Analysis of abnormal steroid hormone production by mass spectrometry imaging and targeted proteomics

• Project/Area Number: 17K09887
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Endocrinology
• Research Institution: Nara Medical University
• Principal Investigator: Hatano Osamu 奈良県立医科大学, 医学部, 講師 (40164850)
• Co-Investigator(Kenkyū-buntansha): 竹田 浩之 愛媛大学, プロテオサイエンスセンター, 准教授 (40609393) ; 大西 健 茨城県立医療大学, 保健医療学部, 名誉教授 (50152195) ; 竹森 洋 岐阜大学, 工学部, 教授 (90273672) ; 三宅 牧人 奈良県立医科大学, 医学部, 講師 (80601400)
• Project Period (FY): 2017-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: ステロイドホルモン / 塩誘導キナーゼ / AlphaScreen / コムギ胚芽無細胞タンパク質合成 / 卵巣 / 排卵 / ステロイドホルモン産生 / AlphaScreen 法 / 質量分析 / 標的プロテオミクス / Salt-Inducible Kinases / ステロイドホルモン産生組織 / コムギ胚芽無細胞タンパク質合成系 / 質量分析イメージング / 塩誘導性キナーゼ / SIK / ステロイド合成酵素 / プロテオミクス / 副腎 / イメージング / 合成酵素

Outline of Final Research Achievements

Salt-inducible kinases (SIK1,2,3) were initially isolated as factors affecting steroid hormone production and later shown to be involved in diverse phenotypic abnormalities. In this study, in order to elucidate the signalling mechanism of Salt-inducible kinases, we attempted to comprehensively identify proteins that bind to SIK1,2,3 by AlphaScreen method using human protein arrays. In addition, the role of SIKs in the ovulatory function of the ovary was analysed using SIK2- and SIK3-deficient mice. It was found that SIK3 positively regulates ovulation, while SIK2 conversely suppresses ovulatory function.

Academic Significance and Societal Importance of the Research Achievements

哺乳動物におけるステロイドホルモンは、生殖機能、糖代謝、電解質代謝などに関与する。本研究では塩誘導キナーゼ(SIK2,3)欠損マウスの解析により、SIK3 欠損が卵巣の排卵機能異常（無排卵や多嚢胞性卵巣症候群(PCOS）様）に関与することが示された。すなわち、正常な SIK3 は排卵機能に促進的に、又、SIK2 は逆に排卵機能に抑制的に働くことが明らかになった。これらSIK種の違いによる機能発現機構を明らかにするために、SIK各種と相互作用する結合因子の同定を行っており、SIK2,SIK3 の機能阻害や促進による卵巣機能異常/促進に伴う不妊などの治療に役立つと期待される。

