Establishment of in vivo model of anemia by shRNA transgene controlled by erythroid-lineage specific promoter

2017 Fiscal Year Annual Research Report

• Project/Area Number: 15K09447
• Research Institution: Tohoku University
• Principal Investigator: 藤原 亨 東北大学, 大学病院, 講師 (60333796)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 赤血球分化 / 貧血 / 転写因子

Outline of Annual Research Achievements

本研究においては、赤芽球分化に関わる転写因子群の機能解析に焦点をおき、テトラサイクリン発現誘導系を用いてマウス赤血球特異的にshRNAを効果的かつ可逆的に発現させる、in vivo遺伝子ノックダウン解析系を確立することを目指す。
平成29年度までに赤血球系の転写因子であるGATA-1プロモーター制御下にリバーステトラサイクリン制御性トランス活性化因子(rtTA)を発現させるトランスジェニックマウスを作製しており、rtTAが赤芽球に高発現している事を確認した。平成29年度はテトラサイクリン応答因子(TRE)制御下にLmo2特異的shRNAを発現させるトランスジェニックマウスを作成し、前者のマウスと掛け合わせることにより目的のマウス(Tg-EGFP-Lmo2 shRNA::Tg-G1HRD-rtTA)を作製した。現在なお、その形質を解析中である。
LMO2以外の赤血球系転写因子として、サラセミア、ポルフィリン症、Hereditary dyserythropoietic anemiaなどの様々な貧血関連疾患と深く関わるFOG1 (Friend of GATA-1)にも着目し、平成29年度中に機能解析を行った(Fujiwara et al. Biochemical and Biophysical Research Communications. 2017)。今後、本転写因子も赤血球系特異的ノックダウン系に利用する予定である。

