DSCAM/PAK1 pathway suppression reverses neurogenesis deficits in iPSC-derived cerebral organoids from patients with Down syndrome
Open accessAbstract
Down syndrome (DS), caused by trisomy of chromosome 21, occurs in 1 of every 800 live births. Early defects in cortical development likely account for the cognitive impairments in DS, although the underlying molecular mechanism remains elusive. Here, we performed histological assays and unbiased single-cell RNA-Seq (scRNA-Seq) analysis on cerebral organoids derived from 4 euploid cell lines and from induced pluripotent stem cells (iPSCs) from 3 individuals with trisomy 21 to explore cell-type-specific abnormalities associated with DS during early brain development. We found that neurogenesis was significantly affected, given the diminished proliferation and decreased expression of layer II and IV markers in cortical neurons in the subcortical regions; this may have been responsible for the reduced size of the organoids. Furthermore, suppression of the DSCAM/PAK1 pathway, which showed enhanced activity in DS, using CRISPR/Cas9, CRISPR interference (CRISPRi), or small-molecule inhibitor treatment reversed abnormal neurogenesis, thereby increasing the size of organoids derived from DS iPSCs. Our study demonstrates that 3D cortical organoids developed in vitro are a valuable model of DS and provide a direct link between dysregulation of the DSCAM/PAK1 pathway and developmental brain defects in DS.
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Results from induced pluripotent stem cell studies of neurogenesis are discordant: the generation of neurons from induced pluripotent stem cells has been reported in some studies to be impaired and in others not to be impaired, which may be explained by technical differences in the cell lines and protocols used.
“However, results from these studies are discordant: the generation of neurons from iPSCs has been reported in some studies to be impaired74,77–79 and in others not to be impaired75,80,81, which may be explained by technical differences in the cell lines and protocols used (see Box 1).”
From Consequences of trisomy 21 for brain development in Down syndrome (2024) · cites this paper as [77]
Split into 2 atoms — 1 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 1 unverifiable. Expand the claim to see per-atom verdicts.
Experimental — accuracy not yet measuredGenome-wide dysregulation of gene expression in neurons likely leads to over-expression and/or under-expression of key genes that regulate synaptogenesis.
“Genome-wide dysregulation of gene expression (Box 3) in neurons likely leads to over-expression and/or under-expression of key genes that regulate synaptogenesis, but specific candidates have yet to be identified74,77,81,143.”
From Consequences of trisomy 21 for brain development in Down syndrome (2024) · cites this paper as [77]
The Introduction (page unavailable) reports both upregulated and downregulated genes in Down syndrome cerebral organoids, supporting the gene-expression component. It links those changes chiefly to neurogenesis and nervous-system development, not to genes regulating synaptogenesis; it does not establish that the expression changes cause synaptogenesis deficits in neurons. (LLM, conf=0.82)
Experimental — accuracy not yet measuredAnalyses of neural progenitor cells arising from trisomy 21 induced pluripotent stem cells have provided conflicting data, with some suggesting that apoptosis is increased and others seeing no effect.
“Analyses of NPCs arising from T21 iPSCs have provided conflicting data, with some suggesting that apoptosis is increased74,79,95 and others seeing no effect75,77,81,96.”
From Consequences of trisomy 21 for brain development in Down syndrome (2024) · cites this paper as [77]
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Biomedical applications of organoids in genetic diseases
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Human Brain Organoids Model Abnormal Prenatal Neural Development Induced by Thermal Stimulation
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iPSCs-Derived Neurons and Brain Organoids from Patients
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Chromosomal and cellular therapeutic approaches for Down syndrome: A research update
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Trisomy 21-driven metabolite alterations are linked to cellular injuries in Down syndrome
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Self-citationThe sodium channel SCN2A regulates cortical excitatory and inhibitory neurogenesis
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Self-citationBrain organoids and genome editing: A new era in understanding human brain development and disorders
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Cellularity Defects Are Not Ubiquitous in the Brains of Fetuses With Down Syndrome
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Self-citationModeling Down syndrome neurodevelopment with isogenic cerebral organoidsPreprint
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Spatiotemporal expression of Rest in the brain of Ts1Cje mouse model of Down syndromePreprint
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- Authors
- Tang, X.-Y., Xu, L., Wang, J., Hong, Y., Wang, Y., Zhu, Q., Wang, D., Zhang, X.-Y., Liu, C.-Y., Fang, K.-H., Han, X., Wang, S., Wang, X., Xu, M., Bhattacharyya, A., Guo, X., Lin, M., & Liu, Y.
- Affiliations
- State Key Laboratory of Reproductive Medicine (CN)
- Nanjing Medical University (CN)
- Xiamen University (CN)
- University of Wisconsin–Madison (US)
- Corresponding author
- Guo, Xing
- Lin, Mingyan
- Liu, Yan
- ORCID
- Venue
- Journal of Clinical Investigation
- Published
- 2021-06-15
- Volume
- 131
- Issue
- 12
- ISSN
- 0021-9738 (ISSN-L)
- 1558-8238
- PMC
- PMC8203468
- OpenAlex
- W3158718113
- Cited by
- 143
- References
- 64
- Citations by year
- 2026: 16 · 2025: 29 · 2024: 28 · 2023: 34 · 2022: 30 · 2021: 6
- FWCI
- 21.23
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Investigation (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Chinese Academy of Sciences (XDA16010306)
- Government of Jiangsu Province
- National Key Research and Development Program of China (2019YFA0802703)
- Topics
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Down syndrome and intellectual disability research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Keywords
- Down syndrome, Dscam, PAK1, neurogenesis, brain organoids, iPSC-derived organoids, cortical development, CRISPR interference, small molecule inhibitors, cognitive impairment, in vitro disease model
- MeSH terms
- Cell Line, Cerebral Cortex, Down Syndrome, Humans, Organoids, Signal Transduction, Cell Adhesion Molecules, p21-Activated Kinases, Neurogenesis, Induced Pluripotent Stem Cells
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