Analysis of insulin resistance through DGKz and development of anti-diabetic medicine

• Project/Area Number: 24380152
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Applied animal science
• Research Institution: The University of Tokyo
• Principal Investigator: Hakuno Fumihiko 東京大学, 農学生命科学研究科, 助教 (30282700)
• Co-Investigator(Kenkyū-buntansha): ITO Akihiro 独立行政法人理化学研究所, 吉田化学遺伝学研究室, 専任研究員 (40391859)
• Co-Investigator(Renkei-kenkyūsha): TAKAHASHI Shin-Ichiro 東京大学, 大学院農学生命科学研究科, 准教授 (00197146)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,070,000 (Direct Cost: ¥13,900,000、Indirect Cost: ¥4,170,000); Fiscal Year 2014: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000); Fiscal Year 2013: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: インスリン / 糖尿病 / 糖輸送担体 / ジアシルグリセロールキナーゼ / インスリン抵抗性 / 低分子化合物 / ジアルシグリセロールキナーゼ / インスリン受容体基質 / ジアシルグリセロールキナーゼζ / 高糖尿病薬 / PIP5K / 糖輸送担体（GLUT4) / 細胞膜移行 / 2型糖尿病 / 糖取り込み

Outline of Final Research Achievements

We showed that DGKζ regulates insulin-induced GLUT4 translocation to PM in a canonical insulin-signal independent manner. In addition, DGKζ-associated protein, PIP5K1α modulates GLUT4 translocation in a similar mechanism as DGKζ.
By using ELISA kit, we succeeded to identify around 30 small compounds which inhibit the interaction between IRS-1 and DGKζ. Addition of one of these compounds, Compound A, inhibited the interaction between IRS-1 and DGKζ in HEK293T cells. Moreover Compound A addition recovered TNF-α-induced insulin resistance in 3T3-L1 adipocytes.

Research Products

• [Int'l Joint Research] Univ. of Applied Science Upper Austria(オーストリア)
• [Int'l Joint Research] University of Michigan/Albert Einstein College of Medicine(米国)
• [Int'l Joint Research] Karolinska Institute(スウェーデン)
• [Int'l Joint Research] Cajal Institute(スペイン)
• [Journal Article] The Novel Functions of High-Molecular-Mass Complexes Containing Insulin Receptor Substrates in Mediation and Modulation of Insulin-Like Activities: Emerging Concept of Diverse Functions by IRS-Associated Proteins. (2015)
  • Author(s): Hakuno F, Fukushima T, Yoneyama Y, Kamei H, Ozoe A, Yoshihara H, Yamanaka D, Shibano T, Sone-Yonezawa M, Yu BC, Chida K, Takahashi S.
  • Journal Title: Front Endocrinol (Lausanne). Volume: 6 Pages: 73-73
  • DOI: 10.3389/fendo.2015.00073
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Tumor Necrosis Factor (TNF)-induced Repression of GKAP42 Protein Levels through cGMP-dependent Kinase (cGK)-I Causes Insulin Resistance in 3T3-L1 Adipocytes (2015)
  • Author(s): Ando Y, Shinozawa Y, Iijima Y, Yu B-C, Sone M, Ooi Y, Watanaka Y, Chida K, Hakuno F, Takahashi S-I
  • Journal Title: J Biol Chem Volume: 290 Issue: 9 Pages: 5881-5892
  • DOI: 10.1074/jbc.m114.624759
  • Peer Reviewed
• [Journal Article] Analysis of insulin receptor substrates signaling dynamics on microstructured surfaces. (2015)
  • Author(s): Lanzerstorfer P, Yoneyama Y, Hakuno F, Muller U, Hoglinger O, Takahashi SI, Weghuber J
  • Journal Title: FEBS J. Volume: 282 Issue: 6 Pages: 987-1005
  • DOI: 10.1111/febs.13213
  • Peer Reviewed
• [Journal Article] Nedd4-induced mono-ubiquitination of IRS-2 enhances IGF signaling and mitogenic activity. (2015)
  • Author(s): Toshiaki Fukushima, Hidehito Yoshihara, Haruka Furuta, Fumihiko Hakuno, Jing Luan, Cunming Duan, Yasushi Saeki, Keiji Tanaka, Shun-Ichiro Iemura, TohruNatsume, Kazuhiro Chida, Yusuke Nakatsu, Hideaki Kamata, Tomoichiro Asano, and Shin-Ichiro Takahashi
  • Journal Title: Nature communications Volume: 6 Issue: 1 Pages: 6780-6780
  • DOI: 10.1038/ncomms7780
  • NAID: 120006348510
  • Peer Reviewed / Open Access
• [Journal Article] Aspp2 negatively regulates body growth but not developmental timing by modulating IRS signaling in zebrafish embryos. (2014)
  • Author(s): Liu C, Luan J, Bai Y, Li Y, Lu L, Liu Y, Hakuno F, Takahashi S, Duan C, Zhou J.