Research Products

• [Journal Article] A defined culture method enabling the establishment of ring sideroblasts from induced pluripotent cells of X-linked sideroblastic anemia. (2018)
  • Author(s): Hatta S, Fujiwara T, Yamamoto T, Saito K, Kamata M, Tamai Y, Kawamata S, Harigae H
  • Journal Title: Haematologica Volume: - Pages: -
  • DOI: 10.3324/haematol.2017.179770
  • Peer Reviewed
• [Journal Article] Establishment of a Screening System to Identify Novel GATA-2 Transcriptional Regulators (2018)
  • Author(s): Ohashi K, Fujiwara T, Onodera K, Saito Y, Ichikawa S, Kobayashi M, Okitsu Y, Fukuhara N, Onishi Y, Harigae H.
  • Journal Title: The Tohoku Journal of Experimental Medicine Volume: 244(1) Pages: 41-52
  • DOI: 10.1620/tjem.244.41
  • Peer Reviewed / Open Access
• [Journal Article] Dynamics of absorption, metabolism, and excretion of 5-aminolevulinic acid in human intestinal Caco-2 cells (2017)
  • Author(s): Saito K, Fujiwara T, Ota U, Hatta S, Ichikawa S, Kobayashi M, Okitsu Y, Fukuhara N, Onishi Y, Ishizuka M, Tanaka T, Harigae H
  • Journal Title: Biochemistry and Biophysics Reports Volume: 11 Pages: 105-111
  • DOI: 10.1016/j.bbrep.2017.07.006
  • Peer Reviewed
• [Journal Article] Effects of in vivo deletion of GATA2 in bone marrow stromal cells (2017)
  • Author(s): Hasegawa S, Fujiwara T, Okitsu Y, Kato H, Sato Y, Fukuhara N, Onishi Y, Shimizu R, Yamamoto M, Harigae H
  • Journal Title: Experimental Hematology Volume: 56 Pages: 31-45
  • DOI: 10.1016/j.exphem.2017.08.004
  • Peer Reviewed
• [Journal Article] A novel heterozygous ALAS2 mutation in a female with macrocytic sideroblastic anemia resembling myelodysplastic syndrome with ring sideroblasts: a case report and literature review. (2017)
  • Author(s): Fujiwara T, Fukuhara N, Ichikawa S, Kobayashi M, Okitsu Y, Onishi Y, Furuyama K, Harigae H
  • Journal Title: Annals of Hematology Volume: 96(11) Pages: 1955-1957
  • DOI: 10.1007/s00277-017-3106-7
  • Peer Reviewed
• [Journal Article] Forced FOG1 expression in erythroleukemia cells: Induction of erythroid genes and repression of myelo-lymphoid transcription factor PU.1. (2017)
  • Author(s): Fujiwara T, Sasaki K, Saito K, Hatta S, Ichikawa S, Kobayashi M, Okitsu Y, Fukuhara N, Onishi Y, Harigae H.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 485(2) Pages: 380-387
  • DOI: 10.1016/j.bbrc.2017.02.068
  • Peer Reviewed
• [Journal Article] GATA transcription factors: basic principles and related human disorders. (2017)
  • Author(s): Fujiwara T
  • Journal Title: The Tohoku Journal of Experimental Medicine Volume: 242(2) Pages: 83-91
  • DOI: 10.1620/tjem.242.83.
  • Peer Reviewed / Open Access
• [Journal Article] Impact of TET2 deficiency on iron metabolism in erythroblasts. (2017)
  • Author(s): Inokura K, Fujiwara T, Saito K, Iino T, Hatta S, Okitsu Y, Fukuhara N, Onishi Y, Ishizawa K, Shimoda K, Harigae H.
  • Journal Title: Experimental Hematology Volume: 49 Pages: 56-67
  • DOI: 10.1016/j.exphem.2017.01.002
  • Peer Reviewed
• [Journal Article] An Lysophosphatidic Acid Receptors 1 and 3 Axis Governs Cellular Senescence of Mesenchymal Stromal Cells and Promotes Growth and Vascularization of Multiple Myeloma. (2017)
  • Author(s): Kanehira M, Fujiwara T, Nakajima S, Okitsu Y, Onishi Y, Fukuhara N, Ichinohasama R, Okada Y, Harigae H.
  • Journal Title: Stem Cells Volume: 35(3) Pages: 739-753
  • DOI: 10.1002/stem.2499
  • Peer Reviewed
• [Presentation] Generation of Induced Pluripotent Stem Cell-Derived Erythroblasts From A Patient With X-Linked Sideroblastic Anemia (2017)
  • Author(s): Shunsuke Hatta, Tohru Fujiwara, Takako Yamamoto, Mayumi Kamata, Yoshiko Tamai, Yukiko Nakamura, Shin Kameoka, Hideo Harigae
  • Organizer: Seventh Congress of the International Biolron Society
  • Int'l Joint Research
• [Presentation] Exploring the Mechanism of Fog1-Dependent Transcriptional Regulation in Erythroid Cells (2017)
  • Author(s): Tohru Fujiwara, Katsuyuki Sasaki, Kei Saito, Shunsuke Hatta, Satoshi Ichikawa, Masahiro Kobayashi, Yoko Okitsu, Noriko Fukuhara, Yasushi Onishi, Hideo Harigae
  • Organizer: European Hematology Association 22nd Congress
  • Int'l Joint Research
• [Presentation] ALAS2遺伝子変異を認めた大球性鉄芽球性貧血の一例 (2017)
  • Author(s): 藤原亨、福原規子、猪俣美津恵、大西康、古山和道、張替秀郎
  • Organizer: 第41回日本鉄バイオサイエンス学会学術集会
• [Presentation] Exploring the mechanism of FOG1-dependent transcriptional regulation in erythroid cells (2017)
  • Author(s): Tohru Fujiwara, Katsuyuki Sasaki, Kei Saito, Shunsuke Hatta, Satoshi Ichikawa, Masahiro Kobayashi, Yoko Okitsu, Noriko Fukuhara, Yasushi Onishi, Hideo Harigae
  • Organizer: 第79回日本血液学会学術集会
• [Remarks] 東北大学 血液・免疫病学分野
  • URL: http://www.rh.med.tohoku.ac.jp/index.html