Research Products

• [Int'l Joint Research] University of Illinois at Chicago(米国)
• [Int'l Joint Research] University of Illinois(米国)
• [Journal Article] Visualization of mitophagy using LysoKK, a 7-nitro-2,1,3-benzoxadiazole-(arylpropyl)benzylamine derivative (2022)
  • Author(s): Takemori H, Koga K, Kawaguchi K, Furukawa S, Ito S, Imaishi J, Watanabe M, Maeda M, Mizoguchi M, Oh-Hashi K, Hirata Y, Furuta K
  • Journal Title: Mitochondrion Volume: 62 Pages: 176-180
  • DOI: 10.1016/j.mito.2021.12.004
  • Peer Reviewed
• [Journal Article] Zebrafish as a new model for rhododendrol-induced leukoderma. (2021)
  • Author(s): Hayazaki M, Hatano O, Shimabayashi S, Akiyama T, Takemori H, Hamamoto A.
  • Journal Title: Pigment Cell Melanoma Res. Volume: 34(6) Issue: 6 Pages: 1029-1038
  • DOI: 10.1111/pcmr.13005
  • Peer Reviewed
• [Journal Article] A chalcone derivative suppresses TSLP induction in mice and human keratinocytes through binding to BET family proteins (2021)
  • Author(s): Segawa Ryosuke、Takeda Hiroyuki、Yokoyama Takeshi、Ishida Momoha、Miyata Chihiro、Saito Taiji、Ishihara Ryosuke、Nakagita Tomoya、Sasano Yusuke、Kanoh Naoki、Iwabuchi Yoshiharu、Mizuguchi Mineyuki、Hiratsuka Masahiro、Hirasawa Noriyasu
  • Journal Title: Biochemical Pharmacology Volume: 194 Pages: 114819-114819
  • DOI: 10.1016/j.bcp.2021.114819
  • Peer Reviewed
• [Journal Article] The new live imagers MitoMM1/2 for mitochondrial visualization. (2021)
  • Author(s): Maeda M, Suzuki M, Takashima S, Sasaki T, Oh-Hashi K, Takemori H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 562 Pages: 50-54
  • DOI: 10.1016/j.bbrc.2021.05.040
  • Peer Reviewed
• [Journal Article] Salt Inducible Kinases Are Critical Determinants of Female Fertility. (2020)
  • Author(s): Armouti M, Winston N, Hatano O, Hobeika E, Hirshfeld-Cytron J, Liebermann J, Takemori H, Stocco C.
  • Journal Title: Endocrinology Volume: in press Issue: 7 Pages: 1-13
  • DOI: 10.1210/endocr/bqaa069
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Engineered membrane protein antigens successfully induce antibodies against extracellular regions of claudin-5. (2018)
  • Author(s): Hashimoto Y, et al.
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 8383-8383
  • DOI: 10.1038/s41598-018-26560-9
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Autoantibody Profiling Using Human Autoantigen Protein Array and AlphaScreen. (2018)
  • Author(s): Takeda-H
  • Journal Title: Methods Mol Biol. Volume: 1868 Pages: 93-112
  • DOI: 10.1007/978-1-4939-8802-0_10
  • ISBN: 9781493988013, 9781493988020
• [Journal Article] CP5 system, for simple and highly efficient protein purification with a C-terminal designed mini tag (2017)
  • Author(s): Takeda Hiroyuki、Zhou Wei、Kido Kohki、Suno Ryoji、Iwasaki Takahiro、Kobayashi Takuya、Sawasaki Tatsuya
  • Journal Title: PLOS ONE Volume: 12 Issue: 5 Pages: e0178246-e0178246
  • DOI: 10.1371/journal.pone.0178246
  • Peer Reviewed / Open Access
• [Presentation] CLEM imaging of rhododendrol-induced leukoderma in zebrafish and catalytic oxidation of Japanese cedar wood. (2020)
  • Author(s): Osamu Hatano, Masumi Hayazaki, Akie Hamamoto, Hiroshi Takemori, Hikaru Takaya, Takafumi Shano, Ken Ohnishi, Masaharu Nakamura.
  • Organizer: ABiS Symposium: Forefront and Future of Electron Microscopic Imaging, Okazaki, Japan
  • Int'l Joint Research
• [Presentation] ステロイドホルモンの質量分析イメージング解析とラット副腎皮質の再生 (2018)
  • Author(s): 秦野 修、磯崎勝弘、竹森 洋、西村裕志、竹田浩之、岩崎哲史、大西 健.
  • Organizer: 第1回再生学異分野融合研究会 (岡崎市)
• [Presentation] SALDIと誘導体化によるステロイドホルモンのイオン化法の改良と質量分析イメージングへの応用 (2017)
  • Author(s): 秦野 修、磯崎勝弘、竹森 洋、大西 健、岩崎哲史、片桐昌直、川崎英也、一柳優子、荒川隆一
  • Organizer: 第68回イオン反応研究会/第157回質量分析関西談話会/第6回イオン移動度研究会・合同研究会
• [Book] 創薬研究のためのスクリーニング学実践テキスト 第3章-7. AlphaScreen による HTS (2022)
  • Author(s): 竹田浩之
  • Total Pages: 5
  • Publisher: 羊土社
  • ISBN: 9784758122580