  • Journal Title: Gen Comp Endocrinol Volume: 197 Pages: 82-91
  • DOI: 10.1016/j.ygcen.2013.12.006
  • Peer Reviewed
• [Journal Article] Steroid hormones are novel nucleoside transport inhibitors by competition with nucleosides for their transporters. (2014)
  • Author(s): Kaneko M, Hakuno F, Kamei H, Yamanaka D, Chida K, Minami S, Coe RI, Takahashi S-I.
  • Journal Title: Bicohem Biophysic Res Commun. Volume: 443 Issue: 2 Pages: 505-510
  • DOI: 10.1016/j.bbrc.2013.11.132
  • Peer Reviewed
• [Journal Article] Insulin receptor substrate-1 associates with small nucleolar RNA which contributes to ribosome biogenesis. (2014)
  • Author(s): Ozoe A, Sone M, Fukushima T, Kataoka N, Chida K, Asano T, Hakuno F, Takahashi S-I.
  • Journal Title: Frontiers in Endocrinology Volume: 5 Pages: 24-24
  • DOI: 10.3389/fendo.2014.00024
  • Peer Reviewed / Open Access
• [Journal Article] The AP-1 complex regulates intracellular localization of insulin receptor substrate 1, which is required for insulin-like growth factor I-dependent cell proliferation (2013)
  • Author(s): Yosuke Yoneyama
  • Journal Title: Molecular and Cellular Biology Volume: Volume 33 Issue: 10 Pages: 1991-2003
  • DOI: 10.1128/mcb.01394-12
  • Peer Reviewed
• [Journal Article] Insulin Receptor Substrate-1 (IRS-1) Forms a Ribonucleoprotein Complex Associated with Polysomes (2013)
  • Author(s): Ozoe A, Sone M, Fukushima T, Kataoka N, Arai T, Chida K, Asano T, Hakuno F, Takahashi S-I
  • Journal Title: FEBS Lett. Volume: 587(15) Issue: 15 Pages: 2319-24
  • DOI: 10.1016/j.febslet.2013.05.066
  • Peer Reviewed
• [Journal Article] Insulin injection restores increased insulin receptor substrate (IRS)-2 protein during short-term protein restriction but does not affect reduced insulin-like growth factor (IGF)-I mRNA or increased triglyceride accumulation in the liver of rats. (2013)
  • Author(s): Ozaki Y, Takeda T, Akiyama-Akanishi N, Hakuno F, Toyoshima Y, Takahashi S-I, Takenaka A.
  • Journal Title: Biosci. Biotech. Biochem. Volume: 79 Issue: 1 Pages: 47-53
  • DOI: 10.1007/s12562-012-0570-9
  • Peer Reviewed
• [Journal Article] AP-1 complex regulates intracellular localization of insulin receptor substrate-1 required for insulin-like growth factor-I-dependent cell proliferation (2013)
  • Author(s): Yoneyama Y, Matsuo M, Take K, Kabuta T, Chida K, Hakuno F, Takahashi SI,
  • Journal Title: Mol. Cell. Biol.
