Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Altered proliferation and networks in neural cells derived from idiopathic autistic individuals

Molecular Psychiatry volume 22, pages 820–835 (2017)Cite this article

Subjects

Abstract

Autism spectrum disorders (ASD) are common, complex and heterogeneous neurodevelopmental disorders. Cellular and molecular mechanisms responsible for ASD pathogenesis have been proposed based on genetic studies, brain pathology and imaging, but a major impediment to testing ASD hypotheses is the lack of human cell models. Here, we reprogrammed fibroblasts to generate induced pluripotent stem cells, neural progenitor cells (NPCs) and neurons from ASD individuals with early brain overgrowth and non-ASD controls with normal brain size. ASD-derived NPCs display increased cell proliferation because of dysregulation of a β-catenin/BRN2 transcriptional cascade. ASD-derived neurons display abnormal neurogenesis and reduced synaptogenesis leading to functional defects in neuronal networks. Interestingly, defects in neuronal networks could be rescued by insulin growth factor 1 (IGF-1), a drug that is currently in clinical trials for ASD. This work demonstrates that selection of ASD subjects based on endophenotypes unraveled biologically relevant pathway disruption and revealed a potential cellular mechanism for the therapeutic effect of IGF-1.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Piven J, Palmer P, Jacobi D, Childress D, Arndt S . Broader autism phenotype: evidence from a family history study of multiple-incidence autism families. Am J Psychiatry 1997; 154: 185–190.

    Article  CAS  PubMed  Google Scholar 

  2. Ronald A, Happe F, Bolton P, Butcher LM, Price TS, Wheelwright S et al. Genetic heterogeneity between the three components of the autism spectrum: a twin study. J Am Acad Child Adolesc Psychiatry 2006; 45: 691–699.

    Article  PubMed  Google Scholar 

  3. Garber K . Neuroscience. Autism's cause may reside in abnormalities at the synapse. Science 2007; 317: 190–191.

    Article  CAS  PubMed  Google Scholar 

  4. Marchetto MC, Carromeu C, Acab A, Yu D, Yeo GW, Mu Y et al. A model for neural development and treatment of Rett syndrome using human induced pluripotent stem cells. Cell 2010; 143: 527–539.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Sanders SJ, Murtha MT, Gupta AR, Murdoch JD, Raubeson MJ, Willsey AJ et al. De novo mutations revealed by whole-exome sequencing are strongly associated with autism. Nature 2012; 485: 237–241.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Voineagu I, Wang X, Johnston P, Lowe JK, Tian Y, Horvath S et al. Transcriptomic analysis of autistic brain reveals convergent molecular pathology. Nature 2011; 474: 380–384.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Chow ML, Pramparo T, Winn ME, Barnes CC, Li HR, Weiss L et al. Age-dependent brain gene expression and copy number anomalies in autism suggest distinct pathological processes at young versus mature ages. PLoS Genet 2012; 8: e1002592.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Parikshak NN, Luo R, Zhang A, Won H, Lowe JK, Chandran V et al. Integrative functional genomic analyses implicate specific molecular pathways and circuits in autism. Cell 2013; 155: 1008–1021.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Willsey AJ, Sanders SJ, Li M, Dong S, Tebbenkamp AT, Muhle RA et al. Coexpression networks implicate human midfetal deep cortical projection neurons in the pathogenesis of autism. Cell 2013; 155: 997–1007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Courchesne E, Campbell K, Solso S . Brain growth across the life span in autism: age-specific changes in anatomical pathology. Brain Res 2011; 1380: 138–145.

    Article  CAS  PubMed  Google Scholar 

  11. Courchesne E, Redcay E, Morgan JT, Kennedy DP . Autism at the beginning: microstructural and growth abnormalities underlying the cognitive and behavioral phenotype of autism. Dev Psychopathol 2005; 17: 577–597.

