Genome-wide copy number variation study associates metabotropic glutamate receptor gene networks with attention deficit hyperactivity disorder

Abstract

Attention deficit hyperactivity disorder (ADHD) is a common, heritable neuropsychiatric disorder of unknown etiology. We performed a whole-genome copy number variation (CNV) study on 1,013 cases with ADHD and 4,105 healthy children of European ancestry using 550,000 SNPs. We evaluated statistically significant findings in multiple independent cohorts, with a total of 2,493 cases with ADHD and 9,222 controls of European ancestry, using matched platforms. CNVs affecting metabotropic glutamate receptor genes were enriched across all cohorts (P = 2.1 × 10−9). We saw GRM5 (encoding glutamate receptor, metabotropic 5) deletions in ten cases and one control (P = 1.36 × 10−6). We saw GRM7 deletions in six cases, and we saw GRM8 deletions in eight cases and no controls. GRM1 was duplicated in eight cases. We experimentally validated the observed variants using quantitative RT-PCR. A gene network analysis showed that genes interacting with the genes in the GRM family are enriched for CNVs in 10% of the cases (P = 4.38 × 10−10) after correction for occurrence in the controls. We identified rare recurrent CNVs affecting glutamatergic neurotransmission genes that were overrepresented in multiple ADHD cohorts.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: A deletion directly affecting GRM5 that is exclusive to cases with ADHD and that was replicated in the IMAGE and PUWMa studies.
Figure 2: GRM receptor gene interaction networks affected in ADHD.

References

  1. 1

    Derks, E.M. et al. Genetic and environmental influences on the relation between attention problems and attention deficit hyperactivity disorder. Behav. Genet. 38, 11–23 (2008).

    Article  Google Scholar 

  2. 2

    Wood, A.C., Rijsdijk, F., Saudino, K.J., Asherson, P. & Kuntsi, J. High heritability for a composite index of children's activity level measures. Behav. Genet. 38, 266–276 (2008).

    Article  Google Scholar 

  3. 3

    Haberstick, B.C. et al. Genetic and environmental contributions to retrospectively reported DSM-IV childhood attention deficit hyperactivity disorder. Psychol. Med. 38, 1057–1066 (2008).

    CAS  Article  Google Scholar 

  4. 4

    Glessner, J.T. et al. Autism genome-wide copy number variation reveals ubiquitin and neuronal genes. Nature 459, 569–573 (2009).

    CAS  Article  Google Scholar 

  5. 5

    Wang, K. et al. Common genetic variants on 5p14.1 associate with autism spectrum disorders. Nature 459, 528–533 (2009).

    CAS  Article  Google Scholar 

  6. 6

    Pinto, D. et al. Functional impact of global rare copy number variation in autism spectrum disorders. Nature 466, 368–372 (2010).

    CAS  Article  Google Scholar 

  7. 7

    Franke, B., Neale, B.M. & Faraone, S.V. Genome-wide association studies in ADHD. Hum. Genet. 126, 13–50 (2009).

    CAS  Article  Google Scholar 

  8. 8

    Neale, B.M. et al. Genome-wide association scan of attention deficit hyperactivity disorder. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 147B, 1337–1344 (2008).

    CAS  Article  Google Scholar 

  9. 9

    Lasky-Su, J. et al. Genome-wide association scan of quantitative traits for attention deficit hyperactivity disorder identifies novel associations and confirms candidate gene associations. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 147B, 1345–1354 (2008).

    CAS  Article  Google Scholar 

  10. 10

    Lesch, K.P. et al. Molecular genetics of adult ADHD: converging evidence from genome-wide association and extended pedigree linkage studies. J Neural. Transm. 115, 1573–1585 (2008).

    CAS  Article  Google Scholar 

  11. 11

    Williams, N.M. et al. Rare chromosomal deletions and duplications in attention-deficit hyperactivity disorder: a genome-wide analysis. Lancet 376, 1401–1408 (2010)Epub 2010 Sep 29.

    CAS  Article  Google Scholar 

  12. 12

    Elia, J. et al. Rare structural variants found in attention-deficit hyperactivity disorder are preferentially associated with neurodevelopmental genes. Mol. Psychiatry 15, 637–646 (2010).

    CAS  Article  Google Scholar 

  13. 13

    Elia, J., Gai, X., Hakonarson, H. & White, P.S. Structural variations in attention-deficit hyperactivity disorder. Lancet 377, 377–378 (2011).

    Article  Google Scholar 

  14. 14

    Wang, K. et al. PennCNV: an integrated hidden Markov model designed for high-resolution copy number variation detection in whole-genome SNP genotyping data. Genome Res. 17, 1665–1674 (2007).

