at the Leptin Gene Locus and Cord Blood Leptin Levels INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Gagne-Ouellet, V., Breton, E., Thibeault, K., Fortin, C., Cardenas, A., Guerin, R., Perron, P., Hivert, M., Bouchard, L. 2020; 21 (1)


    Changes in fetal DNA methylation (DNAm) of the leptin (LEP) gene have been associated with exposure to maternal hyperglycemia, but their links with childhood obesity risk are still unclear. We investigated the association between maternal hyperglycemia, placental LEP DNAm (25 5'-C-phosphate-G-3' (CpG) sites), neonatal leptinemia, and adiposity (i.e., BMI and skinfold thickness (ST) (subscapular (SS) + triceps (TR) skinfold measures, and the ratio of SS:TR) at 3-years-old, in 259 mother-child dyads, from Gen3G birth cohort. We conducted multivariate linear analyses adjusted for gestational age at birth, sex of the child, age at follow-up, and cellular heterogeneity. We assessed the causal role of DNAm in the association between maternal glycemia and childhood outcomes, using mediation analysis. We found three CpGs associated with neonatal leptinemia (p ≤ 0.002). Of these, cg05136031 and cg15758240 were also associated with BMI (β = -2.69, p = 0.05) and fat distribution (β = -0.581, p = 0.05) at 3-years-old, respectively. Maternal glycemia was associated with DNAm at cg15758240 (β = -0.01, p = 0.04) and neonatal leptinemia (β = 0.19, p = 0.004). DNAm levels at cg15758240 mediates 0.8% of the association between maternal glycemia and neonatal leptinemia (p < 0.001). Our results support that DNAm regulation of the leptin pathway in response to maternal glycemia might be involved in programming adiposity in childhood.

    View details for DOI 10.3390/ijms21010329

    View details for Web of Science ID 000515378000329

    View details for PubMedID 31947745

    View details for PubMedCentralID PMC6982090

  • Association of Periconception Paternal Body Mass Index With Persistent Changes in DNA Methylation of Offspring in Childhood JAMA NETWORK OPEN Noor, N., Cardenas, A., Rifas-Shiman, S. L., Pan, H., Dreyfuss, J. M., Oken, E., Hivert, M., James-Todd, T., Patti, M., Isganaitis, E. 2019; 2 (12): e1916777


    While prenatal nutrition and maternal obesity are recognized as important contributors to epigenetic changes and childhood obesity, the role of paternal obesity in the epigenome of offspring has not been well studied.To test whether periconception paternal body mass index (BMI) is associated with DNA methylation patterns in newborns, to examine associations between maternal and paternal BMI and the epigenome of offspring, and to examine persistence of epigenetic marks at ages 3 and 7 years.Project Viva is a prebirth cohort study of mothers and children including 2128 live births that enrolled mothers from April 1999 to July 2002 and followed offspring to adolescence. This study analyzed the subset of participants with available data on paternal BMI and DNA methylation in offspring blood in the newborn period, at age 3 years, and at age 7 years. Data were analyzed from July 2017 to October 2019.The primary exposure was paternal periconception BMI; associations were adjusted for maternal prepregnancy BMI and stratified according to maternal BMI above or below 25.The primary outcome was genome-wide DNA methylation patterns in offspring blood collected at birth, age 3 years, and age 7 years.A total of 429 father-mother-infant triads were included. The mean (SD) periconception paternal BMI was 26.4 (4.0) and mean maternal prepregnancy BMI was 24.5 (5.2); 268 fathers had BMI greater than or equal to 25 (mean [SD], 28.5 [3.3]) and 161 had BMI less than 25 (mean [SD], 22.8 [1.8]). Paternal BMI greater than or equal to 25 was associated with increased offspring birth weight compared with paternal BMI less than 25 (mean [SD] z score, 0.38 [0.91] vs 0.11 [0.96]; P = .004). Cord blood DNA methylation at 9 CpG sites was associated with paternal BMI independent of maternal BMI (q < .05). Methylation at cg04763273, between TFAP2C and BMP7, decreased by 5% in cord blood with every 1-unit increase in paternal BMI (P = 3.13 × 10-8); hypomethylation at this site persisted at ages 3 years and 7 years. Paternal BMI was associated with methylation at cg01029450 in the promoter region of the ARFGAP3 gene; methylation at this site was also associated with lower infant birth weight (β = -0.0003; SD = 0.0001; P = .03) and with higher BMI z score at age 3 years.In this study, paternal BMI was associated with DNA methylation, birth weight, and childhood BMI z score in offspring.

    View details for DOI 10.1001/jamanetworkopen.2019.16777

    View details for Web of Science ID 000512329100063

    View details for PubMedID 31880793

    View details for PubMedCentralID PMC6991200

  • Prenatal lead exposure and childhood executive function and behavioral difficulties in project viva NEUROTOXICOLOGY Fruh, V., Rifas-Shiman, S. L., Amarasiriwardena, C., Cardenas, A., Bellinger, D. C., Wise, L. A., White, R. F., Wright, R. O., Oken, E., Henn, B. 2019; 75: 105-115


    Lead is an established neurotoxicant and early life exposure to lead is associated with detrimental impacts on IQ and several neurobehavioral domains. Less is known, however, about effects of prenatal lead exposure below 5 μg/dL on executive function and on social, emotional and self-regulatory behaviors in childhood.To examine the association between prenatal lead exposure and childhood executive function and social, emotional and self-regulatory behaviors.We included 1006 mother-child pairs from the Project Viva prospective pre-birth cohort. We measured prenatal maternal lead in second-trimester erythrocytes. In mid-childhood (median 7.7 years), parents and teachers rated executive function related behaviors using the Behavior Rating Inventory of Executive Function (BRIEF) and behavioral difficulties using the Strengths and Difficulties Questionnaire (SDQ). We used multivariable linear regression models adjusted for maternal, paternal, and child characteristics and metal co-exposures.Mean maternal erythrocyte lead concentration was 1.2 μg/dL (interquartile range [IQR] 0.8-1.5 μg /dL), equivalent to approximately 0.4 μg/dL in whole blood. In adjusted models, associations with parent and teacher-rated scales were largely null, although effect estimates were consistently positive, suggesting worse scores with increasing lead levels. For an IQR increase in lead, BRIEF Global Executive Composite (GEC) was 0.73 (95% CI: -0.06, 1.52) points higher for parent-rated scores and 0.42 (95% CI: -0.39, 1.23) points higher for teacher-rated scores. Associations were strongest for parent-rated BRIEF plan/organize (β = 0.85; 95% CI: 0.12, 1.59) and shift (β = 0.88; 95% CI: 0.01, 1.75) subscales, as well as the SDQ emotional problems subscale (β = 0.18; 95% CI: 0.03, 0.33).In this cohort with lead levels commonly experienced by U.S. women, there were few statistically significant associations with childhood executive function and behavior. However, there was a trend of worse neurobehavioral scores with increasing prenatal lead concentrations, in particular for childhood emotional problems and capacity to plan/organize and shift. Our results highlight the importance of continuing efforts to eliminate lead exposure in the general population.

    View details for DOI 10.1016/j.neuro.2019.09.006

    View details for Web of Science ID 000499764600010

    View details for PubMedID 31513824

    View details for PubMedCentralID PMC6842061

  • Cardenas et al. Reply to "DNA Methylation and Prenatal Exposures" AMERICAN JOURNAL OF EPIDEMIOLOGY Cardenas, A., Lutz, S. M., Everson, T. M., Hivert, M. 2019; 188 (11): 1890-1891

    View details for DOI 10.1093/aje/kwz183

    View details for Web of Science ID 000504115700004

    View details for PubMedID 31647094

    View details for PubMedCentralID PMC7304507

  • Mediation by Placental DNA Methylation of the Association of Prenatal Maternal Smoking and Birth Weight AMERICAN JOURNAL OF EPIDEMIOLOGY Cardenas, A., Lutz, S. M., Everson, T. M., Perron, P., Bouchard, L., Hivert, A. 2019; 188 (11): 1878-1886


    Prenatal maternal smoking is a risk factor for lower birth weight. We performed epigenome-wide association analyses of placental DNA methylation (DNAm) at 720,077 cytosine-phosphate-guanine (CpG) sites and prenatal maternal smoking among 441 mother-infant pairs (2010-2014) and evaluated whether DNAm mediates the association between smoking and birth weight using mediation analysis. Mean birth weight was 3,443 (standard deviation, 423) g, and 38 mothers (8.6%) reported smoking at a mean of 9.4 weeks of gestation. Prenatal maternal smoking was associated with a 175-g lower birth weight (95% confidence interval (CI): -305.5, -44.8) and with differential DNAm of 71 CpGs in placenta, robust to latent-factor adjustment reflecting cell types (Bonferroni-adjusted P < 6.94 × 10-8). Of the 71 CpG sites, 7 mediated the association between prenatal smoking and birth weight (on MDS2, PBX1, CYP1A2, VPRBP, WBP1L, CD28, and CDK6 genes), and prenatal smoking × DNAm interactions on birth weight were observed for 5 CpG sites. The strongest mediator, cg22638236, was annotated to the PBX1 gene body involved in skeletal patterning and programming, with a mediated effect of 301-g lower birth weight (95% CI: -543, -86) among smokers but no mediated effect for nonsmokers (β = -38 g; 95% CI: -88, 9). Prenatal maternal smoking might interact with placental DNAm at specific loci, mediating the association with lower infant birth weight.

    View details for DOI 10.1093/aje/kwz184

    View details for Web of Science ID 000504115700002

    View details for PubMedID 31497855

    View details for PubMedCentralID PMC6825837

  • Associations of prenatal or infant exposure to acetaminophen or ibuprofen with mid-childhood executive function and behaviour PAEDIATRIC AND PERINATAL EPIDEMIOLOGY Rifas-Shiman, S. L., Cardenas, A., Hivert, M., Tiemeier, H., Bertoldi, A. D., Oken, E. 2020; 34 (3): 287-298


    Over-the-counter analgesics during pregnancy or infancy may be related to neurobehavioural problems in children, but little is known about effects of different analgesic types, dosage, and timing.Examine associations of acetaminophen and ibuprofen use during pregnancy and infancy with executive function and behaviour problems in children.We included 1225 mother-child pairs from Project Viva, a pre-birth cohort study. We assessed prenatal acetaminophen and ibuprofen use in early and mid-pregnancy and infant use in the first year of life using questionnaires. Parents and classroom teachers assessed child behaviours in mid-childhood (median 8 years), using the Behavior Rating Inventory of Executive Function (BRIEF) and the Strengths and Difficulties Questionnaire (SDQ), with higher scores indicating worse functioning for both. We examined associations of acetaminophen and ibuprofen use during pregnancy and infancy with mid-childhood neurobehavioural outcomes using linear regression models adjusted for potential confounders.During pregnancy, 46.1% of mothers used acetaminophen ≥10 times and 18.4% used any ibuprofen. In the first year, 65.3% and 39.6% of infants received acetaminophen and ibuprofen ≥6 times, respectively. Higher (≥10 vs <10 times) prenatal acetaminophen (β 1.64 points; 95% confidence interval [CI] 0.59, 2.68) and any ibuprofen (β 1.56, 95% CI 0.19, 2.92) were associated with higher parent-rated BRIEF global scores. Patterns of association were linear across categories and were similar for other parent- and teacher-rated outcomes. Infancy exposure (≥6 vs <6 times) to acetaminophen (β 1.69, 95% CI 0.51, 2.87) and ibuprofen (β 1.40, 95% CI 0.25, 2.55) were associated with higher parent-rated BRIEF GEC scores but associations with teacher-rated scores were weaker.Prenatal and early-life exposure to acetaminophen and ibuprofen were associated with poorer executive function and behaviour in childhood. These findings highlight the need for further research on the mechanisms through which analgesics may act on fetal and child brain development.

