Letter | Published:

CHD8 haploinsufficiency results in autistic-like phenotypes in mice

Nature volume 537, pages 675679 (29 September 2016) | Download Citation


Autism spectrum disorder (ASD) comprises a range of neurodevelopmental disorders characterized by deficits in social interaction and communication as well as by restricted and repetitive behaviours1. ASD has a strong genetic component with high heritability. Exome sequencing analysis has recently identified many de novo mutations in a variety of genes in individuals with ASD2,3, with CHD8, a gene encoding a chromatin remodeller, being most frequently affected4,5,6,7,8. Whether CHD8 mutations are causative for ASD and how they might establish ASD traits have remained unknown. Here we show that mice heterozygous for Chd8 mutations manifest ASD-like behavioural characteristics including increased anxiety, repetitive behaviour, and altered social behaviour. CHD8 haploinsufficiency did not result in prominent changes in the expression of a few specific genes but instead gave rise to small but global changes in gene expression in the mouse brain, reminiscent of those in the brains of patients with ASD. Gene set enrichment analysis revealed that neurodevelopment was delayed in the mutant mouse embryos. Furthermore, reduced expression of CHD8 was associated with abnormal activation of RE-1 silencing transcription factor (REST), which suppresses the transcription of many neuronal genes. REST activation was also observed in the brains of humans with ASD, and CHD8 was found to interact physically with REST in the mouse brain. Our results are thus consistent with the notion that CHD8 haploinsufficiency is a highly penetrant risk factor for ASD, with disease pathogenesis probably resulting from a delay in neurodevelopment.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


Primary accessions


Data deposits

Sequencing data have been deposited in the DDBJ sequence read archive under accession number DRA003116.


  1. 1.

    & Advances in autism genetics: on the threshold of a new neurobiology. Nat. Rev. Genet. 9, 341–355 (2008)

  2. 2.

    et al. De novo mutations revealed by whole-exome sequencing are strongly associated with autism. Nature 485, 237–241 (2012)

  3. 3.

    et al. De novo gene disruptions in children on the autistic spectrum. Neuron 74, 285–299 (2012)

  4. 4.

    et al. Sporadic autism exomes reveal a highly interconnected protein network of de novo mutations. Nature 485, 246–250 (2012)

  5. 5.

    et al. Sequencing chromosomal abnormalities reveals neurodevelopmental loci that confer risk across diagnostic boundaries. Cell 149, 525–537 (2012)

  6. 6.

    et al. Patterns and rates of exonic de novo mutations in autism spectrum disorders. Nature 485, 242–245 (2012)

  7. 7.

    et al. Multiplex targeted sequencing identifies recurrently mutated genes in autism spectrum disorders. Science 338, 1619–1622 (2012)

  8. 8.

    et al. Disruptive CHD8 mutations define a subtype of autism early in development. Cell 158, 263–276 (2014)

  9. 9.

    & CHD proteins: a diverse family with strong ties. Biochem. Cell Biol. 85, 463–476 (2007)

  10. 10.

    & The Chd family of chromatin remodelers. Mutat. Res. 618, 30–40 (2007)

  11. 11.

    , , & CHD8 is an ATP-dependent chromatin remodeling factor that regulates β-catenin target genes. Mol. Cell. Biol. 28, 3894–3904 (2008)

  12. 12.

    et al. CHD8 suppresses p53-mediated apoptosis through histone H1 recruitment during early embryogenesis. Nat. Cell Biol. 11, 172–182 (2009)

  13. 13.

