Williams syndrome is a genetic neurodevelopmental disorder characterized by an uncommon hypersociability and a mosaic of retained and compromised linguistic and cognitive abilities. Nearly all clinically diagnosed individuals with Williams syndrome lack precisely the same set of genes, with breakpoints in chromosome band 7q11.23 (refs 1, 2, 3, 4, 5). The contribution of specific genes to the neuroanatomical and functional alterations, leading to behavioural pathologies in humans, remains largely unexplored. Here we investigate neural progenitor cells and cortical neurons derived from Williams syndrome and typically developing induced pluripotent stem cells. Neural progenitor cells in Williams syndrome have an increased doubling time and apoptosis compared with typically developing neural progenitor cells. Using an individual with atypical Williams syndrome6,7, we narrowed this cellular phenotype to a single gene candidate, frizzled 9 (FZD9). At the neuronal stage, layer V/VI cortical neurons derived from Williams syndrome were characterized by longer total dendrites, increased numbers of spines and synapses, aberrant calcium oscillation and altered network connectivity. Morphometric alterations observed in neurons from Williams syndrome were validated after Golgi staining of post-mortem layer V/VI cortical neurons. This model of human induced pluripotent stem cells8 fills the current knowledge gap in the cellular biology of Williams syndrome and could lead to further insights into the molecular mechanism underlying the disorder and the human social brain.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    et al. VI. Genome structure and cognitive map of Williams syndrome. J. Cogn. Neurosci. 12 (Suppl. 1), 89–107 (2000)

  2. 2.

    et al. Neural basis of genetically determined visuospatial construction deficit in Williams syndrome. Neuron 43, 623–631 (2004)

  3. 3.

    , , , & Bridging cognition, the brain and molecular genetics: evidence from Williams syndrome. Trends Neurosci . 22, 197–207 (1999)

  4. 4.

    , , , & The neurocognitive profile of Williams syndrome: a complex pattern of strengths and weaknesses. J. Cogn. Neurosci. 12 (Suppl. 1), 7–29 (2000)

  5. 5.

    , , & “Everybody in the world is my friend” hypersociability in young children with Williams syndrome. Am. J. Med. Genet. A 124, 263–273 (2004)

  6. 6.

    et al. Is it Williams syndrome? GTF2IRD1 implicated in visual-spatial construction and GTF2I in sociability revealed by high resolution arrays. Am. J. Med. Genet. A 149A, 302–314 (2009)

  7. 7.

    et al. An atypical deletion of the Williams–Beuren syndrome interval implicates genes associated with defective visuospatial processing and autism. J. Med. Genet. 44, 136–143 (2007)

  8. 8.

    , & Modeling neurodevelopmental disorders using human neurons. Curr. Opin. Neurobiol. 22, 785–790 (2012)

  9. 9.

    et al. Hemizygosity at the elastin locus in a developmental disorder, Williams syndrome. Nature Genet . 5, 11–16 (1993)

  10. 10.

    et al. Defining the social phenotype in Williams syndrome: a model for linking gene, the brain, and behavior. Dev. Psychopathol. 20, 1–35 (2008)

  11. 11.

    et al. A model for neural development and treatment of Rett syndrome using human induced pluripotent stem cells. Cell 143, 527–539 (2010)

  12. 12.

    et al. Feeder-free derivation of induced pluripotent stem cells from human immature dental pulp stem cells. Cell Transplant . 20, 1707–1719 (2011)

  13. 13.

    et al. 7q11.23 dosage-dependent dysregulation in human pluripotent stem cells affects transcriptional programs in disease-relevant lineages. Nature Genet . 47, 132–141 (2015)

  14. 14.

    et al. frizzled 9 is expressed in neural precursor cells in the developing neural tube. Dev. Genes Evol. 211, 453–457 (2001)

  15. 15.

    et al. Hippocampal and visuospatial learning defects in mice with a deletion of frizzled 9, a gene in the Williams syndrome deletion interval. Development 132, 2917–2927 (2005)

  16. 16.

    , & SiRNA of frizzled-9 suppresses proliferation and motility of hepatoma cells. Int. J. Oncol. 35, 861–866 (2009)

  17. 17.

    et al. Efficient differentiation of human pluripotent stem cells to endothelial progenitors via small-molecule activation of WNT signaling. Stem Cell Rep . 3, 804–816 (2014)

  18. 18.

    et al. Wnt/beta-catenin/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway. Mol. Cell. Biol. 22, 1172–1183 (2002)

  19. 19.

    et al. Wnt-mediated down-regulation of Sp1 target genes by a transcriptional repressor Sp5. J. Biol. Chem. 282, 1225–1237 (2007)

  20. 20.

    et al. A network of genetic repression and derepression specifies projection fates in the developing neocortex. Proc. Natl Acad. Sci. USA 109, 19071–19078 (2012)

  21. 21.

    et al. The Fezf2–Ctip2 genetic pathway regulates the fate choice of subcortical projection neurons in the developing cerebral cortex. Proc. Natl Acad. Sci. USA 105, 11382–11387 (2008)

  22. 22.

    , , , & The determination of projection neuron identity in the developing cerebral cortex. Curr. Opin. Neurobiol. 18, 28–35 (2008)

  23. 23.

    & Increased dendritic spine densities on cortical projection neurons in autism spectrum disorders. Brain Res . 1309, 83–94 (2010)

  24. 24.

    , & Orchestrating neuronal differentiation: patterns of Ca2+ spikes specify transmitter choice. Trends Neurosci . 27, 415–421 (2004)

  25. 25.

    et al. 3D pattern of brain abnormalities in Williams syndrome visualized using tensor-based morphometry. Neuroimage 36, 1096–1109 (2007)

  26. 26.

    & Frequentist prediction intervals and predictive distributions. Biometrika 92, 529–542 (2005)

  27. 27.

    et al. Transcriptome comparison of human neurons generated using induced pluripotent stem cells derived from dental pulp and skin fibroblasts. PLoS ONE 8, e75682 (2013)

  28. 28.

    , & Systematic optimization of human pluripotent stem cells media using Design of Experiments. Sci. Rep. 5, 9834 (2015)

  29. 29.

    , , & A. affy—analysis of Affymetrix GeneChip data at the probe level. Bioinformatics 20, 307–315 (2004)

  30. 30.

    & Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25, 402–408 ( 2001)

  31. 31.

    et al. Differential L1 regulation in pluripotent stem cells of humans and apes. Nature 503, 525–529 (2013)

  32. 32.

    , & WebGestalt: an integrated system for exploring gene sets in various biological contexts. Nucleic Acids Res . 33, W741–W748 (2005)

  33. 33.

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

  34. 34.

    et al. Single-cell transcriptomics reveals a population of dormant neural stem cells that become activated upon brain injury. Cell Stem Cell 17, 329–340 (2015)

  35. 35.

    et al. Methods for qPCR gene expression profiling applied to 1440 lymphoblastoid single cells. Methods 59, 71–79 (2013)

  36. 36.

    et al. Transcriptional and translational heterogeneity among neonatal mouse spermatogonia. Biol. Reprod. 92, 54 (2015)

  37. 37.

    et al. Regional dendritic and spine variation in human cerebral cortex: a quantitative golgi study. Cereb. Cortex 11, 558–571 (2001)

  38. 38.

    et al. Spatial organization of neurons in the frontal pole sets humans apart from great apes. Cereb. Cortex 21, 1485–1497 (2011)

  39. 39.

    Structural abnormalities of the cerebral cortex in human chromosomal aberrations: a Golgi study. Brain Res . 44, 625–629 (1972)

  40. 40.

    , , & Abnormal neuronal development in the visual cortex of the human fetus and infant with down’s syndrome. A quantitative and qualitative Golgi study. Brain Res . 225, 1–21 (1981)

  41. 41.

    , & Dendritic arborization in the human fetus and infant with the trisomy 18 syndrome. Brain Res. Dev. Brain Res. 54, 291–294 (1990)

  42. 42.

    Prenatal and early postnatal ontogenesis of the human motor cortex: a golgi study. II. The basket-pyramidal system. Brain Res . 23, 185–191 (1970)

  43. 43.

    , , & Perinatal growth of prefrontal layer III pyramids in Down syndrome. Pediatr. Neurol. 27, 36–38 (2002)

  44. 44.

    et al. Quantitative analysis of cortical pyramidal neurons after corpus callosotomy. Ann. Neurol. 54, 126–130 (2003)

  45. 45.

    A reliable Golgi-Kopsch modification. Brain Res. Bull. 4, 127–129 (1979)

  46. 46.

    , & The Golgi rapid method in clinical neuropathology: the morphologic consequences of suboptimal fixation. J. Neuropathol. Exp. Neurol. 37, 13–33 (1978)

  47. 47.

    & A quantitative dendritic analysis of Wernicke’s area in humans. I. Lifespan changes. J. Comp. Neurol. 327, 83–96 (1993)

  48. 48.

    , & The metric analysis of three-dimensional dendritic tree patterns: a methodological review. J. Neurosci. Methods 18, 127–151 (1986)

  49. 49.

    et al. PROMO: Real-time prospective motion correction in MRI using image-based tracking. Magn. Reson. Med. 63, 91–105 (2010)

  50. 50.

    et al. Prospective motion correction of high-resolution magnetic resonance imaging data in children. Neuroimage 53, 139–145 (2010)

  51. 51.

    et al. Prospective motion correction improves diagnostic utility of pediatric MRI scans. Pediatr. Radiol. 41, 1578–1582 (2011)

  52. 52.

    et al. Reliability in multi-site structural MRI studies: effects of gradient non-linearity correction on phantom and human data. Neuroimage 30, 436–443 (2006)

  53. 53.

    , & Cortical surface-based analysis. I. Segmentation and surface reconstruction. Neuroimage 9, 179–194 (1999)

  54. 54.

    , & Cortical surface-based analysis. II: Inflation, flattening, and a surface-based coordinate system. Neuroimage 9, 195–207 (1999)

  55. 55.

    et al. Sequence-independent segmentation of magnetic resonance images. Neuroimage 23 (Suppl 1), S69–S84 (2004)

  56. 56.

    et al. Automatically parcellating the human cerebral cortex. Cereb. Cortex 14, 11–22 (2004)

  57. 57.

    & Measuring the thickness of the human cerebral cortex from magnetic resonance images. Proc. Natl Acad. Sci. USA 97, 11050–11055 (2000)

  58. 58.

    et al. An automated labeling system for subdividing the human cerebral cortex on MRI scans into gyral based regions of interest. Neuroimage 31, 968–980 (2006)

  59. 59.

    , , & Automatic parcellation of human cortical gyri and sulci using standard anatomical nomenclature. Neuroimage 53, 1–15 (2010)

Download references


This work was supported by grants from the California Institute for Regenerative Medicine (CIRM) TR2-01814 and TR4-06747, the National Institutes of Health (NIH) through P01 NICHD033113, NIH Director’s New Innovator Award Program 1-DP2-OD006495-01, R01MH094753, R01MH103134, U19MH107367, U19MH106434, R01MH095741, a National Alliance for Research on Schizophrenia and Depression (NARSAD) Independent Investigator Grant to A.R.M., grants from the Bob and Mary Jane Engman, the JPB Foundation, Paul G. Allen Family Foundation, the Leona M. and Harry B. Helmsley Charitable Trust grant 2012-PG-MED002, Annette C. Merle-Smith, the G. Harold & Leila Y. Mathers Foundation, the Royal Thai Government Scholarship to T.C., a CIRM postdoctoral fellowship to C.A.T., the Rita L. Atkinson Graduate fellowship to B.H.-M and the University of California San Diego Kavli Institute for Brain and Mind. Human tissue was obtained from the University of Maryland Brain and Tissue Bank, which is a brain and tissue repository of the NIH NeuroBioBank. We acknowledge K. Jepsen for the DNA bead arrays and members of the Willert laboratory for assistance with the Wnt pathway experiments. We thank all the participants and their families.

Author information

Author notes

    • Thanathom Chailangkarn
    •  & Cleber A. Trujillo

    These authors contributed equally to this work.

    • Lisa Stefanacci



  1. University of California San Diego, School of Medicine, UCSD Stem Cell Program, Department of Pediatrics/Rady Children’s Hospital San Diego, La Jolla, California 92037, USA

    • Thanathom Chailangkarn
    • , Cleber A. Trujillo
    • , Beatriz C. Freitas
    • , Roberto H. Herai
    • , Lisa Stefanacci
    • , Sarah Romero
    •  & Alysson R. Muotri
  2. University of California San Diego, School of Medicine, Department of Cellular & Molecular Medicine, La Jolla, California 92037, USA

    • Thanathom Chailangkarn
    • , Cleber A. Trujillo
    • , Beatriz C. Freitas
    • , Roberto H. Herai
    • , Lisa Stefanacci
    • , Sarah Romero
    •  & Alysson R. Muotri
  3. Center for Academic Research and Training in Anthropogeny (CARTA), La Jolla, California 92093, USA

    • Thanathom Chailangkarn
    • , Cleber A. Trujillo
    • , Beatriz C. Freitas
    • , Roberto H. Herai
    • , Lisa Stefanacci
    • , Sarah Romero
    •  & Alysson R. Muotri
  4. National Center for Genetic Engineering and Biotechnology (BIOTEC), Virology and Cell Technology Laboratory, Pathum Thani 12120, Thailand

    • Thanathom Chailangkarn
  5. University of California San Diego, Department of Anthropology, La Jolla, California 92093, USA

    • Branka Hrvoj-Mihic
    • , Lisa Stefanacci
    • , Kari L. Hanson
    •  & Katerina Semendeferi
  6. Graduate Program in Health Sciences, School of Medicine, Pontifícia Universidade Católica do Paraná (PUCPR), Curitiba, Paraná, Brazil

    • Roberto H. Herai
  7. The Salk Institute for Biological Studies, Laboratory of Genetics, La Jolla, California 92037, USA

    • Diana X. Yu
    • , Maria C. Marchetto
    • , Cedric Bardy
    • , Lauren McHenry
    •  & Fred H. Gage
  8. University of California San Diego, Multimodal Imaging Laboratory, La Jolla, California 92093, USA

    • Timothy T. Brown
    • , Anders M. Dale
    •  & Eric Halgren
  9. University of California San Diego, School of Medicine, Department of Neurosciences, La Jolla, California 92093, USA

    • Timothy T. Brown
    • , M. Colin Ard
    •  & Eric Halgren
  10. University of California San Diego, Center for Human Development, La Jolla, California 92093, USA

    • Timothy T. Brown
  11. SAHMRI Mind & Brain Theme, Laboratory for Human Neurophysiology and Genetics, Flinders University School of Medicine, Adelaide, South Australia 5000, Australia

    • Cedric Bardy
  12. The Salk Institute for Biological Studies, Laboratory for Cognitive Neuroscience, La Jolla, California 92037, USA

    • Anna Järvinen
    • , Yvonne M. Searcy
    • , Michelle DeWitt
    • , Wenny Wong
    • , Philip Lai
    •  & Ursula Bellugi
  13. Colorado College, Department of Psychology, Colorado Springs, Colorado 80903, USA

    • Bob Jacobs
  14. University of California San Diego, School of Medicine, Department of Radiology, La Jolla, California 92093, USA

    • Anders M. Dale
  15. University of California San Diego, Department of Cognitive Science, La Jolla, California 92093, USA

    • Anders M. Dale
  16. University of Utah, Department of Pediatrics, Salt Lake City, Utah 84108, USA

    • Li Dai
    •  & Julie R. Korenberg
  17. University of Utah, The Brain Institute, Salt Lake City, Utah 84108, USA

    • Li Dai
    •  & Julie R. Korenberg
  18. University of California San Diego, Kavli Institute for Brain and Mind, La Jolla, California 92093, USA

    • Fred H. Gage
    • , Eric Halgren
    • , Katerina Semendeferi
    •  & Alysson R. Muotri
  19. University of California San Diego, School of Medicine, Neuroscience Graduate Program, La Jolla, California 92093, USA

    • Katerina Semendeferi
    •  & Alysson R. Muotri


  1. Search for Thanathom Chailangkarn in:

  2. Search for Cleber A. Trujillo in:

  3. Search for Beatriz C. Freitas in:

  4. Search for Branka Hrvoj-Mihic in:

  5. Search for Roberto H. Herai in:

  6. Search for Diana X. Yu in:

  7. Search for Timothy T. Brown in:

  8. Search for Maria C. Marchetto in:

  9. Search for Cedric Bardy in:

  10. Search for Lauren McHenry in:

  11. Search for Lisa Stefanacci in:

  12. Search for Anna Järvinen in:

  13. Search for Yvonne M. Searcy in:

  14. Search for Michelle DeWitt in:

  15. Search for Wenny Wong in:

  16. Search for Philip Lai in:

  17. Search for M. Colin Ard in:

  18. Search for Kari L. Hanson in:

  19. Search for Sarah Romero in:

  20. Search for Bob Jacobs in:

  21. Search for Anders M. Dale in:

  22. Search for Li Dai in:

  23. Search for Julie R. Korenberg in:

  24. Search for Fred H. Gage in:

  25. Search for Ursula Bellugi in:

  26. Search for Eric Halgren in:

  27. Search for Katerina Semendeferi in:

  28. Search for Alysson R. Muotri in:


A.R.M., T.C. and C.A.T. designed the experiments and wrote the manuscript with input from K.S. and all authors. T.C. processed DPCs, generated and characterized iPSCs, NPCs and neurons, and performed cell number, proliferation, and apoptosis experiments as well as FZD9 knockdown and overexpression and statistical analysis. C.A.T. performed C1 single-cell analyses, synaptic quantification, calcium imaging, cell density experiments, live neuronal morphology analysis and statistical analysis. B.C.F. performed MEA recording, PCR for retrovirus silencing and Wnt pathway gene-expression analysis. B.C.F. and S.E.R. prepared astrocytes for co-culture experiments, NPC characterization by flow cytometry and CHIR 98014 experiments. K.S. designed all morphometry experiments with B.H.-M. and B.J., and co-wrote the manuscript to link the various levels of investigation from the whole-brain imaging findings to the cellular level. L.S. prepared Golgi staining for post-mortem neurons with help from K.L.H. and B.J. B.H.-M. obtained morphometric data on iPSC-derived neurons and post-mortem neurons. D.X.Y., M.C.M., C.A.T. and L.M. performed calcium transient experiments and statistical analysis. T.T.B. performed brain scan and statistical analysis with help from A.M.D. C.B. performed electrophysiological tests. M.D., W.W., P.L. and Y.M.S. performed neurocognitive and social tests. A.J., Y.M.S., and M.C.A. performed analyses and interpretation of social/neurocognitive tests. R.H.H. performed bioinformatics analysis. L.D. and J.R.K. confirmed deletion of all cells from participants with WS who donated them for reprogramming. E.H., U.B., F.H.G., K.S. and A.R.M. edited the manuscript for publication.

Corresponding authors

Correspondence to Katerina Semendeferi or Alysson R. Muotri.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Note 1, Supplementary Tables 1-13 and Supplementary References.

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.