    View details for DOI 10.1111/ppe.12596

    View details for Web of Science ID 000491720900001

    View details for PubMedID 31637744

    View details for PubMedCentralID PMC7170759

  • Editorial: Exposure to Endocrine-Disrupting Chemicals and Cardiometabolic Disease: A Developmental Origins Approach FRONTIERS IN PUBLIC HEALTH Perng, W., Goodrich, J. M., Cardenas, A., Watkins, D. J. 2019; 7: 288

    View details for DOI 10.3389/fpubh.2019.00288

    View details for Web of Science ID 000496841000001

    View details for PubMedID 31649913

    View details for PubMedCentralID PMC6794340

  • Adequate Prenatal Maternal Folate-An Additional Intervention Strategy Among Populations Affected by Prenatal Lead Exposure? JAMA NETWORK OPEN Sanchez-Guerra, M., Cardenas, A., Osorio-Yanez, C. 2019; 2 (10): e1912334
  • Socioeconomic status and DNA methylation from birth through mid-childhood: a prospective study in Project Viva EPIGENOMICS Laubach, Z. M., Perng, W., Cardenas, A., Rifas-Shiman, S. L., Oken, E., DeMeo, D., Litonjua, A. A., Duca, R., Godderis, L., Baccarelli, A., Hivert, M. 2019; 11 (12): 1413-1427


    Aim: We investigated associations of prenatal socioeconomic status (SES) with DNA methylation at birth, and to explore persistence of associations into early (∼3 years) and mid-childhood (∼7 years) among 609 mother-child pairs in a Boston-area prebirth cohort. Materials & methods: First, we created a prenatal SES index comprising individual- and neighborhood-level metrics and examined associations of low (lowest 10%) versus high (upper 90%) SES with genome-wide DNA methylation in cord blood via the Infinium HumanMethylation450 BeadChip. Next, we evaluated persistence of associations detected in cord blood with DNA methylation of the same CpG sites measured in peripheral leukocytes in early- and mid-childhood. Results & conclusion: Low prenatal SES was associated with methylation at CpG sites near ACSF3, TNRC6C-AS1, MTMR4 and LRRN4. The relationship with LRRN4 persisted into early childhood.

    View details for DOI 10.2217/epi-2019-0040

    View details for Web of Science ID 000498631700006

    View details for PubMedID 31509016

    View details for PubMedCentralID PMC6802709

  • Relationships of Long-Term Smoking and Moist Snuff Consumption With a DNA Methylation Age Relevant Smoking Index: An Analysis in Buccal Cells NICOTINE & TOBACCO RESEARCH Nwanaji-Enwerem, J. C., Cardenas, A., Chai, P. R., Weisskopf, M. G., Baccarelli, A. A., Boyer, E. W. 2019; 21 (9): 1267-1273


    Currently, there is no widely accepted, non-self-report measure that simultaneously reflects smoking behaviors and is molecularly informative of general disease processes. Recently, researchers developed a smoking index (SI) using nucleated blood cells and a multi-tissue DNA methylation-based predictor of chronological age and disease (DNA methylation age [DNAm-age]). To better understand the utility of this novel SI in readily accessible cell types, we used buccal cell DNA methylation to examine SI relationships with long-term tobacco smoking and moist snuff consumption.We used a publicly available dataset composed of buccal cell DNA methylation values from 120 middle-aged men (40 long-term smokers, 40 moist snuff consumers, and 40 nonsmokers). DNAm-age (353-CpGs) and SI (66-CpGs) were calculated using CpG sites measured using the Illumina HumanMethylation450 BeadChip. We estimated associations of tobacco consumption habits with both SI and DNAm-age using linear regression models adjusted for chronological age, race, and methylation technical covariates.In fully adjusted models with nonsmokers as the reference, smoking (β = 1.08, 95% CI = 0.82 to 1.33, p < .0001) but not snuff consumption (β = .06, 95% CI = -0.19 to 0.32, p = .63) was significantly associated with SI. SI was an excellent predictor of smoking versus nonsmoking (area under the curve = 0.92, 95% CI = 0.85 to 0.98). Four DNAm-age CpGs were differentially methylated between smokers and nonsmokers including cg14992253 [EIF3I], which has been previously shown to be differentially methylated with exposure to long-term fine-particle air pollution (PM2.5).The 66-CpG SI appears to be a useful tool for measuring smoking-specific behaviors in buccal cells. Still, further research is needed to broadly confirm our findings and SI relationships with DNAm-age.Our findings demonstrate that this 66-CpG blood-derived SI can reflect long-term tobacco smoking, but not long-term snuff consumption, in buccal cells. This evidence will be useful as the field works to identify an accurate non-self-report smoking biomarker that can be measured in an easily accessible tissue. Future research efforts should focus on (1) optimizing the relationship of the SI with DNAm-age so that the metric can maximize its utility as a tool for understanding general disease processes, and (2) determining normal values for the SI CpGs so that the measure is not as study sample specific.

    View details for DOI 10.1093/ntr/nty156

    View details for Web of Science ID 000493386500015

    View details for PubMedID 30053132

    View details for PubMedCentralID PMC6941707

  • Associations of Perfluoroalkyl and Polyfluoroalkyl Substances With Incident Diabetes and Microvascular Disease DIABETES CARE Cardenas, A., Hivert, M., Gold, D. R., Hauser, R., Kleinman, K. P., Lin, P. D., Fleisch, A. F., Calafat, A. M., Ye, X., Webster, T. F., Horton, E. S., Oken, E. 2019; 42 (9): 1824-1832


    Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are suspected endocrine disruptors widely detected across populations. We examine the extent to which PFASs are associated with diabetes incidence and microvascular disease. Secondarily, we tested whether a lifestyle intervention modifies associations and decreases concentrations.We analyzed data from a prospective cohort of 957 participants from the Diabetes Prevention Program (DPP) trial and Diabetes Prevention Program Outcomes Study (DPPOS). At baseline, participants were randomized to an intensive lifestyle intervention of diet, physical activity, and behavior modification or a placebo medication. We quantified plasma concentrations of six PFASs at baseline and 2 years after randomization. Participants were monitored for ∼15 years, repeatedly tested for diabetes, and evaluated for microvascular disease at the end of the follow-up.A doubling in baseline branched perfluorooctanoic acid concentration was associated with a 14% increase in diabetes risk for the placebo (hazard ratio [HR] 1.14, 95% CI 1.04, 1.25) but not in the lifestyle intervention group (HR 1.01, 95% CI 0.92, 1.11, P interaction = 0.11). Mean change in plasma baseline branched perfluorooctanoic acid concentration was greater for the placebo (0.96 ng/mL; 95% CI 0.71, 1.22) compared with the lifestyle intervention group (0.31 ng/mL; 95% CI 0.14, 0.48) 2 years after randomization. Each doubling in N-ethyl-perfluorooctane sulfonamido acetic acid was associated with 17% greater odds of prevalent microvascular disease (OR 1.17, 95% CI 1.05, 1.31), and a similar association was observed for perfluorodimethylhexane sulfonic acid (OR 1.18, 95% CI 1.04, 1.35), regardless of treatment.Some plasma PFASs were associated with diabetes and microvascular disease. Our results suggest that exercise and diet may attenuate the diabetogenic association of PFASs.

    View details for DOI 10.2337/dc18-2254

    View details for Web of Science ID 000481900200037

    View details for PubMedID 31296647

    View details for PubMedCentralID PMC6702604

  • Per- and polyfluoroalkyl substances and blood lipid levels in pre-diabetic adults-longitudinal analysis of the diabetes prevention program outcomes study ENVIRONMENT INTERNATIONAL Lin, P. D., Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Fleisch, A. F., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2019; 129: 343-353


    Exposure to per- and polyfluoroalkyl substances (PFASs) may interfere with lipid regulation. However, most previous studies were cross-sectional with the risk of reverse causation, suggesting a need for long-term prospective studies. We examined the relationship of baseline plasma PFAS concentrations with repeated measures of blood lipids. We included 888 prediabetic adults from the Diabetes Prevention Program (DPP) and DPP Outcomes Study, who had measurements of 6 plasma PFAS concentrations at baseline (1996-1999) and repeated measures of blood lipids over 15 years of follow-up, and were initially randomized to placebo or a lifestyle intervention. We used linear regression to examine cross-sectional associations of PFAS concentrations and lipid levels at baseline, and evaluated prospective risks of hypercholesterolemia and hypertriglyceridemia using Cox proportional hazard models, and tested for effect modification by study arm. Participants (65.9% female, 57.0% White, 65.9% aged 40-59 years) had comparable PFAS concentrations [e.g., median (IQR) perfluorooctanoic acid (PFOA) 4.9 ng/mL (3.2)] with the general U.S. population in 1999-2000. We observed higher total cholesterol at baseline per doubling of PFOA (β: 6.1 mg/dL, 95% CI: 3.1, 9.04), perfluorohexane sulfonic acid (PFHxS, β: 2.2 mg/dL, 95% CI: 0.2, 4.3), and perfluorononanoic acid (PFNA, β: 2.9 mg/dL, 95% CI: 0.7, 5.0). Prospectively, baseline concentrations of several PFASs, including PFOA, PFOS, PFHxS and PFNA, predicted higher risks of incident hypercholesterolemia and hypertriglyceridemia, but only in the placebo group and not the lifestyle intervention group. For example, participants in the placebo group with PFOA concentration > median (4.9 ng/mL) were almost twice as likely (HR: 1.90, 95% CI: 1.25, 2.88) to develop hypertriglyceridemia compared to those ≤median. Findings suggest adverse effects of some PFASs on lipid profiles in prediabetic adults. However, the detrimental effect was attenuated with a lifestyle intervention.

    View details for DOI 10.1016/j.envint.2019.05.027

    View details for Web of Science ID 000470239200034

    View details for PubMedID 31150976

    View details for PubMedCentralID PMC6570418

  • The nasal methylome as a biomarker of asthma and airway inflammation in children NATURE COMMUNICATIONS Cardenas, A., Sordillo, J. E., Rifas-Shiman, S. L., Chung, W., Liang, L., Coull, B. A., Hivert, M., Lai, P. S., Forno, E., Celedon, J. C., Litonjua, A. A., Brennan, K. J., DeMeo, D. L., Baccarelli, A. A., Oken, E., Gold, D. R. 2019; 10: 3095


    The nasal cellular epigenome may serve as biomarker of airway disease and environmental response. Here we collect nasal swabs from the anterior nares of 547 children (mean-age 12.9 y), and measure DNA methylation (DNAm) with the Infinium MethylationEPIC BeadChip. We perform nasal Epigenome-Wide Association analyses (EWAS) of current asthma, allergen sensitization, allergic rhinitis, fractional exhaled nitric oxide (FeNO) and lung function. We find multiple differentially methylated CpGs (FDR < 0.05) and Regions (DMRs; ≥ 5-CpGs and FDR < 0.05) for asthma (285-CpGs), FeNO (8,372-CpGs; 191-DMRs), total IgE (3-CpGs; 3-DMRs), environment IgE (17-CpGs; 4-DMRs), allergic asthma (1,235-CpGs; 7-DMRs) and bronchodilator response (130-CpGs). Discovered DMRs annotated to genes implicated in allergic asthma, Th2 activation and eosinophilia (EPX, IL4, IL13) and genes previously associated with asthma and IgE in EWAS of blood (ACOT7, SLC25A25). Asthma, IgE and FeNO were associated with nasal epigenetic age acceleration. The nasal epigenome is a sensitive biomarker of asthma, allergy and airway inflammation.

    View details for DOI 10.1038/s41467-019-11058-3

    View details for Web of Science ID 000475295700002

    View details for PubMedID 31300640

    View details for PubMedCentralID PMC6625976

  • Human aging DNA methylation signatures are conserved but accelerated in cultured fibroblasts EPIGENETICS Sturm, G., Cardenas, A., Bind, M., Horvath, S., Wang, S., Wang, Y., Hagg, S., Hirano, M., Picard, M. 2019; 14 (10): 961-976


    Aging is associated with progressive and site-specific changes in DNA methylation (DNAm). These global changes are captured by DNAm clocks that accurately predict chronological age in humans but relatively little is known about how clocks perform in vitro. Here we culture primary human fibroblasts across the cellular lifespan (~6 months) and use four different DNAm clocks to show that age-related DNAm signatures are conserved and accelerated in vitro. The Skin & Blood clock shows the best linear correlation with chronological time (r = 0.90), including during replicative senescence. Although similar in nature, the rate of epigenetic aging is approximately 62x times faster in cultured cells than in the human body. Consistent with in vivo data, cells aged under hyperglycemic conditions exhibit an approximately three years elevation in baseline DNAm age. Moreover, candidate gene-based analyses further corroborate the conserved but accelerated biological aging process in cultured fibroblasts. Fibroblasts mirror the established DNAm topology of the age-related ELOVL2 gene in human blood and the rapid hypermethylation of its promoter cg16867657, which correlates with a linear decrease in ELOVL2 mRNA levels across the lifespan. Using generalized additive modeling on twelve timepoints across the lifespan, we also show how single CpGs exhibit loci-specific, linear and nonlinear trajectories that reach rates up to -47% (hypomethylation) to +23% (hypermethylation) per month. Together, these high-temporal resolution global, gene-specific, and single CpG data highlight the conserved and accelerated nature of epigenetic aging in cultured fibroblasts, which may constitute a system to evaluate age-modifying interventions across the lifespan.