    CAS  Article  Google Scholar 

  15. 15

    Colella, S. et al. QuantiSNP: an Objective Bayes Hidden-Markov Model to detect and accurately map copy number variation using SNP genotyping data. Nucleic Acids Res. 35, 2013–2025 (2007).

    CAS  Article  Google Scholar 

  16. 16

    Zhou, K. et al. Meta-analysis of genome-wide linkage scans of attention deficit hyperactivity disorder. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 147B, 1392–1398 (2008).

    CAS  Article  Google Scholar 

  17. 17

    Kent, W.J. et al. The human genome browser at UCSC. Genome Res. 12, 996–1006 (2002).

    CAS  Article  Google Scholar 

  18. 18

    Shannon, P. et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 13, 2498–2504 (2003).

    CAS  Article  Google Scholar 

  19. 19

    Taniura, H., Sanada, N., Kuramoto, N. & Yoneda, Y. A metabotropic glutamate receptor family gene in Dictyostelium discoideum. J. Biol. Chem. 281, 12336–12343 (2006).

    CAS  Article  Google Scholar 

  20. 20

    Conn, P.J. & Pin, J. Phamacology and functions of metabotropic glutamate receptors. Annu. Rev. Pharmacol. Toxicol. 37, 205–237 (1997).

    CAS  Article  Google Scholar 

  21. 21

    Berthele, A. et al. Expression of metabotropic glutamate receptor subtype mRNA (mGluR1–8) in human cerebellum. Neuroreport 10, 3861–3867 (1999).

    CAS  Article  Google Scholar 

  22. 22

    Koob, G.F., Sanna, P.P. & Bloom, F.E. Neuroscience of addiction. Neuron 21, 467–476 (1998).

    CAS  Article  Google Scholar 

  23. 23

    Cryan, J.F. et al. Antidepressant and anxiolytic-like effects in mice lacking the group III metabotropic glutamate receptor mGluR7. Eur. J. Neurosci. 17, 2409–2417 (2003).

    Article  Google Scholar 

  24. 24

    Makoff, A., Pillinga, C., Harrington, K. & Emson, P. Human metabotropic glutamate receptor type 7: Molecular cloning and mRNA distribution in the CNS. Brain Res. Mol. Brain Res. 40, 165–170 (1996).

    CAS  Article  Google Scholar 

  25. 25

    Turic, D. et al. Follow-up of genetic linkage findings on chromosome 16p13: evidence of association of N-methyl-D aspartate glutamate receptor 2A gene polymorphism with ADHD. Mol. Psychiatry 9, 169–173 (2004).

    CAS  Article  Google Scholar 

  26. 26

    Mick, E. & Faraone, S.V. Genetics of attention deficit hyperactivity disorder. Child Adolesc. Psychiatr. Clin. N. Am. 17, 261–284 (2008).

    Article  Google Scholar 

  27. 27

    Turic, D. et al. A family based study implicates solute carrier family 1-member 3 (SLC1A3) gene in attention-deficit/hyperactivity disorder. Biol. Psychiatry 57, 1461–1466 (2005).

    CAS  Article  Google Scholar 

  28. 28

    Elia, J. et al. Candidate gene analysis in an on-going genome-wide association study of attention-deficit hyperactivity disorder: suggestive association signals in ADRA1A. Psychiatr. Genet. 19, 134–141 (2009).

    Article  Google Scholar 

  29. 29

    Mick, E., Neale, B., Middleton, F.A., McGough, J.J. & Faraone, S.V. Genome-wide association study of response to methylphenidate in 187 children with attention-deficit/hyperactivity disorder. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 147B, 1412–1418 (2008).

    CAS  Article  Google Scholar 

  30. 30

    Dorval, K.M. et al. Association of the glutamate receptor subunit gene GRIN2B with attention-deficit/hyperactivity disorder. Genes Brain Behav. 6, 444–452 (2007).

    CAS  Article  Google Scholar 

  31. 31

    Jin, Z., Zang, Y.F., Zeng, Y.W., Zhang, L. & Wang, Y.F. Striatal neuronal loss or dysfunction and choline rise in children with attention-deficit hyperactivity disorder: a 1H-magnetic resonance spectroscopy study. Neurosci. Lett. 315, 45–48 (2001).

    CAS  Article  Google Scholar 

  32. 32

    MacMaster, F.P., Carrey, N., Sparkes, S. & Kusumakar, V. Proton spectroscopy in medication-free pediatric attention-deficit/hyperactivity disorder. Biol. Psychiatry 53, 184–187 (2003).

