Abstract
Attention-deficit and hyperactivity disorder (ADHD) is a common childhood disorder with a substantial genetic component. However, the extent to which epigenetic mechanisms play a role in the etiology of the disorder is unknown. We performed epigenome-wide association studies (EWAS) within the Pregnancy And Childhood Epigenetics (PACE) Consortium to identify DNA methylation sites associated with ADHD symptoms at two methylation assessment periods: birth and school age. We examined associations of both DNA methylation in cord blood with repeatedly assessed ADHD symptoms (age 4–15 years) in 2477 children from 5 cohorts and of DNA methylation at school age with concurrent ADHD symptoms (age 7–11 years) in 2374 children from 9 cohorts, with 3 cohorts participating at both timepoints. CpGs identified with nominal significance (p < 0.05) in either of the EWAS were correlated between timepoints (ρ = 0.30), suggesting overlap in associations; however, top signals were very different. At birth, we identified nine CpGs that predicted later ADHD symptoms (p < 1 × 10–7), including ERC2 and CREB5. Peripheral blood DNA methylation at one of these CpGs (cg01271805 in the promoter region of ERC2, which regulates neurotransmitter release) was previously associated with brain methylation. Another (cg25520701) lies within the gene body of CREB5, which previously was associated with neurite outgrowth and an ADHD diagnosis. In contrast, at school age, no CpGs were associated with ADHD with p < 1 × 10−7. In conclusion, we found evidence in this study that DNA methylation at birth is associated with ADHD. Future studies are needed to confirm the utility of methylation variation as biomarker and its involvement in causal pathways.
| Original language | English |
|---|---|
| Article number | 398 |
| Pages (from-to) | 1-11 |
| Number of pages | 11 |
| Journal | Translational Psychiatry |
| Volume | 10 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - Dec 2020 |
Funding
We thank all the children and families who took part in this study, as well as the support of hospitals, midwives, and pharmacies. ALSPAC: We are grateful to the whole ALSPAC team, which includes interviewers, computer and laboratory technicians, clerical workers, research scientists, volunteers, managers, receptionists, and nurses. The UK Medical Research Council (MRC) and Wellcome (Grant ref: 102215/2/13/2) and the University of Bristol provide core support for ALSPAC. This publication is the work of the authors and E.W. will serve as guarantors for the contents of this paper. A comprehensive list of grant funding is available on the ALSPAC website (http://www.bristol.ac.uk/ alspac/external/documents/grant-acknowledgements.pdf). Methylation data in the ALSPAC cohort were generated as part of the UK BBSRC funded (BB/ I025751/1 and BB/I025263/1) Accessible Resource for Integrated Epigenomic Studies (ARIES, http://www.ariesepigenomics.org.uk). E.B. is supported by an ESRC grant ES/S013229/1. D.C., G.S., and C.L.R. work in a department supported by the UK MRC (MC_UU_00011/5). G.S. is further supported by an MRC New Investigator Research Grant (MR/S009310/1) and the European Joint Programming Initiative “A Healthy Diet for a Healthy Life” (JPI HDHL, NutriPROGRAM project, MRC, MR/S036520/1). GENR: The Generation R Study is conducted by the Erasmus Medical Center in close collaboration with the Erasmus University Rotterdam, Faculty of Social Sciences, the Municipal Health Service Rotterdam area, the Rotterdam Homecare Foundation, Rotterdam, and the Stichting Trombosedienst & Artsenlaboratorium Rijnmond (STAR-MDC), Rotterdam. We gratefully acknowledge the contribution of general practitioners, hospitals, midwives, and pharmacies in Rotterdam. The generation and management of the Illumina 450K methylation array data (EWAS data) for the Generation R Study was executed by the Human Genotyping Facility of the Genetic Laboratory of the Department of Internal Medicine, Erasmus MC, the Netherlands. We thank Mr. Michael Verbiest, Ms. Mila Jhamai, Ms. Sarah Higgins, Mr. Marijn Verkerk, and Dr. Lisette Stolk for their help in creating the EWAS database. We thank Dr. A. Teumer for his work on the quality control and normalization scripts. The general design of the Generation R Study is