    View details for DOI 10.1080/15592294.2019.1626651

    View details for Web of Science ID 000472413200001

    View details for PubMedID 31156022

    View details for PubMedCentralID PMC6691995

  • Epigenetic age acceleration is associated with allergy and asthma in children in Project Viva JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY Peng, C., Cardenas, A., Rifas-Shiman, S. L., Hivert, M., Gold, D. R., Platts-Mills, T. A., Lin, X., Oken, E., Avila, L., Celedon, J. C., Weiss, S. T., Baccarelli, A. A., Litonjua, A. A., DeMeo, D. L. 2019; 143 (6): 2263-+


    Epigenetic clocks have been suggested to capture one feature of the complexity between aging and the epigenome. However, little is known about the epigenetic clock in childhood allergy and asthma.We sought to examine associations of DNA methylation age (DNAmAge) and epigenetic age acceleration with childhood allergy and asthma.We calculated DNAmAge and age acceleration at birth, early childhood, and midchildhood based on the IlluminaHumanMethylation450BeadChip in Project Viva. We evaluated epigenetic clock associations with allergy and asthma using covariate-adjusted linear and logistic regressions. We attempted to replicate our findings in the Genetics of Asthma in Costa Rica Study.At midchildhood (mean age, 7.8 years) in Project Viva, DNAmAge and age acceleration were cross-sectionally associated with greater total serum IgE levels and greater odds of atopic sensitization. Every 1-year increase in intrinsic epigenetic age acceleration was associated with a 1.22 (95% CI, 1.07-1.39), 1.17 (95% CI, 1.03-1.34), and 1.29 (95% CI, 1.12-1.49) greater odds of atopic sensitization and environmental and food allergen sensitization. DNAmAge and extrinsic epigenetic age acceleration were also cross-sectionally associated with current asthma at midchildhood. DNAmAge and age acceleration at birth and early childhood were not associated with midchildhood allergy or asthma. The midchildhood association between age acceleration and atopic sensitization were replicated in an independent data set.Because the epigenetic clock might reflect immune and developmental components of biological aging, our study suggests pathways through which molecular epigenetic mechanisms of immunity, development, and maturation can interact along the age axis and associate with childhood allergy and asthma by midchildhood.

    View details for DOI 10.1016/j.jaci.2019.01.034

    View details for Web of Science ID 000470113200029

    View details for PubMedID 30738172

    View details for PubMedCentralID PMC6556426

  • Maternal corticotropin-releasing hormone is associated with LEP DNA methylation at birth and in childhood: an epigenome-wide study in Project Viva INTERNATIONAL JOURNAL OF OBESITY Tian, F., Rifas-Shiman, S. L., Cardenas, A., Baccarelli, A. A., DeMeo, D. L., Litonjua, A. A., Rich-Edwards, J. W., Gillman, M. W., Oken, E., Hivert, M. 2019; 43 (6): 1244-1255


    Corticotropin-releasing hormone (CRH) plays a central role in regulating the secretion of cortisol which controls a wide range of biological processes. Fetuses overexposed to cortisol have increased risks of disease in later life. DNA methylation may be the underlying association between prenatal cortisol exposure and health effects. We investigated associations between maternal CRH levels and epigenome-wide DNA methylation of cord blood in offsprings and evaluated whether these associations persisted into mid-childhood.We investigated mother-child pairs enrolled in the prospective Project Viva pre-birth cohort. We measured DNA methylation in 257 umbilical cord blood samples using the HumanMethylation450 Bead Chip. We tested associations of maternal CRH concentration with cord blood cells DNA methylation, adjusting the model for maternal age at enrollment, education, maternal race/ethnicity, maternal smoking status, pre-pregnancy body mass index, parity, gestational age at delivery, child sex, and cell-type composition in cord blood. We further examined the persistence of associations between maternal CRH levels and DNA methylation in children's blood cells collected at mid-childhood (n = 239, age: 6.7-10.3 years) additionally adjusting for the children's age at blood drawn.Maternal CRH levels are associated with DNA methylation variability in cord blood cells at 96 individual CpG sites (False Discovery Rate <0.05). Among the 96 CpG sites, we identified 3 CpGs located near the LEP gene. Regional analyses confirmed the association between maternal CRH and DNA methylation near LEP. Moreover, higher maternal CRH levels were associated with higher blood-cell DNA methylation of the promoter region of LEP in mid-childhood (P < 0.05, β = 0.64, SE = 0.30).In our cohort, maternal CRH was associated with DNA methylation levels in newborns at multiple loci, notably in the LEP gene promoter. The association between maternal CRH and LEP DNA methylation levels persisted into mid-childhood.

    View details for DOI 10.1038/s41366-018-0249-0

    View details for Web of Science ID 000470098400012

    View details for PubMedID 30464231

    View details for PubMedCentralID PMC6529291

  • Altered cord blood mitochondrial DNA content and pregnancy lead exposure in the PROGRESS cohort ENVIRONMENT INTERNATIONAL Sanchez-Guerra, M., Peng, C., Trevisi, L., Cardenas, A., Wilson, A., Osorio-Yanez, C., Niedzwiecki, M. M., Zhong, J., Svensson, K., Teresa Acevedo, M., Solano-Gonzalez, M., Amarasiriwardena, C. J., Estrada-Gutierrez, G., Brennan, K. M., Schnaas, L., Just, A. C., Laue, H. E., Wright, R. J., Maria Tellez-Rojo, M., Wright, R. O., Baccarelli, A. A. 2019; 125: 437-444


    Lead (Pb) crosses the placenta and can cause oxidative stress, reduced fetal growth and neurological problems. The principal source of oxidative stress in human cells is mitochondria. Therefore, disruption of normal mitochondrial function during pregnancy may represent a primary mechanism behind the adverse effects of lead. We sought to assess the association of Pb exposure during pregnancy with mitochondrial DNA (mtDNA) content, a sensitive marker of mitochondrial function, in cord blood.This study comprised mother-infant pairs from the Programming Research in Obesity, Growth, Environment and Social Stressors (PROGRESS) study, a prospective birth-cohort that enrolled 1050 pregnant women from Mexico City who were receiving prenatal care between December 2007 and July 2011. Quantitative PCR was used to calculate relative MtDNA content (mitochondrial-to-nuclear DNA ratio (mtDNA/nDNA)) in cord blood. Lead concentrations in both maternal blood (2nd and 3rd trimester and at delivery day) and in cord blood were measured by ICP-MS. Multivariable regression models adjusting for multiple confounders were fitted with 410 mother-infant pairs for whom complete data for mtDNA content, lead levels, and covariates were available.Maternal blood Pb measured in the second (mean 3.79 μg/dL, SD 2.63; β = 0.059, 95% CI 0.008, 0.111) and third trimester (mean 3.90 μg/dL; SD 2.84; β = 0.054, 95% CI 0.002, 0.107) during pregnancy and PB in cord blood (mean 3.50 μg/dL, SD 2.59; β = 0.050, 95% CI 0.004; 0.096) were associated with increased cord blood mtDNA content (mean 1.46, SD 0.44). In two-way interaction analyses, cord blood Pb marginally interacted with gestational age leading to an increase in mtDNA content for pre-term births (Benjamini-Hochberg False Discovery Rate correction; BH-FDR = 0.08).This study shows that lead exposure in pregnancy alters mtDNA content in cord blood; therefore, alteration of mtDNA content might be a mechanism underlying the toxicity of lead.

    View details for DOI 10.1016/j.envint.2019.01.077

    View details for Web of Science ID 000459005200041

    View details for PubMedID 30753999

    View details for PubMedCentralID PMC6391888

  • Epigenome-wide association study reveals methylation pathways associated with childhood allergic sensitization EPIGENETICS Peng, C., Van Meel, E. R., Cardenas, A., Rifas-Shiman, S. L., Sonawane, A. R., Glass, K. R., Gold, D. R., Platts-Mills, T. A., Lin, X., Oken, E., Hivert, M., Baccarelli, A. A., De Jong, N. W., Felix, J. F., Jaddoe, V. W., Duijts, L., Litonjua, A. A., DeMeo, D. L. 2019; 14 (5): 445-466


    Epigenetic mechanisms integrate both genetic variability and environmental exposures. However, comprehensive epigenome-wide analysis has not been performed across major childhood allergic phenotypes. We examined the association of epigenome-wide DNA methylation in mid-childhood peripheral blood (Illumina HumanMethyl450K) with mid-childhood atopic sensitization, environmental/inhalant and food allergen sensitization in 739 children in two birth cohorts (Project Viva-Boston, and the Generation R Study-Rotterdam). We performed covariate-adjusted epigenome-wide association meta-analysis and employed pathway and regional analyses of results. Seven-hundred and five methylation sites (505 genes) were significantly cross-sectionally associated with mid-childhood atopic sensitization, 1411 (905 genes) for environmental and 45 (36 genes) for food allergen sensitization (FDR<0.05). We observed differential methylation across multiple genes for all three phenotypes, including genes implicated previously in innate immunity (DICER1), eosinophilic esophagitis and sinusitis (SIGLEC8), the atopic march (AP5B1) and asthma (EPX, IL4, IL5RA, PRG2, SIGLEC8, CLU). In addition, most of the associated methylation marks for all three phenotypes occur in putative transcription factor binding motifs. Pathway analysis identified multiple methylation sites associated with atopic sensitization and environmental allergen sensitization located in/near genes involved in asthma, mTOR signaling, and inositol phosphate metabolism. We identified multiple differentially methylated regions associated with atopic sensitization (8 regions) and environmental allergen sensitization (26 regions). A number of nominally significant methylation sites in the cord blood analysis were epigenome-wide significant in the mid-childhood analysis, and we observed significant methylation - time interactions among a subset of sites examined. Our findings provide insights into epigenetic regulatory pathways as markers of childhood allergic sensitization.

    View details for DOI 10.1080/15592294.2019.1590085

    View details for Web of Science ID 000464645500001

    View details for PubMedID 30876376

    View details for PubMedCentralID PMC6557548

  • Prenatal maternal antidepressants, anxiety, and depression and offspring DNA methylation: epigenome-wide associations at birth and persistence into early childhood CLINICAL EPIGENETICS Cardenas, A., Faleschini, S., Hidalgo, A., Rifas-Shiman, S. L., Baccarelli, A. A., DeMeo, D. L., Litonjua, A. A., Neumann, A., Felix, J. F., Jaddoe, V. V., El Marroun, H., Tiemeier, H., Oken, E., Hivert, M., Burris, H. H. 2019; 11: 56


    Maternal mood disorders and their treatment during pregnancy may have effects on the offspring epigenome. We aim to evaluate associations of maternal prenatal antidepressant use, anxiety, and depression with cord blood DNA methylation across the genome at birth and test for persistence of associations in early and mid-childhood blood DNA.A discovery phase was conducted in Project Viva, a prospective pre-birth cohort study with external replication in an independent cohort, the Generation R Study. In Project Viva, pregnant women were recruited between 1999 and 2002 in Eastern Massachusetts, USA. In the Generation R Study, pregnant women were recruited between 2002 and 2006 in Rotterdam, the Netherlands. In Project Viva, 479 infants had data on maternal antidepressant use, anxiety, depression, and cord blood DNA methylation, 120 children had DNA methylation measured in early childhood (~ 3 years), and 460 in mid-childhood (~ 7 years). In the Generation R Study, 999 infants had data on maternal antidepressants and cord blood DNA methylation. The prenatal antidepressant prescription was obtained from medical records. At-mid pregnancy, symptoms of anxiety and depression were assessed with the Pregnancy-Related Anxiety Scale and the Edinburgh Postnatal Depression Scale in Project Viva and with the Brief Symptom Inventory in the Generation R Study. Genome-wide DNA methylation was measured using the Infinium HumanMethylation450 BeadChip in both cohorts.In Project Viva, 2.9% (14/479) pregnant women were prescribed antidepressants, 9.0% (40/445) experienced high pregnancy-related anxiety, and 8.2% (33/402) reported symptoms consistent with depression. Newborns exposed to antidepressants in pregnancy had 7.2% lower DNA methylation (95% CI, - 10.4, - 4.1; P = 1.03 × 10-8) at cg22159528 located in the gene body of ZNF575, and this association replicated in the Generation R Study (β = - 2.5%; 95% CI - 4.2, - 0.7; P = 0.006). In Project Viva, the association persisted in early (β = - 6.2%; 95% CI - 10.7, - 1.6) but not mid-childhood. We observed cohort-specific associations for maternal anxiety and depression in Project Viva that did not replicate.The ZNF575 gene is involved in transcriptional regulation but specific functions are largely unknown. Given the widespread use of antidepressants in pregnancy, as well as the effects of exposure to anxiety and depression, implications of potential fetal epigenetic programming by these risk factors and their impacts on development merit further investigation.