    Article  Google Scholar 

  33. 33

    Courvoisie, H., Hooper, S.R., Fine, C., Kwock, L. & Castillo, M. Neurometabolic functioning and neuropsychological correlates in children with ADHD-H: preliminary findings. J. Neuropsychiatry Clin. Neurosci. 16, 63–69 (2004).

    CAS  Article  Google Scholar 

  34. 34

    Carrey, N. et al. Metabolite changes resulting from treatment in children with ADHD: a 1H-MRS study. Clin. Neuropharmacol. 26, 218–221 (2003).

    Article  Google Scholar 

  35. 35

    Gainetdinov, R.R. et al. Role of serotonin in the paradoxical calming effect of psychostimulants on hyperactivity. Science 283, 397–401 (1999).

    CAS  Article  Google Scholar 

  36. 36

    Gainetdinov, R.R., Mohn, A.R., Bohn, L.M. & Caron, M.G. Glutamatergic modulation of hyperactivity in mice lacking the dopamine transporter. Proc. Natl. Acad. Sci. USA 98, 11047–11054 (2001).

    CAS  Article  Google Scholar 

  37. 37

    Masuo, Y., Ishido, M., Morita, M. & Oka, S. Effects of neonatal 6-hydroxydopamine lesion on the gene expression profile in young adult rats. Neurosci. Lett. 335, 124–128 (2002).

    CAS  Article  Google Scholar 

  38. 38

    Miyamoto, K. et al. Involvement of enhanced sensitivity of N-methyl-D-aspartate receptors in vulnerability of developing cortical neurons to methylmercury neurotoxicity. Brain Res. 901, 252–258 (2001).

    CAS  Article  Google Scholar 

  39. 39

    Russell, V., Allie, S. & Wiggins, T. Increased noradrenergic activity in prefrontal cortex slices of an animal model for attention-deficit hyperactivity disorder–the spontaneously hypertensive rat. Behav. Brain Res. 117, 69–74 (2000).

    CAS  Article  Google Scholar 

  40. 40

    Russell, V.A. Dopamine hypofunction possibly results from a defect in glutamate-stimulated release of dopamine in the nucleus accumbens shell of a rat model for attention deficit hyperactivity disorder–the spontaneously hypertensive rat. Neurosci. Biobehav. Rev. 27, 671–682 (2003).

    CAS  Article  Google Scholar 

  41. 41

    DasBanerjee, T. et al. A comparison of molecular alterations in environmental and genetic rat models of ADHD: a pilot study. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 147B, 1554–1563 (2008).

    CAS  Article  Google Scholar 

  42. 42

    Sagvolden, T. et al. The spontaneously hypertensive rat model of ADHD–the importance of selecting the appropriate reference strain. Neuropharmacology 57, 619–626 (2009).

    CAS  Article  Google Scholar 

  43. 43

    Del Bo, R. et al. DPP6 gene variability confers increased risk of developing sporadic amyotrophic lateral sclerosis in Italian patients. J. Neurol. Neurosurg. Psychiatry 79, 1085 (2008).

    CAS  PubMed  Google Scholar 

  44. 44

    Cronin, S., Tomik, B., Bradley, D.G., Slowik, A. & Hardiman, O. Screening for replication of genome-wide SNP associations in sporadic ALS. Eur. J. Hum. Genet. 17, 213–218 (2009).

    CAS  Article  Google Scholar 

  45. 45

    Marshall, C.R. et al. Structural variation of chromosomes in autism spectrum disorder. Am. J. Hum. Genet. 82, 477–488 (2008).

    CAS  Article  Google Scholar 

  46. 46

    Lesch, K.P. et al. Genome-wide copy number variation analysis in ADHD: association with neuropeptide Y gene dosage in an extended pedigree. Mol. Psychiatry 16, 491–503 (2011).

    CAS  Article  Google Scholar 

  47. 47

    Oades, R.D., Daniels, R. & Rascher, W. Plasma neuropeptide Y levels, monoamine metabolism, electrolyte excretion, and drinking behavior in children with attention-deficit hyperactivity-disorder (ADHD). Psychiatry Res. 80, 177–186 (1998).

    CAS  Article  Google Scholar 

  48. 48

    Renström, F. et al. Replication and extension of genome-wide association study results for obesity in 4923 adults from northern Sweden. Hum. Mol. Genet. 18, 1489–1496 (2009).

    Article  Google Scholar 

  49. 49

    Kessler, R.C. et al. Patterns and predictors of attention-deficit/hyperactivity disorder persistence into adulthood: results from the national comorbidity survey replication. Biol. Psychiatry 57, 1442–1451 (2005).