    Article  PubMed  Google Scholar 

  12. Hazlett HC, Poe MD, Gerig G, Styner M, Chappell C, Smith RG et al. Early brain overgrowth in autism associated with an increase in cortical surface area before age 2 years. Arch Gen Psychiatry 2011; 68: 467–476.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Courchesne E, Karns CM, Davis HR, Ziccardi R, Carper RA, Tigue ZD et al. Unusual brain growth patterns in early life in patients with autistic disorder: an MRI study. Neurology 2001; 57: 245–254.

    Article  CAS  PubMed  Google Scholar 

  14. Courchesne E, Carper R, Akshoomoff N . Evidence of brain overgrowth in the first year of life in autism. JAMA 2003; 290: 337–344.

    Article  PubMed  Google Scholar 

  15. Shen MD, Nordahl CW, Young GS, Wootton-Gorges SL, Lee A, Liston SE et al. Early brain enlargement and elevated extra-axial fluid in infants who develop autism spectrum disorder. Brain 2013; 136 (Pt 9): 2825–2835.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Courchesne E, Mouton PR, Calhoun ME, Semendeferi K, Ahrens-Barbeau C, Hallet MJ et al. Neuron number and size in prefrontal cortex of children with autism. JAMA 2011; 306: 2001–2010.

    Article  CAS  PubMed  Google Scholar 

  17. Stoner R, Chow ML, Boyle MP, Sunkin SM, Mouton PR, Roy S et al. Patches of disorganization in the neocortex of children with autism. N Engl J Med 2014; 370: 1209–1219.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Takahashi K, Yamanaka S . Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006; 126: 663–676.

    Article  CAS  PubMed  Google Scholar 

  19. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 2007; 318: 1917–1920.

    Article  CAS  PubMed  Google Scholar 

  20. Freitas BC, Trujillo CA, Carromeu C, Yusupova M, Herai RH, Muotri AR . Stem cells and modeling of autism spectrum disorders. Exp Neurol 2012; 260: 33–43.

    Article  PubMed  Google Scholar 

  21. Shcheglovitov A, Shcheglovitova O, Yazawa M, Portmann T, Shu R, Sebastiano V et al. SHANK3 and IGF1 restore synaptic deficits in neurons from 22q13 deletion syndrome patients. Nature 2013; 503: 267–271.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Pasca SP, Portmann T, Voineagu I, Yazawa M, Shcheglovitov A, Pasca AM et al. Using iPSC-derived neurons to uncover cellular phenotypes associated with Timothy syndrome. Nat Med 2011; 17: 1657–1662.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Beltrao-Braga PC, Pignatari GC, Russo FB, Fernandes IR, Muotri AR . In-a-dish: induced pluripotent stem cells as a novel model for human diseases. Cytometry A 2013; 83: 11–17.

    Article  CAS  PubMed  Google Scholar 

  24. Griesi-Oliveira K, Acab A, Gupta AR, Sunaga DY, Chailangkarn T, Nicol X et al. Modeling non-syndromic autism and the impact of TRPC6 disruption in human neurons. Mol Psychiatry 2015; 20: 1350–1365.

    Article  CAS  PubMed  Google Scholar 

  25. Gmitrowicz A, Kucharska A . Developmental disorders in the fourth edition of the American classification: diagnostic and statistical manual of mental disorders (DSM IV — optional book). Psychiatr Pol 1994; 28: 509–521.

    CAS  PubMed  Google Scholar 

  26. Sanders SJ, Ercan-Sencicek AG, Hus V, Luo R, Murtha MT, Moreno-De-Luca D et al. Multiple recurrent de novo CNVs, including duplications of the 7q11.23 Williams Syndrome Region, are strongly associated with autism. Neuron 2011; 70: 863–885.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Li H, Durbin R . Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 2009; 25: 1754–1760.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N et al. The sequence alignment/map format and SAMtools. Bioinformatics 2009; 25: 2078–2079.

    Article  PubMed  PubMed Central  Google Scholar 

  29. McEvilly RJ, de Diaz MO, Schonemann MD, Hooshmand F, Rosenfeld MG . Transcriptional regulation of cortical neuron migration by POU domain factors. Science 2002; 295: 1528–1532.

