enChIPによる遺伝子座特異的ヒストン修飾酵素複合体とエネルギー代謝機構の解明

2015 Fiscal Year Annual Research Report

• Project/Area Number: 26560395
• Research Institution: The University of Tokyo
• Principal Investigator: 酒井 寿郎 東京大学, 先端科学技術研究センター, 教授 (80323020)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 生活習慣病 / エピゲノム / ヒストン修飾酵素

Outline of Annual Research Achievements

前年度の進捗と研究推進可能性の実務的考察に基づき、DNA切断活性のある3xFLAG-Cas9-NLSを一過性に発現させ熱産生遺伝子Ucp1もしくはAdrb1の遠隔にある活性化エンハンサー領域を欠失させる細胞を樹立してproof of conceptの実験を行うことを優先する推進方策をとった。
CRISPR-CAS9の系で、β1アドレナリン受容体 (Adrb1) の遠隔エンハンサー領域 (-44kb) を欠失させることを目的に、この領域で核内受容体型転写因子PPARγが結合する部位を挟むようにガイドRNAを作製し、2つの異なるガイドRNAをCRISPR-CAS9に組み込んだ発現ベクターを不死化褐色脂肪細胞にトランスフェクションを行った。細胞のソーティングを行い、シングルクローンを選び出した。父方母方の2つのアレルがともに欠失しているクローン5個を約100個のシングルクローンスクリーニングから得ることに成功した。
これに先立ち、不死化脂肪細胞はシングルクローン化し、脂質染色で一様に染まる分化度の良いクローンを選んで上記のプロセスを施行した。
結果、遠隔のエンハンサー領域を欠失した褐色脂肪細胞はAdrb1のβアドレナリン（βAR）刺激に伴う遺伝子発現が減少し、我々がNat Commun (2015) 6, 7052に報告したβAR刺激依存的にAdrb1領域のクロマチンの高次構造変化により急速に転写が制御されることを証明した。

