Cited by 567Neurogenesis and neuroplasticity mechanismsPluripotent Stem Cells ResearchFetal and Pediatric Neurological Disorders

Human iPSC-Derived Cerebral Organoids Model Cellular Features of Lissencephaly and Reveal Prolonged Mitosis of Outer Radial Glia

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Bershteyn, Marina, Nowakowski, Tomasz J., Pollen, Alex A., Di Lullo, Elizabeth, Nene, Aishwarya, Wynshaw-Boris, Anthony, Kriegstein, Arnold R.·2017·Cell Stem Cell

Abstract

Classical lissencephaly is a genetic neurological disorder associated with mental retardation and intractable epilepsy, and Miller-Dieker syndrome (MDS) is the most severe form of the disease. In this study, to investigate the effects of MDS on human progenitor subtypes that control neuronal output and influence brain topology, we analyzed cerebral organoids derived from control and MDS-induced pluripotent stem cells (iPSCs) using time-lapse imaging, immunostaining, and single-cell RNA sequencing. We saw a cell migration defect that was rescued when we corrected the MDS causative chromosomal deletion and severe apoptosis of the founder neuroepithelial stem cells, accompanied by increased horizontal cell divisions. We also identified a mitotic defect in outer radial glia, a progenitor subtype that is largely absent from lissencephalic rodents but critical for human neocortical expansion. Our study, therefore, deepens our understanding of MDS cellular pathogenesis and highlights the broad utility of cerebral organoids for modeling human neurodevelopmental disorders.

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Crossref · 10.1016/j.stem.2016.12.007

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  • Time-lapse confocal microscopy of organoid cells displaying the characteristic mitotic behaviour of oRGs provided further evidence of their identity.

    “Time-lapse confocal microscopy of organoid cells displaying the characteristic mitotic behaviour of oRGs provided further evidence of their identity49,50, and single-cell transcriptome sequencing has been used to confirm the presence of cells with oRG gene expression signatures50.”
  • Miller-Dieker syndrome organoids also showed severe apoptosis of neuroepithelial stem cells in the ventricular-like zone and a mitotic defect in outer radial glia.

    “The organoids also displayed severe apoptosis of neuroepithelial stem cells in the ventricular-like zone and a mitotic defect in oRGs50.”
  • In some cases brain organoids have reproduced disease phenotypes that were also observed in mouse models.

    “In some instances, brain organoids have recapitulated disease phenotypes observed in mouse models, validating both models50.”
  • Cerebral organoids generated from iPSCs of individuals with Miller-Dieker syndrome displayed several developmental phenotypes reported in lissencephaly mouse models, including dysregulation of the neuroepithelial stem cell mitotic spindle and neuronal migration defects.

    “iPSCs derived from individuals with MDS were used to generate cerebral organoids that exhibited several developmental phenotypes reported in lissencephaly mouse models, including dysregulation of the neuroepithelial stem cell mitotic spindle and neuronal migration defects50.”
  • Protocols using inducible transcription factor expression, especially from a genomic safe-harbor locus, can substantially reduce differentiation variability.

    “A strategy that can substantially reduce variability in differentiation is the use of protocols in which differentiation is driven by the inducible expression of transcription factors – in particular when the expression cassette for the inducible transcription factors is integrated in a defined genomic safe-harbor locus in iPSCs109.”

    From CRISPR-based functional genomics for neurological disease (2020) · cites this paper as [109]

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Authors
Bershteyn, M., Nowakowski, T. J., Pollen, A. A., Di Lullo, E., Nene, A., Wynshaw-Boris, A., & Kriegstein, A. R.
Affiliations
  • University of California, San Francisco (US)
  • California Institute of Technology (US)
Corresponding author
  • Bershteyn, Marina
  • Wynshaw-Boris, Anthony
  • Kriegstein, Arnold R.
Venue
Cell Stem Cell
Publisher
Elsevier BV
Published
2017-04
Volume
20
Issue
4
Pages
435-449.e4
ISSN
  • 1875-9777 (ISSN-L)
  • 1934-5909
OpenAlex
W2576459186
Cited by
566
References
90
Citations by year
2026: 19 · 2025: 34 · 2024: 55 · 2023: 57 · 2022: 87 · 2021: 70 · 2020: 87 · 2019: 68 · 2018: 58 · 2017: 31
FWCI
28.22
Type
journal article
Language
en
Open access
Locations
  • Cell stem cell (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion)
Funders
  • California Institute for Regenerative Medicine
  • CIRM (TG2-01153, GCIR-06673)
  • National Institute for Neurological Disorders and Stroke (5K99NS088572)
  • NIH (NS075998, MH105989)
Topics
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Pediatrics, Perinatology and Child Health › Fetal and Pediatric Neurological Disorders
Keywords
lissencephaly, Miller-Dieker syndrome, brain organoids, induced pluripotent stem cells, outer radial glia, mitosis, neuroepithelial stem cells, chromosomal deletion, neurodevelopmental disorders, apoptosis
MeSH terms
Adult, Cell Movement, Epithelium, Female, Humans, Infant, Infant, Newborn, Male, Middle Aged, Mitosis, Neuroglia, Neurons, Organoids, Apoptosis, Cytokinesis, Cerebrum, Lissencephaly, Classical Lissencephalies and Subcortical Band Heterotopias, Induced Pluripotent Stem Cells, Chromosome Duplication

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