    Article  Google Scholar 

  50. 50

    Potkin, S.G. et al. FBIRN. A genome-wide association study of schizophrenia using brain activation as a quantitative phenotype. Schizophr. Bull. 35, 96–108 (2009).

    Article  Google Scholar 

  51. 51

    Wang, X., Bao, X., Pal, R., Agbas, A. & Michaelis, E.K. Transcriptomic responses in mouse brain exposed to chronic excess of the neurotransmitter glutamate. BMC Genomics 11, 360 (2010).

    Article  Google Scholar 

  52. 52

    de Lanerolle, N.C., Eid, T. & Lee, T.S. Genomic expression in the epileptogenic hippocampus and psychiatric co-morbidities. Curr. Psychiatry Rev. 6, 135–144 (2010).

    CAS  Article  Google Scholar 

  53. 53

    Ule, J. et al. An RNA map predicting Nova-dependent splicing regulation. Nature 444, 580–586 (2006).

    CAS  Article  Google Scholar 

  54. 54

    Ule, J. et al. Nova regulates brain-specific splicing to shape the synapse. Nat. Genet. 37, 844–852 (2005).

    CAS  Article  Google Scholar 

  55. 55

    Murias, M., Swanson, J.M. & Srinivasan, R. Functional connectivity of frontal cortex in healthy and ADHD children reflected in EEG coherence. Cereb. Cortex 17, 1788–1799 (2007).

    Article  Google Scholar 

  56. 56

    Wang, L. et al. Altered small-world brain functional networks in children with attention-deficit/hyperactivity disorder. Hum. Brain Mapp. 30, 638–649 (2009).

    CAS  Article  Google Scholar 

  57. 57

    Diskin, S.J. et al. Adjustment of genomic waves in signal intensities from whole-genome SNP genotyping platforms. Nucleic Acids Res. 36, e126 (2008).

    Article  Google Scholar 

  58. 58

    Dennis, G. Jr. et al. DAVID: Database for Annotation, Visualization, and Integrated Discovery. Genome Biol. 4, P3 (2003).

    Article  Google Scholar 

Download references

Acknowledgements

We thank all the children with ADHD and their families who participated in this study and all the control subjects who donated blood samples to The Children's Hospital of Philadelphia (CHOP) for genetic research purposes. We thank the technical staff at the Center for Applied Genomics, CHOP for generating the genotypes used in this study and the medical assistants, and nursing and medical staff who recruited the subjects. We thank the Center for Biomedical Informatics for bioinformatics support. We also thank S. Kristinsson, L.A. Hermannsson and A. Krisbjörnsson for their software design and contribution to the study.

We thank the IMAGE, IMAGE II and PUWMa consortium investigators and the NIMH for making the genotype data available. Funding support for International Multi-Center ADHD Genetics (IMAGE) and IMAGE II Projects was provided by US National Institutes of Health (NIH) grant R01MH62873 to S.V.F., and the genotyping of samples was provided through the Genetic Association Information Network (GAIN). The dataset used for the IMAGE analyses described in this manuscript were obtained from the database of Genotype and Phenotype (dbGaP), which is found at http://www.ncbi.nlm.nih.gov/gap, through dbGaP accession numbers phs000016.v2.p2 (ADHD IMAGE), phs000020.v2.p1 (depression) and phs000019.v1.p1 (psoriasis). Samples and associated phenotype data for the GAIN Major Depression: Stage 1 Genome-wide Association In Population Based Samples Study (principal investigator, P.F. Sullivan, University of North Carolina) were provided by D.I. Boomsma and E. de Geus, Vrije Universiteit Amsterdam (principal investigators, Netherlands Twin Register); B.W. Penninx, Vrije Universiteit Medical Center Amsterdam; F. Zitman, Leiden University Medical Center, Leiden; and W. Nolen, University Medical Center Groningen (principal investigators and site principal investigators, Netherlands Study of Depression and Anxiety). Samples and associated phenotype data for the Collaborative Association Study of Psoriasis were provided by J.T. Elder (University of Michigan, Ann Arbor, Michigan), G.G. Krueger (University of Utah, Salt Lake City, Utah), A. Bowcock (Washington University, St. Louis, Missouri) and G.R. Abecasis (University of Michigan, Ann Arbor, Michigan). Samples and associated phenotype data for the International Multi-Center ADHD Genetics Project were provided by the following investigators: S.V.F. (principal investigator), R.J.L.A., P.A., J.S., R.P.E., B.F., M. Gill, A. Miranda, F. Mulas, R.D.O., H.R., A. Rothenberger, T.B., J. Buitelaar, E. Sonuga-Barke and H.-C.S. (site principal investigators), M. Daly, C. Lange, N. Laird, J. Su and B. Neale (statistical analysis team). Samples and associated phenotype data were accessed through an authorized data access request by H.H. and S.F.A.G. Data collection for the PUWMa sample was supported by NIH grants to S.V.F., J. Biederman, S. Smalley, R. Todd and A.A.T. GWAS genotyping of the PUWMa samples was completed by Genizon and was provided through a grant for genotyping services to S.V.F. The PUWMa consortium represents a Pfizer-funded collaboration among the University of California Los Angeles, Washington University and Massachussetts General Hospital. We thank M.C. O'Donovan, M. Gill, M.J. Owen, P.A. Holmans, A. Thapar, B.M. Neale and A. Miranda for contributing DNA samples and phenotypes to the study and for editing the manuscript. We thank G. DePalma, T. Töpner, A. Guiney and H. Zhang for providing samples for the qRT-PCR CNV validation.