Research Products

• [Journal Article] The KDM3A-KLF2-IRF4 axis maintains myeloma cell survival (2016)
  • Author(s): Ohguchi H, Hideshima T, Bhasin MK, Gorgun GT, Santo L, Cea M, Mimura N, Suzuki R, Tai Y-T, Carrasco RD, Raje N, Richardson PG, Harigae H, Sanda T, Sakai J, Anderson KC
  • Journal Title: Nature Communications Volume: 7 Pages: 10258
  • DOI: 10.1038/ncomms10258
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] JMJD1A is a signal-sensing scaffold that regulates acute chromatin dynamics via SWI/SNF association for thermogenesis (2015)
  • Author(s): Abe Y, Rozqie R, Matsumura Y, Kawamura T, Nakaki R, Tsurutani Y, Tanimura-Inagaki K, Shiono A, Magoori K, Nakamura K, Ogi S, Kajimura S, Kimura H, Tanaka T, Fukami K, Osborne TF, Kodama T, Aburatani H, Inagaki T, Sakai J.
  • Journal Title: Nature Communications Volume: 6 Pages: 7052
  • DOI: 10.1038/ncomms8052
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Transcriptome Analysis of K-877 (a Novel Selective PPARα Modulator (SPPARMα))-Regulated Genes in Primary Human Hepatocytes and the Mouse Liver. (2015)
  • Author(s): Raza-Iqbal S, Tanaka T, Anai M, Inagaki T, Matsumura Y, Ikeda K, Taguchi A, Gonzalez FJ, Sakai J, Kodama T.
  • Journal Title: Journal of Atherosclerosis and Thrombosis Volume: 22 Pages: 754-772
  • DOI: 10.5551/jat.28720
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Analysis of the subcellular localization of the human histone methyltransferase SETDB1 (2015)
  • Author(s): Tachibana K, Gotoh E, Kawamata N, Ishimoto K, Uchihara Y, Iwanari H, Sugiyama A, Kawamura T, Mochizuki Y, Tanaka T, Sakai J, Hamakubo T, Kodama T, Doi T.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 465 Pages: 725-731
  • DOI: 10.1016/j.bbrc.2015.08.065
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] H3K4/H3K9me3 Bivalent Chromatin Domains Targeted by Lineage-Specific DNA Methylation Pauses Adipocyte Differentiation (2015)
  • Author(s): Matsumura Y, Nakaki R, Inagaki T, Yoshida A, Kano Y, Kimura H, Tanaka T, Tsutsumi S, Nakao M, Doi T, Fukami K, Osborne TF, Kodama T, Aburatani H, Sakai J
  • Journal Title: Molecular Cell Volume: 60 Pages: 584-596
  • DOI: 10.1016/j.molcel.2015.10.025
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] H3K4/H3K9me3 Bivalent Chromatin Domains Targeted by Lineage-Specific DNA Methylation Pauses Adipogenesis. (2016)
  • Author(s): Yoshihiro Matsumura, Ryo Nakaki, Takeshi Inagaki, Ayano Yoshida, Yuka Kano, Hiroshi Kimura, Toshiya Tanaka, Shuichi Tsutsumi, Mitsuyoshi Nakao, Takefumi Doi, Kiyoko Fukami, Timothy F. Osborne, Tatsuhiko Kodama, Hiroyuki Aburatani, and Juro Sakai
  • Organizer: The Keystone Symposia -Chromatin and Epigenetics
  • Place of Presentation: Whistler, Canada
  • Year and Date: 2016-03-01
  • Int'l Joint Research
• [Presentation] Metabolic regulation of Adipocyte differentiation by isocitrate dehydrogenease and Jmjc histone (2015)
  • Author(s): 酒井寿郎, Eko F.Ariyant, 曽我朋義, 稲垣毅, 松村欣宏
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
  • Invited
• [Presentation] 細胞系譜特異的DNAメチル化により形成されるH3K4/H3K9me3クロマチンドメインは脂肪細胞分化を停止させる (2015)
  • Author(s): 松村欣宏, 吉田文乃, 仲木竜, 鹿野優佳, 稲垣毅, 油谷浩幸, 酒井寿郎
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
  • Invited
• [Presentation] SETDY Regulates C/EBP alpha Transcription during Adipogenesis (2015)
  • Author(s): 馬郡健太, 山﨑あゆむ, 大木翔太郎, 松村欣宏, 稲垣毅, 酒井寿郎
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
• [Presentation] Metabolic Regulation of Histone Demethylation and Glycolysis Gene Expression by Isocitrate Dehydrogenase 3 during Adipogenesis. (2015)
  • Author(s): Eko Fuji Ariyanto, Yoshihiro Matsumura, Tomoyoshi Soga, Takeshi Inagaki and Juro Sakai
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
• [Presentation] ヒストンメチル化酵素SETDB1のユビキチン化修飾 (2015)
  • Author(s): 鹿野優佳, 松村欣宏, 吉田文乃, 柳茂, 稲垣毅, 酒井寿郎
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
• [Presentation] Functional Analysis of the Transcriptional Regulation by Enhancers of the Thermogenesis Related Genes (2015)
  • Author(s): 小山一貴, 阿部陽平, 松村欣宏, 稲垣毅, 酒井寿郎
  • Organizer: 第38回日本分子生物学会年会・第88回生化学会大会合同大会
  • Place of Presentation: 神戸
  • Year and Date: 2015-12-03
• [Presentation] JMJD1A is a Mediator of Long-Range Genomic Interactions to Regulate Metabolic Gene Expression (2015)
  • Author(s): Takeshi Inagaki, Yohei Abe, Royhan Rozqie, Yoshihiro Matsumura, Takeshi Kawamura, Tatsuhiko Kodama, Hiroyuki Aburatani, Juro Sakai
  • Organizer: Keystone Symposia Conference;Diabetes
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 2015-10-26
  • Int'l Joint Research
• [Presentation] Regulation of Higher-order Chromatin Structure during Thermogenesis in Brown Adipocytes (2015)
  • Author(s): Takeshi Inagaki, Yohei Abe, Royhan Rozqie, Yoshihiro Matsumura, Shingo Kajimura, Juro Sakai
  • Organizer: The 46th NIPS International Symposium
  • Place of Presentation: Nagoya
  • Year and Date: 2015-10-03
  • Int'l Joint Research / Invited
• [Presentation] PKA dependent phospho-switch on JMJD1A regulates higher-order chromatin dynamics via SWI/SNF and PPARγ association for thermogenesis. (2015)
  • Author(s): Yohei Abe, Royhan Rozqie, Yoshihiro Matsumura, Takeshi Kawamura, Ryo Nakaki, Shotaro Ogi, Toshiya Tanaka, Tatsuhiko Kodama, Hiroyuki Aburatani, Takeshi Inagaki, Juro Sakai
  • Organizer: EMBO Conference on Nuclear Receptors
  • Place of Presentation: Ajaccio, Corsica, France
  • Year and Date: 2015-09-24 – 2015-09-28
  • Int'l Joint Research
• [Presentation] JMJD1A Regulates Metabolic Gene Expression by Mediating Long-Range Genomic Interactions in Response to β-Adrenergic Stimulation under Cold Environment (2015)
  • Author(s): Inagaki, T., Abe, Y., Rozqie, R., Matsumura, Y., Kawamura, T., Nakaki, R., Kodama, T., Aburatani, H., Sakai, J.
  • Organizer: THE 40th NAITO CONFERENCE ON Epigenetics-From Histone Code to Therapeutic Strategy
  • Place of Presentation: Sapporo
  • Year and Date: 2015-09-17
• [Presentation] Lineage-specific gene-body DNA methylation and non-canonical H3K4/H3K9me3 bivalent chromatin domains maintain adipocyte lineage (2015)
  • Author(s): Yoshihiro Matsumura, Ayano Yoshida, Ryo Nakaki, Kenichi Wakabayashi, Takeshi Inagaki, Kiyoko Fukami, Hiroyuki Aburatani, Juro Sakai
  • Organizer: THE 40th NAITO CONFERENCE ON Epigenetics-From Histone Code to Therapeutic Strategy
  • Place of Presentation: Sapporo
  • Year and Date: 2015-09-15 – 2015-09-18
• [Presentation] β-Adrenergic induced phosphorylation of JMJD1A drives energy expenditure via SWI/SNF chromatin remodeler in Brown Adipocytes (2015)
  • Author(s): Sakai, J.
  • Organizer: The 47th Annual Scientific Meeting of the Japan Atherosclerosis Society
  • Place of Presentation: Sendai
  • Year and Date: 2015-07-10
  • Invited
• [Presentation] アディポバイオロジーを制御するヒストン脱メチル化酵素JMJD1Aの修飾シグナル (2015)
  • Author(s): 稲垣毅, 阿部陽平, Rozqie Royhan, 松村欣宏, 梶村真吾, 児玉龍彦, 油谷浩幸, 酒井寿郎
  • Organizer: 第58回日本糖尿病学会年次学術集会
  • Place of Presentation: 下関
  • Year and Date: 2015-05-22
  • Invited
• [Presentation] A Role of Isocitrate Dehydrogenase 3 in Regulation of Epigenome and Transcription during Adipogenesis (2015)
  • Author(s): Juro Sakai, Yoshihiro Matsumura, Ariyanto Fuji Eko, Takeshi Inagaki
  • Organizer: 12th Asian Congress of Nutrirtion
  • Place of Presentation: Yokohama
  • Year and Date: 2015-05-16
  • Int'l Joint Research / Invited
• [Presentation] 核内受容体PPARγとシグナル感知脱メチル化酵素によるエネルギー代謝制御機構 (2015)
  • Author(s): 酒井寿郎
  • Organizer: 第88回内分泌学会総会
  • Place of Presentation: 東京
  • Year and Date: 2015-04-25
  • Invited
• [Presentation] 脂肪細胞における新規エピゲノム制御機構 (2015)
  • Author(s): 稲垣毅, 阿部陽平, Rozqie Royhan, 松村欣宏, 川村猛, 仲木竜, 鶴谷悠也, 谷村恭子, 田中十志也, 児玉龍彦, 油谷浩幸, 酒井寿郎
  • Organizer: 第52回日本臨床分子医学会学術集会
  • Place of Presentation: 京都
  • Year and Date: 2015-04-11
  • Invited
• [Remarks] 東京大学先端科学技術研究センター酒井研究室/原著論文
  • URL: http://www.mm.rcast.u-tokyo.ac.jp/publications/pubs.html