Development of a novel gene therapy method via CRISPR/Cas9 mediated in vivo genome editing

• Project/Area Number: 15K18330
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Neurophysiology / General neuroscience
• Research Institution: Gunma University
• Principal Investigator: Konno Ayumu 群馬大学, 大学院医学系研究科, 講師 (40509048)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 遺伝子治療 / アデノ随伴ウイルスベクター / CRISPR/Cas9 / ゲノム編集 / 脊髄小脳失調症 / AAV

Outline of Final Research Achievements

In this study, we aimed to develop a novel gene therapy method for spinocerebellar ataxia type 1 with CRISPR/Cas9 mediated in vivo genome editing using a gene delivery by adeno-associated virus vector. Although we examined SpCas9 and SaCas9 for in vivo knockout, we could not observe the amelioration of an ataxia in a mouse model of spinocerebellar ataxia type 1. Further investigations for higher efficiencies of a gene expression and/or a gene editing are needed.

Research Products

• [Int'l Joint Research] University of Oxford(United Kingdom)
• [Int'l Joint Research] Trustees of Dartmouth College/University Tenessee/Max Planck Florida Institute(米国)
• [Journal Article] Intravenous administration of the adeno-associated virus-PHP.B capsid fails to upregulate transduction efficiency in the marmoset brain (2018)
  • Author(s): Matsuzaki Yasunori、Konno Ayumu、Mochizuki Ryuta、Shinohara Yoichiro、Nitta Keisuke、Okada Yukihiro、Hirai Hirokazu
  • Journal Title: Neuroscience Letters Volume: 665 Pages: 182-188
  • DOI: 10.1016/j.neulet.2017.11.049
  • Peer Reviewed
• [Journal Article] Elavl3 is essential for the maintenance of Purkinje neuron axons (2018)
  • Author(s): Ogawa Yuki、Kakumoto Kyoko、Yoshida Tetsu、Kuwako Ken-ichiro、Miyazaki Taisuke、Yamaguchi Junji、Konno Ayumu、Hata Junichi、Uchiyama Yasuo、Hirai Hirokazu、Watanabe Masahiko、Darnell Robert B.、Okano Hideyuki、Okano Hirotaka James
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 2722-2722
  • DOI: 10.1038/s41598-018-21130-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Viral Vector-Based Evaluation of Regulatory Regions in the Neuron-Specific Enolase (NSE) Promoter in Mouse Cerebellum In Vivo (2017)
  • Author(s): Shinohara Yoichiro、Ohtani Toshinori、Konno Ayumu、Hirai Hirokazu
  • Journal Title: The Cerebellum Volume: 16 Issue: 5-6 Pages: 913-922
  • DOI: 10.1007/s12311-017-0866-5
  • Peer Reviewed
• [Journal Article] Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging (2017)
  • Author(s): Harada Kazuki、Ito Motoki、Wang Xiaowen、Tanaka Mika、Wongso Devina、Konno Ayumu、Hirai Hirokazu、Hirase Hajime、Tsuboi Takashi、Kitaguchi Tetsuya
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 7351-7351
  • DOI: 10.1038/s41598-017-07820-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Minimal Purkinje Cell-Specific PCP2/L7 Promoter Virally Available for Rodents and Non-human Primates (2017)
  • Author(s): Nitta Keisuke、Matsuzaki Yasunori、Konno Ayumu、Hirai Hirokazu
  • Journal Title: Molecular Therapy - Methods & Clinical Development Volume: 6 Pages: 159-170
  • DOI: 10.1016/j.omtm.2017.07.006
  • Peer Reviewed / Open Access
• [Journal Article] Regulatory connection between the expression level of classical protein kinase C and pruning of climbing fibers from cerebellar Purkinje cells (2017)
  • Author(s): Takahashi Nobutaka、Shuvaev Anton N.、Konno Ayumu、Matsuzaki Yasunori、Watanave Masashi、Hirai Hirokazu
  • Journal Title: Journal of Neurochemistry Volume: 143 Issue: 6 Pages: 660-670
  • DOI: 10.1111/jnc.14239
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Inflammation-induced reversible switch of the neuron-specific enolase promoter from Purkinje neurons to Bergmann glia. (2016)
  • Author(s): Sawada Y, Konno A, Nagaoka J, Hirai H
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 27758-27758
  • DOI: 10.1038/srep27758
  • Peer Reviewed / Open Access
• [Journal Article] Fluorescent-based evaluation of chaperone-mediated autophagy and microautophagy activities in cultured cells. (2016)
  • Author(s): Sato M, Seki T, Konno A, Hirai H, Kurauchi Y, Hisatsune A, Katsuki H.