    View details for DOI 10.1186/s13148-019-0653-x

    View details for Web of Science ID 000462907800001

    View details for PubMedID 30925934

    View details for PubMedCentralID PMC6441191

  • Nutrition, DNA Methylation, and Developmental Origins of Cardiometabolic Disease: A Signal Systems Approach Handbook of Nutrition, Diet, and Epigenetics Laubach, Z. M., Faulk, C. D., Cardenas, A., Perng, W. Springer, Cham. 2019
  • Cross sectional association of arsenic and seroprevalence of hepatitis B infection in the United States (NHANES 2003-2014) ENVIRONMENTAL RESEARCH Cardenas, A., Smit, E., Welch, B. M., Bethel, J., Kile, M. L. 2018; 166: 570-576


    Arsenic alters immunological parameters including antibody formation and antigen-driven T-cell proliferation.We evaluated the cross-sectional relationship between urinary arsenic and the seroprevalence of hepatitis B (HBV) infection in the United States using data from six pooled cycles of the National Health and Nutrition Examination Survey (2003-2014, N = 12,447).Using serological data, participants were classified as susceptible, immune due to vaccination, or immune due to past natural infection. We used multinomial logistic regression to evaluate the association between urinary DMA and HBV classification. A sensitivity analysis using total urinary arsenic (TUA) was also conducted. Both DMA and TUA were adjusted for arsenobetaine using a residual regression method RESULTS: A 1-unit increase in the natural logarithm (ln) of DMA was associated with 40% greater adjusted odds of having immunity due to natural infection compared to being susceptible (Odds Ratio [aOR]: 1.40, 95% Confidence Intervals [CI] 1.15, 1.69), 65% greater odds of having immunity due to a natural infection (aOR: 1.65, 95% CI: 1.34, 2.04) and 18% greater odds of being susceptible (aOR: 1.18, 95% CI: 1.05, 1.33) compared to being immune due to vaccination after adjusting for creatinine, age, sex, race, income, country of birth, BMI, survey cycle, serum cotinine, recent seafood intake, and self-reported HBV immunization status.In the U.S. general public, higher urinary arsenic levels were associated with a greater odds of having a serological classification consistent with a past natural hepatitis B infection after adjusting for other risk factors. Additionally, higher urinary arsenic levels were linked to a greater odds of not receiving hepatitis B vaccinations. Given the cross-sectional nature of this analysis, more research is needed to test the hypothesis that environmentally relevant exposure to arsenic modulates host susceptibility to hepatitis B virus.

    View details for DOI 10.1016/j.envres.2018.06.023

    View details for Web of Science ID 000445318200063

    View details for PubMedID 29966877

  • Residential Proximity to Major Roadways at Birth, DNA Methylation at Birth and Midchildhood, and Childhood Cognitive Test Scores: Project Viva (Massachusetts, USA) ENVIRONMENTAL HEALTH PERSPECTIVES Peng, C., den Dekker, M., Cardenas, A., Rifas-Shiman, S. L., Gibson, H., Agha, G., Harris, M. H., Coull, B. A., Schwartz, J., Litonjua, A. A., DeMeo, D. L., Hivert, M., Gilman, M. W., Sagiv, S. K., de Kluizenaar, Y., Felix, J. F., Jaddoe, V. W., Oken, E., Duijts, L., Gold, D. R., Baccarelli, A. 2018; 126 (9): 97006


    Epigenetic variability is hypothesized as a regulatory pathway through which prenatal exposures may influence child development and health.We sought to examine the associations of residential proximity to roadways at birth and epigenome-wide DNA methylation. We also assessed associations of differential methylation with child cognitive outcomes.We estimated residential proximity to roadways at birth using a geographic information system (GIS) and cord blood methylation using Illumina's HumanMethylation450-array in 482 mother-child pairs in Project Viva. We identified individual CpGs associated with residential-proximity-to-roadways at birth using robust linear regression [[Formula: see text]]. We also estimated association between proximity-to-roadways at birth and methylation of the same sites in blood samples collected at age 7-11 y ([Formula: see text]). We ran the same analyses in the Generation R Study for replication ([Formula: see text]). In Project Viva, we investigated associations of differential methylation at birth with midchildhood cognition using linear regression.Living closer to major roadways at birth was associated with higher cord blood (and-more weakly-midchildhood blood) methylation of four sites in LAMB2. For each halving of residential-proximity-to-major-roadways, we observed a 0.82% increase in DNA methylation at cg05654765 [95% confidence interval (CI): (0.54%, 1.10%)], 0.88% at cg14099457 [95% CI: (0.56%, 1.19%)], 0.19% at cg03732535 [95% CI: (0.11%, 0.28)], and 1.08% at cg02954987 [95% CI: (0.65%, 1.51%)]. Higher cord blood methylation of these sites was associated with lower midchildhood nonverbal cognitive scores. Our results did not replicate in the Generation R Study.Our discovery results must be interpreted with caution, given that they were not replicated in a separate cohort. However, living close to major roadways at birth was associated with cord blood methylation of sites in LAMB2-a gene known to be linked to axonal development-in our U.S. cohort. Higher methylation of these sites associated with lower nonverbal cognitive scores at age 7-11 y in the same children.

    View details for DOI 10.1289/EHP2034

    View details for Web of Science ID 000449118800004

    View details for PubMedID 30226399

    View details for PubMedCentralID PMC6375460

  • Association of Perfluoroalkyl and Polyfluoroalkyl Substances With Adiposity JAMA NETWORK OPEN Cardenas, A., Hauser, R., Gold, D. R., Kleinman, K. P., Hivert, M., Fleisch, A. F., Lin, P. D., Calafat, A. M., Webster, T. F., Horton, E. S., Oken, E. 2018; 1 (4): e181493


    Perfluoroalkyl and polyfluoroalkyl substances (PFASs) are ubiquitous synthetic chemicals that are suspected endocrine disruptors.To determine the extent to which PFASs are associated with increases in weight and body size and evaluate whether a lifestyle intervention modifies this association.This prospective cohort study included 957 individuals who participated in the Diabetes Prevention Program trial, conducted from July 1996 to May 2001, and the Diabetes Prevention Program Outcomes Study, conducted from September 2002 to January 2014. Statistical analysis was conducted from September 1, 2017, to May 25, 2018.The initial lifestyle intervention consisted of training in diet, physical activity, and behavior modification, with the major goals of achieving 7% weight loss with subsequent maintenance and a minimum of 150 minutes per week of physical activity. Participants randomized to placebo received standard information about diet and exercise. A total of 6 plasma PFASs were quantified at baseline and 2 years after randomization, means were calculated from baseline and year 2 concentrations, and means were summed to assess total PFAS burden.Weight, waist circumference, and hip girth were measured at baseline and at scheduled visits.Of the 957 participants, 625 (65.3%) were women and 731 participants (76.4%) were between 40 and 64 years of age; 481 participants were randomized to the lifestyle intervention and 476 participants were randomized to the placebo arm. The PFAS concentrations were not different by treatment arm and were similar to concentrations reported for the US population in 1999-2000. The association of PFAS and weight change differed by treatment. Each doubling in total PFAS concentration was associated with an increase of 1.80 kg (95% CI, 0.43-3.17 kg; P = .01) from baseline to 9 years after randomization for the placebo group but not the lifestyle intervention group (-0.59 kg; 95% CI, -1.80 to 0.62 kg; P = .34). Similarly, each doubling in PFAS was associated with a 1.03-cm increase in hip girth in the Diabetes Prevention Program trial for the placebo group (95% CI, 0.18-1.88 cm; P = .02) but not the lifestyle intervention group (-0.09 cm; 95% CI, -0.82 to 0.63 cm; P = .80). No associations were observed for changes in mean waist circumference.Among adults at high risk for diabetes, higher plasma PFAS concentration was associated with increases in weight and hip girth over time, but a lifestyle intervention attenuated these associations. Diet and exercise may mitigate the obesogenic effects of environmental Identifier: NCT00004992 and NCT00038727.

    View details for DOI 10.1001/jamanetworkopen.2018.1493

    View details for Web of Science ID 000452643200010

    View details for PubMedID 30646133

    View details for PubMedCentralID PMC6324277

  • Placental DNA Methylation Adaptation to Maternal Glycemic Response in Pregnancy DIABETES Cardenas, A., Gagne-Ouellet, V., Allard, C., Brisson, D., Perron, P., Bouchard, L., Hivert, M. 2018; 67 (8): 1673-1683


    Maternal hyperglycemia during pregnancy is associated with excess fetal growth and adverse perinatal and developmental outcomes. Placental epigenetic maladaptation may underlie these associations. We performed an epigenome-wide association study (>850,000 CpG sites) of term placentas and prenatal maternal glycemic response 2-h post oral glucose challenge at 24-30 weeks of gestation among 448 mother-infant pairs. Maternal 2-h glycemia postload was strongly associated with lower DNA methylation of four CpG sites (false discovery rate [FDR] q <0.05) within the phosphodiesterase 4B gene (PDE4B). Additionally, three other individual CpG sites were differentially methylated relative to maternal glucose response within the TNFRSF1B, LDLR, and BLM genes (FDR q <0.05). DNA methylation correlated with expression of its respective genes in placental tissue at three out of four independent identified loci: PDE4B (r = 0.31, P < 0.01), TNFRSF1B (r = -0.24, P = 0.013), and LDLR (r = 0.32, P < 0.001). In an independent replication cohort (N = 65-108 samples), results were consistent in direction but not significantly replicated among tested CpG sites in PDE4B and TNFRSF1B Our study provides evidence that maternal glycemic response during pregnancy is associated with placental DNA methylation of key inflammatory genes whose expression levels are partially under epigenetic control.

    View details for DOI 10.2337/db18-0123

    View details for Web of Science ID 000439347300020

    View details for PubMedID 29752424

  • Epigenome-wide association study of Iota serum immunoglobulin E in children: a life course approach CLINICAL EPIGENETICS Peng, C., Cardenas, A., Rifas-Shiman, S. L., Hivert, M., Gold, D. R., Platts-Mills, T. A., Lin, X., Oken, E., Baccarelli, A. A., Litonjua, A. A., DeMeo, D. L. 2018; 10: 55


    IgE-mediated sensitization may be epigenetically programmed in utero, but early childhood environment may further alter complex traits and disease phenotypes through epigenetic plasticity. However, the epigenomic footprint underpinning IgE-mediated type-I hypersensitivity has not been well-understood, especially under a longitudinal early-childhood life-course framework.We used epigenome-wide DNA methylation (IlluminaHumanMethylation450 BeadChip) in cord blood and mid-childhood peripheral blood to investigate pre- and post-natal methylation marks associated with mid-childhood (age 6.7-10.2) total serum IgE levels in 217 mother-child pairs in Project Viva-a prospective longitudinal pre-birth cohort in eastern Massachusetts, USA. We identified methylation sites associated with IgE using covariate-adjusted robust linear regressions.Nineteen methylation marks in cord blood were associated with IgE in mid-childhood (FDR < 0.05) in genes implicated in cell signaling, growth, and development. Among these, two methylation sites (C7orf50, ZAR1) remained robust after the adjustment for the change in DNA methylation from birth to mid-childhood (FDR < 0.05). An analysis of the change in methylation between cord blood and mid-childhood DNA (Δ-DNAm) identified 395 methylation marks in 272 genes associated with mid-childhood IgE (FDR < 0.05), with multiple sites located within ACOT7 (4 sites), EPX (5 sites), EVL (3 sites), KSR1 (4 sites), ZFPM1 (3 sites), and ZNF862 (3 sites). Several of these methylation loci were previously associated with asthma (ADAM19, EPX, IL4, IL5RA, and PRG2).This study identified fetally programmed and mid-childhood methylation signals associated with mid-childhood IgE. Epigenetic priming during fetal development and early childhood likely plays an important role in IgE-mediated type-I hypersensitivity.

