DSCAM/PAK1 pathway suppression reverses neurogenesis deficits in iPSC-derived cerebral organoids from patients with Down syndrome
DSCAM/PAK1 pathway suppression reverses neurogenesis deficits in iPSC-derived cerebral organoids from patients with Down syndrome
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DOI：
10.1172/jci135763
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发表时间：
2021-06-15
影响因子：
15.9
通讯作者：
Liu, Yan
中科院分区：
文献类型：
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作者：
Tang, Xiao-Yan;Xu, Lei;Liu, Yan
唐氏综合症（DS）是由21染色体三体造成的，发生在每800个活产中的1例中。皮质发育的早期缺陷可能解释了DS中的认知障碍，尽管基本的分子机制仍然难以捉摸。在这里，我们对源自4种本质细胞系的大脑器官以及来自3个具有Trisome 21个个体的诱导多能干细胞（IPSC）的大脑器官进行了组织学测定和无偏的单细胞RNA-seq（SCRNA-SEQ）分析，以探索细胞型 - 型细胞型干细胞（IPSC）早期大脑发育过程中与DS相关的特定异常。我们发现，鉴于皮质下区域皮质神经元中II和IV层标记的增殖减少和表达降低，神经发生受到了显着影响。这可能是导致器官尺寸减小的原因。此外，使用CRISPR/CAS9，CRISPR干扰（CRISPRI）或小分子抑制剂治疗抑制DSCAM/PAK1途径在DS中显示出增强的活性。我们的研究表明，体外开发的3D皮质器官是DS的宝贵模型，并在DS中DSCAM/PAK1途径的失调与DS中的发育脑缺陷之间提供了直接联系。
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.