  • Journal Title: Genes to Cells Volume: 21 Issue: 8 Pages: 861-873
  • DOI: 10.1111/gtc.12390
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Viral Vector-Based Dissection of Marmoset GFAP Promoter in Mouse and Marmoset Brains. (2016)
  • Author(s): Shinohara Y, Konno A, Takahashi N, Matsuzaki Y, Kishi S, Hirai H
  • Journal Title: PLoS ONE Volume: 11 Issue: 8 Pages: e0162023-e0162023
  • DOI: 10.1371/journal.pone.0162023
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] FMRP Expression Levels in Mouse CNS Neurons Determine Behavioral Phenotype. (2016)
  • Author(s): Arsenault J, Gholizadeh S, Niibori Y, Pacey LK, Halder SK, Koxhioni E, Konno A, Hirai H, Hampson DR
  • Journal Title: Human Gene Therapy Volume: 27 Issue: 12 Pages: 982-996
  • DOI: 10.1089/hum.2016.090
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Fusion of Human Fetal Mesenchymal Stem Cells with “Degenerating” Cerebellar Neurons in Spinocerebellar Ataxia Type 1 Model Mice. (2016)
  • Author(s): Huda F, Fan Y, Suzuki M, Konno A, Matsuzaki Y, Takahashi N, Chan JK, Hirai H
  • Journal Title: PLoS ONE Volume: 11 Issue: 11 Pages: e0164202-e0164202
  • DOI: 10.1371/journal.pone.0164202
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Transduction Profile of the Marmoset Central Nervous System Using Adeno-Associated Virus Serotype 9 Vectors (2016)
  • Author(s): Matsuzaki Y, Konno A, Mukai R, Honda F, Hirato M, Yoshimoto Y, Hirai H.
  • Journal Title: Molecular Neurobiology Volume: in press Issue: 3 Pages: 1-16
  • DOI: 10.1007/s12035-016-9777-6
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Calcium imaging reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain (2016)
  • Author(s): H Monai, M Ohkura, M Tanaka, Y Oe, A Konno, H Hirai, K Mikoshiba, S Itohara, J Nakai, Y Iwai & H Hirase
  • Journal Title: Nature communications Volume: 7 Issue: 1 Pages: 11100-11100
  • DOI: 10.1038/ncomms11100
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Plasticity of the developmentally arrested staggerer cerebellum in response to exogenous RORα. (2015)
  • Author(s): Iizuka A, Matsuzaki Y, Konno A, Hirai H.
  • Journal Title: Brain Structure and Function Volume: in press Issue: 6 Pages: 1-11
  • DOI: 10.1007/s00429-015-1077-9
  • Peer Reviewed
• [Presentation] In vivo evaluation of the shortened-WPRE in the cerebellum (2017)
  • Author(s): Ayumu Konno, Kazuma Hachisu, Hirokazu Hirai
  • Organizer: 第40回日本神経科学大会
• [Presentation] アデノ随伴ウイルスによる遺伝子導入を介したリピート病モデル動物の作出と治療法の開発 (2017)
  • Author(s): 今野 歩, 渡邊 将, 星野 千秋, 篠原 洋一郎, 松﨑 泰教, 平井 宏和
  • Organizer: 2017年度生命科学系学会合同年次大会（ConBio2017）
• [Presentation] Generation of animal models of spinocerebellar ataxia by AAV9-mediated gene delivery (2016)
  • Author(s): Ayumu Konno, Yasunori Matsuzaki & Hirokazu Hirai
  • Organizer: 第93回日本生理学会大会
  • Place of Presentation: 札幌コンベンションセンター/北海道札幌市
  • Year and Date: 2016-03-22
• [Presentation] AAV9 vector-mediated production of animal models of spinocerebellar ataxia (2016)
  • Author(s): Ayumu Konno, Yasunori Matsuzaki & Hirokazu Hirai
  • Organizer: International Society of Radiation Neurobiology 6th Conference
  • Place of Presentation: 良順会館（長崎大学医学部坂本キャンパス内）/長崎県長崎市
  • Year and Date: 2016-02-12
  • Int'l Joint Research
• [Presentation] A marmoset model of spinocerebellar ataxia type 3 generated using AAV vectors (2015)
  • Author(s): Ayumu Konno, Yasunori Matsuzaki, Yoichiro Shinohara, and Hirokazu Hirai
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場/展示場・兵庫県神戸市
  • Year and Date: 2015-07-28
• [Remarks] 群馬大学大学院医学系研究科脳神経再生医学Web site
  • URL: http://synapse.dept.med.gunma-u.ac.jp/
• [Remarks] 赤色 cAMP 可視化蛍光タンパク質センサーの開発
  • URL: http://www.c.u-tokyo.ac.jp/info/news/topics/files/20170810pressrelease.pdf
• [Remarks] 群馬大学大学院医学系研究科 脳神経再生医学分野
  • URL: http://synapse.dept.med.gunma-u.ac.jp/
• [Remarks] 群馬大学大学院医学系研究科脳神経再生医学分野
  • URL: http://synapse.dept.med.gunma-u.ac.jp/
• [Remarks] 微弱な電気刺激が脳を活性化する仕組みを解明
  • URL: http://www.riken.jp/pr/press/2016/20160322_1/