    Article  CAS  PubMed  Google Scholar 

  30. Wang K, Li M, Hakonarson H . ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res 2010; 38: e164.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 2007; 131: 861–872.

    Article  CAS  PubMed  Google Scholar 

  32. Voineagu I . Gene expression studies in autism: moving from the genome to the transcriptome and beyond. Neurobiol Dis 2012; 45: 69–75.

    Article  CAS  PubMed  Google Scholar 

  33. Chen Y, Huang WC, Sejourne J, Clipperton-Allen AE, Page DT . Pten mutations alter brain growth trajectory and allocation of cell types through elevated beta-catenin signaling. J Neurosci 2015; 35: 10252–10267.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Stein JL, de la Torre-Ubieta L, Tian Y, Parikshak NN, Hernandez IA, Marchetto MC et al. A quantitative framework to evaluate modeling of cortical development by neural stem cells. Neuron 2014; 83: 69–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Belinson H, Nakatani J, Babineau BA, Birnbaum RY, Ellegood J, Bershteyn M et al. Prenatal beta-catenin/Brn2/Tbr2 transcriptional cascade regulates adult social and stereotypic behaviors. Mol Psychiatry 2016 (in press).

  36. Rubenstein JL . Three hypotheses for developmental defects that may underlie some forms of autism spectrum disorder. Curr Opin Neurol 2010; 23: 118–123.

    Article  PubMed  Google Scholar 

  37. Zikopoulos B, Barbas H . Altered neural connectivity in excitatory and inhibitory cortical circuits in autism. Front Hum Neurosci 2013; 7: 609.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Sugitani Y, Nakai S, Minowa O, Nishi M, Jishage K, Kawano H et al. Brn-1 and Brn-2 share crucial roles in the production and positioning of mouse neocortical neurons. Genes Dev 2002; 16: 1760–1765.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Niethammer M, Kim E, Sheng M . Interaction between the C terminus of NMDA receptor subunits and multiple members of the PSD-95 family of membrane-associated guanylate kinases. J Neurosci 1996; 16: 2157–2163.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Takamori S, Rhee JS, Rosenmund C, Jahn R . Identification of a vesicular glutamate transporter that defines a glutamatergic phenotype in neurons. Nature 2000; 407: 189–194.

    Article  CAS  PubMed  Google Scholar 

  41. Hahamy A, Behrmann M, Malach R . The idiosyncratic brain: distortion of spontaneous connectivity patterns in autism spectrum disorder. Nat Neurosci 2015; 18: 302–309.

    Article  CAS  PubMed  Google Scholar 

  42. Davis G, Plaisted-Grant K . Low endogenous neural noise in autism. Autism 2014; 19: 351–356.

    Article  PubMed  Google Scholar 

  43. Oldham MC, Konopka G, Iwamoto K, Langfelder P, Kato T, Horvath S et al. Functional organization of the transcriptome in human brain. Nat Neurosci 2008; 11: 1271–1282.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Konopka G, Bomar JM, Winden K, Coppola G, Jonsson ZO, Gao F et al. Human-specific transcriptional regulation of CNS development genes by FOXP2. Nature 2009; 462: 213–217.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Carper RA, Moses P, Tigue ZD, Courchesne E . Cerebral lobes in autism: early hyperplasia and abnormal age effects. Neuroimage 2002; 16: 1038–1051.

    Article  PubMed  Google Scholar 

  46. Sparks BF, Friedman SD, Shaw DW, Aylward EH, Echelard D, Artru AA et al. Brain structural abnormalities in young children with autism spectrum disorder. Neurology 2002; 59: 184–192.

    Article  CAS  PubMed  Google Scholar 

  47. Hazlett HC, Poe M, Gerig G, Smith RG, Provenzale J, Ross A et al. Magnetic resonance imaging and head circumference study of brain size in autism: birth through age 2 years. Arch Gen Psychiatry 2005; 62: 1366–1376.