    View details for DOI 10.1186/s13148-018-0488-x

    View details for Web of Science ID 000430759600002

    View details for PubMedID 29692868

    View details for PubMedCentralID PMC5905182

  • Trends in urinary arsenic among the US population by drinking water source: Results from the National Health and Nutritional Examinations Survey 2003-2014 ENVIRONMENTAL RESEARCH Welch, B., Smit, E., Cardenas, A., Hystad, P., Kile, M. L. 2018; 162: 8-17


    In 2001, the United States revised the arsenic maximum contaminant level for public drinking water systems from 50µg/L to 10µg/L. This study aimed to examine temporal trends in urinary arsenic concentrations in the U.S. population from 2003 to 2014 by drinking water source among individuals aged 12 years and older who had no detectable arsenobetaine - a biomarker of arsenic exposure from seafood intake.We examined data from 6 consecutive cycles of the National Health and Nutrition Examination Survey (2003-2014; N=5848). Total urinary arsenic (TUA) was calculated by subtracting arsenobetaine's limit of detection and detectable arsenocholine from total arsenic. Additional sensitivity analyses were conducted using a second total urinary arsenic index (TUA2, calculated by adding arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid). We classified drinking water source using 24-h dietary questionnaire data as community supply (n=3427), well or rain cistern (n=506), and did not drink tap water (n=1060).Geometric means (GM) of survey cycles were calculated from multivariate regression models adjusting for age, gender, race/ethnicity, BMI, income, creatinine, water source, type of water consumed, recent smoking, and consumption of seafood, rice, poultry, and juice. Compared to 2003-2004, adjusted TUA was 35.5% lower in 2013-2014 among the general U.S.Stratified analysis by smoking status indicated that the trend in lower TUA was only consistent among non-smokers. Compared to 2003-2004, lower adjusted TUA was observed in 2013-2014 among non-smoking participants who used community water supplies (1.98 vs 1.16µg/L, p<0.001), well or rain cistern users (1.54 vs 1.28µg/L, p<0.001) and who did not drink tap water (2.24 vs 1.53µg/L, p<0.001). Sensitivity analyses showed consistent results for participants who used a community water supplier and to a lesser extent those who did not drink tap water. However, the sensitivity analysis showed overall exposure stayed the same or was higher among well or rain cistern users. Finally, the greatest decrease in TUA was among participants within the highest exposure percentiles (e.g. 95th percentile had 34% lower TUA in 2013/2014 vs 2003/2004, p<0.001).Overall, urinary arsenic levels in the U.S. population declined over a 12-year period that encompassed the adoption of the revised Arsenic Rule. The most consistent trends in declining exposure were observed among non-smoking individuals using public community water systems. These results suggest regulation and prevention strategies to reduce arsenic exposures in the U.S. may be succeeding.

    View details for DOI 10.1016/j.envres.2017.12.012

    View details for Web of Science ID 000426325300002

    View details for PubMedID 29272814

    View details for PubMedCentralID PMC5811395

  • Cumulative exposure to environmental pollutants during early pregnancy and reduced fetal growth: the Project Viva cohort ENVIRONMENTAL HEALTH Rokoff, L. B., Rifas-Shiman, S. L., Coull, B. A., Cardenas, A., Calafat, A. M., Ye, X., Gryparis, A., Schwartz, J., Sagiv, S. K., Gold, D. R., Oken, E., Fleisch, A. F. 2018; 17: 19


    Reduced fetal growth is associated with perinatal and later morbidity. Prenatal exposure to environmental pollutants is linked to reduced fetal growth at birth, but the impact of concomitant exposure to multiple pollutants is unclear. The purpose of this study was to examine interactions between early pregnancy exposure to cigarette smoke, traffic pollution, and select perfluoroalkyl substances (PFASs) on birth weight-for-gestational age (BW/GA).Among 1597 Project Viva mother-infant pairs, we assessed maternal cigarette smoking by questionnaire, traffic pollution at residential address by black carbon land use regression model, and plasma concentration of select PFASs in early pregnancy. We calculated sex-specific BW/GA z-scores, an index of fetal growth, from national reference data. We fit covariate-adjusted multi-pollutant linear regression models and examined interactions between exposures, using a likelihood-ratio test to identify a best-fit model.Two hundred six (13%) mothers smoked during pregnancy. Mean [standard deviation (SD)] for black carbon was 0.8 (0.3) μg/m3, perfluorooctane sulfonate (PFOS) was 29.1 (16.5) ng/mL, and BW/GA z-score was 0.19 (0.96). In the best-fit model, BW/GA z-score was lower in infants of mothers exposed to greater black carbon [- 0.08 (95% CI: -0.15, - 0.01) per interquartile range (IQR)]. BW/GA z-score (95% CI) was also lower in infants of mothers who smoked [- 0.09 (- 0.23, 0.06)] or were exposed to greater PFOS [- 0.03 (- 0.07, 0.02) per IQR], although confidence intervals crossed the null. There were no interactions between exposures. In secondary analyses, instead of PFOS, we examined perfluorononanoate (PFNA) [mean (SD): 0.7 (0.4) ng/mL], a PFAS more closely linked to lower BW/GA in our cohort. The best-fit multi-pollutant model included positive two-way interactions between PFNA and both black carbon and smoking (p-interactions = 0.03).Concurrent prenatal exposures to maternal smoking, black carbon, and PFOS are additively associated with lower fetal growth, whereas PFNA may attenuate associations of smoking and black carbon with lower fetal growth. It is important to examine interactions between multiple exposures in relation to health outcomes, as effects may not always be additive and may shed light on biological pathways.

    View details for DOI 10.1186/s12940-018-0363-4

    View details for Web of Science ID 000425789500003

    View details for PubMedID 29458383

    View details for PubMedCentralID PMC5819079

  • DNA methylation in blood as a mediator of the association of mid-childhood body mass index with cardio-metabolic risk score in early adolescence EPIGENETICS Huang, J. V., Cardenas, A., Colicino, E., Schooling, C., Rifas-Shiman, S. L., Agha, G., Zheng, Y., Hou, L., Just, A. C., Litonjua, A. A., DeMeo, D. L., Lin, X., Oken, E., Hivert, M., Baccarelli, A. A. 2018; 13 (10-11): 1072-1087


    Obesity is associated with higher cardio-metabolic risk even in childhood and adolescence; whether this association is mediated by epigenetic mechanisms remains unclear. We examined the extent to which mid-childhood body mass index (BMI) z-score (median age 7.7 years) was associated with cardio-metabolic risk score in early adolescence (median age 12.9 years) via mid-childhood DNA methylation among 265 children in the Project Viva. We measured DNA methylation in leukocytes using the Infinium Human Methylation450K BeadChip. We assessed mediation CpG-by-CpG using epigenome-wide association analyses, high-dimensional mediation analysis, and natural effect models. We observed mediation by mid-childhood DNA methylation at 6 CpGs for the association between mid-childhood BMI z-score and cardio-metabolic risk score in early adolescence in the high-dimensional mediation analysis (accounting for 10% of the total effect) and in the natural effect model (β = 0.04, P = 3.2e-2, accounting for 13% of the total effect). The natural direct effect of BMI z-score on cardio-metabolic risk score was still evident (β = 0.27, P = 1.1e-25). We also observed mediation by mid-childhood DNA methylation at 5 CpGs that was in the opposite direction from the total effect (natural effect model: β = -0.04, P = 2.0e-2). Mediation in different directions implies a complex role of DNA methylation in the association between BMI and cardio-metabolic risk and needs further investigation. Future studies with larger sample size and greater variability in cardio-metabolic risk will further help elucidate the role of DNA methylation for cardio-metabolic risk.

    View details for DOI 10.1080/15592294.2018.1543503

    View details for Web of Science ID 000456104500005

    View details for PubMedID 30412002

    View details for PubMedCentralID PMC6342073

  • DNA methylation in cord blood as mediator of the association between prenatal arsenic exposure and gestational age EPIGENETICS Bozack, A. K., Cardenas, A., Quamruzzaman, Q., Rahman, M., Mostofa, G., Christiani, D. C., Kile, M. L. 2018; 13 (9): 923-940


    Prenatal arsenic exposure is associated with adverse birth outcomes and disease risk later in life, which could be mediated through epigenetic dysregulation. We evaluated the association between arsenic and gestational age (GA) that was mediated through DNA methylation (DNAm) using data from a Bangladeshi birth cohort. Arsenic exposure was measured in maternal drinking water at ≤16 weeks GA and maternal toenails collected ≤1 month postpartum. Cord blood DNAm was measured using Infinium HumanMethylation450 arrays (n = 44, discovery phase). Top loci identified in the discovery phase were then pyrosequenced in a second group (n = 569, validation phase). Structural equation models (SEM) evaluated the direct and indirect effects of arsenic and DNAm on GA. In the discovery phase, arsenic was associated with differential DNAm of 139 loci that were associated with GA (P < 1.10X10-6; |β regression|>0.10). Each doubling in water arsenic concentration decreased GA by 2 days, which was fully mediated through the main principal component of the top-ten CpGs (P < 0.001). In the validation phase, there were direct and indirect effects of miR214-3 and MCC DNAm on GA. In an adjusted SEM model, mediation of the association between arsenic and GA by miR124-3 was borderline significant (P = 0.061). This study therefore identified DNAm at specific loci in cord blood that mediated the effect of arsenic exposure on GA. Specifically, prenatal arsenic exposure was associated with lower methylation of miR124-3 that mediated the exposure-response of arsenic on GA. Future research should evaluate if these epigenetic changes are persistent and associated with disease risk.

    View details for DOI 10.1080/15592294.2018.1516453

    View details for Web of Science ID 000450445600003

    View details for PubMedID 30175652

    View details for PubMedCentralID PMC6284783

  • Plasma Concentrations of Per- and Polyfluoroalkyl Substances at Baseline and Associations with Glycemic Indicators and Diabetes Incidence among High-Risk Adults in the Diabetes Prevention Program Trial ENVIRONMENTAL HEALTH PERSPECTIVES Cardenas, A., Gold, D. R., Hauser, R., Kleinman, K. P., Hivert, M., Calafat, A. M., Ye, X., Webster, T. F., Horton, E. S., Oken, E. 2017; 125 (10): 107001


    Several per- and polyfluoroalkyl substances (PFAS) are ubiquitous anthropogenic pollutants almost universally detected in humans. Experimental evidence indicates that PFAS alter glucose metabolism and insulin secretion. However, epidemiological studies have yielded inconsistent results.We sought to examine associations between plasma PFAS concentrations, glycemic indicators, and diabetes incidence among high-risk adults.Within the Diabetes Prevention Program (DPP), a trial for the prevention of type 2 diabetes among high-risk individuals, we quantified baseline plasma concentrations of nine PFAS among 957 participants randomized to a lifestyle intervention or placebo. We evaluated adjusted associations for plasma PFAS concentrations with diabetes incidence and key glycemic indicators measured at baseline and annually over up to 4.6 y.Plasma PFAS concentrations were similar to those reported in the U.S. population in 1999-2000. At baseline, in cross-sectional analysis, a doubling in plasma perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid (PFOA) concentrations was associated with higher homeostatic model assessment of insulin resistance (HOMA-IR) [βPFOS=0.39; 95% confidence interval (CI): 0.13, 0.66; βPFOA=0.64; 95% CI: 0.34, 0.94], β-cell function (HOMA-β) (βPFOS=9.62; 95% CI: 1.55, 17.70; βPFOA=15.93; 95% CI: 6.78, 25.08), fasting proinsulin (βPFOS=1.37 pM; 95% CI: 0.50, 2.25; βPFOA=1.71 pM; 95% CI: 0.72, 2.71), and glycated hemoglobin (HbA1c) (βPFOS=0.03%; 95% CI: 0.002, 0.07; βPFOA=0.04%; 95% CI: 0.001, 0.07). There was no strong evidence of associations between plasma PFAS concentrations and diabetes incidence or prospective changes in glycemic indicators during the follow-up period.At baseline, several PFAS were cross-sectionally associated with small differences in markers of insulin secretion and β-cell function. However, there was limited evidence suggesting that PFAS concentrations are associated with diabetes incidence or changes in glycemic indicators during the follow-up period.

    View details for DOI 10.1289/EHP1612

    View details for Web of Science ID 000413793300013

    View details for PubMedID 28974480

    View details for PubMedCentralID PMC5933403

  • Prenatal Exposure to Mercury: Associations with Global DNA Methylation and Hydroxymethylation in Cord Blood and in Childhood. Environmental health perspectives Cardenas, A., Rifas-Shiman, S. L., Godderis, L., Duca, R. C., Navas-Acien, A., Litonjua, A. A., DeMeo, D. L., Brennan, K. J., Amarasiriwardena, C. J., Hivert, M. F., Gillman, M. W., Oken, E., Baccarelli, A. A. 2017; 125 (8): 087022


    Mercury is a global pollutant, and prenatal exposure is associated with adverse health effects. To date, no studies have evaluated the association between prenatal mercury exposure and DNA hydroxymethylation, an epigenetic modification important for tissue differentiation and embryonic development.We sought to evaluate the association between prenatal mercury exposure and offspring global DNA methylation and hydroxymethylation at birth and test for persistence of the association in childhood.Within Project Viva, a U.S. prebirth cohort, we examined associations of maternal second trimester red blood cell mercury (RBC-Hg) concentrations with global 5-hydroxymethylcytosine (%-5hmC) and 5-methylcytosine (%-5mC) DNA content in blood collected at birth (n=306), early childhood (n=68; 2.9 to 4.9 y), and midchildhood (n=260; 6.7 to 10.5 y).Median prenatal RBC-Hg concentration was 3.23μg/g [interquartile range (IQR)=3.29]. At birth, median cord blood %-5mC, %-5hmC, and their ratio were 4.95%, 0.22%, and 24.37, respectively. The mean adjusted difference [95% confidence interval (CI)] of blood %-5hmC for a doubling in prenatal RBC-Hg concentration was -0.013% (-0.029, 0.002), -0.031% (-0.056, -0.006), and 0.005% (-0.007, 0.018) at birth, early, and midchildhood, respectively. The corresponding relative adjusted change in the genomic ratio of %-5mC to %-5hmC for a doubling in prenatal RBC-Hg concentration was 4.70% (0.04, 9.58), 22.42% (7.73, 39.11), and 0.73% (-4.18, 5.88) at birth, early, and midchildhood, respectively. No associations were present between prenatal maternal RBC-Hg and %-5mC at any time point.Prenatal mercury exposure was associated with lower %-5hmC genomic content and a corresponding increase in the ratio of %-5mC to %-5hmC in cord blood. This association was persistent in early but not midchildhood blood. Our results demonstrate the potential malleability of epigenetic modifications associated with mercury exposure in utero.

