公募シンポジウム
シングルセル解析から見えてきた神経変性・神経精神疾患の細胞特性 |
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| 7月6日（木） 14:20-16:20 Room C
1SY⑤-1
アルツハイマー病モデルマウス脳を用いた疾患病理に付随するグリア細胞の探索
Exploration of glial cells associated with disease pathology using Alzheimer's disease model mouse brains
永田 健一1, 笹栗 弘貴2, 橋本 翔子3, 西道 隆臣2, 木山 博資1
1. 名古屋大学 大学院医学系研究科, 2. 理化学研究所 脳神経科学研究センター, 3. 滋賀医科大学 創発的研究センター
Kenichi Nagata1, Hiroki Sasaguri2, Shoko Hashimoto3, Takaomi C. Saido2, Hiroshi Kiyama1
1. Nagoya University, Graduate School of Medicine, 2. RIKEN, Center for Brain Science, 3. Shiga University of Medical Science, Medical Innovation Research Center
Alzheimer’s disease (AD) is the most common type of dementia. Before neurodegeneration, amyloid beta and tau are broadly accumulated in the AD brain. With the technical advances in single-cell RNA-seq, disease-associated microglia (DAM) have been reported to be associated with amyloid pathology. However, it remains unclear whether DAM, emerging after amyloid beta accumulation, change their states after tau accumulation and propagation. In this study, we performed single-cell RNA-seq in two groups of AD model mice: AppNL-G-F/MAPT (dKI) and human brain-derived tau-injected dKI (Tau-dKI). Cell nuclei were collected from the hippocampus of 24-month-old knock-in type AD model mice, and applied to the 10x Genomics platform for library preparation. Sequencing analyses discriminated five microglial subclusters. Among them, cluster 1 and cluster 3 were judged as DAM. The ratio of cluster 3 was significantly higher in Tau-dKI compared with dKI, on the other hand, that of cluster 1 was reduced in Tau-dKI. Both subclusters express a number of known DAM markers with different patterns as well as unique cluster markers. Subsequent histological analyses confirmed the expression of the identified markers in DAM surrounding the amyloid plaque. Our results raise the possibility that tau pathology induces further alteration of microglial states during the progression of AD. | | 7月6日（木） 14:20-16:20 Room C
1SY⑤-2
Rod-shapedミクログリアの遺伝子発現プロファイリングによるキャラクタリゼーション
Characterization of rod-shaped microglia by gene expression profiling
橋本 翔子1,2, 松葉 由紀夫1,2, 永田 健一3, 西道 隆臣2
1. 滋賀医科大学 創発的研究センター, 2. 理化学研究所 脳神経科学研究センター, 3. 名古屋大学大学院 医学系研究科
Shoko Hashimoto1,2, Yukio Matsuba1,2, Kenichi Nagata3, Takaomi Saido2
1. Medical Innovation Research Center, Shiga University of Medical Science, 2. RIKEN, Center for Brain Science, 3. Nagoya University, Graduate School of Medicine
When microglia become activated under the pathological condition, it changes its morphology and plays a neuroprotective or neurotoxic role. We found “rod-shaped microglia”, whose cell body is elongated, under oxidative stress.Glutathione, an important antioxidant in the brain, is decreased with ageing and in neurodegenerative diseases including Alzheimer's disease (AD). To elucidate the effect of glutathione loss on AD progression, we analyzed the AD pathologies in brain specific-conditional knockout mouse of glutamyl-cysteine ligase catalytic subunit (GCLC). GCLC-KO showed severe neuroinflammation from 2 months old, and brain atrophy caused by neuronal cell death from 6 months old. Interestingly, we observed “rod-shaped” microglia in the cortex and the hippocampal CA1 in young GCLC-KO. It appeared in 2-months old and disappeared by 6-7-months old, and almost all of them was lying in one direction: from layer V toward outside in cortex. To characterize rod-shaped microglia, expressions of some genes were evaluated by in situ hybridization. A part of rod-shaped microglia expressed cst7, a marker of disease associated microglia (DAM), suggesting that rod-shaped microglia are one of DAM-like activated microglia. We carried out single cell(sc) RNA-seq using GCLC-KO and will proceed the characterization of rod-shaped microglia with the result of scRNA-seq. | | 7月6日（木） 14:20-16:20 Room C
