Investigation of kidney microenvironment by cell reconstruction in vivo

• Project/Area Number: 16K15214
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General medical chemistry
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: KANAI Masami 東京医科歯科大学, 統合研究機構, 教授 (70321883)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2018: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
• Keywords: 再生 / ジフテリア毒素 / 腎臓 / ヒト化マウス / 細胞再構築

Outline of Final Research Achievements

By use of the combination of human diphtheria toxin (DT) and its receptor, it is possible to kill the certain cells in vivo. We have succeed to development of transgenic lines to replace host cells to recipient new transgenic mose (Tg) line. We have tried to the combination of ROSA26-iDTR and Ltf (Lactoferrin)-iCre Tg. Ltf is expressed in seminal vesicles, epididymis and epithelial cells of the vas deferens in 60-day-old mature males. Administration of diphtheria toxin in this line made it possible to replace these epithelial cells after sexual maturation. We have succeeded in creating a system that allows xenogeneic cell transplantation using a new DT line.

Academic Significance and Societal Importance of the Research Achievements

精細管は管状の閉鎖空間で、比較的免疫寛容を受け易い環境を有する。AMH-Treck Tgマウス(GFP KI)や、Ltf/ROSA26-iDTRトランスジェニックマウスに中胚葉系上皮様細胞などの移植条件を決定し、その後、免疫寛容を呈するNODマウスへ遺伝背景のバッククロスすることで、ヒトiPS由来細胞異種細胞の移入しいては、xenotransplantationの確立へと展開させることが可能となる。将来的には、ヒトiPS細胞から腎各種細胞へのin vivo分化系の探索の糸口となり、その結果、社会的意義が大きいと思われる。

Research Products

• [Journal Article] Molecular and genetic characterization of partial masculinization in embryonic ovaries grafted into male nude mice.Molecular and genetic characterization of partial masculinization in embryonic ovaries grafted into male nude mice. (2019)
  • Author(s): Miura K, Harikae K, Nakaguchi M, Imaimatsu K, Hiramatsu R, Tomita A, Hirate Y, Kanai-Azuma M, Kurohmaru M, Ogura A, Kanai Y.
  • Journal Title: PLos One Volume: 14(3) Issue: 3 Pages: e0212367-e0212367
  • DOI: 10.1371/journal.pone.0212367
  • Peer Reviewed / Open Access
• [Journal Article] Essential roles of L-type amino acid transporter 1 in syncytiotrophoblast development by presenting fusogenic 4F2hc. (2017)
  • Author(s): Ohgaki R, Ohmori T, Hara S, Nakagomi S, Kanai-Azuma M, Kaneda-Nakashima K, Okuda S, Nagamori S, Kanai Y.
  • Journal Title: Mol Cell Biol. Volume: 印刷中 Issue: 11
  • DOI: 10.1128/mcb.00427-16
  • Peer Reviewed / Open Access
• [Journal Article] In vivo dynamics of GFRα1-positive spermatogonia stimulated by GDNF signals using a bead transplantation assay (2016)
  • Author(s): Uchida A, Kishi K, Aiyama Y, Miura K, Takase HM, Suzuki H, Kanai-Azuma M, Iwamori T, Kurohmaru M, Tsunekawa N, Kanai Y
  • Journal Title: Biochem Biophys Res Commun Volume: 476 Issue: 4 Pages: 546-552
  • DOI: 10.1016/j.bbrc.2016.05.160
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] TRECK法を用いた子宮内膜上皮置換法の確立 (2018)
  • Author(s): 中野有紀、平手良和、上村麻実、金井克晃、金井正美
  • Organizer: 第161回日本獣医学会
• [Presentation] アミノ酸トランスポーターLAT1は膜融合関連因子4F2hcの細胞膜提示による胎盤合胞体性栄養膜の形成に必須である (2017)
  • Author(s): 大垣隆一、大森崇弘、原早織、中込咲綾、金井正美、兼田（中島）加珠子、奥田傑、永森収志、金井好克
  • Organizer: 第12回トランスポーター研究会
• [Presentation] ライブ手術を併用し動物数の削減に配慮した心臓外科手術トレーニングプログラム構築の試み (2016)
  • Author(s): 大内克洋、水野友裕、大井啓司、八島正文、八丸剛、長岡英気、藤原立樹、黒木秀仁、田崎大、竹下斉史、木下亮二、遠藤衆、金井正美、木村太郎、荒井裕国
  • Organizer: 第63回日本実験動物学会
  • Place of Presentation: ミューザ川崎(神奈川県川崎市）
  • Year and Date: 2016-05-18
• [Book] 獣医発生学 第二版 第１章分担 (2019)
  • Author(s): 金井正美
  • Total Pages: 434
  • Publisher: 学窓社
• [Remarks]
  • URL: http://www.tmd-cea.jp/eam/
• [Remarks] 疾患モデル解析学分野HP
  • URL: http://www.tmd-cea.jp/eam/