  • Peer Reviewed
• [Journal Article] Nexjlin, a cardiomyopathy-associated F-actin binding protein, binds and regulates IRS1 signaling in skeletal muscle cells. (2012)
  • Author(s): Lee A, Hakuno F, Northcott P, Pessin JE, Rozakis Adcock M
  • Journal Title: PLoS One Volume: 8 Issue: 1 Pages: e55634-e55634
  • DOI: 10.1371/journal.pone.0055634
  • Peer Reviewed
• [Journal Article] Phosphatidylinsitol 3-kinase (PI3K) activity bound to insulin-like growth factor-I (IGF-I) receptor, which is continuously sustained by IGF-I stimulation, is required for IGF-I-induced cell proliferation. (2012)
  • Author(s): Fukushima T, Nakamura Y, Yamanaka D, Shibano T, Chida K, Minami S, Asano T, Hakuno F, Takahashi S-I.
  • Journal Title: J. Biol. Chem. Volume: 287 Issue: 35 Pages: 29713-29721
  • DOI: 10.1074/jbc.m112.393074
  • Peer Reviewed
• [Journal Article] Insulin/insulin-like growth factor (IGF) stimulation abrogates an association between a deubiquitinating enzyme USP7 and insulin receptor substrates (IRSs) followed by proteasomal degradation of IRSs (2012)
  • Author(s): Yoshihara H, Fukushima T, Hakuno F, Saeki Y, Tanaka K, Ito A, Yoshida M, Iemura S, Natsume T, Asano T, Chida K, Girnita L, Takahashi S-I
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 423 Issue: 1 Pages: 122-127
  • DOI: 10.1016/j.bbrc.2012.05.093
  • Peer Reviewed
• [Journal Article] Novel repressor regulates insulin sensitivity through interaction with Foxo1 (2012)
  • Author(s): Nakae J, Cao Y, Hakuno F, Takemori H, Kawano Y, Sekioka R, Abe T, Kiyonari H, Tanaka T, Sakai J, Takahashi SI, Itoh H
  • Journal Title: EMBO J. Volume: 31 Issue: 10 Pages: 2275-2295
  • DOI: 10.1038/emboj.2012.97
  • Peer Reviewed
• [Journal Article] Phosphatidylinositol 3-kinase binding protein, PITKAP/XB130 is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells. (2012)
  • Author(s): Yamanaka D, Akama T, Fukushima T, Nedachi T, Kawasaki C, Chida K, Minami S, Suzuki K, Hakuno F, Takahashi S-I.
  • Journal Title: Mol Endocrinol Volume: 26 Issue: 6 Pages: 1043-1055
  • DOI: 10.1210/me.2011-1349
  • Peer Reviewed
• [Presentation] 巨大なシグナル分子複合体IRSomeがインスリン活性の調節に果たす新しい役割 (2015)
  • Author(s): 高橋伸一郎、伯野史彦、安藤康年
  • Organizer: 第３８回日本分子生物学会年会／第８８回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-02
  • Invited
• [Presentation] The protein or compound that inhibits interaction between DGKζ and IRS-1 restore the TNF-α-induced suppression of glucose uptake in 3T3-L1 adipocytes. (2015)
  • Author(s): Yu BC, Liu TY, Ando Y, Ito A, Yoshida M, Hakuno F , Chida K, Takahashi S-I
  • Organizer: 第３８回日本分子生物学会年会／第８８回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-02
• [Presentation] GLUT4のセリンリン酸化が糖膜透過能に果たす役割 (2015)
  • Author(s): 渡部千尋、久保真実子、米澤（曽根）芽里、伯野史彦、高橋伸一郎
  • Organizer: 第３８回日本分子生物学会年会／第８８回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-02
• [Presentation] Insulin receptor substrates- associated proteins which are related with insulin resistance (2015)
  • Author(s): Bu-Chin Yu, Fumihiko Hakuno, Shin-Ichiro Takahashi
  • Organizer: International Seminar: Current boundaries of the expanding landscape of insulin-like activities and beyond
  • Place of Presentation: Spain, Madrid
  • Year and Date: 2015-09-24
  • Int'l Joint Research
• [Presentation] Negative feedback regulation of IGF-I receptor internalization by insulin receptor substrates (2015)
  • Author(s): Fumihiko Hakuno, Yousuke Yoneyama, Shin-Ichiro Takahashi
  • Organizer: International Seminar: Current boundaries of the expanding landscape of insulin-like activities and beyond
  • Place of Presentation: Spain, Madrid
  • Year and Date: 2015-09-24
  • Int'l Joint Research
• [Presentation] Diacylglycerol kinase ζ (DGKζ) and PIP5K1α cooperatively enhanced insulin-induced glucose uptake in 3T3-L1 adipocytes (2015)
  • Author(s): Hakuno F, Liu TY, Kakino M, Fujimoto H, Ando Y, Chida K, Takahashi SI.
