一般口演

一般口演2

2021/9/30　11:00～12:00　オンデマンド D会場 O2-1 繊毛病関連蛋白質のhTERT-RPE1細胞における機能解析 Functional analysis of the ciliopathy-associated proteins in hTERT-RPE1 cells ^○銀生卓瑪¹, Ko Miyoshi¹, Sarina Han¹, Yuanyuan Qin¹, Genki Amano¹, Takeshi Yoshimura¹, Taiichi Katayama¹ 1.大阪大学・連合小児発達学研究科・分子生物遺伝学研究領域, 2.Department of Bifunctional Imaging, Medical Photonics Research Center, Hamamatsu University School of Medicine ^○Zhuoma Yinsheng¹, Ko Miyoshi¹, Sarina Han¹, Yuanyuan Qin¹, Genki Amano¹, Takeshi Yoshimura¹, Taiichi Katayama¹ 1.Department of Child Development and Molecular Brain Science, United Graduate School of Child Development, Osaka University, 2.Department of Bifunctional Imaging, Medical Photonics Research Center, Hamamatsu University School of Medicine Impaired function of primary cilia results in pleiotropic disorders collectively termed ciliopathies. Joubert syndrome (JBTS) is a ciliopathy characterized by retinal degeneration and brain malformation, while mechanism by which mutations of causative genes such as ARL13B, INPP5E, RPGRIP1L and CEP290 result in JBTS is poorly understood. The ARL13B and INPP5E proteins localize to the ciliary membrane, while the RPGRIP1L and CEP290 proteins localize to the proximal region of the cilium, termed the transition zone, and form a complex that functions as a ciliary gatekeeper. In this study, RPGRIP1L-knock out (KO) and CEP290-KO hTERT-RPE1 cells were established by genome editing. Primary cilia of these KO cells were longer than that of control cells. Further, the localization of the ARL13B and INPP5E proteins at the ciliary membrane was profoundly reduced in the KO cells. The exogenous expression of RPGRIP1L in RPGRIP1L-KO cells rescued the defect. Our finding suggests that the RPGRIP1L and CEP290 proteins contribute to the maintenance of primary cilia at the proper length and the localization of ARL13B and INPP5E at the ciliary membrane. I have no COI with regard to our presentation.		2021/9/30　11:00～12:00　オンデマンド D会場 O2-2 運動ニューロンにおけるRNA結合タンパク質Quakingの役割 An RNA-binding protein Quaking safeguards motor neuron function through pre-mRNA processing ^○矢野佳芳¹, Takako Furukawa¹, Akihide Koyama³, Chihiro Nakamoto⁴, Kenji Sakimura⁴, Osamu Onodeara⁵, Hirohide Takebayashi¹, Hideyuki Okano², Masato Yano^1,2 1.Division of Neurobiology and Anatomy, Niigata University Graduate School of Medical and Dental Sciences, 2.Department of Physiology, Keio University School of Medicine, 3.Department of Legal Medicine, Niigata University Graduate School of Medical and Dental Sciences, 4.Department of Animal Model Development, Brain Research Institute, Niigata University, 5.Department of Neurology, Brain Research Institute, Niigata University ^○Yoshika Hayakawa-Yano¹, Takako Furukawa¹, Akihide Koyama³, Chihiro Nakamoto⁴, Kenji Sakimura⁴, Osamu Onodeara⁵, Hirohide Takebayashi¹, Hideyuki Okano², Masato Yano^1,2 1.Division of Neurobiology and Anatomy, Niigata University Graduate School of Medical and Dental Sciences, 2.Department of Physiology, Keio University School of Medicine, 3.Department of Legal Medicine, Niigata University Graduate School of Medical and Dental Sciences, 4.Department of Animal Model Development, Brain Research Institute, Niigata University, 5.Department of Neurology, Brain Research Institute, Niigata University Emerging evidence has revealed that many RNA binding proteins (RBPs) link to dysregulation of RNA metabolism in motor neuron diseases, such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA). However, the molecular mechanisms underlying the vulnerability of the motor neuron are yet to be elucidated. We have been focusing on the roles of cell-specific expressing RBPs to understand how they generate cell-specific transcriptomics and function in CNS. In this study, we searched for RBPs that generate the motor neuron-ness through the post-transcriptional network, related to MND/ALS pathology. Here we show the molecular mechanism and cellular functions of one such an RBP, Qki5 that generates motor neuron-ness. Immunohistochemical analysis and single-cell transcriptome analysis using human iPS-derived motor neuron revealed that Qki5 is specifically expressed in the motor neurons among the other types of neurons, in mouse and human spinal cord. Interestingly, RNAseq analysis of NSC-34 cells, cortical neuron, and OPC revealed that Qki5 plays a crucial role to generate the motor neuron-specific transcriptome through the pre-mRNA splicing. By using mouse genetics, motor neuron-specific ablation of Qki protein causes neurodegeneration with TDP-43 proteinopathy in the young adult mouse. To further investigate Qki5 function, we used comprehensive approaches, RNAseq combined with HITS-CLIP and discovered novel Qki5-dependent alternative splicing events including dysregulation of the ALS causative genes, which could be relevant for the maintenance of motor neuron and stress response pathway. Finally, we will discuss the roles of Qki5 protein in RNA regulation to safeguard the motor neuron under various stress conditions and a link to neurodegenerative diseases.