    View details for DOI 10.1289/EHP1467

    View details for PubMedID 28934725

    View details for PubMedCentralID PMC5783674

  • Cord blood DNA methylation and adiposity measures in early and mid-childhood CLINICAL EPIGENETICS Kresovich, J. K., Zheng, Y., Cardenas, A., Joyce, B. T., Rifas-Shiman, S. L., Oken, E., Gillman, M. W., Hivert, M., Baccarelli, A. A., Hou, L. 2017; 9: 86


    Excess adiposity in childhood is associated with numerous adverse health outcomes. As this condition is difficult to treat once present, identification of risk early in life can help inform and implement strategies to prevent the onset of the condition. We performed an epigenome-wide association study to prospectively investigate the relationship between cord blood DNA methylation and adiposity measurements in childhood.We measured genome-wide DNA methylation from 478 children in cord blood and measured overall and central adiposity via skinfold caliper measurements in early (range 3.1-3.3 years) and mid-childhood (age range 7.3-8.3 years) and via dual X-ray absorptiometry (DXA) in mid-childhood. Final models were adjusted for maternal age at enrollment, pre-pregnancy body mass index, education, folate intake during pregnancy, smoking during pregnancy, and gestational weight gain, and child sex, race/ethnicity, current age, and cord blood cell composition.We identified four promoter proximal CpG sites that were associated with adiposity as measured by subscapular (SS) and triceps (TR) ratio (SS:TR) in early childhood, in the genes KPRP, SCL9A10, MYLK2, and PRLHR. We additionally identified one gene body CpG site associated with early childhood SS + TR on PPAPDC1A; this site was nominally associated with SS + TR in mid-childhood. Higher methylation at one promoter proximal CpG site in MMP25 was also associated with SS:TR in mid-childhood. In regional analyses, methylation at an exonal region of GFPT2 was positively associated with SS:TR in early childhood. Finally, we identified regions of two long, non-coding RNAs which were associated with SS:TR (LOC100049716) and fat-free mass index (LOC102723493) in mid-childhood.This analysis identified novel CpG loci associated with adiposity outcomes. However, our results suggest little consistency across the various adiposity outcomes tested, particularly among the more accurate DXA measurements of body composition. We recommend using caution when interpreting these associations.

    View details for DOI 10.1186/s13148-017-0384-9

    View details for Web of Science ID 000408024500003

    View details for PubMedID 28814982

    View details for PubMedCentralID PMC5558655

  • Prenatal Exposure to Mercury: Associations with Global DNA Methylation and Hydroxymethylation in Cord Blood and in Childhood ENVIRONMENTAL HEALTH PERSPECTIVES Cardenas, A., Rifas-Shiman, S. L., Godderis, L., Duca, R., Navas-Acien, A., Litonjua, A. A., DeMeo, D. L., Brennan, K. J., Amarasiriwardena, C. J., Hivert, M., Gillman, M. W., Oken, E., Baccarelli, A. A. 2017; 125 (8)

    View details for DOI 10.1289/EHP1467

    View details for Web of Science ID 000461491100002

  • Exposure to Low Levels of Lead in Utero and Umbilical Cord Blood DNA Methylation in Project Viva: An Epigenome-Wide Association Study ENVIRONMENTAL HEALTH PERSPECTIVES Wu, S., Hivert, M., Cardenas, A., Zhong, J., Rifas-Shiman, S. L., Agha, G., Colicino, E., Just, A. C., Amarasiriwardena, C., Lin, X., Litonjua, A. A., DeMeo, D. L., Gillman, M. W., Wright, R. O., Oken, E., Baccarelli, A. A. 2017; 125 (8): 087019


    Early-life exposure to lead is associated with deficits in neurodevelopment and with hematopoietic system toxicity. DNA methylation may be one of the underlying mechanisms for the adverse effects of prenatal lead on the offspring, but epigenome-wide methylation data for low levels of prenatal lead exposure are lacking.We investigated the association between prenatal maternal lead exposure and epigenome-wide DNA methylation in umbilical cord blood nucleated cells in Project Viva, a prospective U.S.-based prebirth cohort with relatively low levels of lead exposure.Among 268 mother-infant pairs, we measured lead concentrations in red blood cells (RBC) from prenatal maternal blood samples, and using HumanMethylation450 Bead Chips, we measured genome-wide methylation levels at 482,397 CpG loci in umbilical cord blood and retained 394,460 loci after quality control. After adjustment for batch effects, cell types, and covariates, we used robust linear regression models to examine associations of prenatal lead exposure with DNA methylation in cord blood at epigenome-wide significance level [false discovery rate (FDR)<0.05].The mean [standard deviation (SD)] maternal RBC lead level was 1.22 (0.63) μg/dL. CpG cg10773601 showed an epigenome-wide significant negative association with prenatal lead exposure (-1.4% per doubling increase in lead exposure; p=2.3×10-7) and was annotated to C-Type Lectin Domain Family 11, Member A (CLEC11A), which functions as a growth factor for primitive hematopoietic progenitor cells. In sex-specific analyses, we identified more CpGs with FDR<0.05 among female infants (n=38) than among male infants (n=2). One CpG (cg24637308), which showed a strong negative association with prenatal lead exposure among female infants (-4.3% per doubling increase in lead exposure; p=1.1×10-06), was annotated to Dynein Heavy Chain Domain 1 gene (DNHD1) which is highly expressed in human brain. Interestingly, there were strong correlations between blood and brain methylation for CpG (cg24637308) based on another independent set of samples with a high proportion of female participants.Prenatal low-level lead exposure was associated with newborn DNA methylation, particularly in female infants.

    View details for DOI 10.1289/EHP1246

    View details for Web of Science ID 000413790600029

    View details for PubMedID 28858830

    View details for PubMedCentralID PMC5783669

  • Maternal intake of fried foods and risk of gestational diabetes mellitus ANNALS OF EPIDEMIOLOGY Osorio-Yanez, C., Gelaye, B., Qiu, C., Bao, W., Cardenas, A., Enquobahrie, D. A., Williams, M. A. 2017; 27 (6): 384-390


    We examined the relationship of maternal periconceptional (i.e., before conception and early pregnancy) intake of fried foods with gestational diabetes mellitus (GDM) risk.In a prospective birth cohort in Seattle and Tacoma, Washington State, USA, we assessed maternal periconceptional fried food intake using a food frequency questionnaire among 3414 participants. We used multivariable generalized linear regression models to derive estimates of relative risks (RRs; and 95% confidence intervals, 95% CIs) of GDM in relation to the intake of different types of fried foods (i.e., fried fish, fried chicken, fried potatoes, chips, and donuts).A total of 169 GDM incident cases were identified in this cohort (4.96%). Compared with no fried fish intake, fried fish intake >1 servings/month was associated with 68% higher GDM risk (adjusted RR and 95% CI; 1.68 [1.16, 2.45]; Ptrend = .019). After adjusting for confounders, the RRs (95% CI) of GDM relative to fried chicken intake were 1.0, 1.44 (0.98, 2.09), and 1.81 (1.22, 2.70) for none, ≤1 and > 1 servings/month intake of fried chicken, respectively (Ptrend = .002). Dietary intake of fried potatoes, snack chips or donuts was not significantly associated with higher GDM risk. Limitations of our study include the lack of information about frying methods and the intake of fried foods at home and away from home.Regular intake of fried fish and fried chicken are associated with elevated GDM risk.

    View details for DOI 10.1016/j.annepidem.2017.05.006

    View details for Web of Science ID 000404807800004

    View details for PubMedID 28641758

    View details for PubMedCentralID PMC5578760

  • Persistent DNA methylation changes associated with prenatal mercury exposure and cognitive performance during childhood SCIENTIFIC REPORTS Cardenas, A., Rifas-Shiman, S. L., Agha, G., Hivert, M., Litonjua, A. A., DeMeo, D. L., Lin, X., Amarasiriwardena, C. J., Oken, E., Gillman, M. W., Baccarelli, A. A. 2017; 7: 288


    Prenatal exposure to mercury, a known neurotoxic metal, is associated with lower cognitive performance during childhood. Disruption of fetal epigenetic programming could explain mercury's neurodevelopmental effects. We screened for epigenome-wide methylation differences associated with maternal prenatal blood mercury levels in 321 cord blood DNA samples and examined the persistence of these alterations during early (n = 75; 2.9-4.9 years) and mid-childhood (n = 291; 6.7-10.5 years). Among males, prenatal mercury levels were associated with lower regional cord blood DNA methylation at the Paraoxonase 1 gene (PON1) that persisted in early childhood and was attenuated in mid-childhood blood. Cord blood methylation at the PON1 locus predicted lower cognitive test scores measured during early childhood. Methylation at the PON1 locus was associated with PON1 expression in an independent set of cord blood samples. The observed persistent epigenetic disruption of the PON1 gene may modulate mercury toxicity in humans and might serve as a biomarker of exposure and disease susceptibility.

    View details for DOI 10.1038/s41598-017-00384-5

    View details for Web of Science ID 000396847900049

    View details for PubMedID 28325913

    View details for PubMedCentralID PMC5428306

  • Cross-sectional study of social behaviors in preschool children and exposure to flame retardants ENVIRONMENTAL HEALTH Lipscomb, S. T., McClelland, M. M., MacDonald, M., Cardenas, A., Anderson, K. A., Kile, M. L. 2017; 16: 23


    Children are exposed to flame retardants from the built environment. Brominated diphenyl ethers (BDE) and organophosphate-based flame retardants (OPFRs) are associated with poorer neurocognitive functioning in children. Less is known, however, about the association between these classes of compounds and children's emotional and social behaviors. The objective of this study was to determine if flame retardant exposure was associated with measurable differences in social behaviors among children ages 3-5 years.We examined teacher-rated social behaviors measured using the Social Skills Improvement Rating Scale (SSIS) and personal exposure to flame retardants in children aged 3-5 years who attended preschool (n = 72). Silicone passive samplers worn for 7 days were used to assess personal exposure to 41 compounds using gas chromatography-mass spectrophotometer. These concentrations were then summed into total BDE and total OPFR exposure prior to natural log transformation. Separate generalized additive models were used to evaluate the relationship between seven subscales of the SSIS and lnΣBDE or lnΣOPFR adjusting for other age, sex, adverse social experiences, and family context.All children were exposed to a mixture of flame retardant compounds. We observed a dose dependent relationship between lnΣOPFR and two subscales where children with higher exposures were rated by their preschool teachers as having less responsible behavior (p = 0.07) and more externalizing behavior problems (p = 0.03). Additionally, children with higher lnΣBDE exposure were rated by teachers as less assertive (p = 0.007).We observed a cross-sectional association between children's exposure to flame retardant compounds and teacher-rated social behaviors among preschool-aged children. Children with higher flame retardant exposures exhibited poorer social skills in three domains that play an important role in a child's ability to succeed academically and socially.