  • Organizer: Gordon Conference
  • Place of Presentation: Ventura in USA
  • Year and Date: 2015-03-08 – 2015-03-12
• [Presentation] Roles of Insulin Receptor Substrate Complexes Containing Rabankyrin-5 in Insulin-Like Growth Factor-I-Induced Macropinocytosis. (2013)
  • Author(s): Takayuki Yano, Yosuke Yoneyama, Toshiaki Matsui, Hiroshi Okajima, Kazuhiro Chida, Fumihiko Hakuno, Shin-Ichiro Takahashi
  • Organizer: ENDO2013
  • Place of Presentation: San Francisco
• [Presentation] インスリン受容体基質（IRS）-1のアセチル化酵素と脱アセチル化酵素の特定 (2013)
  • Author(s): 藤枝久美子、米山鷹介、伊藤明博、吉田稔、千田和広、伯野史彦、高橋伸一郎
  • Organizer: 第65回日本細胞生物学会大会
  • Place of Presentation: 名古屋
• [Presentation] Constitutive expression suppressed cell invasion, leading to inhibition of myogenesis (2013)
  • Author(s): Fumihiko Hakuno.
  • Organizer: INTERNATIONAL OPEN WORKSHOP “Emerging Paradigms of Insulin-Like Activities in Physiology & Disease: From Pathophysiology to Targeted Therapy”
  • Place of Presentation: Karolinska Institute, Stockholm
  • Invited
• [Presentation] Constitutive IRS1 expression suppressed cell invasion and migration, leading to inhibition of myogenesis in L6 myoblasts. (2013)
  • Author(s): Fumihiko Hakuno, Yosuke Yoneyama, Kazuhiro Chida, Shin-Ichiro Takahashi.
  • Organizer: American Society of Cell Biology
  • Place of Presentation: New Orleans, USA
• [Presentation] Roles of insulin receptor substrate-1 interacting with clathrin adaptor AP-2 complex at submembranous actin cytoskeleton (2013)
  • Author(s): Yosuke Yoeneyama, Kazuhiro Chida, Fumihiko Hakuno , Shin-Ichiro Takahashi.
  • Organizer: American Society of Cell Biology
  • Place of Presentation: New Orleans, USA
• [Presentation] Diacylglycerol kinase ζ , a novel binding partner of insulin receptor substrates, negatively modulated GLUT4 translocation in 3T3-L1 adipocytes. (2012)
  • Author(s): Hakuno F, Ando Y, Kakino M, Fujimoto H, Chida K, Takahashi SI
  • Organizer: 6rh GRS-IGF 2012
  • Place of Presentation: Munich, Germany
  • Year and Date: 2012-10-19
• [Book] 酸化ストレスとインスリン様活性の相互作用が健康寿命延伸に果たす役割 (2013)
  • Author(s): 高橋伸一郎、金子元、竹中麻子、東祐輔、尾添淳文、伯野史彦
  • Total Pages: 8
  • Publisher: 医学のあゆみ
• [Book] 解説：インスリン様活性と高齢化社会で克服すべき疾病 (2013)
  • Author(s): 高橋伸一郎、伯野史彦、亀井宏泰、Leonard Girnita、Ignacio Torres-Aleman、東祐輔、福嶋俊明、柴野卓志、尾添淳文、山中大介
  • Total Pages: 11
  • Publisher: 化学と生物
• [Remarks] ２型糖尿病の原因の一つであるインスリン抵抗性発症機構の一部を解明
  • URL: http://www.a.u-tokyo.ac.jp/topics/2015/20150409-1.html
• [Remarks] 東京大学大学院 農学生命科学研究科 応用動物科学専攻 動物細胞制御学研究室
  • URL: http://endo.ar.a.u-tokyo.ac.jp/
• [Remarks] 動物細胞制御学研究室
  • URL: http://endo.ar.a.u-tokyo.ac.jp/
• [Remarks]
  • URL: http://endo.ar.a.u-tokyo.ac.jp/index0.html