Analysis of mice modified with genes controlling the amplitude of circadian rhythm by CRISPR / Cas9 method

• Project/Area Number: 16K15193
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental physiology(including physical medicine and nutritional physiology)
• Research Institution: Saitama Medical University
• Principal Investigator: IKEDA MASAAKI 埼玉医科大学, 医学部, 教授 (80232198)
• Research Collaborator: KUMAGAI megumi ; NAKAJIMA yoshihiro
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: 時計遺伝子 / 概日リズム / Bmal1 / 視交叉上核 / ゲノム編集 / うつ病 / うつ病モデル / CRISPR/Cas9 / 遺伝子ノックダウン / 振幅 / 周期 / 気分障害 / リズム振幅

Outline of Final Research Achievements

Postmortem brain studies reported that the circadian rhythmic expression of clock genes was attenuated in the dorsolateral prefrontal cortex and anterior cingulate cortex (Li et al, PNAS 2013). This report suggested that depression has circadian rhythm dysfunction in the brain. The goal of this study was to create a depression model mouse with reduced amplitude of circadian rhythm in the brain region associated with the cause of depression. As a preparatory step for creating a model mouse, knockout cells of SRC1 of the p160 family, a factor that enhances the amplitude of the circadian rhythm, were created. The SRC1 knockout cells were shown to reduce the amplitude of Bmal1 and Per2 promoter rhythms.

Academic Significance and Societal Importance of the Research Achievements

概日リズムの機能が中枢神経系で減弱することは、感情障害の発症要因となることが想定されている。私たちは概日リズムの振幅を制御する因子を探索している。その過程でｐ160因子が振幅の増強に関わっていることを発見した。今回、この因子の発現を増強あるいは低下させたモデル細胞を作出し、概日リズム振幅の増減が細胞機能に及ぼす影響を解析した。今後はｐ160の機能を改変し、時計遺伝子発現リズムの振幅をうつ病発症と関連するとされる領域の神経細胞で減弱化させたマウスを作出し、中枢神経系におけるリズム振幅減弱化と感情障害との関連を解析する計画であり、これらの成果はうつ病の解明や新しい治療法開発につながると考えている。

Research Products

• [Journal Article] Cell-based screen identifies a new potent and highly selective CK2 inhibitor for modulation of circadian rhythms and cancer cell growth. (2019)
  • Author(s): Oshima T, Niwa Y, Kuwata K, Srivastava A, Hyoda T, Tsuchiya Y, Kumagai M, Tsuyuguchi M, Tamaru T, Sugiyama A, Ono N, Zolboot N, Aikawa Y, Oishi S, Nonami A, Arai F, Hagihara S, Yamaguchi J, Tama F, Kunisaki Y, Yagita K, Ikeda M, Kinoshita T, Kay SA, Itami K, Hirota T.
  • Journal Title: Science Advance Volume: 23 Issue: 1 Pages: 9060-9060
  • DOI: 10.1126/sciadv.aau9060
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Evaluation of Teneligliptin Effects on Transcriptional Activity of PPARγ in Cell-Based Assays (2018)
  • Author(s): Takenaka Yasuhiro、Inoue Ikuo、Nakano Takanari、Ikeda Masaaki、Kakinuma Yoshihiko、Ikegami Yuichi、Shimada Akira、Noda Mitsuhiko
  • Journal Title: Journal of Nippon Medical School Volume: 85 Issue: 2 Pages: 95-101
  • DOI: 10.1272/jnms.2018_85-15
  • NAID: 130006731178
  • ISSN: 1345-4676, 1347-3409
  • Year and Date: 2018-04-15
  • Peer Reviewed / Open Access
• [Journal Article] Regulation of molecular clock oscillations and phagocytic activity via muscarinic Ca2+ signaling in human retinal pigment epithelial cells (2017)
  • Author(s): Ikarashi R, Akechi H, Kanda Y, Ahmad A, Takeuchi K, Morioka E, Sugiyama T, Ebisawa T, Ikeda M & Ikeda M
  • Journal Title: Scientific Reports Volume: ７ Issue: 1 Pages: 44175-44175
  • DOI: 10.1038/srep44175
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Cellular and molecular basis of chronotherapy for cancer (2019)
  • Author(s): Masaaki Ikeda, Megumi Kumagai, Yasutsuna Sasaki, Yoshihiro Nakajima, Ken-Ichi Fujita
  • Organizer: The 9th Federation of the Asian and Oceanian Physiological Societies Congr
  • Int'l Joint Research / Invited
• [Presentation] The 20th anniversary of the discovery of mammalian clock genes (2018)
  • Author(s): Masaaki Ikeda, Megumi Kumagai
  • Organizer: 第94回 日本理学会大会
  • Invited
• [Presentation] Molecular and cellular basis of chronotherapy for cancer (2018)
  • Author(s): Masaaki Ikeda
  • Organizer: 7th Swiss Chronobiology Meeting
  • Int'l Joint Research / Invited
• [Presentation] ノーベル賞研究に貢献した時計遺伝子BMAL1の発見 (2018)
  • Author(s): 池田 正明
  • Organizer: 第５回時間栄養科学研究会
  • Invited
• [Presentation] がん分子標的薬の時間治療に向けて基盤解明 がん分子標的薬の時間治療に向けて基盤解明 がん分子標的薬の時間治療に向けて基盤解明 (2018)
  • Author(s): 池田 正明 , 熊谷 恵 , 中島 芳浩 , 佐々木 康綱 , 藤田 健一
  • Organizer: 第95回日本生理学会大会, 2018年3月30日（高松）
  • Invited
• [Presentation] 時計遺伝子発見と進歩- 発見20年を振り返って (2017)
  • Author(s): 池田正明、熊谷 恵
  • Organizer: 第94回日本生理学会大会
  • Place of Presentation: 浜松市
  • Year and Date: 2017-03-30
  • Invited
• [Presentation] 時計遺伝子発見と進歩-発見20年を振り返って (2017)
  • Author(s): 池田正明, 熊谷恵
  • Organizer: 第94回日本生理学会大会, 2017年3月30日（浜松）
  • Invited
• [Presentation] 概日時計の分子機構 (2017)
  • Author(s): 池田正明, 熊谷恵, 中島芳浩
  • Organizer: 第44回日本毒性学会学術年会, 2017年7月10日（横浜）
  • Invited
• [Presentation] Role for p160 coactivators in amplitude of circadian oscillation (2017)
  • Author(s): Kumagai M, Nakajima Y, Ikeda M
  • Organizer: Europian Biological Rhythms Society XV Cogress, July 30-August 3, 2017, Amsterdam
  • Int'l Joint Research
• [Presentation] 時計遺伝子の概日リズム性発現の振幅とコアクティベータの関与 (2016)
  • Author(s): 熊谷 恵、千葉康、中島芳浩、池田正明
  • Organizer: 第23回日本時間生物学会学術大会
  • Place of Presentation: 名古屋市
  • Year and Date: 2016-11-12
• [Remarks] 時計遺伝子Bma1
  • URL: http://www.saitama-med.ac.jp/uinfo/seiri2/Masaaki%20Ikeda2.html