    , & Histone H1 recruitment by CHD8 is essential for suppression of the Wnt-β-catenin signaling pathway. Mol. Cell. Biol. 32, 501–512 (2012)

  14. 14.

    et al. Early embryonic death in mice lacking the β-catenin-binding protein Duplin. Mol. Cell. Biol. 24, 8386–8394 (2004)

  15. 15.

    et al. Abnormal behavior in a chromosome-engineered mouse model for human 15q11-13 duplication seen in autism. Cell 137, 1235–1246 (2009)

  16. 16.

    et al. Dysfunction in GABA signalling mediates autism-like stereotypies and Rett syndrome phenotypes. Nature 468, 263–269 (2010)

  17. 17.

    et al. Shank3 mutant mice display autistic-like behaviours and striatal dysfunction. Nature 472, 437–442 (2011)

  18. 18.

    et al. Autistic-like behaviour in Scn1a+/− mice and rescue by enhanced GABA-mediated neurotransmission. Nature 489, 385–390 (2012)

  19. 19.

    et al. Absence of CNTNAP2 leads to epilepsy, neuronal migration abnormalities, and core autism-related deficits. Cell 147, 235–246 (2011)

  20. 20.

    et al. CHD8 regulates neurodevelopmental pathways associated with autism spectrum disorder in neural progenitors. Proc. Natl Acad. Sci. USA 111, E4468–E4477 (2014)

  21. 21.

    et al. The autism-associated chromatin modifier CHD8 regulates other autism risk genes during human neurodevelopment. Nat. Commun. 6, 6404 (2015)

  22. 22.

    et al. Transcriptomic analysis of autistic brain reveals convergent molecular pathology. Nature 474, 380–384 (2011)

  23. 23.

    et al. Integrative functional genomic analyses implicate specific molecular pathways and circuits in autism. Cell 155, 1008–1021 (2013)

  24. 24.

    et al. Synaptic, transcriptional and chromatin genes disrupted in autism. Nature 515, 209–215 (2014)

  25. 25.

    et al. Spatio-temporal transcriptome of the human brain. Nature 478, 483–489 (2011)

  26. 26.

    , , & de novo convergence of autism genetics and molecular neuroscience. Trends Neurosci. 37, 95–105 (2014)

  27. 27.

    , , , & REST and its corepressors mediate plasticity of neuronal gene chromatin throughout neurogenesis. Cell 121, 645–657 (2005)

  28. 28.

    , & RE-1 silencing transcription factor (REST): a regulator of neuronal development and neuronal/endocrine function. Cell Tissue Res. 359, 99–109 (2015)

  29. 29.

    et al. REST regulates distinct transcriptional networks in embryonic and neural stem cells. PLoS Biol. 6, e256 (2008)

  30. 30.

    et al. Coexpression networks implicate human midfetal deep cortical projection neurons in the pathogenesis of autism. Cell 155, 997–1007 (2013)

  31. 31.

    , & Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting. Cell 73, 1155–1164 (1993)

  32. 32.

    et al. Establishment by the rat lymph node method of epitope-defined monoclonal antibodies recognizing the six different α chains of human type IV collagen. Histochem. Cell Biol. 104, 267–275 (1995)

  33. 33.

    et al. VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases. Genes Dev. 18, 3055–3065 (2004)

  34. 34.

    , , , & The identification of peroxisome proliferator-activated receptor alpha-independent effects of oleoylethanolamide on intestinal transit in mice. Neurogastroenterol. Motil. 21, 420–429 (2009)

  35. 35.

    et al. Deficiency of schnurri-2, an MHC enhancer binding protein, induces mild chronic inflammation in the brain and confers molecular, neuronal, and behavioral phenotypes related to schizophrenia. Neuropsychopharmacology 38, 1409–1425 (2013)

  36. 36.

    , , , & Comprehensive behavioral analysis of cluster of differentiation 47 knockout mice. PLoS One 9, e89584 (2014)

  37. 37.

    , & Behavioral phenotypes of genetic mouse models of autism. Genes Brain Behav. 15, 7–26 (2016)

  38. 38.

    , , , & T-maze forced alternation and left-right discrimination tasks for assessing working and reference memory in mice. J. Vis. Exp. 60, 3300 (2012)

  39. 39.

