Elucidation of Zfat function in fetal erythropoiesis using a conditional knockout mouse

• Project/Area Number: 15K08166
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Fukuoka University
• Principal Investigator: DOI Keiko 福岡大学, 医学部, 講師 (10341538)
• Research Collaborator: Shirasawa Senji ; Tsunoda Toshiyuki ; Koyanagi Midori
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 赤血球 / 転写因子 / ZFAT

Outline of Final Research Achievements

ZFAT, originally identified as a candidate susceptibility gene for autoimmune thyroid disease, has been reported to be involved in primitive hematopoiesis and T cell development. However, whether or not Zfat is involved in definitive erythropoiesis in the fetal liver during mammalian development remains unknown. Here, we analyzed the role of Zfat during mouse fetal erythropoiesis in the fetal liver using tamoxifen-inducible CreERT2 Zfat-deficient mice. Zfat-deficient mice exhibit moderate anemia with small and pale fetal liver through a decreased number of erythroblasts by E12.5. The apoptosis sensitivity in fetal liver erythroid progenitors was enhanced by Zfat-deficiency ex vivo. Moreover, Zfat knockdown partially inhibited the CD71-/lowTer119- to CD71highTer119- transition of fetal liver erythroid progenitors with the impairment in the elevation of CD71 expression. These results thus demonstrate that Zfat plays a critical role for erythropoiesis in the fetal liver.

Academic Significance and Societal Importance of the Research Achievements

本研究でのマウス個体におけるZFATの機能的役割を解明するアプローチは、ZFATの重要な機能制御ネットワークを明らかにすることが予想され、また生命維持機構における重要な転写制御ネットワークにおいて新たな知見を付加することが期待される。また、コンディショナルZFAT欠損マウスの表現型解析からは、胎児発生期の血球系分化におけるZFATの機能的役割が解明できることが予想され、免疫系の制御機構の破綻に起因する自己免疫疾患を含む免疫関連疾患とZFATの機能的役割との関連性から病因・病態の解明、さらには創薬ターゲットの探索による先駆的な治療法の開発への応用へ繋げることが期待される。

Research Products

• [Journal Article] Zfat expression in ZsGreen reporter gene knoc;in mice: Implications for a novel function of Zfat in definitive erythropoiesis. (2018)
  • Author(s): Tsunoda T, Doi K, Ishikura S, Luo H, Nishi K, Matsuzaki H, Koyanagi M, Tanaka Y, Okamura T, Shirasawa S
  • Journal Title: Int J Mol Med. Volume: 42 Pages: 2595-2603
  • DOI: 10.3892/ijmm.2018.3806
  • Peer Reviewed / Open Access
• [Journal Article] Molecular mechanisms of transcriptional regulation by the nuclear zinc-finger protein Zfat in T cells. (2016)
  • Author(s): Ishikura S, Tsunoda T, Nakabayashi K, Doi K, Koyanagi M, Hayashi K, Kawai T, Tanaka Y, Iwaihara Y, Luo H, Nishi K, Okamura T, Shirasawa S.
  • Journal Title: Biochim Biophys Acta. Volume: 1859(11) Issue: 11 Pages: 1398-1410
  • DOI: 10.1016/j.bbagrm.2016.08.010
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The Nuclear Zinc Finger Protein Zfat Maintains FoxO1 Protein Levels in Peripheral T Cells by Regulating the Activities of Autophagy and the Akt Signaling Pathway. (2016)
  • Author(s): Ishikura S, Iwaihara Y, Tanaka Y, Luo H, Nishi K, Doi K, Koyanagi M, Okamura T, Tsunoda T, Shirasawa S.
  • Journal Title: J Biol Chem. Volume: 291(29) Issue: 29 Pages: 15282-91
  • DOI: 10.1074/jbc.m116.723734
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Zfat-deficient CD4⁺ CD8⁺ double-positive thymocytes are susceptible to apoptosis with deregulated activation of p38 and JNK. (2015)
  • Author(s): Ishikura S, Ogawa M, Doi K, Matsuzaki H, Iwaihara Y, Tanaka Y, Tsunoda T, Hideshima H, Okamura T, Shirasawa S.
  • Journal Title: J Cell Biochem. Volume: 16(1) Issue: 1 Pages: 149-57
  • DOI: 10.1002/jcb.24954
  • Peer Reviewed / Open Access
• [Journal Article] Marked Reduction in FoxO1 Protein by its Enhanced Proteasomal Degradation in Zfat-deficient Peripheral T-Cells. (2015)
  • Author(s): Iwaihara Y, Ishikura S, Doi K, Tsunoda T, Fujimoto T, Okamura T, Shirasawa S.
  • Journal Title: Anticancer Res. Volume: 35 Pages: 4419-4423
  • Peer Reviewed
• [Presentation] 末梢T細胞のTCR刺激時における転写制御分子Zfatの機能メカニズムの解明 (2018)
  • Author(s): 土井佳子、石倉周平、角田俊之、小柳緑、田中陽子、中林一彦、白澤専二
  • Organizer: 日本人類遺伝学会第63回大会
• [Presentation] T細胞における転写制御分子ZFATの分子機構の解明 (2017)
  • Author(s): 土井佳子、石倉周平、角田俊之、小柳緑、田中陽子、中林一彦、白澤専二
  • Organizer: 日本人類遺伝学会第62回大会
• [Presentation] Identification of the genes regulated by ZFAT in T cells through ChIP-seq analysis (2016)
  • Author(s): Keiko Doi, Toshiyuki Tsunoda, Midori Koyanagi, Shuhei Ishikura, Yoko Tanaka, Kazuhiko Nakabayashi, Senji Shirasawa
  • Organizer: 第13回国際人類遺伝学会
  • Place of Presentation: 国立京都国際会館（京都府京都市左京区）
  • Int'l Joint Research
• [Presentation] T細胞における免疫系転写制御分子Zfatの転写制御標的分子の同定 (2015)
  • Author(s): 土井佳子、角田俊之、小柳緑、石倉周平、田中陽子、白澤専二
  • Organizer: 第１９回バイオ治療法研究会学術集会
  • Place of Presentation: 東京理科大学森戸記念館(東京都新宿区)
  • Year and Date: 2015-12-05
• [Presentation] ChIP-seq解析によるT細胞におけるZFATの転写制御標的分子の同定 (2015)
  • Author(s): 土井佳子、角田俊之、小柳緑、石倉周平、田中陽子、中林一彦、白澤専二
  • Organizer: 日本人類遺伝学会第60回大会
  • Place of Presentation: 京王プラザホテル(東京都新宿区)
  • Year and Date: 2015-10-15