made possible by financial support from Erasmus Medical Center, Rotterdam, Erasmus University Rotterdam, the Netherlands Organization for Health Research and Development (ZonMw), and the Ministry of Health, Welfare and Sport. The EWAS data were funded by a grant from the Netherlands Genomics Initiative (NGI)/Netherlands Organisation for Scientific Research (NWO) Netherlands Consortium for Healthy Aging (NCHA; project nr. 050-060-810), by funds from the Genetic Laboratory of the Department of Internal Medicine, Erasmus MC, and by a grant from the National Institute of Child and Human Development (R01HD068437). A.N. and H.T. are supported by a grant of the Dutch Ministry of Education, Culture, and Science and the Netherlands Organization for Scientific Research (NWO grant No. 024.001.003, Consortium on Individual Development). A.N. is also supported by a Canadian Institutes of Health Research team grant. The work of H.T. is further supported by a NWO-VICI grant (NWO-ZonMW: 016.VICI.170.200). M.v.I. and M.B.-K. were supported by the Netherlands Organization for Scientific Research (SPINOZA, VICI), and M.B.-K. was supported by the European Research Council (AdG 669249). J.F.F. has received funding from the European Joint Programming Initiative “A Healthy Diet for a Healthy Life” (JPI HDHL, NutriPROGRAM project, ZonMw the Netherlands no.529051022). This project received funding from the European Union’s Horizon 2020 research and innovation program (733206, LifeCycle; 633595, DynaHEALTH). GLAKU: We thank all the research nurses, research assistants, and laboratory personnel involved in the GLAKU study. The study has been supported by Academy of Finland, University of Helsinki, Hope and Optimism Initiative, Finnish Foundation for Pediatric Research, Sigrid Juselius Foundation, Jalmari and Rauha Ahokas Foundation, Signe and Ane Gyllenberg Foundation, Yrjo Jahnsson Foundation, Juho Vainio Foundation, Emil Aaltonen Foundation, and Ministry of Education and Culture, Finland. INMA: A full roster of the INMA Project Investigators can be found at http:// www.proyectoinma.org/presentacion-inma/listado-investigadores/en_listado-investigadores.html. S.A. is funded by a Juan de la Cierva-Incorporación fellowship (IJCI-2017-34068) awarded by the Spanish Ministerio de Economía, Industria y Competitividad. M.G. is funded by a Miguel Servet fellowship (MS13/00054, CP18/00018) awarded by the Spanish Institute of Health Carlos III. ISGlobal is a member of the CERCA Programme, Generalitat de Catalunya. J.J. is funded by a Miguel Servet fellowship (MS14/00108, CP14/00108) awarded by the Spanish Institute of Health Carlos III. HELIX: The authors would like to thank all practitioners and researchers in the six countries who took part in this study. The authors would like to thank Sonia Brishoual, Angelique Serre, and Michele; Grosdenier (Poitiers Biobank, CRB BB-0033-00068, Poitiers, France) for biological sample management; and Professor Frederic Millot (Principal Investigator), Elodie Migault, Manuela Boue, and Sandy Bertin (Clinical Investigation Center, Inserm. CIC1402, CHU de Poitiers, Poitiers, France) for planning and investigational actions. The authors would like to thank Veronique Ferrand-Rigalleau, Céline Leger, and Noella Gorry (CHU de Poitiers, Poitiers, France) for administrative assistance (EDEN). The authors would like to thank Silvia Fochs, Nuria Pey, Cecilia Persavente, and Susana Gross for field work, sample management, and overall management in INMA. The authors would like to thank Georgia Chalkiadaki and Danai Feida for biological sample management, to Eirini Michalaki, Mariza Kampouri, Anny Kyriklaki, and Minas Iakovidis for field study performance and to Maria Fasoulaki for administrative assistance (RHEA). The authors would also like to thank Ingvild Essén for thorough field work, Heidi Marie Nordheim for biological sample management, and the Norwegian Mother, Father and Child cohort study (MoBa) administrative unit. The MoBa cohort study is supported by the Norwegian Ministry of Health and Care Services and the Ministry of Education and Research. The contribution of the Spanish National Genotyping Center (CEGEN-PRB2) is also acknowledged. NEST: The NEST cohort has been supported by the National Institute of Environmental Health Sciences (R01ES016772 [to C.H.], R21ES014947 [to C.H.], P30ES025128 [to C.H., D.D.J., R.M.], R01MD011746 [to C.H., S.K.M.], P30ES011961 pilot project [to S.K.M.], P01ES022831 [to S.K.M., B.F.F.]), the US Environmental Protection Agency (RD-83543701 [to S.K.M., B.F.F.]), the National Institute of Diabetes and Digestive and Kidney Diseases (R01DK085173 [to C.H., S.K.M.]), the National Institute of Aging (R21AG041048 [to B.F.F.]), and the Duke Cancer Institute [to C.H., S.K.M.]