The study was supported by an Institutional Development Award to the Center for Applied Genomics from CHOP (H.H.), which funded all of the discovery genome-wide genotyping for this study. This work was additionally supported in part by NIH grant K23MH066275 (J.E.), University of Pennsylvania National Center for Research Resources Clinical and Translational Science Awards grant UL1-RR-024134 (J.E.), by a Research Development Award from the Cotswold Foundation (H.H. and S.F.A.G.) and by US Department of Health and Human Services grant 1RC2MH089924 (H.H.). N.T., T.S. and J.D.B. received support from the Seaver Foundation and a Conte Center for Neuroscience of Mental Disorders grant from the NIMH (P50MH066392).

Sample and phenotype data collection of parts of the IMAGE II cohort was supported by the Deutsche Forschungsgemeinschaft (KFO 125, SFB 581 and SFB TRR 58 to K.-P.L. and A. Reif, ME 1923/5-1 and ME 1923/5-3 to C.M.F. and J.M.) and the Bundesministerium für Bildung und Forschung (BMBF 01GV0605 to K.-P.L.).

Author information

Affiliations

Authors

Contributions

H.H. and J.E. designed the CHOP study and supervised the data analyses and interpretation. S.V.F., M.G., P.A. and J. Buitelaar designed the IMAGE and IMAGE II studies. S.V.F. designed the PUWMa study and coordinated the analyses for the IMAGE, IMAGE II and PUWMa studies. J.T.G. and K.W. conducted the statistical analyses. C.E.K. and E.C.F. directed the stage 1 genotyping. J.D.B. coordinated the validation analyses. N.T. performed the qRT-PCR validation of the CNVs. J.T.G. and H.H. drafted the manuscript. J.E. collected the CHOP samples. C.R., P.S. and J.L.R. collected the NIMH samples. C.M.F., H.-C.S., A.A.T., A. Reif, A. Rothenberger, B.F., E.O.M., H.R., J. Buitelaar, K.-P.L., L.K., T.B., R.P.E., F.M., R.D.O., J.S., E.S.-B., T.J.R., M.R., J.R., A.W., S.W., J.M., H.P., C.S., S.K.L., S.L.S., J. Biederman, L.K., P.A. and R.J.L.A. collected data for the IMAGE, IMAGE II and PUWMa projects. J. Biederman, E.O.M., S.V.F., S.K.L., S.L.S. and A.A.T. collected samples for the PUWMa study. F.A.M. genotyped the IMAGE II data. H.H. directed and D.H. and J.T.G. performed the gene interaction network and functional enrichment analyses. All authors contributed to the manuscript preparation. S.F.A.G. accessed the public domain data, assisted with the interpretation of the data and edited the manuscript. All other authors contributed samples and/or were involved with data mining and processing.

Corresponding author

Correspondence to Hakon Hakonarson.

Ethics declarations

Competing interests

For the last 3 years, M.R. has been in the speakers' bureau for Janssen-Cilag. In previous years, he was on the speakers' bureau for MEDICE.

Supplementary information

Supplementary Text and Figures

Supplementary Note, Supplementary Figures 1–11 and Supplementary Tables 1–17. (PDF 2931 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Elia, J., Glessner, J., Wang, K. et al. Genome-wide copy number variation study associates metabotropic glutamate receptor gene networks with attention deficit hyperactivity disorder. Nat Genet 44, 78–84 (2012). https://doi.org/10.1038/ng.1013

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing