Abstract

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.

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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 (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

    Deceased.

Affiliations

  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

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Contributions

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.

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https://doi.org/10.1038/nature19067

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