    View details for DOI 10.1186/s12940-017-0224-6

    View details for Web of Science ID 000395975100002

    View details for PubMedID 28274271

    View details for PubMedCentralID PMC5343384

  • Perioperative supplementation of polyunsaturated omega-3 fatty acid for the prevention of atrial fibrillation after cardiothoracic surgery AMERICAN JOURNAL OF HEALTH-SYSTEM PHARMACY Joss, J. D., Hernan, J., Collier, R., Cardenas, A. 2017; 74 (1): E17-E23


    The effect of perioperative supplementation of polyunsaturated omega-3 fatty acid (PUFA) on the frequency of atrial fibrillation (AF) in patients without a history of AF was evaluated.A total of 561 patients were randomized to receive either 1 g of PUFA or placebo twice daily. Treatment was started up to five days before surgery or within 24 hours after surgery, depending on when consent was received. Treatment was continued until the patient's follow-up visit with the cardiologist up to four weeks after surgery. The primary endpoint of the study was AF before hospital discharge. Secondary endpoints included AF within one week after surgery, AF within one month after surgery, length of hospital stay, postoperative bleeding complications, and readmission within one month after surgery.No significant reduction in the risk of AF was observed at hospital discharge (relative risk [RR], 0.98; p = 0.922) or at three weeks after surgery (RR, 0.98; p = 0.844). After restricting the analysis to treatment-adherent patients, the association remained nonsignificant at hospital discharge (RR, 0.90; p = 0.374) and at three weeks after surgery (RR, 0.90; p = 0.330). No significant differences were observed between treatments for rates of readmission, death, and bleeding complications or the length of hospital stay.Perioperative supplementation with PUFA did not decrease the risk of AF in the immediate postoperative period. PUFA also had no effect on the length of hospital stay, postoperative bleeding complications, and readmissions within one month after surgery.

    View details for DOI 10.2146/ajhp150740

    View details for Web of Science ID 000405931700006

    View details for PubMedID 28007717

  • Assessment of Hospital Staff's Knowledge of Osteopathic Manipulative Medicine: A Survey-Based Study JOURNAL OF THE AMERICAN OSTEOPATHIC ASSOCIATION Smith-Kelly, J. B., Cardenas, A. 2016; 116 (12): 764-769


    Research has shown that osteopathic manipulative treatment (OMT) in hospitalized patients can shorten hospital length of stay. However, hospital staff may be unfamiliar with OMT and its use in this setting.To assess a hospital staff's knowledge of osteopathic manipulative medicine (OMM) and investigate whether there is a correlation between job category and knowledge of OMM.The study used a 13-item survey that was developed using SurveyMonkey. A brief description stating the purpose of the survey with a hyperlink to the survey was sent in an e-mail to the employee LISTSERV at Good Samaritan Regional Medical Center. This LISTSERV included all 1933 employees at the medical center. After 10 days the survey was closed and the responses were collected and interpreted by a statistician and the primary investigator. Incomplete surveys were included.A total of 474 employees (24.5%) returned the survey. Of these respondents, 267 (66.9%) responded that OMM could be done in the hospital. Only 97 respondents (24.6%) reported seeing OMM performed in the hospital. Physicians had the highest awareness of OMM (53.7%), compared with all other employees (7%).An overall lack of knowledge exists regarding OMM among hospital staff, especially nonphysician employees, at a medical clinic.

    View details for DOI 10.7556/jaoa.2016.151

    View details for Web of Science ID 000393050400002

    View details for PubMedID 27893142

  • Arsenic exposure and the seroprevalence of total hepatitis A antibodies in the US population: NHANES, 2003-2012 EPIDEMIOLOGY AND INFECTION Cardenas, A., Smit, E., Bethel, J. W., Houseman, E. A., Kile, M. L. 2016; 144 (8): 1641-1651


    We evaluated the association between urinary arsenic and the seroprevalence of total hepatitis A antibodies (total anti-HAV: IgG and IgM) in 11 092 participants aged ⩾6 years using information collected in the US National Health and Nutrition Examination Survey (2003-2012). Multivariate logistic regression models evaluated associations between total anti-HAV and total urinary arsenic defined as the sum of arsenite, arsenate, monomethylarsonate and dimethylarsinate (TUA1). Effect modification by self-reported HAV immunization status was evaluated. Total anti-HAV seroprevalence was 35·1% [95% confidence interval (CI) 33·3-36·9]. Seropositive status was associated with higher arsenic levels and this association was modified by immunization status (P = 0·03). For participants that received ⩾2 vaccine doses or did not know if they had received any doses, a positive dose-response association was observed between increasing TUA1 and odds of total anti-HAV [odds ratio (OR) 1·42, 95% CI 1·11-1·81; and OR 1·75, 95% CI 1·22-2·52], respectively. A positive but not statistically significant association was observed in those who received <2 doses (OR 1·46, 95% CI 0·83-2·59) or no dose (OR 1·12, 95% CI 0·98-1·30). Our analysis indicates that prevalent arsenic exposure was associated with positive total anti-HAV seroprevalence. Further studies are needed to determine if arsenic increases the risk for incident hepatitis A infection or HAV seroconversion.

    View details for DOI 10.1017/S0950268815003088

    View details for Web of Science ID 000375804300008

    View details for PubMedID 26739255

    View details for PubMedCentralID PMC4855991

  • Estimating Effects of Arsenic Exposure During Pregnancy on Perinatal Outcomes in a Bangladeshi Cohort EPIDEMIOLOGY Kile, M. L., Cardenas, A., Rodrigues, E., Mazumdar, M., Dobson, C., Golam, M., Quamruzzaman, Q., Rahman, M., Christiani, D. C. 2016; 27 (2): 173-181


    The relationship between arsenic and birth weight is not well understood. The objective was to evaluate the causal relationship between prenatal arsenic exposure and birth weight considering the potential mediation effects of gestational age and maternal weight gain during pregnancy using structural equation models.A prospectively enrolled cohort of pregnant women was recruited in Bangladesh from 2008 to 2011. Arsenic was measured in personal drinking water at the time of enrollment (gestational age <16 weeks, N = 1,140) and in toenails collected ≤1 month postpartum (N = 624) using inductively coupled plasma mass spectrometry. Structural equation models estimated the direct and indirect effects of arsenic on birth weight with gestational age and maternal weight gain considered as mediating variables.Every unit increase in natural log water arsenic was indirectly associated with decreased birth weight (β = -19.17 g, 95% confidence interval [CI]: -24.64, -13.69) after adjusting for other risk factors. This association was mediated entirely through gestational age (β = -17.37 g, 95% CI: -22.77, -11.98) and maternal weight gain during pregnancy (β = -1.80 g, 95% CI: -3.72, 0.13). When exposure was modeled using toenail arsenic concentrations, similar results were observed. Every increase in natural log toenail arsenic was indirectly associated with decreased birth weight (β = -15.72 g, 95% CI: -24.52, -6.91) which was mediated through gestational age (β = -13.59 g, 95% CI: -22.10, -5.07) and maternal weight gain during pregnancy (β = -2.13 g, 95% CI: -5.24, 0.96).Arsenic exposure during pregnancy was associated with lower birth weight. The effect of arsenic on birth weight appears to be mediated mainly through decreasing gestational age and to a lesser extent by lower maternal weight gain during pregnancy.

    View details for DOI 10.1097/EDE.0000000000000416

    View details for Web of Science ID 000369866900003

    View details for PubMedID 26583609

    View details for PubMedCentralID PMC4733817

  • Validation of a DNA methylation reference panel for the estimation of nucleated cells types in cord blood EPIGENETICS Cardenas, A., Allard, C., Doyon, M., Houseman, E., Bakulski, K. M., Perron, P., Bouchard, L., Hivert, M. 2016; 11 (11): 773-779


    Cord blood is widely used as surrogate tissue in epigenome-wide association studies of prenatal conditions. Cell type composition variation across samples can be an important confounder of epigenome-wide association studies in blood that constitute a mixture of cells. We evaluated a newly developed cord blood reference panel to impute cell type composition from DNA methylation levels, including nucleated red blood cells (nRBCs). We estimated cell type composition from 154 unique cord blood samples with available DNA methylation data as well as direct measurements of nucleated cell types. We observed high correlations between the estimated and measured composition for nRBCs (r = 0.92, R2 = 0.85), lymphocytes (r = 0.77, R2 = 0.58), and granulocytes (r = 0.72, R2 = 0.52), and a moderate correlation for monocytes (r = 0.51, R2 = 0.25) as well as relatively low root mean square errors from the residuals ranging from 1.4 to 5.4%. These results validate the use of the cord blood reference panel and highlight its utility and limitations for epidemiological studies.

    View details for DOI 10.1080/15592294.2016.1233091

    View details for Web of Science ID 000388622200001

    View details for PubMedID 27668573

    View details for PubMedCentralID PMC5221601

  • In utero arsenic exposure and epigenome-wide associations in placenta, umbilical artery, and human umbilical vein endothelial cells EPIGENETICS Cardenas, A., Houseman, E., Baccarelli, A. A., Quamruzzaman, Q., Rahman, M., Mostofa, G., Wright, R. O., Christiani, D. C., Kile, M. L. 2015; 10 (11): 1054-1063


    Exposure to arsenic early in life has been associated with increased risk of several chronic diseases and is believed to alter epigenetic programming in utero. In the present study, we evaluate the epigenome-wide association of arsenic exposure in utero and DNA methylation in placenta (n = 37), umbilical artery (n = 45) and human umbilical vein endothelial cells (HUVEC) (n = 52) in a birth cohort using the Infinium HumanMethylation450 BeadChip array. Unadjusted and cell mixture adjusted associations for each tissue were examined along with enrichment analyses relative to CpG island location and omnibus permutation tests of association among biological pathways. One CpG in artery (cg26587014) and 4 CpGs in placenta (cg12825509; cg20554753; cg23439277; cg21055948) reached a Bonferroni adjusted level of significance. Several CpGs were differentially methylated in artery and placenta when controlling the false discovery rate (q-value<0.05), but none in HUVEC. Enrichment of hypomethylated CpG islands was observed for artery while hypermethylation of open sea regions were present in placenta relative to prenatal arsenic exposure. The melanogenesis pathway was differentially methylated in artery (Max F P < 0.001), placenta (Max F P < 0.001), and HUVEC (Max F P = 0.02). Similarly, the insulin-signaling pathway was differentially methylated in artery (Max F P = 0.02), placenta (Max F P = 0.02), and HUVEC (Max F P = 0.02). Our results show that prenatal arsenic exposure can alter DNA methylation in artery and placenta but not in HUVEC. Further studies are needed to determine if these alterations in DNA methylation mediate the effect of prenatal arsenic exposure and health outcomes later in life.

    View details for DOI 10.1080/15592294.2015.1105424

    View details for Web of Science ID 000366246600007

    View details for PubMedID 26646901

    View details for PubMedCentralID PMC4844206

  • Differential DNA methylation in umbilical cord blood of infants exposed to mercury and arsenic in utero EPIGENETICS Cardenas, A., Koestler, D. C., Houseman, E., Jackson, B. P., Kile, M. L., Karagas, M. R., Marsit, C. J. 2015; 10 (6): 508-515


    Mercury and arsenic are known developmental toxicants. Prenatal exposures are associated with adverse childhood health outcomes that could be in part mediated by epigenetic alterations that may also contribute to altered immune profiles. In this study, we examined the association between prenatal mercury exposure on both DNA methylation and white blood cell composition of cord blood, and evaluated the interaction with prenatal arsenic exposure. A total of 138 mother-infant pairs with postpartum maternal toenail mercury, prenatal urinary arsenic concentrations, and newborn cord blood were assessed using the Illumina Infinium Methylation450 array. White blood cell composition was inferred from DNA methylation measurements. A doubling in toenail mercury concentration was associated with a 2.5% decrease (95% CI: 5.0%, 1.0%) in the estimated monocyte proportion. An increase of 3.5% (95% CI: 1.0, 7.0) in B-cell proportion was observed for females only. Among the top 100 CpGs associated with toenail mercury levels (ranked on P-value), there was a significant enrichment of loci located in North shore regions of CpG islands (P = 0.049), and the majority of these loci were hypermethylated (85%). Among the top 100 CpGs for the interaction between arsenic and mercury, there was a greater than expected proportion of loci located in CpG islands (P = 0.045) and in South shore regions (P = 0.009) and all of these loci were hypermethylated. This work supports the hypothesis that mercury may be contributing to epigenetic variability and immune cell proportion changes, and suggests that in utero exposure to mercury and arsenic, even at low levels, may interact to impact the epigenome.