2021/9/30　11:00～12:00　オンデマンド D会場 O2-3 C9orf72 FTLD/ALSにおけるRAN翻訳の新規調整因子 A novel modulator of RAN translation in C9orf72 FTLD/ALS ^○後藤志帆¹, Kohji Mori¹, Tomoko Yamashita¹, Yuya Kawabe¹, Tsubasa Omi¹, Tesshin Miyamoto¹, Ryota Uozumi¹, Shizuko Kondo¹, Yoshitaka Nagai², Manabu Ikeda¹ 1.大阪大学大学院, 2.Department of Neurology, Kindai University Faculty of Medicine ^○Shiho Gotoh¹, Kohji Mori¹, Tomoko Yamashita¹, Yuya Kawabe¹, Tsubasa Omi¹, Tesshin Miyamoto¹, Ryota Uozumi¹, Shizuko Kondo¹, Yoshitaka Nagai², Manabu Ikeda¹ 1.Department of Psychiatry, Osaka University Graduate School of Medicine, 2.Department of Neurology, Kindai University Faculty of Medicine Frontotemporal lover degeneration (FTLD) is a dementia that typically presents with behavioral disturbance and/or aphasia. FTLD patients has localized atrophy in their frontal and temporal lobes. Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease with muscle weakness and disruption of motor neuron. These two diseases party share disease mechanisms. An abnormally extended GGGGCC repeats in noncoding region of C9orf72 is a genetic cause of FTLD/ALS. This hexanucleotide repeat sequence is translated into dipeptide repeat proteins (DPR) via repeat associated non-ATG (RAN) translation. DPR accumulates in C9-FTLD/ALS patient’s brain and DPR toxicity has been shown in several disease models. Decreasing of DPR through RAN translation inhibition could have potential for novel therapeutics. However, mechanism of RAN translation is still obscure. In this study, we identified TR1 as a novel modulator of RAN translation by using cellular models of C9-FTLD/ALS. First, knockdown of TR1 reduced RAN translation dependent DPR expression. Conversely, overexpression of TR1 enhanced DPR expression. Repeat RNA didn’t increase with TR1 expression. Moreover, inhibition of TR1’s intrinsic activity decreased DPR expression. Next, to verify whether TR1 selectively regulate RAN translation, we performed puromycin incorporation assay that evaluate nascent polypeptides. In this assay, TR1 overexpression increased poly-GA expression more than puromycin labeled nascent polypeptides expression. This result suggests TR1 selectively modulates RAN translation compared with ATG translation. Collectively, our results implicate that regulation of TR1 could have therapeutic potential for C9-FTLD/ALS through modulating RAN translation.		2021/9/30　11:00～12:00　オンデマンド D会場 O2-4 ヒト死後脳における神経細胞とオリゴデンドロサイト系譜細胞のDNAメチル化状態の解析 Differences of DNA methylation status in neurons and oligodendrocyte lineage cells from postmortem human brains ^○林義剛¹, Kazuhiko Nakabayashi², Yasuhiro Go³, Junya Kamiyasuhira¹, Asmaa Abdullah¹, Mohd Zakiyyah¹, Seiji Hitoshi¹ 1.滋賀医科大学　統合臓器生理学, 2.Department of Maternal Fetal Biology, National Center for Child Health and Development, 3.Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences ^○Yoshitaka Hayashi¹, Kazuhiko Nakabayashi², Yasuhiro Go³, Junya Kamiyasuhira¹, Asmaa Abdullah¹, Mohd Zakiyyah¹, Seiji Hitoshi¹ 1.Department of Integrative Physiology, Shiga University of Medical Science, 2.Department of Maternal Fetal Biology, National Center for Child Health and Development, 3.Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences Epigenetic abnormalities in brains were suggested to play a role in the pathogenesis of psychiatric disorders. Although DNA methylation analysis of brain tissues has been performed using cortical gray matter, it is unclear whether the results reflect pathological conditions or differences in cell density. Therefore, cell type-specific analysis is necessary. In this study, we extracted intact cell nuclei from two human postmortem brains and stained with NeuN, a marker for neuronal cells, and Olig2, a marker for oligodendrocyte lineage cells, and sorted the positive cells to extract DNA. Then the methylation status of the DNA was comprehensively analyzed by the Reduced Representation Bisulfite Sequencing method using a next-generation sequencer. Using the analysis data, we performed gene ontology analysis and compared the methylation status of neurons and oligodendrocytes, and found that synaptic signaling, learning, and memory-related genes were hypomethylated in neurons. On the other hand, oligodendrocyte lineage cells showed hypomethylation in genes related to myelin and glial cell differentiation. In a comparison of mature oligodendrocyte-oligodendrocyte progenitor cells, mature oligodendrocytes were hypomethylated in genes related to myelin and myelination, whereas oligodendrocyte progenitor cells, interestingly, were hypomethylated in genes related to neurogenesis and suppression of cell differentiation. We believe that these results provide fundamental data for understanding the development and pathogenesis of the brain.