双相障碍脑类器官的转录组分析与功能特征研究。
Transcriptome analysis and functional characterization of cerebral organoids in bipolar disorder.
机构信息
Center for Genomic Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
Chemical Biology Program, Broad Institute of MIT & Harvard, Cambridge, MA, USA.
出版信息
Genome Med. 2020 Apr 19;12(1):34. doi: 10.1186/s13073-020-00733-6.
BACKGROUND
Reprogramming human induced pluripotent stem cells (iPSCs) from somatic cells and generating three-dimensional brain organoids from these iPSCs provide access to live human neuronal tissue with disease-specific genetic backgrounds.
METHODS
Cerebral organoids were generated from iPSCs of eight bipolar disorder (BPI) patients and eight healthy control individuals. RNA-seq experiments were undertaken using RNA isolated from the cerebral organoids. Functional activity in the cerebral organoids was studied using microelectrode arrays.
RESULTS
RNA-seq data comparing gene expression profiles in the cerebral organoids showed downregulation of pathways involved in cell adhesion, neurodevelopment, and synaptic biology in bipolar disorder along with upregulation of genes involved in immune signaling. The central hub in the network analysis was neurocan (NCAN), which is located in a locus with evidence for genome-wide significant association in BPI. Gene ontology analyses suggested deficits related to endoplasmic reticulum biology in BPI, which was supported by cellular characterization of ER-mitochondria interactions. Functional studies with microelectrode arrays revealed specific deficits in response to stimulation and depolarization in BPI cerebral organoids.
CONCLUSIONS
Our studies in cerebral organoids from bipolar disorder showed dysregulation in genes involved in cell adhesion, immune signaling, and endoplasmic reticulum biology; implicated a central role for the GWAS hit NCAN in the biology of BPI; and showed evidence of deficits in neurotransmission.
背景
将体细胞重编程为人类诱导多能干细胞（iPSCs），并从这些 iPSCs 中生成三维大脑类器官，为获得具有特定疾病遗传背景的活体人类神经元组织提供了途径。
方法
从 8 名双相情感障碍（BPI）患者和 8 名健康对照个体的 iPSCs 中生成大脑类器官。使用从大脑类器官中分离的 RNA 进行 RNA-seq 实验。使用微电极阵列研究大脑类器官中的功能活性。
结果
比较大脑类器官中基因表达谱的 RNA-seq 数据显示，BPI 中与细胞黏附、神经发育和突触生物学相关的途径下调，而与免疫信号相关的基因上调。网络分析的中心枢纽是神经钙黏蛋白（NCAN），它位于 BPI 中具有全基因组显著关联证据的基因座上。基因本体分析表明，BPI 中存在与内质网生物学相关的缺陷，这得到了内质网-线粒体相互作用的细胞特征的支持。使用微电极阵列进行的功能研究表明，BPI 大脑类器官对刺激和去极化的反应存在特定缺陷。
结论
我们在双相情感障碍大脑类器官中的研究表明，与细胞黏附、免疫信号和内质网生物学相关的基因失调；表明 GWAS 命中 NCAN 在 BPI 生物学中起核心作用；并显示神经传递缺陷的证据。
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