    Article  PubMed  Google Scholar 

  48. Courchesne E, Pierce K, Schumann CM, Redcay E, Buckwalter JA, Kennedy DP et al. Mapping early brain development in autism. Neuron 2007; 56: 399–413.

    Article  CAS  PubMed  Google Scholar 

  49. Garcia-Jimenez C, Garcia-Martinez JM, Chocarro-Calvo A, De la Vieja A . A new link between diabetes and cancer: enhanced WNT/beta-catenin signaling by high glucose. J Mol Endocrinol 2014; 52: R51–R66.

    Article  CAS  PubMed  Google Scholar 

  50. Palsgaard J, Emanuelli B, Winnay JN, Sumara G, Karsenty G, Kahn CR . Cross-talk between insulin and Wnt signaling in preadipocytes: role of Wnt co-receptor low density lipoprotein receptor-related protein-5 (LRP5). J Biol Chem 2012; 287: 12016–12026.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Doble BW, Woodgett JR . GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci 2003; 116 (Pt 7): 1175–1186.

    Article  CAS  PubMed  Google Scholar 

  52. Desbois-Mouthon C, Cadoret A, Blivet-Van Eggelpoel MJ, Bertrand F, Cherqui G, Perret C et al. Insulin and IGF-1 stimulate the beta-catenin pathway through two signalling cascades involving GSK-3beta inhibition and Ras activation. Oncogene 2001; 20: 252–259.

    Article  CAS  PubMed  Google Scholar 

  53. Siddle K . Signalling by insulin and IGF receptors: supporting acts and new players. J Mol Endocrinol 2011; 47: R1–10.

    Article  CAS  PubMed  Google Scholar 

  54. Chenn A, Walsh CA . Regulation of cerebral cortical size by control of cell cycle exit in neural precursors. Science 2002; 297: 365–369.

    Article  CAS  PubMed  Google Scholar 

  55. Hirabayashi Y, Itoh Y, Tabata H, Nakajima K, Akiyama T, Masuyama N et al. The Wnt/beta-catenin pathway directs neuronal differentiation of cortical neural precursor cells. Development 2004; 131: 2791–2801.

    Article  CAS  PubMed  Google Scholar 

  56. Munji RN, Choe Y, Li G, Siegenthaler JA, Pleasure SJ . Wnt signaling regulates neuronal differentiation of cortical intermediate progenitors. J Neurosci 2011; 31: 1676–1687.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Cheng CM, Reinhardt RR, Lee WH, Joncas G, Patel SC, Bondy CA . Insulin-like growth factor 1 regulates developing brain glucose metabolism. Proc Natl Acad Sci USA 2000; 97: 10236–10241.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Ciucci F, Putignano E, Baroncelli L, Landi S, Berardi N, Maffei L . Insulin-like growth factor 1 (IGF-1) mediates the effects of enriched environment (EE) on visual cortical development. PLoS One 2007; 2: e475.

    Article  PubMed  PubMed Central  Google Scholar 

  59. Yoshii A, Constantine-Paton M . BDNF induces transport of PSD-95 to dendrites through PI3K-AKT signaling after NMDA receptor activation. Nat Neurosci 2007; 10: 702–711.

    Article  CAS  PubMed  Google Scholar 

  60. Sweatt JD . The neuronal MAP kinase cascade: a biochemical signal integration system subserving synaptic plasticity and memory. J Neurochem 2001; 76: 1–10.

    Article  CAS  PubMed  Google Scholar 

  61. Ozdinler PH, Macklis JD . IGF-I specifically enhances axon outgrowth of corticospinal motor neurons. Nat Neurosci 2006; 9: 1371–1381.

    Article  PubMed  Google Scholar 

  62. Corvin AP, Molinos I, Little G, Donohoe G, Gill M, Morris DW et al. Insulin-like growth factor 1 (IGF1) and its active peptide (1-3)IGF1 enhance the expression of synaptic markers in neuronal circuits through different cellular mechanisms. Neurosci Lett 2012; 520: 51–56.