    Assessing nest building in mice. Nat. Protocols 1, 1117–1119 (2006)

  40. 40.

    et al. The classification of mRNA expression levels by the phosphorylation state of RNAPII CTD based on a combined genome-wide approach. BMC Genomics 12, 516 (2011)

  41. 41.

    , , , & Mapping and quantifying mammalian transcriptomes by RNA-Seq. Nat. Methods 5, 621–628 (2008)

  42. 42.

    et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl Acad. Sci. USA 102, 15545–15550 (2005)

  43. 43.

    , & Erroneous analyses of interactions in neuroscience: a problem of significance. Nat. Neurosci. 14, 1105–1107 (2011)

  44. 44.

    , , , & Controlling the false discovery rate in behavior genetics research. Behav. Brain Res. 125, 279–284 (2001)

  45. 45.

    & Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse. Dev. Biol. 244, 305–318 (2002)

  46. 46.

    et al. The chromatin remodeller CHD8 is required for E2F-dependent transcription activation of S-phase genes. Nucleic Acids Res. 42, 2185–2196 (2014)

Download references


We thank Y. Kita, K. Tsunematsu, K. Maehara, S. Hirata, M. Kato, Y. Nakajo, T. Akasaka, M. Tanaka, Y. Yamada and K. Takeda for technical assistance; as well as K. Tamada for discussion. Computed tomography was supported by the Center for Advanced Instrumental and Educational Support, Faculty of Agriculture, Kyushu University. This study was supported in part by KAKENHI and by a Grant-in-Aid for Scientific Research on Innovative Areas (Comprehensive Brain Science Network) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

Author information


  1. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, Fukuoka 812-8582, Japan

    • Yuta Katayama
    • , Masaaki Nishiyama
    • , Atsuki Kawamura
    •  & Keiichi I. Nakayama
  2. Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-1192, Japan

    • Hirotaka Shoji
    •  & Tsuyoshi Miyakawa
  3. Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, Fukuoka 812-8582, Japan

    • Yasuyuki Ohkawa
  4. Division of Bioinformatics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, Fukuoka 812-8582, Japan

    • Tetsuya Sato
    •  & Mikita Suyama
  5. RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan

    • Toru Takumi


  1. Search for Yuta Katayama in:

  2. Search for Masaaki Nishiyama in:

  3. Search for Hirotaka Shoji in:

  4. Search for Yasuyuki Ohkawa in:

  5. Search for Atsuki Kawamura in:

  6. Search for Tetsuya Sato in:

  7. Search for Mikita Suyama in:

  8. Search for Toru Takumi in:

  9. Search for Tsuyoshi Miyakawa in:

  10. Search for Keiichi I. Nakayama in:


M.N. and A.K. assisted with animal preparation and molecular biology experiments. H.S. and T.M. conducted behavioural studies. Y.O., T.S. and M.S. performed sequencing and data analysis. Y.K. performed all other experiments and data analysis. T.T. interpreted results. K.I.N. coordinated the study and wrote the manuscript. All authors discussed the data and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Masaaki Nishiyama or Keiichi I. Nakayama.

Reviewer Information Nature thanks E. Eichler, C. Powell and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figure 1, which show the original uncropped western or southern blots for Figures 1a, 4e and Extended Data Figures 1c, d, f, 2e, f, 6a, and 10f, h. The black frames denote how the gels were cropped for the final figure. It also contains a Supplementary Discussion and additional references.

Excel files

  1. 1.

    Supplementary Table 1

    This file contains the source data for behavioural tests.

  2. 2.

    Supplementary Table 2

    This file contains statistical analysis of behavioural data.

  3. 3.

    Supplementary Table 3

    This file contains results (MACS P value, expression level, fold change, and P value) for total genes of ChIP-seq or RNA-seq analysis used in this study.

  4. 4.

    Supplementary Table 4

    This file contains gene symbols of original gene sets and complete results of GSEA reported in the manuscript.

About this article

Publication history






Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.