. The contents are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health or the United States Environmental Protection Agency (US EPA). Further, USEPA does not endorse the purchase of any commercial products or services mentioned in the publication. PREDO: The PREDO Study has been funded by the Academy of Finland, EraNet Neuron, EVO (a special state subsidy for health science research), University of Helsinki Research Funds, the Signe and Ane Gyllenberg Foundation, the Emil Aaltonen Foundation, the Finnish Medical Foundation, the Jane and Aatos Erkko Foundation, the Novo Nordisk Foundation, the Päivikki and Sakari Sohlberg Foundation, and the Sigrid Juselius Foundation granted to members of the Predo study board. Methylation assays were funded by the Academy of Finland. The PREDO study would not have been possible without the dedicated contribution of the PREDO study group members: A-K Pesonen (Department of Psychology and Logopedics, University of Helsinki; Finland), A Aitokallio-Tallberg, A-M Henry, VK Hiilesmaa, T Karipohja, R Meri, S Sainio, T Saisto, S Suomalainen-Konig, V-M Ulander, T Vaitilo (Department of Obstetrics and Gynaecology, University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland), L Keski-Nisula, Maija-Riitta Orden (Kuopio University Hospital, Kuopio Finland), E Koistinen, T Walle, R Solja (Northern Karelia Central Hospital, Joensuu, Finland), M Kurkinen (Päijät-Häme Central Hospital, Lahti, Finland), P.Taipale. P Staven (Iisalmi Hospital, Iisalmi, Finland), and J Uotila (Tampere University Hospital, Tampere, Finland). We also thank all the research nurses, research assistants, and laboratory personnel involved in the PREDO study.
| Funders | Funder number |
|---|---|
| Dutch Ministry of Education, Culture, and Science | |
| EraNet Neuron | |
| Erasmus Medical Center, Rotterdam | |
| Finnish Foundation for Pediatric Research | |
| Hope and Optimism Initiative | |
| NWO-VICI | |
| Netherlands Consortium for Healthy Aging | 050-060-810 |
| Netherlands Genomics Initiative | |
| SPINOZA | |
| Spanish Institute of Health Carlos III | |
| UK MRC | MR/S036520/1 |
| University of Helsinki Research Funds | |
| National Institute on Aging | R21AG041048 |
| National Institute of Diabetes and Digestive and Kidney Diseases | R01DK085173 |
| National Institute of Environmental Health Sciences | R01ES016772, R21ES014947, R24ES028531, R01MD011746, P01ES022831, P30ES011961, P30ES025128 |
| National Institute of Child Health and Human Development | R01HD068437 |
| U.S. Environmental Protection Agency | RD-83543701 |
| Helsingin Yliopisto | |
| Suomen Lääketieteen Säätiö | |
| Yrjö Jahnssonin Säätiö | |
| Jalmari ja Rauha Ahokkaan Säätiö | |
| Wellcome Trust | 102215/2/13/2 |
| Horizon 2020 Framework Programme | 733206, 633595 |
| Duke Cancer Institute | |
| Canadian Institutes of Health Research | |
| Medical Research Council | MR/S009310/1, MC_UU_00011/5 |
| Economic and Social Research Council | ES/S013229/1 |
| European Research Council | 529051022, AdG 669249 |
| University of Bristol | |
| ZonMw | |
| Erasmus Universiteit Rotterdam | |
| Academy of Finland | |
| Ministerie van Volksgezondheid, Welzijn en Sport | |
| Erasmus Medisch Centrum | |
| Opetus- ja Kulttuuriministeriö | IJCI-2017-34068 |
| Nederlandse Organisatie voor Wetenschappelijk Onderzoek | 024.001.003 |
| Helse- og Omsorgsdepartementet | |
| Haridus- ja Teadusministeerium | |
| Jane ja Aatos Erkon Säätiö | |
| Juho Vainion Säätiö | |
| Päivikki ja Sakari Sohlbergin Säätiö | |
| Signe ja Ane Gyllenbergin Säätiö | |
| Instituto de Salud Carlos III | MS14/00108, MS13/00054, CP18/00018, CP14/00108 |
| Emil Aaltosen Säätiö | |
| Sigrid Juséliuksen Säätiö | |
| Novo Nordisk Fonden | |
| Ministerio de Asuntos Económicos y Transformación Digital, Gobierno de España |