1SY⑤-3
Identification of risk factors using cellular models of neurodevelopmental disorders
野村 淳1,2, ずこ あみら2, 岸本 恵子2, 六峰 弘晃2, 深津 和美2, 野村 芳子2, るう ぎょうけい2, 中井 信裕1,2, 河野 掌3, しん じぇい3, たくみ とおる1,2,4
1. 神戸大学 医学部, 2. 理研BSI, 3. 理研IMS, 4. 理研BDR
Jun Nomura1,2, Amira Zuko2, Keiko Kishimoto2, Hiroaki Mutsumine2, Kazumi Fukatsu2, Yoshiko Nomura2, Xioxi Liu2, Nobuhiro Nakai1,2, Tsukasa Kouno3, Jay Shin3, Toru Takumi1,2,4
1. School of Medicine, Kobe Univ., 2. Brain Science Institute, RIKEN, 3. Integ. Med. Sci., RIKEN, 4. Biosys. Dyn. Res., RIKEN
Human genetics has identified numerous copy number variations (CNVs) associated with autism spectrum disorders (ASD). However, the lack of standardized biological resources impedes understanding of the cell-type-specific common features of ASD. Here, we establish a biological resource including 65 genetically modified mouse embryonic stem (ES) cell lines as genetic models of ASD. In this study, we performed neural differentiation using 12 representative cell lines, duplication of 1q21.1 (MIM: 612475), deletion of 2p16.3 (MIM: 614332), 3q29 (MIM: 609425), duplication of 7q11.23 (MIM: 613729), deletion of 15q11.2 (MIM: 615656), 15q13.3 (MIM: 612001), 16p11.2 (MIM: 611913), 16p13.2 (MIM: 616863), 17p11.2 (MIM: 182290), 17q12 (MIM: 614527), Xq27.3 (MIM: 300624) and, Xq28 (MIM: 312750) and their comprehensive analyses, including single-cell RNA sequencing, uncover cell-type-specific susceptible pathways. Moreover, we found that a common phenotype in glutamatergic and GABAergic neurons is reduced expression of Upf3b, a core member of the translational termination and nonsense-mediated decay (NMD). These findings emphasize that the dysfunction of translational machinery in the developing neurons can be a possible target of early intervention for ASD. This ES cell model bank becomes an invaluable resource for studies in vitro and in vivo of ASD or other neuropsychiatric disorders. | | 7月6日（木） 14:20-16:20 Room C
1SY⑤-4
統合失調症患者死後脳を使用したシングルセルレベルでのLINE-1新規挿入検出
Detection of novel somatic LINE-1 insertions at single neurons of patients with schizophrenia
文東 美紀
熊本大学院 分子脳科学
Miki Bundo
Dept. of Mol. Brain Sci., Kumamoto Univ., Kumamoto, Japan
Schizophrenia is one of the most serious of all mental illnesses. Although the causes of schizophrenia are largely unknown, previous epidemiologic studies have shown that the genetic factors play an important role.We hypothesized that somatic genomic variants that arise after fertilization and exist only in the brain cells may be associated with the etiology of schizophrenia. We have especially focused on long interspersed nucleotide element (LINE-1). LINE-1 is one of the most abundant human transposable elements with the ability to amplify its own sequence in the human genome. We previously reported that the copy number of LINE-1 was higher in the neurons of schizophrenia patients than in the neurons of healthy controls and these LINE-1 insertions in schizophrenia were enriched near the neuronal genes. However, as each cell in the brain is thought to have a different pattern of LINE-1 insertions, single-cell analysis should be required to describe the complete somatic variations in the brain.To this purpose, we have developed a novel technique named Novel Elements Concentrated-sequence (NECO-seq) for detecting LINE-1 insertions from a single neuronal nuclei and conducted a case-control study. We will review the recent progress of our research. | |
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