    Article  CAS  PubMed  Google Scholar 

  63. Mariani J, Coppola G, Zhang P, Abyzov A, Provini L, Tomasini L et al. FOXG1-dependent dysregulation of GABA/glutamate neuron differentiation in autism spectrum disorders. Cell 2015; 162: 375–390.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the California Institute for Regenerative Medicine (CIRM) TR2-01814 and TR4-06747, the National Institutes of Health through the NIH Director’s New Innovator Award Program (1-DP2-OD006495-01), an R01 MH100175-01 and U19MH107367 from NIMH, the International Rett Syndrome Foundation (IRSF Grant No. 2915); a NARSAD Independent Investigator Award to ARM, and NIMH Autism Center of Excellence Program Project grant (to EC, KP, AW-B and FHG); this work was also supported by the Helmsley Trust, the JPB Foundation, the Engmann Foundation, a grant from the CDMRP Autism Research Program (to AW-B and FHG); a KL2 CTRI (KL2TR00099) to TP and Postdoctoral Translational Fellowship from Autism Speaks to HB.

Author information

Author notes

  1. M C Marchetto and H Belinson: These two authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Genetics, The Salk Institute, La Jolla, CA, USA

    M C Marchetto, K C Vadodaria, A P D Mendes, Y Nunez, H Ghosh, R Wright & F H Gage

  2. Department of Pediatrics, Institute for Human Genetics, University of California, San Francisco, CA, USA

    H Belinson & A Wynshaw-Boris

  3. Department of Neurology, Program in Neurogenetics, Center for Autism Research and Treatment, Semel Institute, David Geffen School of Medicine, University of California, Los Angeles, CA, USA

    Y Tian, J Ou & D H Geschwind

  4. Department of Pediatrics/Rady Children's Hospital San Diego, Department of Cellular and Molecular Medicine, Stem Cell Program, University of California, San Diego, La Jolla, CA, USA

    B C Freitas, P C Beltrao-Braga, C A Trujillo, Y Nunez & A R Muotri

  5. Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, USA

    C Fu & A Wynshaw-Boris

  6. Department of Obstetrics and Surgery, Center for Cellular and Molecular Therapy, University of São Paulo, São Paulo, Brazil

    P C Beltrao-Braga

  7. Department of Neuroscience, University of Rochester School of Medicine and Dentistry, Rochester, NY, USA

    K Padmanabhan

  8. Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY, USA

    K J Brennand

  9. Department of Neurosciences, University of California, San Diego, La Jolla, CA, USA

    K Pierce, L Eichenfield, T Pramparo, L T Eyler, C C Barnes & E Courchesne

Authors
  1. M C Marchetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H Belinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y Tian
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B C Freitas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K C Vadodaria
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. P C Beltrao-Braga
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. C A Trujillo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A P D Mendes
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. K Padmanabhan
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Y Nunez
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. J Ou
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. H Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. R Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. K J Brennand
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. K Pierce
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. L Eichenfield
    View author publications

    You can also search for this author in PubMed Google Scholar

  18. T Pramparo
    View author publications

    You can also search for this author in PubMed Google Scholar

  19. L T Eyler
    View author publications

    You can also search for this author in PubMed Google Scholar

  20. C C Barnes
    View author publications

    You can also search for this author in PubMed Google Scholar

  21. E Courchesne
    View author publications

    You can also search for this author in PubMed Google Scholar

  22. D H Geschwind
    View author publications

    You can also search for this author in PubMed Google Scholar

  23. F H Gage
    View author publications

    You can also search for this author in PubMed Google Scholar

  24. A Wynshaw-Boris
    View author publications

    You can also search for this author in PubMed Google Scholar

  25. A R Muotri
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to A Wynshaw-Boris or A R Muotri.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Molecular Psychiatry website

Supplementary information

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Marchetto, M., Belinson, H., Tian, Y. et al. Altered proliferation and networks in neural cells derived from idiopathic autistic individuals. Mol Psychiatry 22, 820–835 (2017). https://doi.org/10.1038/mp.2016.95

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/mp.2016.95

This article is cited by

Search

Advanced search

Quick links