    View details for DOI 10.1080/15592294.2015.1046026

    View details for Web of Science ID 000355682500007

    View details for PubMedID 25923418

    View details for PubMedCentralID PMC4622995

  • Arsenic Exposure and Prevalence of the Varicella Zoster Virus in the United States: NHANES (2003-2004 and 2009-2010) ENVIRONMENTAL HEALTH PERSPECTIVES Cardenas, A., Smit, E., Houseman, E., Kerkvliet, N. I., Bethel, J. W., Kile, M. L. 2015; 123 (6): 590-596


    Arsenic is an immunotoxicant. Clinical reports observe the reactivation of varicella zoster virus (VZV) in people who have recovered from arsenic poisoning and in patients with acute promyelocytic leukemia that have been treated with arsenic trioxide.We evaluated the association between arsenic and the seroprevalence of VZV IgG antibody in a representative sample of the U.S.We analyzed data from 3,348 participants of the National Health and Nutrition Examination Survey (NHANES) 2003-2004 and 2009-2010 pooled survey cycles. Participants were eligible if they were 6-49 years of age with information on both VZV IgG and urinary arsenic concentrations. We used two measures of total urinary arsenic (TUA): TUA1 was defined as the sum of arsenite, arsenate, monomethylarsonic acid, and dimethylarsinic acid, and TUA2 was defined as total urinary arsenic minus arsenobetaine and arsenocholine.The overall weighted seronegative prevalence of VZV was 2.2% for the pooled NHANES sample. The geometric means of TUA1 and TUA2 were 6.57 μg/L and 5.64 μg/L, respectively. After adjusting for age, sex, race, income, creatinine, and survey cycle, odds ratios for a negative VZV IgG result in association with 1-unit increases in natural log-transformed (ln)-TUA1 and ln-TUA2 were 1.87 (95% CI: 1.03, 3.44) and 1.40 (95% CI: 1.0, 1.97), respectively.In this cross-sectional analysis, urinary arsenic was inversely associated with VZV IgG seroprevalence in the U.S.This finding is in accordance with clinical observations of zoster virus reactivation from high doses of arsenic. Additional studies are needed to confirm the association and evaluate causal mechanisms.

    View details for DOI 10.1289/ehp.1408731

    View details for Web of Science ID 000357296200020

    View details for PubMedID 25636148

    View details for PubMedCentralID PMC4455594

  • Risk behaviors and self-reported illnesses among Pacific Northwest surfers JOURNAL OF WATER AND HEALTH Harding, A. K., Stone, D. L., Cardenas, A., Lesser, V. 2015; 13 (1): 230-242


    Although surfers have high incidental exposure to marine waters, no studies have investigated if surfer risk behaviors (such as surfing during advisories, near an outfall, during a rain event, or use of personal protective equipment) increase or decrease the risk of acquiring waterborne illnesses. We used a web-based survey to assess the association between risk-based behaviors and self-reported illnesses among Pacific Northwest surfers. Commonly reported illnesses include: ear infection or discharge (38%), sore throat or a cough (28%), diarrhea (16%), fever (10.5%), and vomiting (7%). Surfing often during rain events was associated with an increased likelihood of diarrhea (OR = 2.7; 95% CI: 1.4-5.47), sore throat (OR = 1.26; 95% CI: 1.01-2.05), and ear infection (OR = 1.39; 95% CI: 1.01-2.32). Surfing during a health advisory was associated with increased likelihood of diarrhea (OR = 1.94; 95% CI: 1.03-4.64) and sore throat (OR = 2.32; 95% CI: 1.23-4.40). Other behaviors associated with increased illnesses include body surfing, surfing near an outfall, frequency of surfing, and use of ear plugs. Approximately 40% of surfers were unaware if they had surfed during an active health advisory and 29% knowingly surfed during advisories, suggesting the need to engage this population about potential harm and behaviors that may increase health risk.

    View details for DOI 10.2166/wh.2014.231

    View details for Web of Science ID 000355183800022

    View details for PubMedID 25719482

  • Acquired urethral obstruction in New World camelids: 34 cases (1995-2008) AUSTRALIAN VETERINARY JOURNAL Duesterdieck-Zellmer, K. F., Van Metre, D. V., Cardenas, A., Cebra, C. K. 2014; 92 (8): 313-319


    Document the clinical features, short- and long-term outcomes and prognostic factors in New World camelids with acquired urethral obstruction.Retrospective case study.Case data from medical records of 34 New World camelids presenting with acquired urethral obstruction were collected and follow-up information on discharged patients was obtained. Associations with short- and long-term survival were evaluated using Wilcoxon rank-sum tests, exact-logistic regressions and Kaplan-Meier survival curves.Of the 34 New World camelids 23 were intact males and 11 were castrated; 4 animals were euthanased upon presentation, 7 were treated medically and 23 surgically, including urethrotomy, bladder marsupialisation, tube cystostomy alone or combined with urethrotomy, urethrostomy or penile reefing. Necrosis of the distal penis was found in 4 animals and all were short-term non-survivors. Short-term survival for surgical cases was 65%, and 57% for medical cases. Incomplete urethral obstruction at admission and surgical treatment were associated with increased odds of short-term survival. Of 14 records available for long-term follow-up, 6 animals were alive and 8 were dead (median follow-up 4.5 years, median survival time 2.5 years). Recurrence of urethral obstruction was associated with long-term non-survival.Surgically treated New World camelids with incomplete urethral obstruction have the best odds of short-term survival and those with recurrence of urethral obstruction have a poor prognosis for long-term survival.

    View details for DOI 10.1111/avj.12207

    View details for Web of Science ID 000340242000024

    View details for PubMedID 24964920

  • Effect of prenatal arsenic exposure on DNA methylation and leukocyte subpopulations in cord blood EPIGENETICS Kile, M. L., Houseman, E., Baccarelli, A. A., Quamruzzaman, Q., Rahman, M., Mostofa, G., Cardenas, A., Wright, R. O., Christiani, D. C. 2014; 9 (5): 774-782


    Prenatal arsenic exposure is associated with increased risk of disease in adulthood. This has led to considerable interest in arsenic's ability to disrupt fetal programming. Many studies report that arsenic exposure alters DNA methylation in whole blood but these studies did not adjust for cell mixture. In this study, we examined the relationship between arsenic in maternal drinking water collected ≤ 16 weeks gestational age and DNA methylation in cord blood (n = 44) adjusting for leukocyte-tagged differentially methylated regions. DNA methylation was quantified using the Infinium HumanMethylation 450 BeadChip array. Recursively partitioned mixture modeling examined the relationship between arsenic and methylation at 473,844 CpG sites. Median arsenic concentration in water was 12 µg/L (range<1- 510 µg/L). Log 10 arsenic was associated with altered DNA methylation across the epigenome (P = 0.002); however, adjusting for leukocyte distributions attenuated this association (P = 0.013). We also observed that arsenic had a strong effect on the distribution of leukocytes in cord blood. In adjusted models, every log 10 increase in maternal drinking water arsenic exposure was estimated to increase CD8+ T cells by 7.4% (P = 0.0004) and decrease in CD4+ T cells by 9.2% (P = 0.0002). These results show that prenatal exposure to arsenic had an exposure-dependent effect on specific T cell subpopulations in cord blood and altered DNA methylation in cord blood. Future research is needed to determine if these small changes in DNA methylation alter gene expression or are associated with adverse health effects.

    View details for DOI 10.4161/epi.28153

    View details for Web of Science ID 000335397000014

    View details for PubMedID 24525453

    View details for PubMedCentralID PMC4063836

  • Naphthalene Biomarkers and Relationship with Hemoglobin and Hematocrit in White, Black, and Hispanic Adults: Results from the 2003-2004 National Health and Nutrition Examination Survey JOURNAL OF MEDICAL TOXICOLOGY Sudakin, D. L., Smit, E., Cardenas, A., Harding, A. 2013; 9 (2): 133-138


    Naphthalene is an important contaminant in indoor and outdoor air. Acute overexposure can have toxic effects, resulting in hemolysis. There have been no studies evaluating the impact of environmental exposure on red blood cell indices. We examined 1- and 2-hydroxynaphthalene urinary metabolites (NAP1 and NAP2) in non-Hispanic White, non-Hispanic Black, and Mexican-American adults in the USA and their relationship with hemoglobin (Hb) and hematocrit (HCT). Using the 2003-2004 National Health and Nutrition Examination Survey data, weighted generalized linear regression analyses were used to examine the association between Hb (in grams per deciliter) and HCT (in percent) with NAP1 and NAP2 (per 100,000 ng/L). Beta coefficients ± SE are reported. NAP1 and NAP2 were highest in non-Hispanic Blacks, followed by non-Hispanic Whites, and lowest in Mexican-American adults. There was a positive association between NAP1 and Hb (0.39 ± 0.11, p = 0.0034) and HCT (1.14 ± 0.28, p = 0.0009) after adjusting for age, gender, race, education, and smoking. Stratified analysis by smoking showed similar results with the association being stronger for smokers (Hb 0.63 ± 0.23, p = 0.02; HCT 1.43 ± 0.79, p = 0.09) than nonsmokers (Hb 0.34 ± 0.14, p = 0.03; HCT 1.08 ± 0.42, p = 0.02). The association was also stronger for non-Hispanic blacks (Hb 0.54 ± 0.20, p = 0.02; HCT 1.43 ± 0.55, p = 0.02) than for non-Hispanic whites (Hb 0.37 ± 0.18, p = 0.06; HCT 1.20 ± 0.51, p = 0.03) and was not significant for Mexican-Americans (Hb 0.30 ± 1.7, p = 0.10; HCT 0.99 ± 0.52, p = 0.08). NAP2 was not significantly associated with Hb or HCT. The observed disparity in NAP1 and NAP2 levels by race/ethnicity is consistent with published literature. The origin of these differences in exposure is unclear but may reflect differences in environmental exposure as well as genetic susceptibility. The positive association between NAP1 with HCT and Hb is an unexpected finding. Further research is needed to understand the possible biological mechanisms or other explanations for this association.

    View details for DOI 10.1007/s13181-012-0262-y

    View details for Web of Science ID 000443014100003

    View details for PubMedID 23007805

    View details for PubMedCentralID PMC3556176

  • Lin28 regulates BMP4 and functions with Oct4 to affect ovarian tumor microenvironment CELL CYCLE Ma, W., Ma, J., Xu, J., Qiao, C., Branscum, A., Cardenas, A., Baron, A. T., Schwartz, P., Maihle, N. J., Huang, Y. 2013; 12 (1): 88-97


    Emerging evidence suggests that the tumor microenvironment plays a critical role in regulating cancer stem cells (CSCs) and tumor progression through both autocrine and paracrine signaling. Elevated production of bone morphogenetic proteins (BMPs) from human ovarian cancer cells and stroma has been shown to increase CSC proliferation and tumor growth. Here, we report that Lin28, a stem cell factor, binds to BMP4 mRNA in epithelial ovarian carcinoma cells, thereby promoting BMP4 expression at the post-transcriptional level. As co-expression of Lin28 and Oct4 (another stem cell factor) has been implicated in ovarian cancer CSCs, we also determined that high levels of Lin28 are associated with an unfavorable prognosis when co-expressed with high levels of Oct4. Together, these findings uncover a new level of regulation of BMP4 expression and imply a novel Lin28/Oct4/BMP4-mediated mechanism of regulating ovarian tumor cell growth, thus holding potential for the development of new strategies for the diagnosis and treatment of ovarian cancer.

    View details for DOI 10.4161/cc.23028

    View details for Web of Science ID 000313414700019

    View details for PubMedID 23255092

    View details for PubMedCentralID PMC3570521

  • Effect of Native American Fish Smoking Methods on Dietary Exposure to Polycyclic Aromatic Hydrocarbons and Possible Risks to Human Health JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY Forsberg, N. D., Stone, D., Harding, A., Harper, B., Harris, S., Matzke, M. M., Cardenas, A., Waters, K. M., Anderson, K. A. 2012; 60 (27): 6899-6906


    Although it is known that polycyclic aromatic hydrocarbons (PAHs) can be found in smoked meats, little is known about their prevalence in Native American smoked fish. In this work, the effect of traditional Native American fish smoking methods on dietary exposure to PAHs and possible risks to human health has been assessed. Smoking methods considered smoking structure (tipi or shed) and wood type (apple or alder). Neither smoking structure nor wood type accounted for differences in smoked salmon content of 33 PAHs. Carcinogenic and noncarcinogenic PAH loads in traditionally smoked salmon were 40-430 times higher than those measured in commercial products. Dietary exposure to PAHs could result in excess lifetime cancer risks between 1 × 10(-5) and 1 × 10(-4) at a daily consumption rate of 5 g d(-1) and could approach 1 × 10(-2) at 300 g d(-1). Hazard indexes approached 0.005 at 5 g d(-1), or approximately 0.3 at 300 g d(-1). Levels of PAHs present in smoked salmon prepared using traditional Native American methods may pose elevated cancer risks if consumed at high consumption rates over many years.

    View details for DOI 10.1021/jf300978m

    View details for Web of Science ID 000306297800030

    View details for PubMedID 22690788

    View details for PubMedCentralID PMC3567306

  • Soluble HER2 (sHER2) as a prognostic serum biomarker of non-small cell lung cancer Wilson, B., Cardenas, A., Kimbler, K. D., Gal, T., Witt, L. A., Arnold, S. M., Branscum, A. J., Baron, A. T. AMER SOC CLINICAL ONCOLOGY. 2012